白菜三种雄性不育系与保持系花蕾转录组差异分析及三个花粉发育相关基因功能鉴定
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摘要
利用植物雄性不育特性来选育雄性不育系是作物杂种优势利用中简化制种程序、降低制种成本的重要手段。十字花科作物是杂种优势利用最为普遍的一类作物，雄性不育系(简称不育系)是其生产一代杂种的理想系统。但植物雄性不育产生的机理极其复杂，至今尚未完全解开。花粉败育是植物雄性不育发生的表型体现，弄清花粉发育的全过程和分子机理是研究雄性不育的基础和关键所在。花粉发育涉及众多基因的表达调控，从转录组入手分析花粉发育突变体是获得花粉基因动态表达的途径之一，并可获得较大数目的花粉发育相关基因，有助于在整体水平上理解花粉发育的特征和花粉发育的分子调控机制。本文在获得共享同一保持系(核不育两用系的可育株系)的白菜(Brassica campestris L．ssp．chinensis Makino，syn．B．rapa ssp．chinensis)‘矮脚黄’核不育(‘Aijiaohuang’genic male sterility，ajhGMS)两用系‘Bcajh97-01A／B’、‘Polima’核质互作不育(polima genic-cytoplasmic male sterility，polG-CMS)系‘Bcpol97-05A’以及'Ogura'细胞质不育(Ogura cytoplasmic male sterility，oguCMS)系‘Bcogu97-06A’材料体系的基础上，采用扩增互补脱氧核糖核酸片断长度多态性(complementary deoxyribonucleic acid amplified fragment length polymorphism，cDNA-AFLP)及拟南芥基因芯片分析其花蕾转录组差异，对花粉基因表达谱进行系统分析，研究不同雄性不育遗传模式的基因突变导致下游系列基因表达变化的异同，为建立不同雄性不育遗传模式植物花粉基因表达谱的基本框架奠定基础，并鉴定出一批白菜花粉发育相关基因。进而采用反义RNA或RNA干涉(RNAinterference，RNAi)技术，对细胞壁合成与调控相关的两个多聚半乳糖醛酸酶(polygalacturonase，PG)基因BcMF2(Brassica campestris male fertility 2)和BcMF9(Brassica campestris male fertility 9)，以及一个未知功能的新基因BcMF10(Brassica campestris male fertility 10)进行功能验证，从形态学、分子生物学、细胞学等方面入手鉴别三个基因在白菜花粉发育过程中的作用机制。同时，从基因结构、表达特点、作用方式等方面，比较了白菜花粉表达的PG基因家族成员BcMF2、BcMF9和BcMF6(Brassica campestris male fertility 6)的异同。取得的主要结果如下：
(1)形态和细胞学观察发现‘Bcajh97-01A’为雄性减数分裂的胞质分裂突变体mmc，对应基因MMC可能编码特异作用于雄性减数分裂的蛋白质。形态和细胞学观察发现，ajhGMS两用系不育株系‘Bcajh97-01A’与可育株系'Bcajh97-01B'的差异仅表现在花粉花药发育上，‘Bcajh97-01A’小孢子在减数分裂末期的胞质分裂中出现异常，不能形成胞间壁而导致四分体形成及后续的花粉发育各过程受阻，最终引起花粉败育。遗传学分析表明该不育性状由单基因位点控制，因此，‘Bcajh97-01A’实际上是一个受单基因位点控制的雄性减数分裂的胞质分裂突变体mmc(male meiotic cytokinesis)，其对应基因MMC可能编码一个特异与雄性减数分裂核分裂完成后的胞质分裂相关的蛋白质。
(2)利用cDNA-AFLP技术结合拟南芥基因芯片分析白菜不同不育系之间及与其共同保持系之间的花蕾转录组差异，发现不同遗传模式不育系花蕾基因表达有相似之处，但不尽相同。利用cDNA-AFLP技术及拟南芥ATH1基因芯片，对白菜ojhGMS‘Bcajh97-01A’、polG-CMS‘Bcpol97-05A’和oguCMS‘Bcogu97-06A’，以及它们共同的保持系‘Bcajh97-01B’进行了花蕾转录组比较分析，发现不同的不育基因作用下均引起花蕾正常转录组的巨大变化，不育基因下游众多基因的表达受到抑制，也有相当一部分的基因被激活或表达上升，但作用的不育基因不同，引起的下游基因表达变化也不尽相同。与同一保持系‘Bcajh97-01B’相比，检测到在‘Bcajh97-01A’花蕾中上调表达基因93个、下调表达基因158个；在‘Bcpol97-05A’花蕾中上调表达基因174个、下调表达基因212个；在‘Bcogu97-06A’花蕾中上调表达基因196个、下调表达基因242个。其中。在三种不育系中共同下调表达的基因有37个，上调表达的24个；‘Bcajh97-01A’和‘Bcpol97-05A’共同下调和共同下调表达的基因各5个；‘Bcajh97-01A’和‘Bcogu97-06A’共同下调表达基因为70个，共同上调表达基因9个；‘Bcpol97-05A’和‘Bcogu97-06A’共同下调表达基因7O个，共同上调表达的基因45个。
(3)通过差异表达基因功能分类，发现许多花粉花药发育相关基因功能未知，白菜不同不育系与保持系的花蕾转录组差异分析扩充了植物花粉表达或特异表达基因的数目。对在三种不育系与其共同保持系花蕾转录组中检测到的差异表达基因进行功能分类，发现约50％的基因功能未知。基于三种不育系和保持系花蕾的差别仅表现在花药发育和花粉形成上，在不育系和保持系花蕾中差异表达的基因最可能与花粉花药发育相关。我们发现的这些差异表达基因在一定程度上扩充了植物花粉花药表达和特异表达基因的数目。
(4)基于基因功能分类对不同不育系与保持系花蕾基因表达差异进行的分析表明，不同遗传模式花粉花药转录组尽管具有相似的模块，但具有不同的组成特征。对差异表达基因进行功能分类发现，不育系与保持系差异表达的三组基因可分为大致相同的功能类别，但每一功能类别在每组差异基因中所占的比例不尽相同。在三种不育系花蕾中下调均较多的已知功能的基因与转运通道、蛋白质代谢、电子传递和能量途径、防卫和胁迫反应相关，上调均较多的基因与转运通道、转录调控和普通新陈代谢相关。除此之外，‘Bcajh97-01A’与‘Bcogu97-06A’在花粉花药基因表达上具有更大的共性，在两者花蕾中均下调表达的基因中，细胞壁合成与调控基因排在第一位，细胞骨架蛋白及信号转导相关基因也占了较大的比例，而转录相关基因的比例较小；两者下调表达的基因中，蛋白质代谢相关基因均比较多。‘Bcajh97-01A’与‘Bcogu97-06A’的花蕾转录组的组成特征与拟南芥花粉转录组特征较一致。相比之下，‘Bcpol97-05A’花蕾转录组的组成成分与其他两种不育系有较大的差别。突出表现在细胞壁合成与调控基因、细胞骨架蛋白基因以及信号转导相关基因在不育系花蕾中占较大的比例，而保持系与其相比，这三种类别基因的表达并不占优势，反而更少。由此可见，不同遗传模式花粉花药转录组尽管具有相似的模块，但组成特征各异。
(5)时空表达模式分析发现三种不育系与其共同保持系花蕾差异表达的基因具不同的表达动态。对27条在三种不育系和保持系花蕾中差异表达的片段进行了时空表达模式分析，发现大多数差异片段所代表的基因表达在小孢子有丝分裂前后开始被检测到，在花粉成熟过程中继续积累，但在不同组织和发育阶段中的表达水平不尽相同。此外也有在花粉发育早期就检测到表达的基因。我们的研究结果证实了花粉花药发育中的基因表达调控是极其复杂的。在花粉花药发育的不同时期表达或不表达，以及表达水平有序地发生变化都是与花粉花药正常发育、实现其功能所需相对应的。我们的研究为明确白菜花粉花药发育不同时期的基因群作出了有益的探索。
(6)表达和功能分析发现BcMF2可能是一个与花粉内壁发育相关的新PG基因。对本实验室前期从‘Bcajh97-01B’花蕾中分离得到的PG基因BcMF2进行时空表达模式分析，发现其转录本最早在四分体时期的花蕾中开始出现，随后在单核花粉粒时期的花蕾中表达量达到最高，之后，随着花粉发育的进行表达量逐渐降低直至花粉成熟，表明其属于花粉表达“早期”基因。利用反义RNA技术研究BcMF2的功能，发现BcMF2表达受抑制的转基因植株花粉体外正常萌发率大幅度降低。约80％的花粉管生长到一定程度后顶端膨大形成泡状结构，花粉管能穿过柱头组织，但在花柱道中的生长停止。进一步观察显示转基因植株花粉畸形，萌发沟的数目及分布不规则，萌发沟在形成过程中位置和数目紊乱，花粉粒内壁异常增厚。推测BcMF2表达受抑制影响了花粉内壁中果胶的动态代谢平衡而导致萌发沟及花粉粒内壁发育的异常。BcMF2与已知的在花粉发育早期起作用的PG基因没有序列及表达特点的相似性，因此它可能是一个新的在花粉发育早期表达的PG基因。
(7)表达和功能分析发现分离获得的另一个白菜PG基因BcMF9可能参与花粉内壁和外壁的发育。分离获得在可育株‘Bcajh97-01B’花蕾中特异表达的差异片段BBS13／BPO023的全长序列BcMF9，发现该基因具有典型的PG基因结构特征。分子进化分析表明其属于花粉表达或特异表达的PG基因类群。时空表达模式分析显示BcMF9属于花粉表达“晚期”基因，因其表达最早出现在四分小孢子中，在单核小孢子中开始增强，然后一直持续较高水平的表达直到花粉成熟。BcMF9的转录本亦在绒毡层中出现，从四分体时期开始直到绒毡层降解退化，一直在绒毡层细胞中维持很强的表达信号。转反义BcMF9的植株花粉离体正常萌发率明显降低，约81％的花粉萌发时花粉管爆裂。花粉管能穿过柱头组织，但不久便停止生长。扫描电镜和透射电镜观察显示，BcMF9表达受抑制产生外壁网纹浅的畸形花粉，花粉粒萌发沟处内壁增厚异常，萌发沟的位置和数目紊乱，花粉发育晚期外壁的覆盖层和基粒棒部分脱落，含油层外溢，说明BcMF9可能参与花粉壁的发育。此外，发现转反义BcMF9植株花药绒毡层降解速度加快，绒毡层发育发生异常前的小孢子发育正常，而异常后的花粉外壁发育受影响。推测BcMF9表达受抑制可能破坏了绒毡层细胞的程序性死亡过程，启动了另一个死亡途径，加速了绒毡层的解体。绒毡层正常程序性细胞死亡(programmed cell death，PCD)的破坏，导致其向花粉外壁输送物质过程的改变，从而影响了花粉壁的正常发育。但由于BcMF9编码的蛋白在其N端有一段信号肽序列，而它的表达首先出现在四分体时期的绒毡层和小孢子中，且在绒毡层中的表达信号要比在小孢子中强。因此推测其功能发挥还有另一个可能：BcMF9在绒毡层表达，表达产物分泌到花粉囊腔，进而在花粉壁的发育中直接起作用。
(8)比较分析发现PG基因家族成员BcMF2、BcMF6和BcMF9在花粉发育中可能扮演各自不同的角色。从基因序列特征、表达模式特点、基因进化关系及生物学功能等方面比较三个花粉发育相关的PG基因BcMF2、BcMF6和BcMF9的异同，发现尽管3个PG基因的表达所出现的发育阶段一致，但三者在各个发育阶段的表达量及表达变化的趋势各不相同。三者在十字花科物种中的进化特点也不完全一致。3个PG基因的表达受抑制引发了类似的结果，但并不尽相同，说明它们在花粉发育中所起的作用及作用模式可能各异。
(9)分离获得在‘Bcajh97-01B’和‘Bcpol97-05A’花蕾中高水平表达的基因BcMF10，序列特征分析提示其可能与信号转导途径相关，RNAi研究发现BcMF10表达受抑制导致花粉萌发异常。分离获得在‘Bcajh97-01B’和‘Bcpol97-05A’花蕾中高水平表达，但在‘Bcajh97-01A’和‘Bcogu97-06A’花蕾中沉默的基因片段BBS31／BPO079的全长序列BcMF10。利用生物信息学分析发现预测的BcMF10氨基酸序列中具有可能与细胞增殖、分化和PCD相关的蛋白激酶C磷酸化位点，参与调控信号转导相关的蛋白激酶的酪氨酸磷酸化位点，及其它与细胞内信号转导、蛋白定位以及黏附等过程有关的位点。推测BcMF10编码的蛋白可能在花粉发育过程的信号转导中发挥重要作用。利用RT-PCR(reverse transcriptase PCR，反转录PCR)和原位杂交分析了BcMF10的时空表达特点，发现其转录本最早出现在‘Bcajh97-01B’花药发育早期的花粉母细胞和绒毡层细胞中，随后在减数分裂时期的小孢子和绒毡层中表达量进一步提高，之后随着发育的进行，表达量逐渐降低，但表达部位没有改变，在花粉成熟期消失，属于花粉表达“早期”基因。通过构建含BcA9绒毡层特异启动子的BcMF10 RNAi表达载体，利用转基因阻断内源BcMF10的表达，发现75％左右的转基因植株花粉不能正常萌发，约15.6％的花粉能长出花粉管，但花粉管长到一定长度时顶端爆裂，正常萌发率只有10.5％，说明BcMF10表达受抑制影响了花粉的正常萌发。但与通常发现的花粉管一伸出萌发孔即发生爆裂的现象不同，BcMF10表达受抑制植株花粉管能长到一定长度，然后将内容物从顶端喷出，花粉管生长停止，推测BcMF10也可能与花粉壁的发育相关。根据BcMF10预测蛋白质的结构特征及其在早期绒毡层细胞和小孢子中共表达的特点，可以认为其可能作用于细胞壁发育过程中绒毡层与花粉的信号传递过程。
The application of male sterility becomes an important way in heterosis breeding to simplify the seed production procedures and reduce the production cost. The utilization of hererosis is prevalent in cruciferae crops breeding, and the male sterile line is a useful system for their hybrid seed production. However, the occurrence and the mechanism of plant male sterility still remain mysteries that have not been unveiled thoroughly. For male sterility mainly manifests in pollen abortion, to make clear of the whole process of pollen development and the mechanism underlined thus become the basis point for understanding of male sterility. Pollen development is regulated by a complex regulatory network that comprises a wide variety of genes. Transcriptional profiling on the mutant compared with its wild-type is a successful approach to investigate the dynamic gene expression during pollen development and to detect genes involved in this biological process, and ultimately to help to understand the characteristic of pollen development and the corresponding molecular mechanism. In this study, we established three different male sterile types, 'Aijiaohuang' genic male sterility AB line (ajhGMS 'Bcajh97-01A/B'), 'Polima' genic-cytoplasmic male sterility (polG-CMS 'Bcpol97-05A') and 'Ogura' cytoplasmic male sterility (oguCMS 'Bcogu97-06A') in Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis. These male sterile lines share a common maintainer line. Then, we conducted profiling comparisons between flower buds of these male sterile lines and their maintainer line using complementary deoxyribonucleic acid amplified fragment length polymorphism (cDNA-AFLP) technology together with ATH1 genome array analysis. By analyzing the changes in expression pattern of genes acting downstream due to the gene mutation in different male sterile plants, plenty of candidate genes involved in pollen development were identified, and thus laid a foundation for establishing the profiles of gene expression in different types of male sterile lines. After that, three genes were selected for further characterization, including two putative polygalacturonase (PG) genes, BcMF2 {Brassica campestris male fertility 2) and BcMF9 {Brassica campestris male fertility 9) which may be related to cell wall biosynthesis and regulation, and one novel gene, BcMFW {Brassica campestris male fertility 10) with unknown function. Anti-sense RNA or RNA interference (RNAi) technique were used to investigate the function of the three genes, and morphologic, molecular biology as well as cytology analysis were carried through in functional assignment. Comparison of gene structure, expression pattern and biological function of three members, BcMF2, BcMF6 {Brassica campestris male fertility 6) and BcMF9 in PG gene family were also performed in this study. The results obtained are as follows.
(1) Observation on the pollen development in male sterile line 'Bcajh97-01A' suggests that it's a mutant of male meiosis cytokinesis in B. campestris, and the corresponding gene, MMC may encode a protein acting specifically in male meiosis. When observed the morphology and cytology character of the ajhGMS 'Bcajh97-01A/B' we found that the only detectable difference between the male sterile plants 'Bcajh97-01A' and the fertile plants 'Bcajh97-01B' was the formation of mature pollen grains. Just after telophase II, the cytokinesis of male meiosis is disturbed in 'Bcajh97-01A' anther, and the formation of intersporal wall does not occur, which in turn results in the failure of mature pollen formation. Genetics analysis indicated that the male sterility was controlled by a single recessive allele. These results suggest that 'Bcajh97-01A' plant is indeed a mutant of male meiosis cytokinesis, and the corresponding mutated gene MMC may be directly involved in male meiotic cytokinesis.
(2) Investigations on the difference among the flower buds transcriptional profiles of male sterile lines compared with the common maintainer line both by cDNA-AFLP and ATH1 genome array analysis, show that gene expression in the flower buds of these various male sterile types are similar but not entirely identical. We conducted expression profiling comparisons between each male sterile line, ajhGMS 'Bcajh97-01A'.polG-CMS 'Bcpol97-05A' and oguCMS 'Bcogu97-06A' and the common maintainer line of B. campestris both by cDNA-AFLP analysis and hybridizing with the ATH1 genome array from Arabidopsis genome. The outcomes indicate that the mutation of the gene which controls the fertility of each plant have caused dramatic changes in expression pattern of genes acting downstream. Expression of numerous genes are blocked or weakened, otherwise that of some other genes are activated or enhanced. But gene expression changes in different male sterile line are not identical due to the different gene mutation. Compared to the maintainer line 'Bcajh97-01B', 93 genes are up-regulated and 158 are down-regulated in 'Bacjh97-01A', 174 genes are up-regulated and 212 are down-regulated in 'Bcpol97-05A' 196 genes are up-regulated and 242 are down-regulated in 'Bcogu97-06A', and thereinto, 37 genes down-regulated and 24 up-regulated simultaneously in three male sterile types are included. There are also many genes sharing common expression pattern between the male sterile lines, including 5 genes down-regulated and 24 up-regulated simultaneously in 'Bcajh97-01A' and 'Bcpol97-05A' 70 genes down-regulated and 9 up-regulated simultaneously in 'Bcajh97-01A' and 'Bcogu97-06A' 70 genes down-regulated and 45 up-regulated simultaneously in 'Bcpol97-05A' and 'Bcogu97-06A'
(3) Function classification of differentially expressed genes reveals that most of the genes involved in pollen and/ or anther development are function-unknown. Expression profiling comparisons of different male sterile lines and their maintainer line increase the number of pollen and/ or anther-expressed and/ or specific gene. We classified the differentially expressed gene detected in the flower buds transcriptome of the male sterile lines and their common maintainer line into several categories, based on the putative function as well as gene ontology annotations derived from homologies. About 50% of these genes have an unknown or hypothetical function. On account of the fact that each of these male sterile plant only differed from the fertile plants line by the failure of pollen formation and anther development, we conclude that the differentially-expressed genes in the flower buds between the male sterile lines and the maintainer line are most likely to act in pollen and/ or anther development. Those differentially-expressed genes with unknown of hypothetical function may represent new candidate proteins involved in pollen and/ or anther development. If this is true, the number of genes expressed and/ or specifically expressed in pollen and anther will be increased to some extent for the identification of these genes.
(4) Based on the function classification, it is found that pollen and/ or anther transcriptome from different plants with unlike genetic pattern possesses similar 'module' but unique composition. When classified the differentially-expressed genes into functional categories, we found that there were similar categories in three individual groups of differentially-expressed genes derived from the comparison between each male sterile line and its maintainer line. However, the proportions of the same category in three groups are not entirely identical. The major categories of genes down-regulated simultaneously in all the male sterile lines are involved in transporter and channel, protein metabolism, electron transport or energy pathways and defense mechanisms and stress response; up-regulated simultaneously in those are involved in transporter and channel, transcription and general metabolism. Further more, many similarities were found between the gene expression in pollen and/ or anther of 'Bcajh97-01A' and 'Bcogu97-06A' Genes related to cell wall biosynthesis and regulation is ranked in the first place of the down-regulated genes both in 'Bcajh97-01A' and 'Bcogu97-06A' and genes related to cytoskeleton and signal transduction also occupy a higher proportion. Otherwise, few genes involved in transcription have been detected in the down-regulated genes both in 'Bcajh97-01A' and 'Bcogu97-06A' In the up-regulated genes in both of these two transcriptome, those related to protein metabolism account for a higher proportion. The transcriptome composition features of flower buds in 'Bcajh97-01A' as well as in 'Bcogu97-06A' are more similar to that of Arabidopsis pollen. In contrast, there is lesser similarity between the transcriptome of flower buds of 'Bcpol97-05A' and the other two. The most distinguished difference is the proportion of genes involved in cell wall biosynthesis and regulation, cytoskeleton and signal transduction. These categories occupy higher proportion in the male sterile pollen of 'Bcpol97-05A' but a lower proportion in the fertile pollen of 'Bcpol97-05B' Thus it can be seen that though there is similar module in the pollen and/ or anther transcriptome of the male sterile line with different genetic pattern, each of them possesses unique composition.
(5) Spatial and temporal expression pattern analysis demonstrates that each gene related to pollen and/ or anther development has a distinct expression dynamic. We investigated the expression pattern of 27 fragments derived from the transcriptome comparison above. Although most of the transcripts were first detected around the time of microspore mitosis and accumulate continuously as the pollen matured, each transcript had a unique expression level at different developmental stages. The expressions of some other fragments were detectable at the earlier stage of pollen development. These results from the expression pattern analysis validate the complexity of gene expression and regulation during pollen and/ or anther development, and demonstrate that activation or silencing and orderly changes of expression level are consistent with the requirements for normal development of pollen and/ or anther and thus for achieving their function. This study also made a relevant attempt to assign genes to different developmental stages of pollen and anther in B. campestris.
(6) Expression pattern and functional analysis of BcMF2 indicate that it may encode a novel PG involved in pollen intine deposition. Spatial and temporal expression pattern analysis was performed for BcMF2, a gene encoding PG isolated from flower buds of 'Bcajh97-01B' previously. BcMF2 transcript was first detected in the flower buds at stage 3, which corresponded to the tetrad stage of microspore development, though it was comparatively weak. The signal then became stronger up to the flower buds at stage 4, which corresponded to the uninucleate stage, and then declined in the flower buds at stage 5 full of mature pollen grains. This expression pattern considered it an "early" gene in pollen. Functional analysis of BcMF2 by anti-sense technique revealed that it might be involved in pollen wall development. The germination of pollen from transgenic plants with anti-sense BcMF2 was affected. About 80% transgenic pollen formed a balloon-tipped pollen tube during in vitro germination. Though the pollen tube could penetrate through the stigma, it stopped growth in the style. Further investigation on the configuration and the development of transgenic pollen revealed that all the pollens were malformed, and the number of germination furrow as well as its distribution was anomalistic. The intine of transgenic pollen thickened unconventionally during pollen development. It's a hypothesis that the silence of BcMF2 may result in the disturbance in pectin metabolism in the pollen wall and in turn leads to the aborted intine and germination furrow. For there is no similarity found in sequence and expression pattern between BcMF2 and the known PG gene acting at the earlier stage, BcMF2 is considered as a novel gene encoding PG whose transcript is accumulated at the earlier stage.
