温敏核雄性不育水稻育性转换相关基因的克隆及其RNAi植物表达载体的构建
摘要
本研究以具有明显育性转换特征的水稻温敏核雄性不育系Tb7S为材料，通过抑制性消减杂交(Suppression Subtractive Hybridization，SSH)技术筛选减数分裂期的Tb7S幼穗在高温可育和低温不育条件下的差异表达cDNA片段。经SSH富集cDNA片段构建了两个cDNA质粒文库，即cDNA正向消减和反向消减质粒文库。
分别从正向消减和反向消减质粒文库中随机选取39个和35个克隆进行分析。重组质粒EcoR1酶切鉴定表明随机挑取的74克隆中均载有约200-600bp大小的插入片段，进一步的测序表明cDNA片段大小介于200bp-300bp之间的约占41％，100bp-200bp的约24％，300bp-400bp的约23％，400bp以上的约12％，与理论预期值大小相符。在序列分析过程中发现有部分克隆是重复检出的同一基因，也有些克隆是同一基因的不同片段，但在两个cDNA消减文库中的cDNA未发现有相同的序列，说明所进行的SSH消减杂交效率高，同时SSH消减效率鉴定实验结果也证明了这一点。序列分析表明除去重复克隆，74个cDNA克隆代表了61个不同的基因，说明约82％的克隆代表不同的基因，表明cDNA消减群体内部重复拷贝少，不同基因间丰度的差异得到了很好的均衡。对这61个序列进行BLAST检索查询，结果发现有17个为已知基因，约占26％，有44个为未知功能的基因，约为74％。
从测序的克隆中随机选取10个(每个文库5个克隆)，用RT-PCR技术分析这些克隆所代表的基因在Tb7S幼穗处于高温可育和低温不育条件下的差异表达情况。结果鉴定出2个
………………… ．华南热带农业大学博士学位论文…．，……………．·￥
cDNA在高温可育幼穗中特异表达，另有2个cDNA在低温不育幼穗中特异表达，提示这4
个。DNA代表的基因的表达可能与水稻温敏核不育系Tb7S的育性转换相关。
在RT．PCR的鉴定出差异表达。DNA片段的基础上，选取一个未知功能的SZcDNA片段，
用RACE技术克隆其全长CDNA。对扩增的片段进行测序和拼接，结果获得全长为2976hp
的 SZ cDNA，其编码区为 2367hp，此外，还含有 75 hp的 5’－UTR 534hp的 3’－UTR。对推
导的全长氨基酸序列用 3DPSSM Protein Prediction软件和 PredictProtein软件进行分析，结果
提示SZ基因的编码产物可能是一个参与信号转导的调控因子。通过PSORT进行蛋白质定位
信号分析，提示该基因的表达产物含有一段线粒体靶肽（mitochondrial枯吧eting peptide），推
测该基因编码产物可能在干扰线粒体的功能方面起作用。如果该推测成立，暗示三系和两系
水稻之间可能存在某种联系。Southern杂交分析表明在Tb7S基因组中SZ基因为单拷贝基因。
对获得的新基因有必要进行生物学功能的鉴定，而RNAi技术是一种高效的阻抑基因表
达的新技术。本研究成功构建了SZ C DNDNA 片段的RN＊1 植物表达载体
pCAMBIA130lAP人石，Sallnyll和Pmll／BstEll酶切鉴定表明正向和反向特异片段正确
插入表达载体，EcoR I／Sma酶切鉴定表明 ubiquitiin启动子也正确插入。构建的载体含有潮
霉素筛选标记基因，利用农杆菌介导法导入Tb7S愈伤组织，获得了抗性愈伤组织，后续工
作正在进行。
OBJECTIVE: The aim of the research is to isolate the genes associated with fertility transformation of thermo-sensitive genie male sterile rice. METHODS: Two-directional (forward and backward) Suppression Subtractive Hybridization (SSH) was performed on cDNAs of fertile young panicle and sterile young panicle of thermo-sensitive genie male sterile rice (Tb7S), and two Subtracted cDNA libraries were established with the forward and the backward subtracted cDNAs, positive clones from each subtracted cDNA libraries were selected for sequencing and BLAST analysis, the differential expression of 10 clones between fertile young panicle and sterile young panicle of Tb7S were analysed by RT-PCR; RACE technique was used to amplify the 5' cDNA fragment and 3' cDNA fragment of S2; The plant expression vector of pCAMBIA1301-P-A-S was constructed. In pCAMBIA1301-P-A-S, gene-specific sequences in the antisense (A) and sense (s) orientations were linked with a 1300-bp of the GUS gene and controlled by ubiquitin promoter, t
he 400-bp gene-specific sequences in the antisense (A) and sense (s) orientations were inserted in the Sal IIIBgl II and Pml l/Bst Ell sites, the ubiquitin promoter was inserted in the EcoR HSma I site, the callus from tissue cultured young embryo were used as material in agrobacterium-mediated genetic transformation.