(7) Another PG gene named BcMF9 in B. campestris was isolated and characterized as a gene playing roles both in intine and exine formation. BcMF9 was amplified from a fragment, BBS13/BPO023 detected in the flower buds of 'Bcajh97-01B' Alignment against the published sequences revealed the presence of highly conserved domains and features among plant polygalacturonases in the deduced amino acid sequence of BcMF9, and molecular phylogenetic analysis put it into the cluster of pollen-expressed PG. BcMF9 was considered as a "late" gene in pollen, because its expression signal was first detectable in the microspore at the tetrad stage, and became stronger at the uninucleate stage and maintained till pollen matured. Its transcript was also detected in the tapetum cell from tetrad stage to the degeneration of tapetal cells. Functional analysis of BcMF9 by anti-sense technique revealed that it may play roles both in intine and exine formation. The in vitro germination percentage of transgenic pollen was dramatically reduced. About 81% pollen tubes burst during germination. The pollen tube could penetrate through the stigma, but it stopped growth in the style soon after that. Further investigation revealed that silence of BcMF9 led to the produce of malformed pollen with a flat meshwork on the face and anomalous germination furrows. The intine of transgenic pollen thickened unconventionally during the pollen development and at the late stage, the bacula and the tectum fell off from the exine, which subsequently resulted in the overflowing of tryphine. These results strongly indicate that BcMF9 may play an important role in pollen wall formation. Furthermore, we found that degeneration of tapetum was accelerated in the transgenic anther. Microspore development was normal before the abnormity in tepetum occurring, but soon after the abnormity occurring, it was defected in exine formation. We speculate that the silence of BcMF9 disturbes the normal process of programmed cell death (PCD) in the tapetal cells, and starts an alternative cell death process which accelerated the degeneration process. Break of the normal PCD in tapetum may alter the releasing of tapetal wall materials onto the pollen grains and thus affect the complete deposition of pollen wall. Alternatively, BcMF9 may act directly in the formation of pollen wall. It may be expressed in the tepetum and then is secreted into the locule and located on the microspore during the pollen development, for the putative signal peptide in BcMF9 protein and the unique expression pattern of BcMF9.
(8) Comparison of BcMF2, BcMF6 and BcMF9, three members of the PG gene family reveals that they may play different role in pollen development. The gene structure, expression pattern, gene evolution and biological function of three PG genes, BcMF2, BcMF6 and BcMF9 were analyzed. Though it was found that these PG genes accumulated in the same developmental stages, the expression level in each stage and the dynamics of expression of them are different. The evolutions of them in family Cruciferae are not coincident, and the silences of them result in similar but not identical phenotype. These results illuminate that the function and the pattern of gene activity of BcMF2, BcMF6 and BcMF9 may be different.
(9) A novel gene named BcMFlO predominantly expressed in flower buds of 'Bcajh97-01B' and ? 'Bcpol97-05A' but silenced in those of 'Bcajh97-01A' and 'Bcogu97-06A' was isolated and characterized. This gene was amplified from a fragment, BBS31I BPO079 detected in the comparison of flower buds transcriptome of male sterile lines and the maintainer line. Bioinformatical analysis of the sequence revealed several patterns which were involved in protein location, cell adhesion and signal transduction associated with cell proliferation, differentiation, and apoptosis. It is supposed that BcMF10 may represent a new candidate protein involved in signal transduction. RT-PCR and in situ hybridization was performed to investigate the expression pattern of BcMF10, and the result indicated it was an "early" gene in pollen. In 'Bcajh97-01B' BcMF10 has its transcript began to be detectable in pollen mother cell and tapetum at the early developmental stage, and became strong during the meiosis. Then, the expression decreased gradually up to the mature pollen. However, the location of its transcript has not changed. Functional analysis by RNAi technique revealed that silence of BcMF10 blocked the normal germination of pollen. About 75% transgenic pollens could not germinate in vitro, and 15.6% grew with a burst tip. But the burst pollen tube could grow to some extent and then spray the content, which distinguished it from the other burst tube observed generally with an immediate burst soon after germination. We suppose that BcMF10 may encode a protein synthesized in the tapetum and secreted out of the cell which plays a role in the secretion of nutriments or cell wall materials for the pollen grains.
(1)形态和细胞学观察发现‘Bcajh97-01A’为雄性减数分裂的胞质分裂突变体mmc，对应基因MMC可能编码特异作用于雄性减数分裂的蛋白质。形态和细胞学观察发现，ajhGMS两用系不育株系‘Bcajh97-01A’与可育株系'Bcajh97-01B'的差异仅表现在花粉花药发育上，‘Bcajh97-01A’小孢子在减数分裂末期的胞质分裂中出现异常，不能形成胞间壁而导致四分体形成及后续的花粉发育各过程受阻，最终引起花粉败育。遗传学分析表明该不育性状由单基因位点控制，因此，‘Bcajh97-01A’实际上是一个受单基因位点控制的雄性减数分裂的胞质分裂突变体mmc(male meiotic cytokinesis)，其对应基因MMC可能编码一个特异与雄性减数分裂核分裂完成后的胞质分裂相关的蛋白质。
(2)利用cDNA-AFLP技术结合拟南芥基因芯片分析白菜不同不育系之间及与其共同保持系之间的花蕾转录组差异，发现不同遗传模式不育系花蕾基因表达有相似之处，但不尽相同。利用cDNA-AFLP技术及拟南芥ATH1基因芯片，对白菜ojhGMS‘Bcajh97-01A’、polG-CMS‘Bcpol97-05A’和oguCMS‘Bcogu97-06A’，以及它们共同的保持系‘Bcajh97-01B’进行了花蕾转录组比较分析，发现不同的不育基因作用下均引起花蕾正常转录组的巨大变化，不育基因下游众多基因的表达受到抑制，也有相当一部分的基因被激活或表达上升，但作用的不育基因不同，引起的下游基因表达变化也不尽相同。与同一保持系‘Bcajh97-01B’相比，检测到在‘Bcajh97-01A’花蕾中上调表达基因93个、下调表达基因158个；在‘Bcpol97-05A’花蕾中上调表达基因174个、下调表达基因212个；在‘Bcogu97-06A’花蕾中上调表达基因196个、下调表达基因242个。其中。在三种不育系中共同下调表达的基因有37个，上调表达的24个；‘Bcajh97-01A’和‘Bcpol97-05A’共同下调和共同下调表达的基因各5个；‘Bcajh97-01A’和‘Bcogu97-06A’共同下调表达基因为70个，共同上调表达基因9个；‘Bcpol97-05A’和‘Bcogu97-06A’共同下调表达基因7O个，共同上调表达的基因45个。
(3)通过差异表达基因功能分类，发现许多花粉花药发育相关基因功能未知，白菜不同不育系与保持系的花蕾转录组差异分析扩充了植物花粉表达或特异表达基因的数目。对在三种不育系与其共同保持系花蕾转录组中检测到的差异表达基因进行功能分类，发现约50％的基因功能未知。基于三种不育系和保持系花蕾的差别仅表现在花药发育和花粉形成上，在不育系和保持系花蕾中差异表达的基因最可能与花粉花药发育相关。我们发现的这些差异表达基因在一定程度上扩充了植物花粉花药表达和特异表达基因的数目。
(4)基于基因功能分类对不同不育系与保持系花蕾基因表达差异进行的分析表明，不同遗传模式花粉花药转录组尽管具有相似的模块，但具有不同的组成特征。对差异表达基因进行功能分类发现，不育系与保持系差异表达的三组基因可分为大致相同的功能类别，但每一功能类别在每组差异基因中所占的比例不尽相同。在三种不育系花蕾中下调均较多的已知功能的基因与转运通道、蛋白质代谢、电子传递和能量途径、防卫和胁迫反应相关，上调均较多的基因与转运通道、转录调控和普通新陈代谢相关。除此之外，‘Bcajh97-01A’与‘Bcogu97-06A’在花粉花药基因表达上具有更大的共性，在两者花蕾中均下调表达的基因中，细胞壁合成与调控基因排在第一位，细胞骨架蛋白及信号转导相关基因也占了较大的比例，而转录相关基因的比例较小；两者下调表达的基因中，蛋白质代谢相关基因均比较多。‘Bcajh97-01A’与‘Bcogu97-06A’的花蕾转录组的组成特征与拟南芥花粉转录组特征较一致。相比之下，‘Bcpol97-05A’花蕾转录组的组成成分与其他两种不育系有较大的差别。突出表现在细胞壁合成与调控基因、细胞骨架蛋白基因以及信号转导相关基因在不育系花蕾中占较大的比例，而保持系与其相比，这三种类别基因的表达并不占优势，反而更少。由此可见，不同遗传模式花粉花药转录组尽管具有相似的模块，但组成特征各异。
(5)时空表达模式分析发现三种不育系与其共同保持系花蕾差异表达的基因具不同的表达动态。对27条在三种不育系和保持系花蕾中差异表达的片段进行了时空表达模式分析，发现大多数差异片段所代表的基因表达在小孢子有丝分裂前后开始被检测到，在花粉成熟过程中继续积累，但在不同组织和发育阶段中的表达水平不尽相同。此外也有在花粉发育早期就检测到表达的基因。我们的研究结果证实了花粉花药发育中的基因表达调控是极其复杂的。在花粉花药发育的不同时期表达或不表达，以及表达水平有序地发生变化都是与花粉花药正常发育、实现其功能所需相对应的。我们的研究为明确白菜花粉花药发育不同时期的基因群作出了有益的探索。
(6)表达和功能分析发现BcMF2可能是一个与花粉内壁发育相关的新PG基因。对本实验室前期从‘Bcajh97-01B’花蕾中分离得到的PG基因BcMF2进行时空表达模式分析，发现其转录本最早在四分体时期的花蕾中开始出现，随后在单核花粉粒时期的花蕾中表达量达到最高，之后，随着花粉发育的进行表达量逐渐降低直至花粉成熟，表明其属于花粉表达“早期”基因。利用反义RNA技术研究BcMF2的功能，发现BcMF2表达受抑制的转基因植株花粉体外正常萌发率大幅度降低。约80％的花粉管生长到一定程度后顶端膨大形成泡状结构，花粉管能穿过柱头组织，但在花柱道中的生长停止。进一步观察显示转基因植株花粉畸形，萌发沟的数目及分布不规则，萌发沟在形成过程中位置和数目紊乱，花粉粒内壁异常增厚。推测BcMF2表达受抑制影响了花粉内壁中果胶的动态代谢平衡而导致萌发沟及花粉粒内壁发育的异常。BcMF2与已知的在花粉发育早期起作用的PG基因没有序列及表达特点的相似性，因此它可能是一个新的在花粉发育早期表达的PG基因。
(7)表达和功能分析发现分离获得的另一个白菜PG基因BcMF9可能参与花粉内壁和外壁的发育。分离获得在可育株‘Bcajh97-01B’花蕾中特异表达的差异片段BBS13／BPO023的全长序列BcMF9，发现该基因具有典型的PG基因结构特征。分子进化分析表明其属于花粉表达或特异表达的PG基因类群。时空表达模式分析显示BcMF9属于花粉表达“晚期”基因，因其表达最早出现在四分小孢子中，在单核小孢子中开始增强，然后一直持续较高水平的表达直到花粉成熟。BcMF9的转录本亦在绒毡层中出现，从四分体时期开始直到绒毡层降解退化，一直在绒毡层细胞中维持很强的表达信号。转反义BcMF9的植株花粉离体正常萌发率明显降低，约81％的花粉萌发时花粉管爆裂。花粉管能穿过柱头组织，但不久便停止生长。扫描电镜和透射电镜观察显示，BcMF9表达受抑制产生外壁网纹浅的畸形花粉，花粉粒萌发沟处内壁增厚异常，萌发沟的位置和数目紊乱，花粉发育晚期外壁的覆盖层和基粒棒部分脱落，含油层外溢，说明BcMF9可能参与花粉壁的发育。此外，发现转反义BcMF9植株花药绒毡层降解速度加快，绒毡层发育发生异常前的小孢子发育正常，而异常后的花粉外壁发育受影响。推测BcMF9表达受抑制可能破坏了绒毡层细胞的程序性死亡过程，启动了另一个死亡途径，加速了绒毡层的解体。绒毡层正常程序性细胞死亡(programmed cell death，PCD)的破坏，导致其向花粉外壁输送物质过程的改变，从而影响了花粉壁的正常发育。但由于BcMF9编码的蛋白在其N端有一段信号肽序列，而它的表达首先出现在四分体时期的绒毡层和小孢子中，且在绒毡层中的表达信号要比在小孢子中强。因此推测其功能发挥还有另一个可能：BcMF9在绒毡层表达，表达产物分泌到花粉囊腔，进而在花粉壁的发育中直接起作用。
(8)比较分析发现PG基因家族成员BcMF2、BcMF6和BcMF9在花粉发育中可能扮演各自不同的角色。从基因序列特征、表达模式特点、基因进化关系及生物学功能等方面比较三个花粉发育相关的PG基因BcMF2、BcMF6和BcMF9的异同，发现尽管3个PG基因的表达所出现的发育阶段一致，但三者在各个发育阶段的表达量及表达变化的趋势各不相同。三者在十字花科物种中的进化特点也不完全一致。3个PG基因的表达受抑制引发了类似的结果，但并不尽相同，说明它们在花粉发育中所起的作用及作用模式可能各异。
(9)分离获得在‘Bcajh97-01B’和‘Bcpol97-05A’花蕾中高水平表达的基因BcMF10，序列特征分析提示其可能与信号转导途径相关，RNAi研究发现BcMF10表达受抑制导致花粉萌发异常。分离获得在‘Bcajh97-01B’和‘Bcpol97-05A’花蕾中高水平表达，但在‘Bcajh97-01A’和‘Bcogu97-06A’花蕾中沉默的基因片段BBS31／BPO079的全长序列BcMF10。利用生物信息学分析发现预测的BcMF10氨基酸序列中具有可能与细胞增殖、分化和PCD相关的蛋白激酶C磷酸化位点，参与调控信号转导相关的蛋白激酶的酪氨酸磷酸化位点，及其它与细胞内信号转导、蛋白定位以及黏附等过程有关的位点。推测BcMF10编码的蛋白可能在花粉发育过程的信号转导中发挥重要作用。利用RT-PCR(reverse transcriptase PCR，反转录PCR)和原位杂交分析了BcMF10的时空表达特点，发现其转录本最早出现在‘Bcajh97-01B’花药发育早期的花粉母细胞和绒毡层细胞中，随后在减数分裂时期的小孢子和绒毡层中表达量进一步提高，之后随着发育的进行，表达量逐渐降低，但表达部位没有改变，在花粉成熟期消失，属于花粉表达“早期”基因。通过构建含BcA9绒毡层特异启动子的BcMF10 RNAi表达载体，利用转基因阻断内源BcMF10的表达，发现75％左右的转基因植株花粉不能正常萌发，约15.6％的花粉能长出花粉管，但花粉管长到一定长度时顶端爆裂，正常萌发率只有10.5％，说明BcMF10表达受抑制影响了花粉的正常萌发。但与通常发现的花粉管一伸出萌发孔即发生爆裂的现象不同，BcMF10表达受抑制植株花粉管能长到一定长度，然后将内容物从顶端喷出，花粉管生长停止，推测BcMF10也可能与花粉壁的发育相关。根据BcMF10预测蛋白质的结构特征及其在早期绒毡层细胞和小孢子中共表达的特点，可以认为其可能作用于细胞壁发育过程中绒毡层与花粉的信号传递过程。
The application of male sterility becomes an important way in heterosis breeding to simplify the seed production procedures and reduce the production cost. The utilization of hererosis is prevalent in cruciferae crops breeding, and the male sterile line is a useful system for their hybrid seed production. However, the occurrence and the mechanism of plant male sterility still remain mysteries that have not been unveiled thoroughly. For male sterility mainly manifests in pollen abortion, to make clear of the whole process of pollen development and the mechanism underlined thus become the basis point for understanding of male sterility. Pollen development is regulated by a complex regulatory network that comprises a wide variety of genes. Transcriptional profiling on the mutant compared with its wild-type is a successful approach to investigate the dynamic gene expression during pollen development and to detect genes involved in this biological process, and ultimately to help to understand the characteristic of pollen development and the corresponding molecular mechanism. In this study, we established three different male sterile types, 'Aijiaohuang' genic male sterility AB line (ajhGMS 'Bcajh97-01A/B'), 'Polima' genic-cytoplasmic male sterility (polG-CMS 'Bcpol97-05A') and 'Ogura' cytoplasmic male sterility (oguCMS 'Bcogu97-06A') in Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis. These male sterile lines share a common maintainer line. Then, we conducted profiling comparisons between flower buds of these male sterile lines and their maintainer line using complementary deoxyribonucleic acid amplified fragment length polymorphism (cDNA-AFLP) technology together with ATH1 genome array analysis. By analyzing the changes in expression pattern of genes acting downstream due to the gene mutation in different male sterile plants, plenty of candidate genes involved in pollen development were identified, and thus laid a foundation for establishing the profiles of gene expression in different types of male sterile lines. After that, three genes were selected for further characterization, including two putative polygalacturonase (PG) genes, BcMF2 {Brassica campestris male fertility 2) and BcMF9 {Brassica campestris male fertility 9) which may be related to cell wall biosynthesis and regulation, and one novel gene, BcMFW {Brassica campestris male fertility 10) with unknown function. Anti-sense RNA or RNA interference (RNAi) technique were used to investigate the function of the three genes, and morphologic, molecular biology as well as cytology analysis were carried through in functional assignment. Comparison of gene structure, expression pattern and biological function of three members, BcMF2, BcMF6 {Brassica campestris male fertility 6) and BcMF9 in PG gene family were also performed in this study. The results obtained are as follows.
(1) Observation on the pollen development in male sterile line 'Bcajh97-01A' suggests that it's a mutant of male meiosis cytokinesis in B. campestris, and the corresponding gene, MMC may encode a protein acting specifically in male meiosis. When observed the morphology and cytology character of the ajhGMS 'Bcajh97-01A/B' we found that the only detectable difference between the male sterile plants 'Bcajh97-01A' and the fertile plants 'Bcajh97-01B' was the formation of mature pollen grains. Just after telophase II, the cytokinesis of male meiosis is disturbed in 'Bcajh97-01A' anther, and the formation of intersporal wall does not occur, which in turn results in the failure of mature pollen formation. Genetics analysis indicated that the male sterility was controlled by a single recessive allele. These results suggest that 'Bcajh97-01A' plant is indeed a mutant of male meiosis cytokinesis, and the corresponding mutated gene MMC may be directly involved in male meiotic cytokinesis.
(2) Investigations on the difference among the flower buds transcriptional profiles of male sterile lines compared with the common maintainer line both by cDNA-AFLP and ATH1 genome array analysis, show that gene expression in the flower buds of these various male sterile types are similar but not entirely identical. We conducted expression profiling comparisons between each male sterile line, ajhGMS 'Bcajh97-01A'.polG-CMS 'Bcpol97-05A' and oguCMS 'Bcogu97-06A' and the common maintainer line of B. campestris both by cDNA-AFLP analysis and hybridizing with the ATH1 genome array from Arabidopsis genome. The outcomes indicate that the mutation of the gene which controls the fertility of each plant have caused dramatic changes in expression pattern of genes acting downstream. Expression of numerous genes are blocked or weakened, otherwise that of some other genes are activated or enhanced. But gene expression changes in different male sterile line are not identical due to the different gene mutation. Compared to the maintainer line 'Bcajh97-01B', 93 genes are up-regulated and 158 are down-regulated in 'Bacjh97-01A', 174 genes are up-regulated and 212 are down-regulated in 'Bcpol97-05A' 196 genes are up-regulated and 242 are down-regulated in 'Bcogu97-06A', and thereinto, 37 genes down-regulated and 24 up-regulated simultaneously in three male sterile types are included. There are also many genes sharing common expression pattern between the male sterile lines, including 5 genes down-regulated and 24 up-regulated simultaneously in 'Bcajh97-01A' and 'Bcpol97-05A' 70 genes down-regulated and 9 up-regulated simultaneously in 'Bcajh97-01A' and 'Bcogu97-06A' 70 genes down-regulated and 45 up-regulated simultaneously in 'Bcpol97-05A' and 'Bcogu97-06A'
(3) Function classification of differentially expressed genes reveals that most of the genes involved in pollen and/ or anther development are function-unknown. Expression profiling comparisons of different male sterile lines and their maintainer line increase the number of pollen and/ or anther-expressed and/ or specific gene. We classified the differentially expressed gene detected in the flower buds transcriptome of the male sterile lines and their common maintainer line into several categories, based on the putative function as well as gene ontology annotations derived from homologies. About 50% of these genes have an unknown or hypothetical function. On account of the fact that each of these male sterile plant only differed from the fertile plants line by the failure of pollen formation and anther development, we conclude that the differentially-expressed genes in the flower buds between the male sterile lines and the maintainer line are most likely to act in pollen and/ or anther development. Those differentially-expressed genes with unknown of hypothetical function may represent new candidate proteins involved in pollen and/ or anther development. If this is true, the number of genes expressed and/ or specifically expressed in pollen and anther will be increased to some extent for the identification of these genes.
(4) Based on the function classification, it is found that pollen and/ or anther transcriptome from different plants with unlike genetic pattern possesses similar 'module' but unique composition. When classified the differentially-expressed genes into functional categories, we found that there were similar categories in three individual groups of differentially-expressed genes derived from the comparison between each male sterile line and its maintainer line. However, the proportions of the same category in three groups are not entirely identical. The major categories of genes down-regulated simultaneously in all the male sterile lines are involved in transporter and channel, protein metabolism, electron transport or energy pathways and defense mechanisms and stress response; up-regulated simultaneously in those are involved in transporter and channel, transcription and general metabolism. Further more, many similarities were found between the gene expression in pollen and/ or anther of 'Bcajh97-01A' and 'Bcogu97-06A' Genes related to cell wall biosynthesis and regulation is ranked in the first place of the down-regulated genes both in 'Bcajh97-01A' and 'Bcogu97-06A' and genes related to cytoskeleton and signal transduction also occupy a higher proportion. Otherwise, few genes involved in transcription have been detected in the down-regulated genes both in 'Bcajh97-01A' and 'Bcogu97-06A' In the up-regulated genes in both of these two transcriptome, those related to protein metabolism account for a higher proportion. The transcriptome composition features of flower buds in 'Bcajh97-01A' as well as in 'Bcogu97-06A' are more similar to that of Arabidopsis pollen. In contrast, there is lesser similarity between the transcriptome of flower buds of 'Bcpol97-05A' and the other two. The most distinguished difference is the proportion of genes involved in cell wall biosynthesis and regulation, cytoskeleton and signal transduction. These categories occupy higher proportion in the male sterile pollen of 'Bcpol97-05A' but a lower proportion in the fertile pollen of 'Bcpol97-05B' Thus it can be seen that though there is similar module in the pollen and/ or anther transcriptome of the male sterile line with different genetic pattern, each of them possesses unique composition.
(5) Spatial and temporal expression pattern analysis demonstrates that each gene related to pollen and/ or anther development has a distinct expression dynamic. We investigated the expression pattern of 27 fragments derived from the transcriptome comparison above. Although most of the transcripts were first detected around the time of microspore mitosis and accumulate continuously as the pollen matured, each transcript had a unique expression level at different developmental stages. The expressions of some other fragments were detectable at the earlier stage of pollen development. These results from the expression pattern analysis validate the complexity of gene expression and regulation during pollen and/ or anther development, and demonstrate that activation or silencing and orderly changes of expression level are consistent with the requirements for normal development of pollen and/ or anther and thus for achieving their function. This study also made a relevant attempt to assign genes to different developmental stages of pollen and anther in B. campestris.
(6) Expression pattern and functional analysis of BcMF2 indicate that it may encode a novel PG involved in pollen intine deposition. Spatial and temporal expression pattern analysis was performed for BcMF2, a gene encoding PG isolated from flower buds of 'Bcajh97-01B' previously. BcMF2 transcript was first detected in the flower buds at stage 3, which corresponded to the tetrad stage of microspore development, though it was comparatively weak. The signal then became stronger up to the flower buds at stage 4, which corresponded to the uninucleate stage, and then declined in the flower buds at stage 5 full of mature pollen grains. This expression pattern considered it an "early" gene in pollen. Functional analysis of BcMF2 by anti-sense technique revealed that it might be involved in pollen wall development. The germination of pollen from transgenic plants with anti-sense BcMF2 was affected. About 80% transgenic pollen formed a balloon-tipped pollen tube during in vitro germination. Though the pollen tube could penetrate through the stigma, it stopped growth in the style. Further investigation on the configuration and the development of transgenic pollen revealed that all the pollens were malformed, and the number of germination furrow as well as its distribution was anomalistic. The intine of transgenic pollen thickened unconventionally during pollen development. It's a hypothesis that the silence of BcMF2 may result in the disturbance in pectin metabolism in the pollen wall and in turn leads to the aborted intine and germination furrow. For there is no similarity found in sequence and expression pattern between BcMF2 and the known PG gene acting at the earlier stage, BcMF2 is considered as a novel gene encoding PG whose transcript is accumulated at the earlier stage.
(7) Another PG gene named BcMF9 in B. campestris was isolated and characterized as a gene playing roles both in intine and exine formation. BcMF9 was amplified from a fragment, BBS13/BPO023 detected in the flower buds of 'Bcajh97-01B' Alignment against the published sequences revealed the presence of highly conserved domains and features among plant polygalacturonases in the deduced amino acid sequence of BcMF9, and molecular phylogenetic analysis put it into the cluster of pollen-expressed PG. BcMF9 was considered as a "late" gene in pollen, because its expression signal was first detectable in the microspore at the tetrad stage, and became stronger at the uninucleate stage and maintained till pollen matured. Its transcript was also detected in the tapetum cell from tetrad stage to the degeneration of tapetal cells. Functional analysis of BcMF9 by anti-sense technique revealed that it may play roles both in intine and exine formation. The in vitro germination percentage of transgenic pollen was dramatically reduced. About 81% pollen tubes burst during germination. The pollen tube could penetrate through the stigma, but it stopped growth in the style soon after that. Further investigation revealed that silence of BcMF9 led to the produce of malformed pollen with a flat meshwork on the face and anomalous germination furrows. The intine of transgenic pollen thickened unconventionally during the pollen development and at the late stage, the bacula and the tectum fell off from the exine, which subsequently resulted in the overflowing of tryphine. These results strongly indicate that BcMF9 may play an important role in pollen wall formation. Furthermore, we found that degeneration of tapetum was accelerated in the transgenic anther. Microspore development was normal before the abnormity in tepetum occurring, but soon after the abnormity occurring, it was defected in exine formation. We speculate that the silence of BcMF9 disturbes the normal process of programmed cell death (PCD) in the tapetal cells, and starts an alternative cell death process which accelerated the degeneration process. Break of the normal PCD in tapetum may alter the releasing of tapetal wall materials onto the pollen grains and thus affect the complete deposition of pollen wall. Alternatively, BcMF9 may act directly in the formation of pollen wall. It may be expressed in the tepetum and then is secreted into the locule and located on the microspore during the pollen development, for the putative signal peptide in BcMF9 protein and the unique expression pattern of BcMF9.
(8) Comparison of BcMF2, BcMF6 and BcMF9, three members of the PG gene family reveals that they may play different role in pollen development. The gene structure, expression pattern, gene evolution and biological function of three PG genes, BcMF2, BcMF6 and BcMF9 were analyzed. Though it was found that these PG genes accumulated in the same developmental stages, the expression level in each stage and the dynamics of expression of them are different. The evolutions of them in family Cruciferae are not coincident, and the silences of them result in similar but not identical phenotype. These results illuminate that the function and the pattern of gene activity of BcMF2, BcMF6 and BcMF9 may be different.
(9) A novel gene named BcMFlO predominantly expressed in flower buds of 'Bcajh97-01B' and ? 'Bcpol97-05A' but silenced in those of 'Bcajh97-01A' and 'Bcogu97-06A' was isolated and characterized. This gene was amplified from a fragment, BBS31I BPO079 detected in the comparison of flower buds transcriptome of male sterile lines and the maintainer line. Bioinformatical analysis of the sequence revealed several patterns which were involved in protein location, cell adhesion and signal transduction associated with cell proliferation, differentiation, and apoptosis. It is supposed that BcMF10 may represent a new candidate protein involved in signal transduction. RT-PCR and in situ hybridization was performed to investigate the expression pattern of BcMF10, and the result indicated it was an "early" gene in pollen. In 'Bcajh97-01B' BcMF10 has its transcript began to be detectable in pollen mother cell and tapetum at the early developmental stage, and became strong during the meiosis. Then, the expression decreased gradually up to the mature pollen. However, the location of its transcript has not changed. Functional analysis by RNAi technique revealed that silence of BcMF10 blocked the normal germination of pollen. About 75% transgenic pollens could not germinate in vitro, and 15.6% grew with a burst tip. But the burst pollen tube could grow to some extent and then spray the content, which distinguished it from the other burst tube observed generally with an immediate burst soon after germination. We suppose that BcMF10 may encode a protein synthesized in the tapetum and secreted out of the cell which plays a role in the secretion of nutriments or cell wall materials for the pollen grains.
引文
白书农．植物发育生物学，北京：北京大学出版社，2003
布坎南 BB，格鲁依森姆 W，琼斯 RL 主编 (瞿礼嘉，顾红雅，白书农，赵进东，陈章良主译)．植物生物化学与分子生物学，北京：科学出版社，2004
曹家树，曹寿椿，易清明．白菜及相邻类群基因组 DNA 的 RAPD 分析．园艺学报，1995，22：47～52
曹双河，张相岐，张爱民．光 (温) 敏雄性不育的调控机理和分子遗传学研究进展．植物学通报，2005，22(1)：19～26
陈新建，陈占宽，刘国顺，刘鸿先．植物对水分胁迫响应的分子机制与抗逆基因工程的研究进展．热带亚热带植物学报，2000，8(1)：81～90
崔辉梅，曹家树，张明龙，姚祥坦，向珣．白菜和芜菁 Ogura 型雄性不育系与保持系的获得及其细胞学观察．园艺学报，2004，31(4)：467～471
李德葆，周雪平，许建平等．基因工程操作技术，上海：上海科技出版社，1996
利容千．几种农作物雄性不育的细胞学研究．武汉大学学报，1978，83～96
刘乐承，余小林，叶纨芝，向珣，曹家树．白菜雄性不育相关基因 BcMF3 功能的反义 RNA 验证．浙江大学学报 (农业与生命科学版)，2006a，32(5)：473～478
刘乐承，向珣，曹家树．白菜雄性不育相关基因 BcMF4 基因功能的 RNAi 验证．遗传，2006b，28(11)：1428～1434
刘仁虎，赵建伟，肖勇，孟金．拟南芥 cDNA 芯片和油菜-拟南芥比较作图相结合筛选甘蓝型油菜抗菌核病先验基因．中国科学C辑生命科学，2005，35(1)：13～21
刘玉梅．甘蓝显性细胞核雄性不育的细胞学特征、生化基础及其分子标记的研究．博士学位论文，2003
龙欢，姚家玲，涂金星．3 种甘蓝型油菜雄性不育系花药发育的细胞学研究．华中农业大学学报．2005，24：570～575
沈俊儒，吴建勇，张创，刘平武，杨光圣．甘蓝型油菜华油杂6号及其亲本的基因差异表达研究．中国农业科学，2006，39(1)：23～28
孙敬三，钱迎倩．植物细胞学研究方法，北京：科学出版社，1987
王台，丁兆军．减数分裂及其基因研究进展．科学通报，2002，47(4)：241～248
王永勤，曹家树，符庆功，余小林，叶纨芝，向珣．利用 cDNA-AFLP 技术分析白菜核雄性不育两用系的表达差异．中国农业科学，2003，36(5)：557～560
王永勤，余小林，曹家树．白菜小孢子发育相关基因 BcMF3 的分离及其表达特征分析．遗传学报，2004，31(11)： 1302～1308
余小林，曹家树，许立奎，齐文雯，王小静．优化白菜类蔬菜遗传转化体系的研究．浙江大学学报 (农业与生命科学版)，2005，31(5)：529～534
余小林．白菜雄性不育相关基因 CYP86MF 的功能验证及人工不育系的创建．博士学位论文，2002
张强，黄鹂，曹家树．白菜多聚半乳糖醛酸酶基因 BcMF6 的克隆、序列分析及其表达．园艺学报，2007，34(1)：117～124
张强．白菜花粉外壁蛋白基因 BcMF5 和多聚半乳糖醛酸酶基因 BcMF6 的分离及其功能验证．博士学位论文，2006
张弢．十字花科植物 BcMF2 和 BcMF4 同源基因的克隆、表达及进化关系的研究．博士学位论文，2005
Aarts MG，Hodge R, Kalantidis K, Forack D, Wilson ZA, Mulligan BJ, Stiekema WJ, Scott R, Pereira A. The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes. Plant J, 1997, 12(3): 615～623
Agashe B, Prasad CK, Siddiqi I. Identification and analysis of DYAD: a gene required for meiotic chromosome organization and female meiotic progression in Arabidopsis. Development, 2002, 129:3935～3943
Ahmad N, Gupta S, Husain MM, Heiskanen KM, Mukhtar H. Differential antiproliferative and apoptotic response of sanguinarine for cancer cells versus normal cells. Clinical Cancer Research, 2000, 6:1524～1528
Ahmed K, Davis AT, Wang H, Faust RA, Yu S, Tawfic S. Significance of protein kinase CK2 nuclear signaling in neoplasia. J Cell Biochem Suppl, 2000, 35:130～5
Albani D, Altosaar I, Arnison PG, Fabijanski SF. A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus: sequences in its 5_flanking region are conserved in other pollen-specific promoters. Plant Mol Biol, 1991, 16:501～513
Albertsen MC, Palmer RG. A comparative light-and electron-microscopic study of microsporogenesis in male sterile (MS1) and male fertile soybeas (Glycine max (L.) Merr.). Am J Bot, 1979, 66:253～265
Allen RL, Lonsdale DM. Molecular characterization of one of the maize polygalacturonase gene family members which are expressed during late pollen development. Plant J, 1993, 3(2): 261～271
Amagai M, Ariizumi T, Endo M, Hatakeyama K, Kuwata C, Shibata D, Toriyama K, Watanabe M. Identification of anther-specific genes in a cruciferous model plant, Arabidopsis thaliana, by using a combination of Arabidopsis macroarray and mRNA derived from Brassica oleracea. Sex Plant Reprod, 2003, 15:213～220
Araya A, Zabaleta E, Blanc V, Brgu D, Hemould M, M ouras A, Litvak S. RNA editing in plant mitochondria, cytoplasmic male sterility and plant breeding. Electron J Biotech, 1998, 1:31～39
Ariizumi T, Hatakeyama K, Hinata K, Sato S, Kato T, Tabata S, Toriyama K. A novel male-sterile mutant of Arabidopsis thaliana, faceless pollen-1, produces pollen with a smooth surface and an acetolysis-sensitive exine. Plant Mol Biol, 2003, 53:107～116
Ariizumi T, Hatakeyama K, Hinata K, Sato S, Kato T, Tabata S, Toriyama K. The HKM gene, which is identical to the MS1 gene of Arabidopsis thaliana, is essential for primexine formation and exine pattern formation. Sex Plant Reprod, 2005, 18: 1～7
Armstrong SJ, Caryl AP, Jones GH, Christopher F, Franklin H. Asy1, a protein required for meiotic chromosome synapsis, localizes to axis-associated chromatin in Arabidopsis and Brassica. J Cell Sci, 2002, 115: 3645～3655
Asano T, Katagiri H, Takata K, Lin JL, Ishihara H, Inukai K, Tsukuda K, Kikuchi M, Hirano H, Yazaki Y. The role of N-glycosylation of GLUT1 for glucose transport activity. J Biol Chem, 1991, 266: 24632～24636
Athanasiou A, Khosravi D, Tamari F, Shore JS. Characterization and localization of short-specific polygalacturonase in distylous Turnera subulata (Turneraceae). Am J Bot, 2003, 90(5):675～682
Atkinson A, Health R, Simposon R, Clarke A, Anderson M. Proteinase inhibition in Nicotiana alata stigmas are derived from a precursor protein which is.processed into five homologous inhibitors. Plant Cell, 1993, 5:203～213
Azumi Y, Liu D, Zhao D, Li W, Wang G, Hu Y, Ma H. Homolog interaction during meiotic prophase I in Arabidopsis required the SOLO DANCERS gene encoding a novel cyclin-like protein. EMBO J, 2002, 21: 3081～3095
Bachem CWB, van der Hoeven RS, de Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF. Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development. Plant J, 1996, 9:745～753
Bai X, Peirson BN, Dong F, Xue C, Makaroff CA. Isolation and characterization of SYN1, a RAD21-like gene essential for meiosis in Arabidopsis. Plant Cell, 1999, 11: 417～430
Balk J, Leaver CJ. The PET1-CMS mitochondrial mutation in sunflower is associated with premature programmed cell death and cytochrome c release. Plant Cell, 2001, 13: 1803～1818
Beadle GM. A gene in Zea mays for failure of cytokinesis during meiosis. Cytologia, 1932, 3: 142～155
Becket J, Boavida LC, Carneiro J, Haury M, Feijo JA. Transcriptional profiling of Arabidopsis tissues reveals the unique characteristics of the pollen transcriptome. Plant Physiol, 2003, 133: 713～725
Bedinger P. The remarkable biology of pollen. Plant Cell, 1992, 4: 879～887
Bednarska E. The effect of exogenous Ca~(2+) ions on pollen grain germination and pollen tube growth. Sex Plant Reprod, 1989, 2: 53～58
Bergerat A, de Massy B, Gadelle D, Varoutas PC, Nicolas A, Forterre P. An atypical topoisomerase Ⅱ from Archaea with implications for meiotic recombination. Nature, 1997, 386: 414～417
Bhatt AM, Lister C, Page T, Fransz P, Findlay K, Jones GH, Dickinson HG, Dean C. The DIF1 gene of Arabidopsis is required for meiotic chromosome segregation and belongs to the REC8/RAD21 cohesin gene family. Plant J, 1999, 19: 463～472
Bishop DK, Park D, Xu L, Kleckner N. DMC1: a meiosis-specific yeast homolog of E. coli RecA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell, 1992, 69: 439～456
Blackmore S, Barnes SH. Pollen wall development in angiosperms. In Blackmore S, Knox RB, eds. Microspores Evolution and Ontogeny. Academic Press: San Diego, 1990, pp 173～192
Bleuyard JY, White CL The Arabidopsis homologue of Xrcc3 plays an essential role in meiosis. EMBO J, 2004, 23 (2): 439～449
Boisson B, Giglione C, Meinnel T. Unexpected protein families including cell defense components feature in the N-myristoylome of a higher eukaryote. J Biol Chem, 2003, 78: 4341～43429
Bonghi C, Rascio N, Ramina A, Casadoro G. Cellulase and polygalacturonase involvement in the abscission of leaf and explants of peach. Plant Mol. Biol, 1992, 20:839～848
Bonhomme S, Budar F, Ferault M, Pelletier G. A 2.5 kb Ncol fragment of Ogura radish mitochondrial-DNA is correlated with cytoplasmic male sterility in Brassica hybrids. Curr Genet, 1991, 19: 121～127
Bonhomme S, Horlow C, Guyon A, Ferault M, Vezon D, Marchand M, de Laissardiere S, Bechtold N, Pelletier G. Screening for gametophytic mutations in the Versailles collection of Arabidopsis thaliana transformants: first results for two putative male gametophytic mutants. Acta Hort, 1998a, 459: 173-181
Bonhomme S, Horlow C, Vezon D, de Laissaxdiere S, Guyon A, Ferault M, Marchand M, Bechtold N, Pelletier G. T-DNA mediated disruption of essential gametophytic genes in Arabidopsis is unexpectedly rare and cannot be inferred from segregation distortion alone. Mol Gen Genet, 1998b, 260: 444～452
Bonza MC, Morandini P, Luoni L, Geisler M, Palmgren MG. De Michelis MI. At-ACA8 encodes a plasma membrane-localized calcium-ATPase of Arabidopsis with a calmodulin-binding domain at the N terminus. Plant Physiol, 2000, 123: 1495～1506
Borgese N, Aggujaro D, Carrera P, Pietrini G, Bassetti M. A Role for N-myristoylation in protein targeting: NADH-cytochrome b5 reductase requires myristic acid for association with outer mitochondrial but not ER membranes. J Cell Biol, 1996, 135: 1501～1513
Boyes DC, Chen C-H, Tantikanjana T, Esch JJ, Nasrallah JB. Isolation of a Second S-Locus-Related eDNA from Brassica oleracea: Genetic Relationships Between the S Locus and Two Related Loci. Genetics, 1991, 127: 221～228
Brown SM, Crouch ML Characterization of a gene family abundantly expressed in Oenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell, 1990, 2: 263～274
Bussink HID, Buxton F, Visser J. Expression and sequence comparison of the A. niger and A. tubigensis genes encoding polygalacturonase Ⅱ. Curr Genet, 1991, 19: 467～474
Buteau H, Pezet A, Ferrag F, Perrot-Applanat M, Kelly PA, Edery M. N-glycosylation of the prolactin receptor is not required for activation ofgene transcription but is crucial for its cell surface targeting. Mol Endocrinol, 1998, 12: 544～555
Buttner M, Sauer N. Monosaccharide transporters in plants: structure, function and physiology. Biochim Biophys Acta, 2000, 1465: 263～274
Buttner M, Truernit E, Baler K, Scholz-Starke J, Sontheim M, Lauterbach C, Huss VAR, Saner N. AtSTP3, a green leaf-specific, low affinity monosaccharide-H~+ symporter ofArabidopsis thaliana. Plant Cell Environ, 1999, 23: 175～184
Cai X, Dong F, Edelmann RE, Makaroff CA. The Arabidopsis SYN1 cohesin protein is required for sister chromatid arm cohesion and homologous chromosome pairing. J Cell Sci, 2003, 116: 2999～3007
Canales, C, Bhatt A M, Scott R, Dickinson H. EXS, a putative LRR receptor kinase, regulates male germline cell number and tapetal identity and promotes seed development inArabidopsis. Curr Biol, 2002, 12: 1718～1727
Cao J-S, Yu X-L, Ye W-Z, Lu G. Xiang X. Functional analysis of a novel male fertility CYP86MF gene in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). Plant Cell Rep, 2006, 24 (12): 715～723
Caryl AP, Armstrong SJ, Jones GH, Franklin FCH. A homologue of the yeast HOP1 gene is inactivated in the Arabidopsis meiotic mutant asyl. Chromosoma, 2000, 109: 62～71
Caryl AP, Jones GH, Franklin FC. Dissecting plantmeiosis using Arabidopsis thaliana mutants. J Exp Bot, 2003, 54: 25～38
Cerster J, Allard S, Robert LS. Molecular Characterization of Two Brassica napus pollen-expressed genes encoding putative arabinogalactan proteins. Plant Physiol, 1996, 110: 1231～1237
Chaturvedi MM, Kumar A, Darnay BG, Chainy GBN, Agarwal S, Aggarwal BB. Sanguinarine (pseudochelerythrine)is a potent inhibitor of NF-κB activation, IκBa phosphorylation, and degradation. J Biol Chem, 1997, 272: 30129～30134
Chaubal R, Zanella C, Trirrmell MR, Fox TW, Albertsen M C, Bedinger P. Two male-sterile mutants of Zea mays (Poaceae) with an extra cell division in the anther wall. Am J Bot, 2000, 87: 1193～1201
Chaudhury AM, Lavithis M, Taylor PE, Craig S, Singh MB, Signer ER, Knox RB, Dennis ES. Genetic control of male fertility in Arabidopsis thaliana: structural analysis of premeiotic developmental mutants. Sex Plant Reprod, 1994, 7: 17～28
Chay CH, Buehler EG, Thorn JM, Whelan TM, Eiedinger PA. Purification of maize pollen exines and analysis of associated proteins. Plant Physiol, 1992, 100: 756～761
Chen C, Marcus A, Li W, Hu Y, Calzada J-PV, Grossniklaus U, Cyr RJ, Ma H. The Arabidopsis ATK1 gene is required for spindle morphogenesis in male meiosis. Development, 2002a, 129: 2401～2409
Chen W, Provart N, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan S, Zou G, Whithama S, Budworth P, Tan Y, Xie Z, Chen X, Lam S, Kreps J, Harper J, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl J, Wang X, Zhu T. Expression profile matrix of Arabidopsis transcription factor genes implies their putative functions in response to environmental stresses. Plant Cell, 2002b, 14: 559～574
Chen YC, McCormick S. sidecar pollen, an Arabidopsis thaliana male gametophytic mutant with aberrant cell divisions during pollen development, Development, 1996, 122: 3243～3253
Cheung AY, Wu HM, Di Stilto V, Glaven R, Chen C, Wong E, Ogdahl J, Estavillo A.Pollen-pistil interactions in Nicotiana tabacura. Ann Bot, 2000, 85: 29～37
Cheung AY, Chen CYH, Tao LZ, Andreyeva T, Twell D, Wu HM. Regulation of pollen tube growth by Rat-like GTPases. J Experi Bot, 2003, 54 (380): 73～81
Cheung AY, May B, Kawata EE, Gu Q, Wu HM. Characterization of cDNAs for stylar transmitting tissue-specific proline-rich proteins in tobacco. Plant J, 1993, 3:151～160
Cheung AY, Wu HM. Arabinogalactan proteins in plant sexual reproduction. Protoplasma, 1999, 208: 87～98
Cho HT, Kende H, Expression of expansin genes is correlated with growth in deepwater rice. Plant Cell, 1997, 9: 1661～1671
Clement SA, Tan CC, Guo J, Kitta K, Suzuki YJ. Roles of protein kinase C and alpha-tocopherol in regulation of signal transduction for GATA-4 phosphorylation in HL-1 cardiac muscle cells. Free Radic Biol Med, 2002, 32: 341～349.
Cnudde F, Moretti C, Porceddu A, Pezzotti M, Gerats T. Transcript profiling on developing Petunia hybrida floral organs. Sex Plant Reprod, 2003, 16: 77～85
Consiglio F, Conicella C, Monti L, Carputo D. Highlights of meiotic genes in Arabidopsis thaliana. A J B, 2003, 2 (12): 516～520
Cosgrove DJ, Bedinger P, Durachko DM. Group I allergens of grass pollen as cell wall-loosening agents. Proc Nail Acad Sci USA, 1997, 94: 6559～6564
Cosgrove DJ. Loosening of plant cell walls by expansins. Nature, 2000, 407: 321～326
Couteau F. Belzile F, Horlow C, Grandjean O, Vezon D, Doutfiaux MP. Random chromosome segregation without meiotic arrest in both male and female meiocytes of a dmcl mutant of Arabidopsis. Plant Cell, 1999, 11: 1623～1634
Dawe RK, Freeling M. The role of initial cells in maize anther morphogenesis. Development, 1992, 116: 1077～1085
Dellapenna D, Bennett AB. In vitro synthesis and processing of tomato fruit polygatacturonase. Plant Physiol, 1988, 86: 1057～1063
DeRisi JL, lyer VR, Brown PO. Exploring the metabolic and genetic control of gene expression on a genomic scale. Science, 1997, 278: 680～686
Dickinson HG, Elleman CJ. Pollen coatings: chimeric genetics and new functions. Sex Plant Reprod, 2000, 12: 302～309
Dickinson HG. Dry stigmas, water and self-incompatibility in Brassica. Sex Plant Reprod, 1995, 8: 1～10
Ding DX-H, Vera JC, Heaney ML, Golde DW. N-glycosylation of the human granulocyte-macrophage colony-stimulating factor receptor alpha subunit is essential for figand binding and signal transduction. J Biol Chen, 1995, 270: 24580～24584.
Domi'nguez E, Mercado JA. Pollen sporopollenin: degradation and structural elucidation. Sex Plant Reprod, 1999, 12: 171～178
Dong X-Y, Hong Z-L, Sivaramakrishnan M, Mahfouz M, Verma D P S. Callose synthase (CalS5) is required for exine formation during microgametogenesis and for pollen viability in Arabidopsis. Plant J, 2005, 42: 315～328
Doughty J, Dixon S, Hiscock SJ, Willis AC, Parkin IAP, Dickinson HG. PCP-Al, a defensin-like Brassica pollen coat protein that binds the S-locus glycoprotein, is the product of gametophytic gene expression. Plant Cell, 1998, 10: 1333～1347
Doughty J, Hedderson F, McCubbin A, Dickinson HG. Interaction between a coating-borne pepfide of the Brassica pollen grain and stigmatic S (self-incompatibility)-locus-specific glycoprnteins. Proc Natl Acad Sci, USA, 1993, 90: 467～471
Doughty J, Wong HY, Dickinson HG. Cysteine-rich pollen coat proteins (PCPs) and their interactions with stigmatic S (incompatibility) and S-related proteins in Brassica: putative roles in SI and pollination. Ann Bot, 2000, 85 (A): 161～169
Doutriaux MP, Couteau F, Bergounioux C, White C. Isolation and characterization of the RAD51 and DMC1 homologs from Arabidopsis thaliana. Mol Gen Genet, 1998, 257: 283～291
Dubald M, Barakate A, Mandaron P, Mache R. The ubiquitous presence of exopolygalacturonase in maize suggests a fundamental cellular function for this enzyme. Plant J, 1993, 4:781～791
Eady C, Lindsey K, Twell D. The significance of microspore division and division symmetry for vegetative cell-specific transcription and generative cell differentiation, Plant Cell, 1995, 7: 65～74
El-Ghazaly G, Jelsen WA. DeveIopment of wheat (Triticum aeasvum L.) pollen, Ⅱ Histochemical differentiation of wall and Ublsch bodies during development. Amer Bot, 1987, 74 (9): 1396～1418
Fei HM, Sawhney VK. Ultrastructural characterization of male sterile33 (ms33) mutant in Arabidopsis affected in pollen desiccation and maturation. Can J Bot, 2001, 79: 118～129
Fernandez B, Czech MP, Meisner H. Role of protein kinase C in signal attenuation following T cell receptor engagement. J Biol Chem 1999, 274: 20244～20250
Fischer RL, Bennett AB. Role of cell wall hydrolyses in fruit riping. Annu Rev Plant Physiol. Plant Mol Biol, 1991, 42: 675～703
Franklin-Tong N, Bryant J. P4C1-Plant Reproductive Biology. Elsevier Abstracts, 2002: S125～S137
Franklin-Tong VE. Signaling and modulation of pollen tube growth. Plant Cell, 1999, 11: 727～738
Futamura N, Mori H, Kouchi H, Shinohara K. Male flower-specific expression of genes for polygalacturonase, pectin methylesterase and β-1,3-glucanase in a dioecious willow (Salix gilgliana Seemen). Plant Cell Physiol, 2000, 41: 16～26
Gallego ME, White CI. RAD50 function is essential for telomere maintenance in Arabidopsis. Proc Natl Acad Sci USA, 2001, 98: 1711～1716
Gilbert C, Gaudry M, Naccache PH. Rapid priming of calcium mobilization and superoxide anion production in human neutrophils by substimulatory concentrations of phorbol esters: a novel role for protein kinase C and tyrosine phosphorylation in the up-modulation of signal transduction. Cell Signal, 1992, 4: 511～523
Girke T, Todd J, Ruuska S, White J, Benning C, Ohlrogge J. Microarray analysis of develo ping Arabidopsis seeds. Plant Physiol, 2000, 124 (4): 1570～1581
Goldberg RB, Beals TP, Sanders PM. Anther development: Basic principles and practical applications. Plant Cell, 1993, 5: 1217～1229
Golovkin M, Reddy ASN. A calmodulin-binding protein from Arabidopsis has an essential role in pollen germination. Proc Natl Acad Sci USA, 2003,100 (18): 10558～10563
Grelon M, Vezon D, Gendrot G, Pelletier G, AtSPO11-1 is necessary for efficient meiotic recombination in plants. EMBO J, 2001, 20: 589～600
Grini PE, Schnittger A, Schwarz H, Zimmermann I, Schwab B, Jurgensa G, Hulskampa M. Isolation of ethyl methanesulfonate-induced gametophytic mutants in Arabidopsis thaliana by a segregation distortion assay using the multimarker chromosome 1. Genetics, 1999, 151: 849～863
Grobe K, Powell L D. Role of protein kinase C in the phosphorylation of CD33 (Siglec-3) and its effect on lectin activity. Blood, 2002, 99: 3188～3196
Gu Y, Vernoud V, Fu Y, Yang ZB. ROP GTPase regulation of pollen tube growth through the dynamics of tip-localized F-actin. J Experi Bot, 2003, 54(380): 93～101
Guyon VN, Astwood JD, Garner EC, Dunker AK, Taylor LP. Isolation and characterization of cDNAs expressed in the early stages of flavonol-induced pollen germination in petunia. Plant Physiol, 2000, 123: 699～710
Hadfield KA, Bennett AB. Polygalacturonases: many genes in serch of a function. Plant Physiol, 1998, 117: 337～343
Hadfield KA, Rose JKC, Yaver DS, Berka RM, Bennett AB. Polygalacturonase gene expression in ripe melon fruit supports a role for polygalacturonase in ripening-associated pectin disassembly. Plant Physiol, 1998, 117: 363～373
Hanson MR, Bentolila S. Interactions of mitocbondrial and nuclear genes that affect male gametophyte development. Plant Cell, 2004, 16: 154～169
He C, Mascarenhas JP. MEI1, AnArabidopsis gene required for male meiosis: isolation and characterization. Sex Plant Reprod, 1998, 11: 199～207
Heizmann P, Luu DT, Dumas C. Pollen-stigma adhesion in the Brassicaceae. Ann Bot, 2000, 85 (A): 23～28
Hepler PK, Vidali L, Cheung AY. Polarized cell growth in higher plants. Annu Rev Cell Dev Biol, 2001, 17: 159～187
Heslop-Harrison Y, Hesiop-Harrison J, Reger BJ. Pollen tube guidance and the regulation of tube number in Zea mays. Acta Bot Neerl, 1985, 34: 193～211
Heslop-Harrison Y, Reger BJ. Tissue organization, pollen receptivity and pollen tube guidance in normal and mutant stigmas of the grass Pennisetum Pyphoides (Burm) Stap et Hubb. Sex Plant Reprod, 1988, 1: 182～183
Higginson T, Li SF, Roger W, Parish.AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana. Plant J, 2003, 35(2): 177～192
Hiscock SJ, Dewey FM, Coleman JOD, Dickinson HG. Identification and Localization of an active cutinase in the pollen of Brassica napus L. Planta, 1994, 193: 377～384
Hiscock SJ, Doughty J, Willis AC, Dickinson HG. A 7-kDa pollen coating-borne peptide from Brassica napus interacts with S-locus glycuprotein and S-locus-related glycoprotein. Planta, 1995, 196: 367～374
Holdaway-Clarke TL, Feijo JA, Hackett GR, Kunkel JG, Hepler PK. Pollen tube growth and the lintracellular cytosolic calcium gradient oscillate in phase while extracellular calcium influx is delayed. Plant Cell, 1997, 9: 1999～2010
Holstege FC, Jennings EG, Wyrick JJ, Lee TI, Hengartner CJ, Green MR, Golub TR, Lander ES, Young RA. Dissecting the regulatory circuitry of a eukaryotic genome. Cell, 1998, 95: 717～728
Hong HP, Gerster JL, Datla RSS, Albani K, Scoles G. Keller W and Roberts LS. The promoter of a Brassica napus polygalacturonase gene directs pollen expression of beta-glucuroaidase in transgenic Brassica phnts. Plant Cell Rep, 1997, 16: 373～378
Honys D, Twell D. Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol, 2003, 132: 640～652
Honys D, Twell D. Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biology, 2004, 5:R85
Horda CLH, Chena C, DeYoung BJ, Clarke SE, Ma H. The BAM1/BAM2 receptor-like kinases are important regulators of Arabidopsis early anther development. Plant Cell, 2006, 18: 1667～1680
Horvath DP, Schaffer R, West M, Wisman E. Arabidopsis microarrays identify conserved.and differentially expressed genes involved in shoot growth and development from distantly related plant species. Plant J, 2003, 34 (1): 125～134
Horvitz R, Herskowitz I. Mechanisms of asymmetric cell division: Two Bs or not two Bs, that is the question. Cell, 1992, 68: 237～255
Huala SK, Sussex IM. Determination and cell interactions in reproductive meristems. Plant Cell, 1993, 8:1391～1407,
Hughes TR, Marton MJ, Jones AR, Roberts CJ, Stoughton R, Armour CD, Bennett HA, Coffey E, Dai H, He YD. Functional discovery via a compendium of expression profiles. Cell, 2000, 102: 109～126
Hulskamp M, Kopczak SD, Horejsi TF, Kihl BK, Pruitt RE. Identification of genes required for pollen stigma recognition in Arabidopsis thaliana. Plant J, 1995, 8: 703～715
Hulskamp M, Nikesh SP, Grini P, Schneitz K, Zimmermann I, Lolle SJ, Pruitt RE. The STUD gene is required for male-specific cytokinesis after telophase Ⅱ of meiosis in Arabidopsis thaliana. Dev Biol, 1997, 187: 114～124
Ikeda S, Nasrallah JB, Dixit R, Preiss S, Nasrallah ME. An aquaporin-like gene required for the Brassica self-incompatibility response. Science, 1997, 276: 1564～1566
Jakobsen MK, Poulsen LR, Schulz A, Fleurat-Lessard P, Mφller A, Husted S, Schiφtt M, Amtmann A, Palmgren MG. Pollen development and fertilization in Arabidopsis is dependent on the MALE GAMETOGENESIS IMPAIRED ANTHERS gene encoding a Type V P-type ATPase. Genes & Dev, 2005, 19: 2757～2769
Jauh GY, Lord EM. Movement of the tube cell in lily style in the absence of the pollen grain and the spent pollen tube. Sex Plant Reprod, 1995, 8: 168～172
Jefferson RA, Kavanagh TA, Bevan MW. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 1987, 6: 3901～3907
Jenkins ES, Paul W, Coupe SA, Bell SJ, Davies EC, Roberts JA. Characterization of an mRNA encoding a polygalacturonase expressed during pod development in oilseed rape (Brassica napus L.). J Exp Bot, 1996, 47: 111～115
Jiang L, Yang S-L, Xie L-F, Puah CS, Zhang X-Q, Yang W-C, Sundaresan V, Ye D. VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract. Plant Cell, 2005, 17: 584～596
John ME and Petersen MW. Cotton (Gossypium hirsutum L.) pollen-specific polygalaeturonase mRNA: tissue and temporal specificity of its promoter in transgenic tobacco. Plant Mol Biol, 1994, 26(6): 1989～1993
Johnson MA, Preuss D. Plotting a course: Multiple signals guide pollen tubes to their targets. Dev Cell, 2002, 2: 273～281
Johnson SA, McCormick S. Pollen Germinates Precociously in the Anthers of raring.to-go, an Arabidopsis Gametophytic Mutant1. Plant Physiol, 2001, 126:685～695
Kalaitzis P, Solomos T, Tucker ML. Three different polygalacturonases are expressed in tomato leaf and flower abscission, each with a different temporal expression pattern. Plant Physiol, 1997, 113:130～308 Kapoor S, Kobayashi A, Takatsuji H, Silencing of the tapetum-specific zinc finger gene TAZ1 causes premature degeneration of tapetum and pollen abortion in petunia. Plant Cell, 2002, 14: 2353～2367
Kasahara RD, Portereiko MF, Sandakie-Nikolova L, Rabiger DS, Drews GN. MYB98 is required for pollen tube guidance and synergid cell differentiation in arabidopsis. Plant Cell, 2005, 17: 2981～2992
Kaur J, Sebastian J, Siddiqi I. The Arabidopsis-mei2-Like genes pplay a role in meiosis and vegetative growth in Arabidopsis. Plant Cell, 2006, 18: 545～559
Kawaguchi K, Shibuya N. Characterization of arahinogalactan-proteins and a related olignsaccharide in developing rice anthers. In Nothnagel EA, Bacic A, Clarke AE, eds, Cell Develop Biology of Arabinogalactanproteins. Kiuwer Academic/Plenum Publishers: New York, 2000, pp 149～152
Kawakami Y, Kitanra J, Hartman SE, Lowell CA, Siraganian RP, Kawakami T. Regulation of protein ldnase CBI by two protein-tyrosine kinases, Btk and Syk. Proc Natl Acad Sci USA, 2000, 97: 7423～7428
Keeney S, Giroux CN, Kleckner N. Meiosis-specific DNA doublestrand breaks are catalyzed by SPOll, a member of a widely conserved protein family. Cell, 1997, 88: 375～384
Klimyuk VI, Jones JDG. AtDMC1, the Arabidopsis homologue of the yeast DMC1 gene: characterization, transposon-induced allelic variation and meiosis-associated expression. Plant J, 1997, 11:1～14
Lalanne E, Honys D, Johnson A, Borner GHH, Lilley KS, Dupree P, Grossnildaus U, Twell D. SETH1 and SETH2, two components of the glycosylphosphatidylinositol anchor biosynthetic pathway, are required for pollen germination and tube growth in Arabidopsis. Plant Cell, 2004, 16: 229～240
Laser KD, Lersten NR. Anatomy and cytology of microsporogenesis in cytoplasmic male sterile angiosperms. Bot Rev, 1972, 38(3): 425～454
Lee JY, Lee DH. Use of serial analysis of gene expression technology to reveal changes in gene expression in Arabidopsis pollen undergoing cold stress. Plant Physioi, 2003, 132: 517～529
Lennon K, Roy S, Hepler RK, Lord EM. The structure of the transmitting tissue of Arabidopsis thaliana(L.) and the path of pollen tube growth. Sex Plant Reprod, 1998, 11: 49～59
Lennon KA, Lord EM. The in vivo pollen tube cell of Arabidopsis thaliana: Tube cell cytoplasm, wall and cytoskeleton. Protuplasma, 2000, 214: 45～56
Lester DR, Sherman WB, Atwell BJ. Endopolygalacturonase and the Melting Flesh (M) locus in peach. J Amer Soci Horti Sci, 1996, 121, 231～235
Li H, Bacic A, Read SM. Role of a callose synthase zymogen in regulating wall deposition in pollen tubes of Nicotiana alata Link et Otto. Planta, 1999b, 208: 528～538
Li H, Lin Y, Heath RM, Zhu MX, Yang Z. Control of pollen tube tip growth by a Rop GTPase-dependent pathway that leads to the tip-localized calcium influx. Plant Cell, 1999a, 11: 1731～1742
Li H, Wu G, Ware D, Davis KR, Yang Z. Arabidopsis Rho-related GTPases: differential gene expression in pollen and polar localization in fission yeast. Plant Physiol, 1998, 118: 407～417
Li L-B, Chen N, Ramamoorthy S, Chi L, Cui X-N, Wang LC, Reith MEA. The Role of N-glycosylation in function and surface trafficking of the human dopamine transporter. J Biol Chem, 2004, 279: 21012～21020
Li YQ, Mareck A, Faleri C, Moseatelli A, Liu Q, Cresti M. Detectionand localization of pectin methylesterase isoforms in pollen tubes of Nicotiana tabacum L. Planta, 2002, 214: 734-740
Lin R, Hiscott J. A role for casein kinase Ⅱ phosphorylation in the regulation of IRF-1 transcriptional activity. Mol Cell Biochem 1999, 191: 169～180
Lin Y, Yang Z. Inhibition of pollen tube elongation by microinjected anti-Rop1Ps antibodies suggests a crucial role for Rho-type GTPases in the control of tip growth. Plant Cell, 1997, 9:1647～1659
Liu L-C, Cao J-S, Yu X-L, Xiang X, Pei Y-J. Expression of an antisense BcMF3 affects mierosporogenesis and pollen tube growth in Arabidopsis. Agricul Sci in China, 2006, 5(5): 339～345
Liu XC, Jones K, Dickinson HG. DNA synthesis and cytoplasmic differentiation in tapetal cells of normal and cytoplasmic cytoplasmic male sterile lines of Petunia hybrida. Theor Appl Genet, 1987, 74: 846～851
Lobstein E, Guyon A, Ferault M, Twell D, Pelletier G, Bonhomme S. The putative Arabidopsis homolog of yeast vps52p is required for pollen tube elongation, localizes to golgi, and might be involved in vesicle trafficking. Plant Physinl, 2004, 135(3): 1480～1490
Lord EM, Russell SD. Mechanisms of pollination and fertilization in plants. Annu Rev Cell Devel Biol, 2002, 18: 81～105
Lord EM. Adhesion and cell movement during pollination: churches la femme. Trends Plant Sci, 2000, 5: 368～373
Lush WM, Spurck T, Joosten R. Pollen tube guidance by the pistil of a solanaceous plant. Ann Bot, 2000, 85(A): 39～47
Ma J, Duncan D, Morrow DJ, Fernandes J, Walbot V. Transcriptome profiling of maize anthers using genetic ablation to analyze pre-meiotic and tapetal cell types. Plant J, 2007, 50: 637～648
Magnard JL, Yang M, Chen YCS, Leary M, McCormick S. The Arabidopsis gene TARDYASY'NCHRONOUS MEIOSIS is required for the normal pace and synchrony of cell division during male meiosis. Plant Physiol, 2001, 127:1157～1166
Matho R, Trewavas AJ. Localized apical increases of cytosolic free calcium control pollen tube orientation. Plant Cell, 1996, 8: 1935～1949
Mandaokar A, Kumar VD, Amway M, Browse J. Microarray and differential display identify genes involved in jasmonate-dcpendent anther development. Plant Mol Biol, 2003, 52(4): 775～86
Markovic O, Janecek S. Pectin degrading glycoside hydrolases of family 28: sequence-structural features, specificifies and evolution. Protein Eng Des Sel, 2001, 14: 615～631
Marquez J, Seoane-Camba JA, Suarez-Cervera M. Allergenic and antigenic proteins released in the apertural sporoderm during the activation process in grass pollen grains. Sex Plant Repred, 1997, 10: 269～278
Masearenhas JP. Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol, 1990, 41:317～338
Mascarenhas JP. The male gametophyte of flowering plants. Plant Cell, 1989, 1: 657～664
Mathilde G. Ghislaine G. Daniel V, and Georges P. The Arabidopsis MEI1 gene encodes a protein with five BRCT domains that is involved in meiosis-specific DNA repair events independent of SPO11-induced DSBs. Plant J, 2003, 35(4): 465～75
Mayfield JA, Preuss D. Rapid initiation of Arabidop.sis pollination requires the oleosin-domain protein GRP17. Nat Cell Biol. 2000, 2:128～130
McCormick S. Control of male gametophyte development. Plant Cell, 2004, 16:S142～S153
McCormick S. Male gametophyte development. Plant cell, 1993, 5: 1265～1275
McCoy TJ, Smith LY. Genetics, cytology, and crossing behavior of an alfalfa (Medicago sativa) mutant resulting in failure of the postmeiotic cytokinesis. Can J Genet & Cytol, 1983, 25: 390-397
Mele G, Ori N, Sato Y, Hake S. The knottedl-like homeobox gene BREVIPEDICELLUS regulates cell differentiation by modulating metabolic pathways. Genes & Development, 2003, 17: 2088～2093
Mercier R, Armstrong SJ, Horlow C, Jackson NP, Makaroff CA, Vezon D, Pelletier G, Jones GH, Franklin FCH. The meiotic protein SWI1 is required for axial element formation and recombinationinitiation in Arabidopsis. Development, 2003, 130: 3309～3318
Mercier R, Vezon D, Bullier E, Motamayor JC, Sellier A, Lefevre F, Pelletier G, Horlow C. Switch1 (SWI1): a novel protein required for the establishment of sister chromatid cohesion and for bivalent formation at meiosis. Gene Dev, 2001, 15: 1859～1871
Mitsui H, Nakatani K, Yamaguchi-Shinozaki K, Shinozaki K, Nishikawa K, Takahashi H. Sequencing and characterization of the kinesin-related genes katB and katC of Arabidopsis thaliana. Plant Mol Biol, 1994, 25: 865～876
Mitsui H, Yamaguchi-Shinozaki K, Shinozaki K, Nishikawa K, Takahashi H. Identification of a gene family (kat) encoding kinesin-like proteins in Arabidopsis thaliana and the characterization of secondary structure of KatA. Mol Gen Genet, 1993, 238: 362～368
Moon YH, Chen L, Pan RL, Chang HS, Zhu T, Maffeo DM, Sung ZR. EMF genes maintain vegetative development by repressing the flower program in Arabidopsis. Plant Cell, 2003, 15: 681～693
Motamayor JC, Vezon D, Bajon C, Sanvanet A, Grandjean O, Marchand M, Bechtold N, Pelletier G, Horlow C. Switch (swi1), an Arabidopsis thaliana mutant affected in the female meiotic switch. Sex Plant Reprod, 2000, 12: 209～218
Mouline K, Very A-A, Gaymard F, Boucherez J, Pilot G, Devic M, Bouchez D, Thiband J-B, Sentenac H. Pollen tube development and competitive ability are impaired by disruption of a Shaker K~+ channel in Arabidopsis. Genes Dev, 2002, 16(3): 339～350
Moutinho A, Hussey PJ, Trewavas AJ, MalhoR. cAMP acts as a second messenger in pollen tube growth and reorientation. Proc Natl Acad Sci USA, 2001, 98(18): 10481～10486
Mu J-H, Stains JP, Kao T-h. Characterization of a pollen-expressed gene encgding a putative pectin esterase of Petunia inflata. Plant Mol Biol, 1994, 25: 539～54
Muschietti J, Dircks L, Vancanneyt G, McCormick S. LAT52 protein is essential for tomato pollen development: Pollen expressing antisense LAT52 RNA hydrates and germinates abnormally and cannot achieve fertilization. Plant J, 1994, 6: 321～338
Neelam A, Sexton R. Cellulase (endo_-1, 4 glucanase) and cell wall breakdown during anther development in the sweet pea (Lathyrus odoratus L.): isolation and characterization of partial cDNA clones. J Plant Physiol, 1995, 146: 622～628
Niewiadomski P, Knappe S, Geimer S, Fischer K, Schulz B, Unte US, Rosso MG, Ache P, Flugge U, Schneidera A. The Arabidopsis plastidic glucose 6-phosphate/phosphate translocator GPT1 is essential for pollen maturation and embryo sac development. Plant Cell, 2005, 17: 760～775
Niu L, Heaney ML, Vera JC, Golde DW. High-affinity binding to the GM-CSF receptor requires intact N-glycosylation sites in the extracellular domain of the beta subunit. Blood, 2000, 95: 3357～3362
Nonomura KI, Miyoshi K, Eiguchi M, Suzuki T, Miyao A, Hirochika H, Kurata N. The MSP1 Gene is necessary to restrict the number of cells entering into male and female sporogenesis and to initiate anther wall formation in rice. Plant Cell, 2003, 15: 1728～1739
Olsen AN, Mundy J, Skrivcr K. Peptomics, identification of novel cationic Arabidopsis peptides with conserved sequence motifs. In Silico Biology, 2002, 2, 39～44
Orlandi I, Bettiga M, Alberghina L, Vai M. Transcriptional profiling of ubp10 null mutant reveals altered subtelomeric gene expression and insurgence of oxidative stress response. J Biol Chem, 2004, 279(8):6414～6425
Otegui MS, Staehelin LA. 3D Tomographic analysis of post-meiotic cytokinesis during pollen development in Arabidopsis thaliana. Planta, 2004, 218: 501～515
Owen HA, Makaroff CA. Ultrastructure of microsporogenesis and microgametogenesis in Arabidopsis thaliana (L.) Heynh. ecotype Wassilewskija (Brassicaceae). Protoplasma, 1995, 185: 7～21
Pacini E, Franchi E, Hesse M. The tapetum: its form, function and possible phylogeny in rmbryophyta. Plant Systematics and Evolution, 1985, 149: 155～185
Palanivelu R, Brass L, Edlund AF, Preuss D. Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABAlevels. Cell, 2003, 14(1):47～59
Papini A, Mosti S, Brighigna L. Programmed-cell-death events during tapetum development of angiosperms. Protoplasma, 1999, 207: 213～221
Park BS, Kim JS, Kim SH, Park YD. Characterization of a pollen-preferential gene, BAN102, from Chinese cabbage. Plant Cell Rep, 2005, 14: 1～8
Park J, Hill MM, Hess D, Brazil DP, Hofsteenge J, Hemmings BA. Identification of tyrosine phosphorylation sites on 3-phosphoinositide-dependent protein kinase-1 and their role in regulating kinase activity. J Biol Chem, 2001, 276: 7459～37471
Park SK, Howden R, Twell D. The Arabidopsis thaiiana gametophytic mutation gemini pollenl disrupts microspore polarity, division asymmetry and pollen cell fate. Development, 1998, 125(19): 3789～3799
Park SK, Twell D. Novel patterns of ectopic cell plate growth and lipid body distribution in the Arabidopsis gemini pollenl mutant. Plant Physiol, 2001, 126(2): 899～909
Parkin IAP, Gulden SM, Sharpe AG. Lukens L, Trick M, Osborn TC, Lydiate DJ. Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana. Genetics, 2005, 171 (2): 765～781
Parre E, Geitmann A. Pectin and the role of the physical properties of the cell wall in pollen tube growth of Solanum chacoense. Planta, 2005, 220: 582～592
Paxson-Sowders DM, Dodrill CH, Owen HA, Makaroff CA. DEX1, a novel plant protein, is required for exine pattern formation during pollen development in Arabidopsis. Plant Physiol, 2001, 127: 1739～1749
Paxson-Sowders DM, Owen HA, Makaroff CA. A comparative ultrastructural analysis of exine pattern development in wild type Arabidopsis and a mutant defective in pattern formation. Protoplasma, 1997, 198: 53～65
Pennell RI, Jam-fiche L, Kjellbom P, Scofield GN, Peart JM, Roberts K. Developmental regulation of a plasma membrane arabinogalactan protein epitope in oilseed rape flowers. Plant Cell, 1991, 3: 1317～1326
Pennell RI, Knox JP, Scofield GN, Selvendran RR, Roberts K. A family of abundant plasma membrane-associated glycoproteins related to the arabinogalactan proteins is unique to flowering plants. J Cell Biol, 1989, 108: 1967～1977
Pereira LG, Coimbra S, Oliveira H, Monteiro L, Sottomayor M. Expression of arabinogalactan protein genes in pollen tubes of Arabidopsis thaliana. Planta, 2006, 223: 374～380
Petersen M, Brodersen P, Naested H, Andreasson E, Lindhart U, Johansen B, Nielsen HB, Lacy M, Austin MJ, Parker JE, Sharma SB, Klessig DF, Martienssen R, Mattsson O, Jenscn AB, Mundy J. Arabidopsis map kinase 4 negatively regulates systemic acquired resistance. Cell, 2000, 103: 1111～1120
Petersen M, Sander L, Child R, Van Onckelen H, Ulvskov P, Borkhardt B. Isolation and characterisation of a pod dehiscence zone-specific polygalacturonase from Brassica napus. Plant Mol Biol, 1996, 31:517-527
Picton JM, Steer MW. The effects of ruthenium red, lanthanum, fluorescein isothiocyanate and trifluoperazine on vesicle transport, vesicle fusion and tip extension in pollen tubes. Planta, 1985, 163: 20～26
Pierson ES, Miller DD, Callaham DA, Shipley AM, Rivers BA, Cresti M, Hepler P. Pollen tube growth is coupled to the extracellular calcium ion flux and the intracellular calcium gradient: effect of BAPTA-type buffers and hypertonic media. Plant Cell, 1994, 6: 1818～1828
Piffanelli P, Ross JHE, Murphy DJ. Biogenesis and function of the lipidic structures of pollen grains. Sex Plant Reprod, 1998, 11: 65～80
Pina C, Pinto F, Feijo J, Becker J. Gene family analysis of the Arabidopsis pollen transcriptome reveals biological implications for cell growth, division control, and gene expression Regulation. Plant Physiol, 2005, 138: 744～756
Poethig S. Genetic mosaics and cell lineage analysis in plants. Trends Genet, 1989, 5: 273～277
Porat R, Lu P, O'Neill SD. Arabidopsis SKP1, a homologue of a cell cycle regulator gene, is predominantly expressed in meristematic cells. Planta, 1998, 204: 345～351
Pressey R, Reger BJ. Polygalacturonase in pollen from corn and other grasses. Plant Sci, 1989, 59: 57～62
Pressey R. Polygalacturonase in tree pollens. Phytochemistry. 1991, 30: 1753～1755
Preuss D, Lemieux B, Yen G, Davis RW. A conditional sterile mutation eliminates surface components from Arabidopsis pollen and disrupts cell signalling during fertilization. Genes and Development, 1993, 7: 974～985
Procissi A, Guyon A, Pierson ES, Giritch A, Knuiman B, Grandjean O, Tonelli C, Derksen J, Pelletier G, Bonhomme S. K/NKY POLLEN encodes a SABRE-like protein required for tip growth in, Arabidopsis and conserved among eukaryotes. Plant J, 2003, 36 (6): 894～904
Procissi A, Laissardie're SD, Fe'rault M, Vezon D, Pelletier G, Bonhomme S. Five gametophytic mutations affecting pollen development and pollen tube growth in Arabidopsis thaliana. Genetics, 2001, 158: 1773～1783
Raghavan V. A transient accumulation of poly (A) containing RNA in the tapetum of Hyoscyamus niger during microsporogenesis. Dev Biol, 1981, 81: 342～348
Raghavan V. mRNAs and a cloned histone gene are differentially expressed during anther and pollen development in rice (Oryza sativa L.). J Cell Sci, 1989, 92: 217～229
Rato C, Monteiro D, Hepler PK, Malho R. Calmodulin activity and cAMP signalling modulate growth and apical secretion in pollen tubes. Plant J, 2004, 38 (6): 887～897
Redgwell RJ, MacRae EA, Hallett I, Fischer M, Perry J, Harker R. In vivo and in vitro swelling of cell walls during fruit ripening. Planta, 1997, 203: 162～173
Rhee SY, Osborne E, Poindexter PD, Somerville CR. Microspore separation in the quartet 3 mutants of Arabidopsis is impaied by a defect in a developmentally regulated polygalacturonase required for pollen mother cell wall degradation. Plant Physiol, 2003, 133: 1170～1180
Rhee SY, Somerville CR.Tettad pollen formation in quartet mutants of Arabidopsis thaliana is associated with persistence of pectic polysaccharides of the pollen mother cell wall. Plant J, 1998, 15: 79～88
Robert LS, Allard S, Gerster JL, Cass L, Simmonds J. Isolation and characterization of a polygalacturonase gene highly expressed in Brassica napus pollen. Plant Mol Biol, 1993, 23(6): 1273～1278
Rockmill B, Roeder GS. Meiosis in asynaptic yeast. Genetics, 1990, 126: 563～574
Rose JKC, Lee HH, Bennett AB. Expression of a divergent expansin gene is fruit-specific and ripening-regulated. Proc Nail Acad Sci USA, 1997, 94: 5955～5960
Ross KJ, Fransz P, Armstrong SJ, Vizir I, Mulligan B, Franklin FCH, Jones GH. Cytological characterization of four meiotic mutants of Arabidopsis isolated from T-DNA-transformed lines. Chromosome Res, 1997, 5: 551～559
Rozwadowski K, Zhao RH, Jackman L, Huebert T, Burkhart WE, Hemmingsen SM, Greenwood J, Roths SJ. Characterization and immunolocalizatioa of a cytosolic calcium-binding protein from Brassica napus and Arabidopsis pollen. Plant Physiol, 1999, 120: 787～798
Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning. New York: Cold Spring Harbor Laboratory Press, 1989
Sainchez-Moran E, Mercier R, Higgins JD, Armstrong. SJ, Jones GH, Franklin CH. A strategy to investigate the plant meiotic proteome. Cytogenet Genome Research, 2005, 109: 181～189
Sanders PM, Bui AQ, Weterings K, McIntire KN, Hsu Y-C, Lee PY, Truong MT, Beals TP, Goldberg RB. Anther developmental defects in Arabidopsis thaliana male-sterile mutants. Sex Plant Reprod, 1999, 11:297～322
Sato S, Hotta Y, Tabata S. Structural analysis of a RecA-like gene in the genome of Arabidopsis thaliana. DNA Research, 1995, 2:89～93
Schiefthaler U, Balasubramanian S, Sieber P, Chevalier D, Wisman E, Schneitz K. Molecular analysis of NOZZLE, a gene involved in pattern formation and early sporogenesis during sex organ development in Arabidopsis thaliana. Proc Nail Acad Sci USA, 1999, 96:11664～11669
Schiφtt M, Romanowsky SM, Baekgaard L, Jakobsen MK, Palmgren MG, Harper JF. A plant plasma membrane Ca~(2+) pump is required for normal pollen tube growth and fertilization. Proc Natl Acad Sci USA, 2004, 101(25): 9502～9507
Schneidereit A, Scholz-Starke J, Buttner M. Functional characterization and expression analyses of the glucose-specific AtSTP9 monosaccharide transporter in pollen of Arabidopsis. Plant Physiol, 2003, 133: 182～190
Scholz-Starke J, Buttner M, Sauer N. AtSTP6, a new pollen-specific H~+-monosacchadde symporter from Arabidopsis. Plant PhysioI, 2003, 131: 70～77
Schopfer CR, Nasrallah ME, Nasrallah JB. The male determinant of self-incompatibility in Brassica. Science, 1999, 286: 1607～1700
Schwarzacher. Meiosis, recombination and chromosomes: a review of gene isolation and fluorescent in situ hybridization data in plants. J Exp Bot, 2003, 54:11-23
Scott RJ, Spielman M, Dickinsonb HG. Stamen structure and function. Plant Cell, 2004, 16: S46-S60
Sexton R, Campillo ED, Duncan D, Lewis LN. The purification of an anther cellulase(1:4-glucan hydrolase) from Lathyrus odoratus L. and its relationship to the similar enzyme found in abscission zones. Plant Sci, 1990, 67:169-176
Sharp DJ, Rogers GC, Scholey, JM. Microtubule motors in mitosis. Nature, 2000, 407:41-47
Sheridan WF, Golubeva EA, Abrhamova Ⅱ, Golubovskaya IN. The macl mutation alters the developmental fate of the hypodermal cells and their cellular progeny in the maize anther. Genetics, 1999, 153: 933-941
Shinohara A, Ogawa H, Matsuda Y, Ushio N, Ikeo K, Ogawa T. Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and RecA. Nature Genetics, 1993, 4: 239～243
Shukia A.K, Vijayaraghavan MR, Chaudhry B. Biology of pollen. New Delhi: APH Publishing Corporation. 1998
Singh M, Brown GG. Suppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitoehondrial gent region. Plant Cell, 1991, 3: 1349～1362
Sitrit Y, Downie B, Bennett AB, Bradford KJ. A novel exo-polygalacturonase is ass ociated with radicle protrusion in tomato seeds. Plant Physiol, 1996, 111(Supplement): 161～163
Sobotka R, Sakova L, Curn V. Molecular mechanisms of self-incompatibility in Brassica. Curt Issues Mol Biol, 2000, 2 (4): 103～12
Sommer-Knudsen J, Clarke AE, Bacic A. A galactose-dch, cell-wall glycoprotein from styles of Nicotiana alata. Plant J, 1996, 9: 71～83
Sommer-Knudsen J, Lush WM, Bacic A, Clarke AE. Re-evaluation of the role of a transmitting tract-specific glycoprotein on pollen tube growth. Plant J, 1998, 13: 529～535
Southworth D. Exine biochemistry. In S Blackmore, RB Knox, eds, Microspores: Evolution and Ontogeny. Academic Press: London, 1990, pp 193～212
Spielman M, Preuss D, Li FL, Browne WE, Scott R J, Dickinson HG. TETRASPORE is required for male meiotic cytokinesis inArabidopsis thaliana. Development, 1997, 124: 2645～2657
Stevens R, Grelon M, Vezon D, Oh J, Meyer P, Perennes C, Domenichini S, Bergounioux C. A CDC45 homolog in Arabidopsis is essential for meiosis, as shown by RNA interference-induced gene silencing. Plant Cell, 2004, 16: 99～113
Stratilova E, Dzurova M, Markovic O, Jornvall H. An essential tyrosine residue of Aspergillus polygalacturonase. FEBS Lett, 1996, 11, 382 (1-2): 164～166
Swain SM, Muller AI, Singh DP. The gar2 and rga alleles increase the growth of gibberellin-deficient pollen tubes in Arabidopsis, Plant Physiol, 2004, 134: 694～705
Sym M, Engebrecht J, Roeder GS. ZIP1 is a synaptonemal complex protein required for meiotic chromosome synapsis. Cell, 1993, 72: 365～378
Sym M, Roeder GS. Crossover interference is abolished in the absence of a synaptonemal complex protein. Cell, 1994, 79: 283～292
Sze H, Padmanaban S, Cellier F, Honys D, Cheng N-H, Bock KW, je'ro GC, Li X, Twell D, Ward JM, Hirschi KD. Expression patterns of a novel AtCHX gene family highlight potential roles in osmotic adjustment and K~+ Homeostasis in Pollen Development. Plant Physiol, 2004, 136: 2532～2547
Takayama S, Shiba H, Iwano M, Asano K, Hara M, Che FS, Watanabe M, Hinata K, Isogai A. Isolation and characterization of pollen coat proteins of Brassica campestris that interact with S Iocus-related glycoprotein 1 involved in pollen-stigma adhesion. Proc Natl Acad Sci USA, 2000a, 97(7): 3765～3770
Takayama S, Shiba H, Iwano M, Shimosato H, Che F-S, Kai N, Watanabe M, Suzuki G, Hinata K, Isogai A. The pollen determinant of self-incompatibility in Brassica campestris. Proc Natl Acad Sci USA, 2000b, 97: 1920～1925
Takegami MH, Yoshioka M, Tanaka I, Ito M. Characteristics of isolated microsporocytes from liliaceous plants for studies of the meiotic cell cycle in vitro. Plant Cell Physiol, 1981, 22: 1～10
Taylor JE, Tucker GA, Lasslett Y, Smith CJS, Arnold CM, Watson CF, Schuch W, Grierson D, Roberts JA. Polygalacturonase expression during leaf abscission of normal and transgenic plants. Planta, 1990, 183: 133～138
Taylor JE, Webb STJ, Coupe SA, Tucker GA, Roberts JA. Changes in polygalacturonase activity and solubility of polyuronides during ethylene-stimulated leaf abscission in Sambucus nigra. J Exp Bot, 1993, 44: 93～98
Taylor LP, Hepler PK. Pollen germination and tube growth. Annu Rev Plant Physiol Plant Mol Biol, 1997, 48: 461～491
Tebbutt SJ, Rogers HJ, Lonsdale DM. Characterization of a tobacco gene encoding a pollen-specific polygalacturonase. Plant Mol Biol, 1994, 25: 283～297
Tirlapur, UK, Scali M, Moscatelli A, Del Casino C, Cai G, Tiezzi A, Cresti M. Confocal image analysis of spatial variations in immunocytochemically identified calmodulin during pollen hydration, germination and pollen tube tip growth in Nicotiana tabacum L. Zygote, 1994, 2: 63～68
Torki M, Mandaron P, Mache R, Falconer D. Characterization of a ubiquitous expressed gene family encoding polygalacturonase in Arabidopsis thaliana. Gene, 2000, 242: 427～436
Torki M, Mandaron P, Thomas F, Quigley R, Mache F, Falconer D. Differential expression of a polygalacturonase gene family in Arabidopsis thaliana. Mol Gen Genet, 1999, 261: 948～952
Truernit E, Schmid J, Epple P, Illig J, Sauer N. The sink-specific and stress regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors and pathogen challenge. Plant Cell, 1996, 8: 2169～2182
Truernit E, Stadler R, Baier K, Sauer N. A male gametophyte-specific monosaccharide transporter in Arabidopsis. Plant J, 1999, 17: 191～201
Tsuchida M, Manthei ER, Alam T, Knechtle SJ, Hamawy MM. Regulation of T cell receptor-and CD28-induced tyrosine phosphorylation of the focal adhesion tyrosine kinases Pyk2 and Fak by protein kinase C. J Biol Chem, 2000, 275(2): 1344～1350
Tucker GA, Schindler BC, Roberts JA. Flower abscission in mutant tomato plants. Planta, 1984, 160: 164～167
Twell D, Howden R. Mechanisms of asymmetric division and cell-fate determination in developing pollen, in Androgenesis and Haploid Plants. In Chupeau Y, Caboche M, Henry Y, eds, Memory of Jean-Pierre Bourgin, 1998, pp 69～103
Twell D, Park SK, Lalanne E. Asymmetric division and cell-fate determination in developing pollen. Trends Plant Sci, 1998, 3(8): 305～310
Twell D. Pollen developmental biology. In O'Neill SD, Roberts JA, eds, Plant Reproduction. Annu Rev Plant, 2002, 6: 86～153
Unte US, Sorensen AM, Pesaresi P, Gandikota M, Leister D, Saedter H, Huijser P. SPL8, an SBP-Box gene that affects pollen sac development in Arabidopsis. Plant Cell, 2003, 15(4): 1009～1019
Uyenoyama MK. Evolutionary dynamics of self-incompatibility alleles in Brassica. Genetics, 2000, 156: 351～359
van Aelst AC, van Went JL. Ultrastructural immunolocalization of pectin and glycoproteins in Arabidopsis thaliana pollen grains. Protoplasma, 1992, 168: 14～19
Wakeley PR, Rogers HJ, Rozycka M, Greenland AJ, Hussey PJ. A maize pectin methylesterase-like gene, ZmC5, specifically expressed in pollen. Plant Mol Bio, 1998, 37: 187～192
Wang H, Wu H-M, Cheung AY. Development and pollination regulated accumulation and glycosylation of a stylar transmitting tissue-specific proline-rich protein. Plant Cell, 1993, 5:1639～1650
Wang Y-Q, Ye W-Z, Can J-S, Yu X-L, Xiang X, Lu G. Cloning and characterization of the microspore development related gene BcMF2 in Chinese cabbage pak-choi (Brassica campestris L. ssp. chinensis Makino). J Integrative Plant Biol, 2005, 47 (7): 863～872
Wang Y-Q, Yu X-L, Cao J-S. Isolation and characterization of BcMF3, a gene expressed only in maintainer line in Chinese cabbage pak-choi (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee). Acta Genetica Sinica, 2004, 31(11): 1302～1308
Waterkeyn L and Bienfait A. On a possible function of the callosic special wall in Ipomoea purpurea (L) Roth. Grana, 1970, 10: 13～20
Wellmer F, Riechmann JL, Aives-Ferreira M, Meyerowitz EM. Genome-wide analysis of spatial gene expression in Arabidopsis flowers. Plant Cell, 2004, 16: 1314～1326
Wheeler MJ, Franklin-Tong VE, Franklin FCH, The molecular and genetic basis of pollen-pistil interactions, New phytologist, 2001, 151: 565～584
Whittington AT, Vugrek O, Wei KJ, Hasenbein NG, Sugimoto K, Rashbrooke MC, Wasteneys GO. MOR1 is essential for organizing cortical microtubules in plants. Nature, 2001, 411: 610～613
Wilhelmi LK, Preuss D. Self-sterility in Arabidopsis due to defective pollen tube guidance. Science, 1996, 274: 1535～1537
Willing RP, Bashe D, Mascarenhas, JP. An analysis of the quantity and diversity of messenger RNAs from pollen and shoots of Zea mays. Theor Appl Genet, 1988, 75: 751～753
Willing RP, Mascarenhas JP. Analysis of the complexity and diversity of mRNAs from pollen and shoots of Tradescantia. Plant Physiol, 1984, 75: 865～868
Wilson ZA, Morroll SM, Dawson J, Swarup R, Tighe PJ. The Arabidopsis MALE STERILITY1(MS1) gene is a transcriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors. Plant J, 2001, 28: 27～39
Wu H-M, Wang H, Cheung AY. A floral transmitting tissue specific glycoprotein attracts pollen tubes and stimulates their growth. Cell, 1995, 82: 383～393
Wu H-M, Wong E, Ogdahl J, Cheung AY. A pollen tube growth-promoting arabinogalactan protein from Nicotiana alata is similar to the tobacco TTS protein. Plant J, 2000, 22: 165～176
Xu Z, Dooner HK. The Maize aberrant pollen transmission 1 gene is a SABRE/KIP homolog required for pollen tube growth. Genetics, 2006, 172: 1251～1261
Yang CY, Spileman M, Coles JP, Li Y, Ghelani S, Bourdon V, Brown RC, Lemmon BE, Scott RJ, Dickinson HG. TETRASPORE encodes a kinesis required for male meiotic cytokinesis in Arabidopsis. Plant J, 2003b, 34: 229～240
Yang M, Hu Y, Lodhi M, McCombie W R, Ma H. The Atrabidopsis SKP1-LIKE1 gene is essential for male meiosis and may control homologue separation. Proc Natl Acad Sci USA, 1999b, 96: 11416～11421
Yang M, McCormick S. The Arabidopsis MEI1 gene likely encodes a protein with BRCT domains. Sex Plant Reprod, 2002, 14: 355～357
Yang SL, Xie LF, Mao HZ, Puah CS, Yang WC, Jiang LX, Sundaresan V, Ye D. TAPETUM DETERMINANT1 is required for cell specialization in the Arabidopsis anther. Plant Cell, 2003a, 15: 2792～2804
Yang WC, Sundaresan V. Genetics of gametophyte biogenesis in Arabidopsis. Curr Opin Plant Biol, 2000, 3: 53～57
Yang WC, Ye D, Xu J, Sundaresan V. The SPOROCYTELESS gene of Arabidopsis is required for itiation of sporogenesis and encodes a novel nuclear protein. Genes Dev, 1999a, 13: 2108～2117
Yang Z. Signaling tip growth in plants. Curr Opin Plant Biol, 1998, 1: 525～530
Ye W-Z, Cao J-S, Xiang X, Zeng G-W, Molecular cloning and characterization of the genic male sterility related gene CYP86MF in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee). J Hort Sci & Biotech, 2003, 78(3): 319～323
Yu X-L, Cao J-S, Ye W-Z, Wang Y-Q. Construction of an antisense CYP86MF gene plasmid vector and production of a male-sterile Chinese cabbage transformant by the pollen-tube method. J Hort Sci & Biotech, 2004, 79(5): 833～839
Zhao D, Han T, Risseeuw E, Crosby WL, Ma H. Conservation and divergence of ASK1 and ASK2 gene functions during male meiosis in Arabidopsis thaliana. Plant Mol Biol, 2003, 53(1-2): 163～73
Zhao DZ, Wang GF, Speal B, Ma H. The EXCESS MICROSPOROCYTES1 gene encodes a putative leucine-rich repeat receptor protein kinase that controls somatic and reproductive cell fates in the Arabidopsis anther. Genes Dev, 2002, 16: 2021～2031
Zheng Z-L, Yang Z, The Rop GTPase switch turns on polar growth in pollen. Trends Plant Sci, 2000, 5: 298～303
Zinkd GM, Preuss D. Dissecting Arabidopsis pollen-stigma interactions reveals novel mechanisms that confer mating specificity. Ann Bot, 2000, 85(A): 15～21
布坎南 BB，格鲁依森姆 W，琼斯 RL 主编 (瞿礼嘉，顾红雅，白书农，赵进东，陈章良主译)．植物生物化学与分子生物学，北京：科学出版社，2004
曹家树，曹寿椿，易清明．白菜及相邻类群基因组 DNA 的 RAPD 分析．园艺学报，1995，22：47～52
曹双河，张相岐，张爱民．光 (温) 敏雄性不育的调控机理和分子遗传学研究进展．植物学通报，2005，22(1)：19～26
陈新建，陈占宽，刘国顺，刘鸿先．植物对水分胁迫响应的分子机制与抗逆基因工程的研究进展．热带亚热带植物学报，2000，8(1)：81～90
崔辉梅，曹家树，张明龙，姚祥坦，向珣．白菜和芜菁 Ogura 型雄性不育系与保持系的获得及其细胞学观察．园艺学报，2004，31(4)：467～471
李德葆，周雪平，许建平等．基因工程操作技术，上海：上海科技出版社，1996
利容千．几种农作物雄性不育的细胞学研究．武汉大学学报，1978，83～96
刘乐承，余小林，叶纨芝，向珣，曹家树．白菜雄性不育相关基因 BcMF3 功能的反义 RNA 验证．浙江大学学报 (农业与生命科学版)，2006a，32(5)：473～478
刘乐承，向珣，曹家树．白菜雄性不育相关基因 BcMF4 基因功能的 RNAi 验证．遗传，2006b，28(11)：1428～1434
刘仁虎，赵建伟，肖勇，孟金．拟南芥 cDNA 芯片和油菜-拟南芥比较作图相结合筛选甘蓝型油菜抗菌核病先验基因．中国科学C辑生命科学，2005，35(1)：13～21
刘玉梅．甘蓝显性细胞核雄性不育的细胞学特征、生化基础及其分子标记的研究．博士学位论文，2003
龙欢，姚家玲，涂金星．3 种甘蓝型油菜雄性不育系花药发育的细胞学研究．华中农业大学学报．2005，24：570～575
沈俊儒，吴建勇，张创，刘平武，杨光圣．甘蓝型油菜华油杂6号及其亲本的基因差异表达研究．中国农业科学，2006，39(1)：23～28
孙敬三，钱迎倩．植物细胞学研究方法，北京：科学出版社，1987
王台，丁兆军．减数分裂及其基因研究进展．科学通报，2002，47(4)：241～248
王永勤，曹家树，符庆功，余小林，叶纨芝，向珣．利用 cDNA-AFLP 技术分析白菜核雄性不育两用系的表达差异．中国农业科学，2003，36(5)：557～560
王永勤，余小林，曹家树．白菜小孢子发育相关基因 BcMF3 的分离及其表达特征分析．遗传学报，2004，31(11)： 1302～1308
余小林，曹家树，许立奎，齐文雯，王小静．优化白菜类蔬菜遗传转化体系的研究．浙江大学学报 (农业与生命科学版)，2005，31(5)：529～534
余小林．白菜雄性不育相关基因 CYP86MF 的功能验证及人工不育系的创建．博士学位论文，2002
张强，黄鹂，曹家树．白菜多聚半乳糖醛酸酶基因 BcMF6 的克隆、序列分析及其表达．园艺学报，2007，34(1)：117～124
张强．白菜花粉外壁蛋白基因 BcMF5 和多聚半乳糖醛酸酶基因 BcMF6 的分离及其功能验证．博士学位论文，2006
张弢．十字花科植物 BcMF2 和 BcMF4 同源基因的克隆、表达及进化关系的研究．博士学位论文，2005
Aarts MG，Hodge R, Kalantidis K, Forack D, Wilson ZA, Mulligan BJ, Stiekema WJ, Scott R, Pereira A. The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes. Plant J, 1997, 12(3): 615～623
Agashe B, Prasad CK, Siddiqi I. Identification and analysis of DYAD: a gene required for meiotic chromosome organization and female meiotic progression in Arabidopsis. Development, 2002, 129:3935～3943
Ahmad N, Gupta S, Husain MM, Heiskanen KM, Mukhtar H. Differential antiproliferative and apoptotic response of sanguinarine for cancer cells versus normal cells. Clinical Cancer Research, 2000, 6:1524～1528
Ahmed K, Davis AT, Wang H, Faust RA, Yu S, Tawfic S. Significance of protein kinase CK2 nuclear signaling in neoplasia. J Cell Biochem Suppl, 2000, 35:130～5
Albani D, Altosaar I, Arnison PG, Fabijanski SF. A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus: sequences in its 5_flanking region are conserved in other pollen-specific promoters. Plant Mol Biol, 1991, 16:501～513
Albertsen MC, Palmer RG. A comparative light-and electron-microscopic study of microsporogenesis in male sterile (MS1) and male fertile soybeas (Glycine max (L.) Merr.). Am J Bot, 1979, 66:253～265
Allen RL, Lonsdale DM. Molecular characterization of one of the maize polygalacturonase gene family members which are expressed during late pollen development. Plant J, 1993, 3(2): 261～271
Amagai M, Ariizumi T, Endo M, Hatakeyama K, Kuwata C, Shibata D, Toriyama K, Watanabe M. Identification of anther-specific genes in a cruciferous model plant, Arabidopsis thaliana, by using a combination of Arabidopsis macroarray and mRNA derived from Brassica oleracea. Sex Plant Reprod, 2003, 15:213～220
Araya A, Zabaleta E, Blanc V, Brgu D, Hemould M, M ouras A, Litvak S. RNA editing in plant mitochondria, cytoplasmic male sterility and plant breeding. Electron J Biotech, 1998, 1:31～39
Ariizumi T, Hatakeyama K, Hinata K, Sato S, Kato T, Tabata S, Toriyama K. A novel male-sterile mutant of Arabidopsis thaliana, faceless pollen-1, produces pollen with a smooth surface and an acetolysis-sensitive exine. Plant Mol Biol, 2003, 53:107～116
Ariizumi T, Hatakeyama K, Hinata K, Sato S, Kato T, Tabata S, Toriyama K. The HKM gene, which is identical to the MS1 gene of Arabidopsis thaliana, is essential for primexine formation and exine pattern formation. Sex Plant Reprod, 2005, 18: 1～7
Armstrong SJ, Caryl AP, Jones GH, Christopher F, Franklin H. Asy1, a protein required for meiotic chromosome synapsis, localizes to axis-associated chromatin in Arabidopsis and Brassica. J Cell Sci, 2002, 115: 3645～3655
Asano T, Katagiri H, Takata K, Lin JL, Ishihara H, Inukai K, Tsukuda K, Kikuchi M, Hirano H, Yazaki Y. The role of N-glycosylation of GLUT1 for glucose transport activity. J Biol Chem, 1991, 266: 24632～24636
Athanasiou A, Khosravi D, Tamari F, Shore JS. Characterization and localization of short-specific polygalacturonase in distylous Turnera subulata (Turneraceae). Am J Bot, 2003, 90(5):675～682
Atkinson A, Health R, Simposon R, Clarke A, Anderson M. Proteinase inhibition in Nicotiana alata stigmas are derived from a precursor protein which is.processed into five homologous inhibitors. Plant Cell, 1993, 5:203～213
Azumi Y, Liu D, Zhao D, Li W, Wang G, Hu Y, Ma H. Homolog interaction during meiotic prophase I in Arabidopsis required the SOLO DANCERS gene encoding a novel cyclin-like protein. EMBO J, 2002, 21: 3081～3095
Bachem CWB, van der Hoeven RS, de Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF. Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development. Plant J, 1996, 9:745～753
Bai X, Peirson BN, Dong F, Xue C, Makaroff CA. Isolation and characterization of SYN1, a RAD21-like gene essential for meiosis in Arabidopsis. Plant Cell, 1999, 11: 417～430
Balk J, Leaver CJ. The PET1-CMS mitochondrial mutation in sunflower is associated with premature programmed cell death and cytochrome c release. Plant Cell, 2001, 13: 1803～1818
Beadle GM. A gene in Zea mays for failure of cytokinesis during meiosis. Cytologia, 1932, 3: 142～155
Becket J, Boavida LC, Carneiro J, Haury M, Feijo JA. Transcriptional profiling of Arabidopsis tissues reveals the unique characteristics of the pollen transcriptome. Plant Physiol, 2003, 133: 713～725
Bedinger P. The remarkable biology of pollen. Plant Cell, 1992, 4: 879～887
Bednarska E. The effect of exogenous Ca~(2+) ions on pollen grain germination and pollen tube growth. Sex Plant Reprod, 1989, 2: 53～58
Bergerat A, de Massy B, Gadelle D, Varoutas PC, Nicolas A, Forterre P. An atypical topoisomerase Ⅱ from Archaea with implications for meiotic recombination. Nature, 1997, 386: 414～417
Bhatt AM, Lister C, Page T, Fransz P, Findlay K, Jones GH, Dickinson HG, Dean C. The DIF1 gene of Arabidopsis is required for meiotic chromosome segregation and belongs to the REC8/RAD21 cohesin gene family. Plant J, 1999, 19: 463～472
Bishop DK, Park D, Xu L, Kleckner N. DMC1: a meiosis-specific yeast homolog of E. coli RecA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell, 1992, 69: 439～456
Blackmore S, Barnes SH. Pollen wall development in angiosperms. In Blackmore S, Knox RB, eds. Microspores Evolution and Ontogeny. Academic Press: San Diego, 1990, pp 173～192
Bleuyard JY, White CL The Arabidopsis homologue of Xrcc3 plays an essential role in meiosis. EMBO J, 2004, 23 (2): 439～449
Boisson B, Giglione C, Meinnel T. Unexpected protein families including cell defense components feature in the N-myristoylome of a higher eukaryote. J Biol Chem, 2003, 78: 4341～43429
Bonghi C, Rascio N, Ramina A, Casadoro G. Cellulase and polygalacturonase involvement in the abscission of leaf and explants of peach. Plant Mol. Biol, 1992, 20:839～848
Bonhomme S, Budar F, Ferault M, Pelletier G. A 2.5 kb Ncol fragment of Ogura radish mitochondrial-DNA is correlated with cytoplasmic male sterility in Brassica hybrids. Curr Genet, 1991, 19: 121～127
Bonhomme S, Horlow C, Guyon A, Ferault M, Vezon D, Marchand M, de Laissardiere S, Bechtold N, Pelletier G. Screening for gametophytic mutations in the Versailles collection of Arabidopsis thaliana transformants: first results for two putative male gametophytic mutants. Acta Hort, 1998a, 459: 173-181
Bonhomme S, Horlow C, Vezon D, de Laissaxdiere S, Guyon A, Ferault M, Marchand M, Bechtold N, Pelletier G. T-DNA mediated disruption of essential gametophytic genes in Arabidopsis is unexpectedly rare and cannot be inferred from segregation distortion alone. Mol Gen Genet, 1998b, 260: 444～452
Bonza MC, Morandini P, Luoni L, Geisler M, Palmgren MG. De Michelis MI. At-ACA8 encodes a plasma membrane-localized calcium-ATPase of Arabidopsis with a calmodulin-binding domain at the N terminus. Plant Physiol, 2000, 123: 1495～1506
Borgese N, Aggujaro D, Carrera P, Pietrini G, Bassetti M. A Role for N-myristoylation in protein targeting: NADH-cytochrome b5 reductase requires myristic acid for association with outer mitochondrial but not ER membranes. J Cell Biol, 1996, 135: 1501～1513
Boyes DC, Chen C-H, Tantikanjana T, Esch JJ, Nasrallah JB. Isolation of a Second S-Locus-Related eDNA from Brassica oleracea: Genetic Relationships Between the S Locus and Two Related Loci. Genetics, 1991, 127: 221～228
Brown SM, Crouch ML Characterization of a gene family abundantly expressed in Oenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell, 1990, 2: 263～274
Bussink HID, Buxton F, Visser J. Expression and sequence comparison of the A. niger and A. tubigensis genes encoding polygalacturonase Ⅱ. Curr Genet, 1991, 19: 467～474
Buteau H, Pezet A, Ferrag F, Perrot-Applanat M, Kelly PA, Edery M. N-glycosylation of the prolactin receptor is not required for activation ofgene transcription but is crucial for its cell surface targeting. Mol Endocrinol, 1998, 12: 544～555
Buttner M, Sauer N. Monosaccharide transporters in plants: structure, function and physiology. Biochim Biophys Acta, 2000, 1465: 263～274
Buttner M, Truernit E, Baler K, Scholz-Starke J, Sontheim M, Lauterbach C, Huss VAR, Saner N. AtSTP3, a green leaf-specific, low affinity monosaccharide-H~+ symporter ofArabidopsis thaliana. Plant Cell Environ, 1999, 23: 175～184
Cai X, Dong F, Edelmann RE, Makaroff CA. The Arabidopsis SYN1 cohesin protein is required for sister chromatid arm cohesion and homologous chromosome pairing. J Cell Sci, 2003, 116: 2999～3007
Canales, C, Bhatt A M, Scott R, Dickinson H. EXS, a putative LRR receptor kinase, regulates male germline cell number and tapetal identity and promotes seed development inArabidopsis. Curr Biol, 2002, 12: 1718～1727
Cao J-S, Yu X-L, Ye W-Z, Lu G. Xiang X. Functional analysis of a novel male fertility CYP86MF gene in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). Plant Cell Rep, 2006, 24 (12): 715～723
Caryl AP, Armstrong SJ, Jones GH, Franklin FCH. A homologue of the yeast HOP1 gene is inactivated in the Arabidopsis meiotic mutant asyl. Chromosoma, 2000, 109: 62～71
Caryl AP, Jones GH, Franklin FC. Dissecting plantmeiosis using Arabidopsis thaliana mutants. J Exp Bot, 2003, 54: 25～38
Cerster J, Allard S, Robert LS. Molecular Characterization of Two Brassica napus pollen-expressed genes encoding putative arabinogalactan proteins. Plant Physiol, 1996, 110: 1231～1237
Chaturvedi MM, Kumar A, Darnay BG, Chainy GBN, Agarwal S, Aggarwal BB. Sanguinarine (pseudochelerythrine)is a potent inhibitor of NF-κB activation, IκBa phosphorylation, and degradation. J Biol Chem, 1997, 272: 30129～30134
Chaubal R, Zanella C, Trirrmell MR, Fox TW, Albertsen M C, Bedinger P. Two male-sterile mutants of Zea mays (Poaceae) with an extra cell division in the anther wall. Am J Bot, 2000, 87: 1193～1201
Chaudhury AM, Lavithis M, Taylor PE, Craig S, Singh MB, Signer ER, Knox RB, Dennis ES. Genetic control of male fertility in Arabidopsis thaliana: structural analysis of premeiotic developmental mutants. Sex Plant Reprod, 1994, 7: 17～28
Chay CH, Buehler EG, Thorn JM, Whelan TM, Eiedinger PA. Purification of maize pollen exines and analysis of associated proteins. Plant Physiol, 1992, 100: 756～761
Chen C, Marcus A, Li W, Hu Y, Calzada J-PV, Grossniklaus U, Cyr RJ, Ma H. The Arabidopsis ATK1 gene is required for spindle morphogenesis in male meiosis. Development, 2002a, 129: 2401～2409
Chen W, Provart N, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan S, Zou G, Whithama S, Budworth P, Tan Y, Xie Z, Chen X, Lam S, Kreps J, Harper J, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl J, Wang X, Zhu T. Expression profile matrix of Arabidopsis transcription factor genes implies their putative functions in response to environmental stresses. Plant Cell, 2002b, 14: 559～574
Chen YC, McCormick S. sidecar pollen, an Arabidopsis thaliana male gametophytic mutant with aberrant cell divisions during pollen development, Development, 1996, 122: 3243～3253
Cheung AY, Wu HM, Di Stilto V, Glaven R, Chen C, Wong E, Ogdahl J, Estavillo A.Pollen-pistil interactions in Nicotiana tabacura. Ann Bot, 2000, 85: 29～37
Cheung AY, Chen CYH, Tao LZ, Andreyeva T, Twell D, Wu HM. Regulation of pollen tube growth by Rat-like GTPases. J Experi Bot, 2003, 54 (380): 73～81
Cheung AY, May B, Kawata EE, Gu Q, Wu HM. Characterization of cDNAs for stylar transmitting tissue-specific proline-rich proteins in tobacco. Plant J, 1993, 3:151～160
Cheung AY, Wu HM. Arabinogalactan proteins in plant sexual reproduction. Protoplasma, 1999, 208: 87～98
Cho HT, Kende H, Expression of expansin genes is correlated with growth in deepwater rice. Plant Cell, 1997, 9: 1661～1671
Clement SA, Tan CC, Guo J, Kitta K, Suzuki YJ. Roles of protein kinase C and alpha-tocopherol in regulation of signal transduction for GATA-4 phosphorylation in HL-1 cardiac muscle cells. Free Radic Biol Med, 2002, 32: 341～349.
Cnudde F, Moretti C, Porceddu A, Pezzotti M, Gerats T. Transcript profiling on developing Petunia hybrida floral organs. Sex Plant Reprod, 2003, 16: 77～85
Consiglio F, Conicella C, Monti L, Carputo D. Highlights of meiotic genes in Arabidopsis thaliana. A J B, 2003, 2 (12): 516～520
Cosgrove DJ, Bedinger P, Durachko DM. Group I allergens of grass pollen as cell wall-loosening agents. Proc Nail Acad Sci USA, 1997, 94: 6559～6564
Cosgrove DJ. Loosening of plant cell walls by expansins. Nature, 2000, 407: 321～326
Couteau F. Belzile F, Horlow C, Grandjean O, Vezon D, Doutfiaux MP. Random chromosome segregation without meiotic arrest in both male and female meiocytes of a dmcl mutant of Arabidopsis. Plant Cell, 1999, 11: 1623～1634
Dawe RK, Freeling M. The role of initial cells in maize anther morphogenesis. Development, 1992, 116: 1077～1085
Dellapenna D, Bennett AB. In vitro synthesis and processing of tomato fruit polygatacturonase. Plant Physiol, 1988, 86: 1057～1063
DeRisi JL, lyer VR, Brown PO. Exploring the metabolic and genetic control of gene expression on a genomic scale. Science, 1997, 278: 680～686
Dickinson HG, Elleman CJ. Pollen coatings: chimeric genetics and new functions. Sex Plant Reprod, 2000, 12: 302～309
Dickinson HG. Dry stigmas, water and self-incompatibility in Brassica. Sex Plant Reprod, 1995, 8: 1～10
Ding DX-H, Vera JC, Heaney ML, Golde DW. N-glycosylation of the human granulocyte-macrophage colony-stimulating factor receptor alpha subunit is essential for figand binding and signal transduction. J Biol Chen, 1995, 270: 24580～24584.
Domi'nguez E, Mercado JA. Pollen sporopollenin: degradation and structural elucidation. Sex Plant Reprod, 1999, 12: 171～178
Dong X-Y, Hong Z-L, Sivaramakrishnan M, Mahfouz M, Verma D P S. Callose synthase (CalS5) is required for exine formation during microgametogenesis and for pollen viability in Arabidopsis. Plant J, 2005, 42: 315～328
Doughty J, Dixon S, Hiscock SJ, Willis AC, Parkin IAP, Dickinson HG. PCP-Al, a defensin-like Brassica pollen coat protein that binds the S-locus glycoprotein, is the product of gametophytic gene expression. Plant Cell, 1998, 10: 1333～1347
Doughty J, Hedderson F, McCubbin A, Dickinson HG. Interaction between a coating-borne pepfide of the Brassica pollen grain and stigmatic S (self-incompatibility)-locus-specific glycoprnteins. Proc Natl Acad Sci, USA, 1993, 90: 467～471
Doughty J, Wong HY, Dickinson HG. Cysteine-rich pollen coat proteins (PCPs) and their interactions with stigmatic S (incompatibility) and S-related proteins in Brassica: putative roles in SI and pollination. Ann Bot, 2000, 85 (A): 161～169
Doutriaux MP, Couteau F, Bergounioux C, White C. Isolation and characterization of the RAD51 and DMC1 homologs from Arabidopsis thaliana. Mol Gen Genet, 1998, 257: 283～291
Dubald M, Barakate A, Mandaron P, Mache R. The ubiquitous presence of exopolygalacturonase in maize suggests a fundamental cellular function for this enzyme. Plant J, 1993, 4:781～791
Eady C, Lindsey K, Twell D. The significance of microspore division and division symmetry for vegetative cell-specific transcription and generative cell differentiation, Plant Cell, 1995, 7: 65～74
El-Ghazaly G, Jelsen WA. DeveIopment of wheat (Triticum aeasvum L.) pollen, Ⅱ Histochemical differentiation of wall and Ublsch bodies during development. Amer Bot, 1987, 74 (9): 1396～1418
Fei HM, Sawhney VK. Ultrastructural characterization of male sterile33 (ms33) mutant in Arabidopsis affected in pollen desiccation and maturation. Can J Bot, 2001, 79: 118～129
Fernandez B, Czech MP, Meisner H. Role of protein kinase C in signal attenuation following T cell receptor engagement. J Biol Chem 1999, 274: 20244～20250
Fischer RL, Bennett AB. Role of cell wall hydrolyses in fruit riping. Annu Rev Plant Physiol. Plant Mol Biol, 1991, 42: 675～703
Franklin-Tong N, Bryant J. P4C1-Plant Reproductive Biology. Elsevier Abstracts, 2002: S125～S137
Franklin-Tong VE. Signaling and modulation of pollen tube growth. Plant Cell, 1999, 11: 727～738
Futamura N, Mori H, Kouchi H, Shinohara K. Male flower-specific expression of genes for polygalacturonase, pectin methylesterase and β-1,3-glucanase in a dioecious willow (Salix gilgliana Seemen). Plant Cell Physiol, 2000, 41: 16～26
Gallego ME, White CI. RAD50 function is essential for telomere maintenance in Arabidopsis. Proc Natl Acad Sci USA, 2001, 98: 1711～1716
Gilbert C, Gaudry M, Naccache PH. Rapid priming of calcium mobilization and superoxide anion production in human neutrophils by substimulatory concentrations of phorbol esters: a novel role for protein kinase C and tyrosine phosphorylation in the up-modulation of signal transduction. Cell Signal, 1992, 4: 511～523
Girke T, Todd J, Ruuska S, White J, Benning C, Ohlrogge J. Microarray analysis of develo ping Arabidopsis seeds. Plant Physiol, 2000, 124 (4): 1570～1581
Goldberg RB, Beals TP, Sanders PM. Anther development: Basic principles and practical applications. Plant Cell, 1993, 5: 1217～1229
Golovkin M, Reddy ASN. A calmodulin-binding protein from Arabidopsis has an essential role in pollen germination. Proc Natl Acad Sci USA, 2003,100 (18): 10558～10563
Grelon M, Vezon D, Gendrot G, Pelletier G, AtSPO11-1 is necessary for efficient meiotic recombination in plants. EMBO J, 2001, 20: 589～600
Grini PE, Schnittger A, Schwarz H, Zimmermann I, Schwab B, Jurgensa G, Hulskampa M. Isolation of ethyl methanesulfonate-induced gametophytic mutants in Arabidopsis thaliana by a segregation distortion assay using the multimarker chromosome 1. Genetics, 1999, 151: 849～863
Grobe K, Powell L D. Role of protein kinase C in the phosphorylation of CD33 (Siglec-3) and its effect on lectin activity. Blood, 2002, 99: 3188～3196
Gu Y, Vernoud V, Fu Y, Yang ZB. ROP GTPase regulation of pollen tube growth through the dynamics of tip-localized F-actin. J Experi Bot, 2003, 54(380): 93～101
Guyon VN, Astwood JD, Garner EC, Dunker AK, Taylor LP. Isolation and characterization of cDNAs expressed in the early stages of flavonol-induced pollen germination in petunia. Plant Physiol, 2000, 123: 699～710
Hadfield KA, Bennett AB. Polygalacturonases: many genes in serch of a function. Plant Physiol, 1998, 117: 337～343
Hadfield KA, Rose JKC, Yaver DS, Berka RM, Bennett AB. Polygalacturonase gene expression in ripe melon fruit supports a role for polygalacturonase in ripening-associated pectin disassembly. Plant Physiol, 1998, 117: 363～373
Hanson MR, Bentolila S. Interactions of mitocbondrial and nuclear genes that affect male gametophyte development. Plant Cell, 2004, 16: 154～169
He C, Mascarenhas JP. MEI1, AnArabidopsis gene required for male meiosis: isolation and characterization. Sex Plant Reprod, 1998, 11: 199～207
Heizmann P, Luu DT, Dumas C. Pollen-stigma adhesion in the Brassicaceae. Ann Bot, 2000, 85 (A): 23～28
Hepler PK, Vidali L, Cheung AY. Polarized cell growth in higher plants. Annu Rev Cell Dev Biol, 2001, 17: 159～187
Heslop-Harrison Y, Hesiop-Harrison J, Reger BJ. Pollen tube guidance and the regulation of tube number in Zea mays. Acta Bot Neerl, 1985, 34: 193～211
Heslop-Harrison Y, Reger BJ. Tissue organization, pollen receptivity and pollen tube guidance in normal and mutant stigmas of the grass Pennisetum Pyphoides (Burm) Stap et Hubb. Sex Plant Reprod, 1988, 1: 182～183
Higginson T, Li SF, Roger W, Parish.AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana. Plant J, 2003, 35(2): 177～192
Hiscock SJ, Dewey FM, Coleman JOD, Dickinson HG. Identification and Localization of an active cutinase in the pollen of Brassica napus L. Planta, 1994, 193: 377～384
Hiscock SJ, Doughty J, Willis AC, Dickinson HG. A 7-kDa pollen coating-borne peptide from Brassica napus interacts with S-locus glycuprotein and S-locus-related glycoprotein. Planta, 1995, 196: 367～374
Holdaway-Clarke TL, Feijo JA, Hackett GR, Kunkel JG, Hepler PK. Pollen tube growth and the lintracellular cytosolic calcium gradient oscillate in phase while extracellular calcium influx is delayed. Plant Cell, 1997, 9: 1999～2010
Holstege FC, Jennings EG, Wyrick JJ, Lee TI, Hengartner CJ, Green MR, Golub TR, Lander ES, Young RA. Dissecting the regulatory circuitry of a eukaryotic genome. Cell, 1998, 95: 717～728
Hong HP, Gerster JL, Datla RSS, Albani K, Scoles G. Keller W and Roberts LS. The promoter of a Brassica napus polygalacturonase gene directs pollen expression of beta-glucuroaidase in transgenic Brassica phnts. Plant Cell Rep, 1997, 16: 373～378
Honys D, Twell D. Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol, 2003, 132: 640～652
Honys D, Twell D. Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biology, 2004, 5:R85
Horda CLH, Chena C, DeYoung BJ, Clarke SE, Ma H. The BAM1/BAM2 receptor-like kinases are important regulators of Arabidopsis early anther development. Plant Cell, 2006, 18: 1667～1680
Horvath DP, Schaffer R, West M, Wisman E. Arabidopsis microarrays identify conserved.and differentially expressed genes involved in shoot growth and development from distantly related plant species. Plant J, 2003, 34 (1): 125～134
Horvitz R, Herskowitz I. Mechanisms of asymmetric cell division: Two Bs or not two Bs, that is the question. Cell, 1992, 68: 237～255
Huala SK, Sussex IM. Determination and cell interactions in reproductive meristems. Plant Cell, 1993, 8:1391～1407,
Hughes TR, Marton MJ, Jones AR, Roberts CJ, Stoughton R, Armour CD, Bennett HA, Coffey E, Dai H, He YD. Functional discovery via a compendium of expression profiles. Cell, 2000, 102: 109～126
Hulskamp M, Kopczak SD, Horejsi TF, Kihl BK, Pruitt RE. Identification of genes required for pollen stigma recognition in Arabidopsis thaliana. Plant J, 1995, 8: 703～715
Hulskamp M, Nikesh SP, Grini P, Schneitz K, Zimmermann I, Lolle SJ, Pruitt RE. The STUD gene is required for male-specific cytokinesis after telophase Ⅱ of meiosis in Arabidopsis thaliana. Dev Biol, 1997, 187: 114～124
Ikeda S, Nasrallah JB, Dixit R, Preiss S, Nasrallah ME. An aquaporin-like gene required for the Brassica self-incompatibility response. Science, 1997, 276: 1564～1566
Jakobsen MK, Poulsen LR, Schulz A, Fleurat-Lessard P, Mφller A, Husted S, Schiφtt M, Amtmann A, Palmgren MG. Pollen development and fertilization in Arabidopsis is dependent on the MALE GAMETOGENESIS IMPAIRED ANTHERS gene encoding a Type V P-type ATPase. Genes & Dev, 2005, 19: 2757～2769
Jauh GY, Lord EM. Movement of the tube cell in lily style in the absence of the pollen grain and the spent pollen tube. Sex Plant Reprod, 1995, 8: 168～172
Jefferson RA, Kavanagh TA, Bevan MW. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 1987, 6: 3901～3907
Jenkins ES, Paul W, Coupe SA, Bell SJ, Davies EC, Roberts JA. Characterization of an mRNA encoding a polygalacturonase expressed during pod development in oilseed rape (Brassica napus L.). J Exp Bot, 1996, 47: 111～115
Jiang L, Yang S-L, Xie L-F, Puah CS, Zhang X-Q, Yang W-C, Sundaresan V, Ye D. VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract. Plant Cell, 2005, 17: 584～596
John ME and Petersen MW. Cotton (Gossypium hirsutum L.) pollen-specific polygalaeturonase mRNA: tissue and temporal specificity of its promoter in transgenic tobacco. Plant Mol Biol, 1994, 26(6): 1989～1993
Johnson MA, Preuss D. Plotting a course: Multiple signals guide pollen tubes to their targets. Dev Cell, 2002, 2: 273～281
Johnson SA, McCormick S. Pollen Germinates Precociously in the Anthers of raring.to-go, an Arabidopsis Gametophytic Mutant1. Plant Physiol, 2001, 126:685～695
Kalaitzis P, Solomos T, Tucker ML. Three different polygalacturonases are expressed in tomato leaf and flower abscission, each with a different temporal expression pattern. Plant Physiol, 1997, 113:130～308 Kapoor S, Kobayashi A, Takatsuji H, Silencing of the tapetum-specific zinc finger gene TAZ1 causes premature degeneration of tapetum and pollen abortion in petunia. Plant Cell, 2002, 14: 2353～2367
Kasahara RD, Portereiko MF, Sandakie-Nikolova L, Rabiger DS, Drews GN. MYB98 is required for pollen tube guidance and synergid cell differentiation in arabidopsis. Plant Cell, 2005, 17: 2981～2992
Kaur J, Sebastian J, Siddiqi I. The Arabidopsis-mei2-Like genes pplay a role in meiosis and vegetative growth in Arabidopsis. Plant Cell, 2006, 18: 545～559
Kawaguchi K, Shibuya N. Characterization of arahinogalactan-proteins and a related olignsaccharide in developing rice anthers. In Nothnagel EA, Bacic A, Clarke AE, eds, Cell Develop Biology of Arabinogalactanproteins. Kiuwer Academic/Plenum Publishers: New York, 2000, pp 149～152
Kawakami Y, Kitanra J, Hartman SE, Lowell CA, Siraganian RP, Kawakami T. Regulation of protein ldnase CBI by two protein-tyrosine kinases, Btk and Syk. Proc Natl Acad Sci USA, 2000, 97: 7423～7428
Keeney S, Giroux CN, Kleckner N. Meiosis-specific DNA doublestrand breaks are catalyzed by SPOll, a member of a widely conserved protein family. Cell, 1997, 88: 375～384
Klimyuk VI, Jones JDG. AtDMC1, the Arabidopsis homologue of the yeast DMC1 gene: characterization, transposon-induced allelic variation and meiosis-associated expression. Plant J, 1997, 11:1～14
Lalanne E, Honys D, Johnson A, Borner GHH, Lilley KS, Dupree P, Grossnildaus U, Twell D. SETH1 and SETH2, two components of the glycosylphosphatidylinositol anchor biosynthetic pathway, are required for pollen germination and tube growth in Arabidopsis. Plant Cell, 2004, 16: 229～240
Laser KD, Lersten NR. Anatomy and cytology of microsporogenesis in cytoplasmic male sterile angiosperms. Bot Rev, 1972, 38(3): 425～454
Lee JY, Lee DH. Use of serial analysis of gene expression technology to reveal changes in gene expression in Arabidopsis pollen undergoing cold stress. Plant Physioi, 2003, 132: 517～529
Lennon K, Roy S, Hepler RK, Lord EM. The structure of the transmitting tissue of Arabidopsis thaliana(L.) and the path of pollen tube growth. Sex Plant Reprod, 1998, 11: 49～59
Lennon KA, Lord EM. The in vivo pollen tube cell of Arabidopsis thaliana: Tube cell cytoplasm, wall and cytoskeleton. Protuplasma, 2000, 214: 45～56
Lester DR, Sherman WB, Atwell BJ. Endopolygalacturonase and the Melting Flesh (M) locus in peach. J Amer Soci Horti Sci, 1996, 121, 231～235
Li H, Bacic A, Read SM. Role of a callose synthase zymogen in regulating wall deposition in pollen tubes of Nicotiana alata Link et Otto. Planta, 1999b, 208: 528～538
Li H, Lin Y, Heath RM, Zhu MX, Yang Z. Control of pollen tube tip growth by a Rop GTPase-dependent pathway that leads to the tip-localized calcium influx. Plant Cell, 1999a, 11: 1731～1742
Li H, Wu G, Ware D, Davis KR, Yang Z. Arabidopsis Rho-related GTPases: differential gene expression in pollen and polar localization in fission yeast. Plant Physiol, 1998, 118: 407～417
Li L-B, Chen N, Ramamoorthy S, Chi L, Cui X-N, Wang LC, Reith MEA. The Role of N-glycosylation in function and surface trafficking of the human dopamine transporter. J Biol Chem, 2004, 279: 21012～21020
Li YQ, Mareck A, Faleri C, Moseatelli A, Liu Q, Cresti M. Detectionand localization of pectin methylesterase isoforms in pollen tubes of Nicotiana tabacum L. Planta, 2002, 214: 734-740
Lin R, Hiscott J. A role for casein kinase Ⅱ phosphorylation in the regulation of IRF-1 transcriptional activity. Mol Cell Biochem 1999, 191: 169～180
Lin Y, Yang Z. Inhibition of pollen tube elongation by microinjected anti-Rop1Ps antibodies suggests a crucial role for Rho-type GTPases in the control of tip growth. Plant Cell, 1997, 9:1647～1659
Liu L-C, Cao J-S, Yu X-L, Xiang X, Pei Y-J. Expression of an antisense BcMF3 affects mierosporogenesis and pollen tube growth in Arabidopsis. Agricul Sci in China, 2006, 5(5): 339～345
Liu XC, Jones K, Dickinson HG. DNA synthesis and cytoplasmic differentiation in tapetal cells of normal and cytoplasmic cytoplasmic male sterile lines of Petunia hybrida. Theor Appl Genet, 1987, 74: 846～851
Lobstein E, Guyon A, Ferault M, Twell D, Pelletier G, Bonhomme S. The putative Arabidopsis homolog of yeast vps52p is required for pollen tube elongation, localizes to golgi, and might be involved in vesicle trafficking. Plant Physinl, 2004, 135(3): 1480～1490
Lord EM, Russell SD. Mechanisms of pollination and fertilization in plants. Annu Rev Cell Devel Biol, 2002, 18: 81～105
Lord EM. Adhesion and cell movement during pollination: churches la femme. Trends Plant Sci, 2000, 5: 368～373
Lush WM, Spurck T, Joosten R. Pollen tube guidance by the pistil of a solanaceous plant. Ann Bot, 2000, 85(A): 39～47
Ma J, Duncan D, Morrow DJ, Fernandes J, Walbot V. Transcriptome profiling of maize anthers using genetic ablation to analyze pre-meiotic and tapetal cell types. Plant J, 2007, 50: 637～648
Magnard JL, Yang M, Chen YCS, Leary M, McCormick S. The Arabidopsis gene TARDYASY'NCHRONOUS MEIOSIS is required for the normal pace and synchrony of cell division during male meiosis. Plant Physiol, 2001, 127:1157～1166
Matho R, Trewavas AJ. Localized apical increases of cytosolic free calcium control pollen tube orientation. Plant Cell, 1996, 8: 1935～1949
Mandaokar A, Kumar VD, Amway M, Browse J. Microarray and differential display identify genes involved in jasmonate-dcpendent anther development. Plant Mol Biol, 2003, 52(4): 775～86
Markovic O, Janecek S. Pectin degrading glycoside hydrolases of family 28: sequence-structural features, specificifies and evolution. Protein Eng Des Sel, 2001, 14: 615～631
Marquez J, Seoane-Camba JA, Suarez-Cervera M. Allergenic and antigenic proteins released in the apertural sporoderm during the activation process in grass pollen grains. Sex Plant Repred, 1997, 10: 269～278
Masearenhas JP. Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol, 1990, 41:317～338
Mascarenhas JP. The male gametophyte of flowering plants. Plant Cell, 1989, 1: 657～664
Mathilde G. Ghislaine G. Daniel V, and Georges P. The Arabidopsis MEI1 gene encodes a protein with five BRCT domains that is involved in meiosis-specific DNA repair events independent of SPO11-induced DSBs. Plant J, 2003, 35(4): 465～75
Mayfield JA, Preuss D. Rapid initiation of Arabidop.sis pollination requires the oleosin-domain protein GRP17. Nat Cell Biol. 2000, 2:128～130
McCormick S. Control of male gametophyte development. Plant Cell, 2004, 16:S142～S153
McCormick S. Male gametophyte development. Plant cell, 1993, 5: 1265～1275
McCoy TJ, Smith LY. Genetics, cytology, and crossing behavior of an alfalfa (Medicago sativa) mutant resulting in failure of the postmeiotic cytokinesis. Can J Genet & Cytol, 1983, 25: 390-397
Mele G, Ori N, Sato Y, Hake S. The knottedl-like homeobox gene BREVIPEDICELLUS regulates cell differentiation by modulating metabolic pathways. Genes & Development, 2003, 17: 2088～2093
Mercier R, Armstrong SJ, Horlow C, Jackson NP, Makaroff CA, Vezon D, Pelletier G, Jones GH, Franklin FCH. The meiotic protein SWI1 is required for axial element formation and recombinationinitiation in Arabidopsis. Development, 2003, 130: 3309～3318
Mercier R, Vezon D, Bullier E, Motamayor JC, Sellier A, Lefevre F, Pelletier G, Horlow C. Switch1 (SWI1): a novel protein required for the establishment of sister chromatid cohesion and for bivalent formation at meiosis. Gene Dev, 2001, 15: 1859～1871
Mitsui H, Nakatani K, Yamaguchi-Shinozaki K, Shinozaki K, Nishikawa K, Takahashi H. Sequencing and characterization of the kinesin-related genes katB and katC of Arabidopsis thaliana. Plant Mol Biol, 1994, 25: 865～876
Mitsui H, Yamaguchi-Shinozaki K, Shinozaki K, Nishikawa K, Takahashi H. Identification of a gene family (kat) encoding kinesin-like proteins in Arabidopsis thaliana and the characterization of secondary structure of KatA. Mol Gen Genet, 1993, 238: 362～368
Moon YH, Chen L, Pan RL, Chang HS, Zhu T, Maffeo DM, Sung ZR. EMF genes maintain vegetative development by repressing the flower program in Arabidopsis. Plant Cell, 2003, 15: 681～693
Motamayor JC, Vezon D, Bajon C, Sanvanet A, Grandjean O, Marchand M, Bechtold N, Pelletier G, Horlow C. Switch (swi1), an Arabidopsis thaliana mutant affected in the female meiotic switch. Sex Plant Reprod, 2000, 12: 209～218
Mouline K, Very A-A, Gaymard F, Boucherez J, Pilot G, Devic M, Bouchez D, Thiband J-B, Sentenac H. Pollen tube development and competitive ability are impaired by disruption of a Shaker K~+ channel in Arabidopsis. Genes Dev, 2002, 16(3): 339～350
Moutinho A, Hussey PJ, Trewavas AJ, MalhoR. cAMP acts as a second messenger in pollen tube growth and reorientation. Proc Natl Acad Sci USA, 2001, 98(18): 10481～10486
Mu J-H, Stains JP, Kao T-h. Characterization of a pollen-expressed gene encgding a putative pectin esterase of Petunia inflata. Plant Mol Biol, 1994, 25: 539～54
Muschietti J, Dircks L, Vancanneyt G, McCormick S. LAT52 protein is essential for tomato pollen development: Pollen expressing antisense LAT52 RNA hydrates and germinates abnormally and cannot achieve fertilization. Plant J, 1994, 6: 321～338
Neelam A, Sexton R. Cellulase (endo_-1, 4 glucanase) and cell wall breakdown during anther development in the sweet pea (Lathyrus odoratus L.): isolation and characterization of partial cDNA clones. J Plant Physiol, 1995, 146: 622～628
Niewiadomski P, Knappe S, Geimer S, Fischer K, Schulz B, Unte US, Rosso MG, Ache P, Flugge U, Schneidera A. The Arabidopsis plastidic glucose 6-phosphate/phosphate translocator GPT1 is essential for pollen maturation and embryo sac development. Plant Cell, 2005, 17: 760～775
Niu L, Heaney ML, Vera JC, Golde DW. High-affinity binding to the GM-CSF receptor requires intact N-glycosylation sites in the extracellular domain of the beta subunit. Blood, 2000, 95: 3357～3362
Nonomura KI, Miyoshi K, Eiguchi M, Suzuki T, Miyao A, Hirochika H, Kurata N. The MSP1 Gene is necessary to restrict the number of cells entering into male and female sporogenesis and to initiate anther wall formation in rice. Plant Cell, 2003, 15: 1728～1739
Olsen AN, Mundy J, Skrivcr K. Peptomics, identification of novel cationic Arabidopsis peptides with conserved sequence motifs. In Silico Biology, 2002, 2, 39～44
Orlandi I, Bettiga M, Alberghina L, Vai M. Transcriptional profiling of ubp10 null mutant reveals altered subtelomeric gene expression and insurgence of oxidative stress response. J Biol Chem, 2004, 279(8):6414～6425
Otegui MS, Staehelin LA. 3D Tomographic analysis of post-meiotic cytokinesis during pollen development in Arabidopsis thaliana. Planta, 2004, 218: 501～515
Owen HA, Makaroff CA. Ultrastructure of microsporogenesis and microgametogenesis in Arabidopsis thaliana (L.) Heynh. ecotype Wassilewskija (Brassicaceae). Protoplasma, 1995, 185: 7～21
Pacini E, Franchi E, Hesse M. The tapetum: its form, function and possible phylogeny in rmbryophyta. Plant Systematics and Evolution, 1985, 149: 155～185
Palanivelu R, Brass L, Edlund AF, Preuss D. Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABAlevels. Cell, 2003, 14(1):47～59
Papini A, Mosti S, Brighigna L. Programmed-cell-death events during tapetum development of angiosperms. Protoplasma, 1999, 207: 213～221
Park BS, Kim JS, Kim SH, Park YD. Characterization of a pollen-preferential gene, BAN102, from Chinese cabbage. Plant Cell Rep, 2005, 14: 1～8
Park J, Hill MM, Hess D, Brazil DP, Hofsteenge J, Hemmings BA. Identification of tyrosine phosphorylation sites on 3-phosphoinositide-dependent protein kinase-1 and their role in regulating kinase activity. J Biol Chem, 2001, 276: 7459～37471
Park SK, Howden R, Twell D. The Arabidopsis thaiiana gametophytic mutation gemini pollenl disrupts microspore polarity, division asymmetry and pollen cell fate. Development, 1998, 125(19): 3789～3799
Park SK, Twell D. Novel patterns of ectopic cell plate growth and lipid body distribution in the Arabidopsis gemini pollenl mutant. Plant Physiol, 2001, 126(2): 899～909
Parkin IAP, Gulden SM, Sharpe AG. Lukens L, Trick M, Osborn TC, Lydiate DJ. Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana. Genetics, 2005, 171 (2): 765～781
Parre E, Geitmann A. Pectin and the role of the physical properties of the cell wall in pollen tube growth of Solanum chacoense. Planta, 2005, 220: 582～592
Paxson-Sowders DM, Dodrill CH, Owen HA, Makaroff CA. DEX1, a novel plant protein, is required for exine pattern formation during pollen development in Arabidopsis. Plant Physiol, 2001, 127: 1739～1749
Paxson-Sowders DM, Owen HA, Makaroff CA. A comparative ultrastructural analysis of exine pattern development in wild type Arabidopsis and a mutant defective in pattern formation. Protoplasma, 1997, 198: 53～65
Pennell RI, Jam-fiche L, Kjellbom P, Scofield GN, Peart JM, Roberts K. Developmental regulation of a plasma membrane arabinogalactan protein epitope in oilseed rape flowers. Plant Cell, 1991, 3: 1317～1326
Pennell RI, Knox JP, Scofield GN, Selvendran RR, Roberts K. A family of abundant plasma membrane-associated glycoproteins related to the arabinogalactan proteins is unique to flowering plants. J Cell Biol, 1989, 108: 1967～1977
Pereira LG, Coimbra S, Oliveira H, Monteiro L, Sottomayor M. Expression of arabinogalactan protein genes in pollen tubes of Arabidopsis thaliana. Planta, 2006, 223: 374～380
Petersen M, Brodersen P, Naested H, Andreasson E, Lindhart U, Johansen B, Nielsen HB, Lacy M, Austin MJ, Parker JE, Sharma SB, Klessig DF, Martienssen R, Mattsson O, Jenscn AB, Mundy J. Arabidopsis map kinase 4 negatively regulates systemic acquired resistance. Cell, 2000, 103: 1111～1120
Petersen M, Sander L, Child R, Van Onckelen H, Ulvskov P, Borkhardt B. Isolation and characterisation of a pod dehiscence zone-specific polygalacturonase from Brassica napus. Plant Mol Biol, 1996, 31:517-527
Picton JM, Steer MW. The effects of ruthenium red, lanthanum, fluorescein isothiocyanate and trifluoperazine on vesicle transport, vesicle fusion and tip extension in pollen tubes. Planta, 1985, 163: 20～26
Pierson ES, Miller DD, Callaham DA, Shipley AM, Rivers BA, Cresti M, Hepler P. Pollen tube growth is coupled to the extracellular calcium ion flux and the intracellular calcium gradient: effect of BAPTA-type buffers and hypertonic media. Plant Cell, 1994, 6: 1818～1828
Piffanelli P, Ross JHE, Murphy DJ. Biogenesis and function of the lipidic structures of pollen grains. Sex Plant Reprod, 1998, 11: 65～80
Pina C, Pinto F, Feijo J, Becker J. Gene family analysis of the Arabidopsis pollen transcriptome reveals biological implications for cell growth, division control, and gene expression Regulation. Plant Physiol, 2005, 138: 744～756
Poethig S. Genetic mosaics and cell lineage analysis in plants. Trends Genet, 1989, 5: 273～277
Porat R, Lu P, O'Neill SD. Arabidopsis SKP1, a homologue of a cell cycle regulator gene, is predominantly expressed in meristematic cells. Planta, 1998, 204: 345～351
Pressey R, Reger BJ. Polygalacturonase in pollen from corn and other grasses. Plant Sci, 1989, 59: 57～62
Pressey R. Polygalacturonase in tree pollens. Phytochemistry. 1991, 30: 1753～1755
Preuss D, Lemieux B, Yen G, Davis RW. A conditional sterile mutation eliminates surface components from Arabidopsis pollen and disrupts cell signalling during fertilization. Genes and Development, 1993, 7: 974～985
Procissi A, Guyon A, Pierson ES, Giritch A, Knuiman B, Grandjean O, Tonelli C, Derksen J, Pelletier G, Bonhomme S. K/NKY POLLEN encodes a SABRE-like protein required for tip growth in, Arabidopsis and conserved among eukaryotes. Plant J, 2003, 36 (6): 894～904
Procissi A, Laissardie're SD, Fe'rault M, Vezon D, Pelletier G, Bonhomme S. Five gametophytic mutations affecting pollen development and pollen tube growth in Arabidopsis thaliana. Genetics, 2001, 158: 1773～1783
Raghavan V. A transient accumulation of poly (A) containing RNA in the tapetum of Hyoscyamus niger during microsporogenesis. Dev Biol, 1981, 81: 342～348
Raghavan V. mRNAs and a cloned histone gene are differentially expressed during anther and pollen development in rice (Oryza sativa L.). J Cell Sci, 1989, 92: 217～229
Rato C, Monteiro D, Hepler PK, Malho R. Calmodulin activity and cAMP signalling modulate growth and apical secretion in pollen tubes. Plant J, 2004, 38 (6): 887～897
Redgwell RJ, MacRae EA, Hallett I, Fischer M, Perry J, Harker R. In vivo and in vitro swelling of cell walls during fruit ripening. Planta, 1997, 203: 162～173
Rhee SY, Osborne E, Poindexter PD, Somerville CR. Microspore separation in the quartet 3 mutants of Arabidopsis is impaied by a defect in a developmentally regulated polygalacturonase required for pollen mother cell wall degradation. Plant Physiol, 2003, 133: 1170～1180
Rhee SY, Somerville CR.Tettad pollen formation in quartet mutants of Arabidopsis thaliana is associated with persistence of pectic polysaccharides of the pollen mother cell wall. Plant J, 1998, 15: 79～88
Robert LS, Allard S, Gerster JL, Cass L, Simmonds J. Isolation and characterization of a polygalacturonase gene highly expressed in Brassica napus pollen. Plant Mol Biol, 1993, 23(6): 1273～1278
Rockmill B, Roeder GS. Meiosis in asynaptic yeast. Genetics, 1990, 126: 563～574
Rose JKC, Lee HH, Bennett AB. Expression of a divergent expansin gene is fruit-specific and ripening-regulated. Proc Nail Acad Sci USA, 1997, 94: 5955～5960
Ross KJ, Fransz P, Armstrong SJ, Vizir I, Mulligan B, Franklin FCH, Jones GH. Cytological characterization of four meiotic mutants of Arabidopsis isolated from T-DNA-transformed lines. Chromosome Res, 1997, 5: 551～559
Rozwadowski K, Zhao RH, Jackman L, Huebert T, Burkhart WE, Hemmingsen SM, Greenwood J, Roths SJ. Characterization and immunolocalizatioa of a cytosolic calcium-binding protein from Brassica napus and Arabidopsis pollen. Plant Physiol, 1999, 120: 787～798
Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning. New York: Cold Spring Harbor Laboratory Press, 1989
Sainchez-Moran E, Mercier R, Higgins JD, Armstrong. SJ, Jones GH, Franklin CH. A strategy to investigate the plant meiotic proteome. Cytogenet Genome Research, 2005, 109: 181～189
Sanders PM, Bui AQ, Weterings K, McIntire KN, Hsu Y-C, Lee PY, Truong MT, Beals TP, Goldberg RB. Anther developmental defects in Arabidopsis thaliana male-sterile mutants. Sex Plant Reprod, 1999, 11:297～322
Sato S, Hotta Y, Tabata S. Structural analysis of a RecA-like gene in the genome of Arabidopsis thaliana. DNA Research, 1995, 2:89～93
Schiefthaler U, Balasubramanian S, Sieber P, Chevalier D, Wisman E, Schneitz K. Molecular analysis of NOZZLE, a gene involved in pattern formation and early sporogenesis during sex organ development in Arabidopsis thaliana. Proc Nail Acad Sci USA, 1999, 96:11664～11669
Schiφtt M, Romanowsky SM, Baekgaard L, Jakobsen MK, Palmgren MG, Harper JF. A plant plasma membrane Ca~(2+) pump is required for normal pollen tube growth and fertilization. Proc Natl Acad Sci USA, 2004, 101(25): 9502～9507
Schneidereit A, Scholz-Starke J, Buttner M. Functional characterization and expression analyses of the glucose-specific AtSTP9 monosaccharide transporter in pollen of Arabidopsis. Plant Physiol, 2003, 133: 182～190
Scholz-Starke J, Buttner M, Sauer N. AtSTP6, a new pollen-specific H~+-monosacchadde symporter from Arabidopsis. Plant PhysioI, 2003, 131: 70～77
Schopfer CR, Nasrallah ME, Nasrallah JB. The male determinant of self-incompatibility in Brassica. Science, 1999, 286: 1607～1700
Schwarzacher. Meiosis, recombination and chromosomes: a review of gene isolation and fluorescent in situ hybridization data in plants. J Exp Bot, 2003, 54:11-23
Scott RJ, Spielman M, Dickinsonb HG. Stamen structure and function. Plant Cell, 2004, 16: S46-S60
Sexton R, Campillo ED, Duncan D, Lewis LN. The purification of an anther cellulase(1:4-glucan hydrolase) from Lathyrus odoratus L. and its relationship to the similar enzyme found in abscission zones. Plant Sci, 1990, 67:169-176
Sharp DJ, Rogers GC, Scholey, JM. Microtubule motors in mitosis. Nature, 2000, 407:41-47
Sheridan WF, Golubeva EA, Abrhamova Ⅱ, Golubovskaya IN. The macl mutation alters the developmental fate of the hypodermal cells and their cellular progeny in the maize anther. Genetics, 1999, 153: 933-941
Shinohara A, Ogawa H, Matsuda Y, Ushio N, Ikeo K, Ogawa T. Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and RecA. Nature Genetics, 1993, 4: 239～243
Shukia A.K, Vijayaraghavan MR, Chaudhry B. Biology of pollen. New Delhi: APH Publishing Corporation. 1998
Singh M, Brown GG. Suppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitoehondrial gent region. Plant Cell, 1991, 3: 1349～1362
Sitrit Y, Downie B, Bennett AB, Bradford KJ. A novel exo-polygalacturonase is ass ociated with radicle protrusion in tomato seeds. Plant Physiol, 1996, 111(Supplement): 161～163
Sobotka R, Sakova L, Curn V. Molecular mechanisms of self-incompatibility in Brassica. Curt Issues Mol Biol, 2000, 2 (4): 103～12
Sommer-Knudsen J, Clarke AE, Bacic A. A galactose-dch, cell-wall glycoprotein from styles of Nicotiana alata. Plant J, 1996, 9: 71～83
Sommer-Knudsen J, Lush WM, Bacic A, Clarke AE. Re-evaluation of the role of a transmitting tract-specific glycoprotein on pollen tube growth. Plant J, 1998, 13: 529～535
Southworth D. Exine biochemistry. In S Blackmore, RB Knox, eds, Microspores: Evolution and Ontogeny. Academic Press: London, 1990, pp 193～212
Spielman M, Preuss D, Li FL, Browne WE, Scott R J, Dickinson HG. TETRASPORE is required for male meiotic cytokinesis inArabidopsis thaliana. Development, 1997, 124: 2645～2657
Stevens R, Grelon M, Vezon D, Oh J, Meyer P, Perennes C, Domenichini S, Bergounioux C. A CDC45 homolog in Arabidopsis is essential for meiosis, as shown by RNA interference-induced gene silencing. Plant Cell, 2004, 16: 99～113
Stratilova E, Dzurova M, Markovic O, Jornvall H. An essential tyrosine residue of Aspergillus polygalacturonase. FEBS Lett, 1996, 11, 382 (1-2): 164～166
Swain SM, Muller AI, Singh DP. The gar2 and rga alleles increase the growth of gibberellin-deficient pollen tubes in Arabidopsis, Plant Physiol, 2004, 134: 694～705
Sym M, Engebrecht J, Roeder GS. ZIP1 is a synaptonemal complex protein required for meiotic chromosome synapsis. Cell, 1993, 72: 365～378
Sym M, Roeder GS. Crossover interference is abolished in the absence of a synaptonemal complex protein. Cell, 1994, 79: 283～292
Sze H, Padmanaban S, Cellier F, Honys D, Cheng N-H, Bock KW, je'ro GC, Li X, Twell D, Ward JM, Hirschi KD. Expression patterns of a novel AtCHX gene family highlight potential roles in osmotic adjustment and K~+ Homeostasis in Pollen Development. Plant Physiol, 2004, 136: 2532～2547
Takayama S, Shiba H, Iwano M, Asano K, Hara M, Che FS, Watanabe M, Hinata K, Isogai A. Isolation and characterization of pollen coat proteins of Brassica campestris that interact with S Iocus-related glycoprotein 1 involved in pollen-stigma adhesion. Proc Natl Acad Sci USA, 2000a, 97(7): 3765～3770
Takayama S, Shiba H, Iwano M, Shimosato H, Che F-S, Kai N, Watanabe M, Suzuki G, Hinata K, Isogai A. The pollen determinant of self-incompatibility in Brassica campestris. Proc Natl Acad Sci USA, 2000b, 97: 1920～1925
Takegami MH, Yoshioka M, Tanaka I, Ito M. Characteristics of isolated microsporocytes from liliaceous plants for studies of the meiotic cell cycle in vitro. Plant Cell Physiol, 1981, 22: 1～10
Taylor JE, Tucker GA, Lasslett Y, Smith CJS, Arnold CM, Watson CF, Schuch W, Grierson D, Roberts JA. Polygalacturonase expression during leaf abscission of normal and transgenic plants. Planta, 1990, 183: 133～138
Taylor JE, Webb STJ, Coupe SA, Tucker GA, Roberts JA. Changes in polygalacturonase activity and solubility of polyuronides during ethylene-stimulated leaf abscission in Sambucus nigra. J Exp Bot, 1993, 44: 93～98
Taylor LP, Hepler PK. Pollen germination and tube growth. Annu Rev Plant Physiol Plant Mol Biol, 1997, 48: 461～491
Tebbutt SJ, Rogers HJ, Lonsdale DM. Characterization of a tobacco gene encoding a pollen-specific polygalacturonase. Plant Mol Biol, 1994, 25: 283～297
Tirlapur, UK, Scali M, Moscatelli A, Del Casino C, Cai G, Tiezzi A, Cresti M. Confocal image analysis of spatial variations in immunocytochemically identified calmodulin during pollen hydration, germination and pollen tube tip growth in Nicotiana tabacum L. Zygote, 1994, 2: 63～68
Torki M, Mandaron P, Mache R, Falconer D. Characterization of a ubiquitous expressed gene family encoding polygalacturonase in Arabidopsis thaliana. Gene, 2000, 242: 427～436
Torki M, Mandaron P, Thomas F, Quigley R, Mache F, Falconer D. Differential expression of a polygalacturonase gene family in Arabidopsis thaliana. Mol Gen Genet, 1999, 261: 948～952
Truernit E, Schmid J, Epple P, Illig J, Sauer N. The sink-specific and stress regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors and pathogen challenge. Plant Cell, 1996, 8: 2169～2182
Truernit E, Stadler R, Baier K, Sauer N. A male gametophyte-specific monosaccharide transporter in Arabidopsis. Plant J, 1999, 17: 191～201
Tsuchida M, Manthei ER, Alam T, Knechtle SJ, Hamawy MM. Regulation of T cell receptor-and CD28-induced tyrosine phosphorylation of the focal adhesion tyrosine kinases Pyk2 and Fak by protein kinase C. J Biol Chem, 2000, 275(2): 1344～1350
Tucker GA, Schindler BC, Roberts JA. Flower abscission in mutant tomato plants. Planta, 1984, 160: 164～167
Twell D, Howden R. Mechanisms of asymmetric division and cell-fate determination in developing pollen, in Androgenesis and Haploid Plants. In Chupeau Y, Caboche M, Henry Y, eds, Memory of Jean-Pierre Bourgin, 1998, pp 69～103
Twell D, Park SK, Lalanne E. Asymmetric division and cell-fate determination in developing pollen. Trends Plant Sci, 1998, 3(8): 305～310
Twell D. Pollen developmental biology. In O'Neill SD, Roberts JA, eds, Plant Reproduction. Annu Rev Plant, 2002, 6: 86～153
Unte US, Sorensen AM, Pesaresi P, Gandikota M, Leister D, Saedter H, Huijser P. SPL8, an SBP-Box gene that affects pollen sac development in Arabidopsis. Plant Cell, 2003, 15(4): 1009～1019
Uyenoyama MK. Evolutionary dynamics of self-incompatibility alleles in Brassica. Genetics, 2000, 156: 351～359
van Aelst AC, van Went JL. Ultrastructural immunolocalization of pectin and glycoproteins in Arabidopsis thaliana pollen grains. Protoplasma, 1992, 168: 14～19
Wakeley PR, Rogers HJ, Rozycka M, Greenland AJ, Hussey PJ. A maize pectin methylesterase-like gene, ZmC5, specifically expressed in pollen. Plant Mol Bio, 1998, 37: 187～192
Wang H, Wu H-M, Cheung AY. Development and pollination regulated accumulation and glycosylation of a stylar transmitting tissue-specific proline-rich protein. Plant Cell, 1993, 5:1639～1650
Wang Y-Q, Ye W-Z, Can J-S, Yu X-L, Xiang X, Lu G. Cloning and characterization of the microspore development related gene BcMF2 in Chinese cabbage pak-choi (Brassica campestris L. ssp. chinensis Makino). J Integrative Plant Biol, 2005, 47 (7): 863～872
Wang Y-Q, Yu X-L, Cao J-S. Isolation and characterization of BcMF3, a gene expressed only in maintainer line in Chinese cabbage pak-choi (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee). Acta Genetica Sinica, 2004, 31(11): 1302～1308
Waterkeyn L and Bienfait A. On a possible function of the callosic special wall in Ipomoea purpurea (L) Roth. Grana, 1970, 10: 13～20
Wellmer F, Riechmann JL, Aives-Ferreira M, Meyerowitz EM. Genome-wide analysis of spatial gene expression in Arabidopsis flowers. Plant Cell, 2004, 16: 1314～1326
Wheeler MJ, Franklin-Tong VE, Franklin FCH, The molecular and genetic basis of pollen-pistil interactions, New phytologist, 2001, 151: 565～584
Whittington AT, Vugrek O, Wei KJ, Hasenbein NG, Sugimoto K, Rashbrooke MC, Wasteneys GO. MOR1 is essential for organizing cortical microtubules in plants. Nature, 2001, 411: 610～613
Wilhelmi LK, Preuss D. Self-sterility in Arabidopsis due to defective pollen tube guidance. Science, 1996, 274: 1535～1537
Willing RP, Bashe D, Mascarenhas, JP. An analysis of the quantity and diversity of messenger RNAs from pollen and shoots of Zea mays. Theor Appl Genet, 1988, 75: 751～753
Willing RP, Mascarenhas JP. Analysis of the complexity and diversity of mRNAs from pollen and shoots of Tradescantia. Plant Physiol, 1984, 75: 865～868
Wilson ZA, Morroll SM, Dawson J, Swarup R, Tighe PJ. The Arabidopsis MALE STERILITY1(MS1) gene is a transcriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors. Plant J, 2001, 28: 27～39
Wu H-M, Wang H, Cheung AY. A floral transmitting tissue specific glycoprotein attracts pollen tubes and stimulates their growth. Cell, 1995, 82: 383～393
Wu H-M, Wong E, Ogdahl J, Cheung AY. A pollen tube growth-promoting arabinogalactan protein from Nicotiana alata is similar to the tobacco TTS protein. Plant J, 2000, 22: 165～176
Xu Z, Dooner HK. The Maize aberrant pollen transmission 1 gene is a SABRE/KIP homolog required for pollen tube growth. Genetics, 2006, 172: 1251～1261
Yang CY, Spileman M, Coles JP, Li Y, Ghelani S, Bourdon V, Brown RC, Lemmon BE, Scott RJ, Dickinson HG. TETRASPORE encodes a kinesis required for male meiotic cytokinesis in Arabidopsis. Plant J, 2003b, 34: 229～240
Yang M, Hu Y, Lodhi M, McCombie W R, Ma H. The Atrabidopsis SKP1-LIKE1 gene is essential for male meiosis and may control homologue separation. Proc Natl Acad Sci USA, 1999b, 96: 11416～11421
Yang M, McCormick S. The Arabidopsis MEI1 gene likely encodes a protein with BRCT domains. Sex Plant Reprod, 2002, 14: 355～357
Yang SL, Xie LF, Mao HZ, Puah CS, Yang WC, Jiang LX, Sundaresan V, Ye D. TAPETUM DETERMINANT1 is required for cell specialization in the Arabidopsis anther. Plant Cell, 2003a, 15: 2792～2804
Yang WC, Sundaresan V. Genetics of gametophyte biogenesis in Arabidopsis. Curr Opin Plant Biol, 2000, 3: 53～57
Yang WC, Ye D, Xu J, Sundaresan V. The SPOROCYTELESS gene of Arabidopsis is required for itiation of sporogenesis and encodes a novel nuclear protein. Genes Dev, 1999a, 13: 2108～2117
Yang Z. Signaling tip growth in plants. Curr Opin Plant Biol, 1998, 1: 525～530
Ye W-Z, Cao J-S, Xiang X, Zeng G-W, Molecular cloning and characterization of the genic male sterility related gene CYP86MF in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee). J Hort Sci & Biotech, 2003, 78(3): 319～323
Yu X-L, Cao J-S, Ye W-Z, Wang Y-Q. Construction of an antisense CYP86MF gene plasmid vector and production of a male-sterile Chinese cabbage transformant by the pollen-tube method. J Hort Sci & Biotech, 2004, 79(5): 833～839
Zhao D, Han T, Risseeuw E, Crosby WL, Ma H. Conservation and divergence of ASK1 and ASK2 gene functions during male meiosis in Arabidopsis thaliana. Plant Mol Biol, 2003, 53(1-2): 163～73
Zhao DZ, Wang GF, Speal B, Ma H. The EXCESS MICROSPOROCYTES1 gene encodes a putative leucine-rich repeat receptor protein kinase that controls somatic and reproductive cell fates in the Arabidopsis anther. Genes Dev, 2002, 16: 2021～2031
Zheng Z-L, Yang Z, The Rop GTPase switch turns on polar growth in pollen. Trends Plant Sci, 2000, 5: 298～303
Zinkd GM, Preuss D. Dissecting Arabidopsis pollen-stigma interactions reveals novel mechanisms that confer mating specificity. Ann Bot, 2000, 85(A): 15～21