RESULT: The experiment of subtractive efficiency indicated that the subtractive efficiency is high, and the plasmid digestion by EcoR I indicated that all the positive clones picked randomly contained cDNA fragments of 200-600bp. The sequence
analysis demonstrated that picked randomly 74 clones represented 61 different genes, which showed the repeat copies of subtracted population of cDNA fragments were few and the abundance between different genes was normalized. Sequencing and BLAST homology search revealed that 17 clones contain sequences of known gene fragments and 44 possibe novel genes show no sequence homologies with any know sequences in the database. 4 clones of 10 clones picked randomly were identified as differentially expressed genes by RT-PCR, among which 2 clones were expressed specifically in the fertile young panicle of Tb7S and the other 2 clones were expressed specifically in the sterile young panicle of Tb7S. A novel full-length of S2 cDNA containing 2976bp was obtained by RACE technique, which contain the ORF of 2367bp, 5'-UTR of 75bp and 3'-UTR of 534bp. The results of 3DPSSM Protein prediction and protein domain prediction indicated that it might be a regulatory factor involved in signal transduction;. The result of PSORT (P
rediction of Protein Localization Sites) indicates that the protein encoded by S2 has a mitochondrial targeting peptide. Southern Blotting showed that the S2 gene is a single cope gene.The plasmid digestion showed that the 400-bp gene-specific sequences in the antisense (A) and sense (s) orientations were inserted in the Sal 11/Bgl II and Pml l/Bst Ell sites, the ubiquitiin promoter was inserted in the EcoR l/Sma I site of the improved pCAMBIA1301. CONCLUSION: (1) All results confirmed the effectiveness and sensitivity of SSH;
(2) S2 gene is a single copy gene; (3) S2 gene is expressed specifically in the sterile young panicle of Tb7S ,which suggest that S2 gene might be a novel gene associated with fertility transformation of thermo-sensitive genie male sterile rice;
(3) The fact that the protein encoded by S2 cDNA has a mitochondrial targeting peptide suggests that there might be relationship between the two-line rice and three-line rice; (4) The RNAi plant expression vector(pCAMBIA1301A-P-A-S) was constructed successfully. In the pCAMBIA1301A-P-A-S, the gene-specific sequences in the antisense (A) and sense (s) orientations were linked with a fragment of the GUS gene and controlled by the ubiquitin promoter.
分别从正向消减和反向消减质粒文库中随机选取39个和35个克隆进行分析。重组质粒EcoR1酶切鉴定表明随机挑取的74克隆中均载有约200-600bp大小的插入片段，进一步的测序表明cDNA片段大小介于200bp-300bp之间的约占41％，100bp-200bp的约24％，300bp-400bp的约23％，400bp以上的约12％，与理论预期值大小相符。在序列分析过程中发现有部分克隆是重复检出的同一基因，也有些克隆是同一基因的不同片段，但在两个cDNA消减文库中的cDNA未发现有相同的序列，说明所进行的SSH消减杂交效率高，同时SSH消减效率鉴定实验结果也证明了这一点。序列分析表明除去重复克隆，74个cDNA克隆代表了61个不同的基因，说明约82％的克隆代表不同的基因，表明cDNA消减群体内部重复拷贝少，不同基因间丰度的差异得到了很好的均衡。对这61个序列进行BLAST检索查询，结果发现有17个为已知基因，约占26％，有44个为未知功能的基因，约为74％。
从测序的克隆中随机选取10个(每个文库5个克隆)，用RT-PCR技术分析这些克隆所代表的基因在Tb7S幼穗处于高温可育和低温不育条件下的差异表达情况。结果鉴定出2个
………………… ．华南热带农业大学博士学位论文…．，……………．·￥
cDNA在高温可育幼穗中特异表达，另有2个cDNA在低温不育幼穗中特异表达，提示这4
个。DNA代表的基因的表达可能与水稻温敏核不育系Tb7S的育性转换相关。
在RT．PCR的鉴定出差异表达。DNA片段的基础上，选取一个未知功能的SZcDNA片段，
用RACE技术克隆其全长CDNA。对扩增的片段进行测序和拼接，结果获得全长为2976hp
的 SZ cDNA，其编码区为 2367hp，此外，还含有 75 hp的 5’－UTR 534hp的 3’－UTR。对推
导的全长氨基酸序列用 3DPSSM Protein Prediction软件和 PredictProtein软件进行分析，结果
提示SZ基因的编码产物可能是一个参与信号转导的调控因子。通过PSORT进行蛋白质定位
信号分析，提示该基因的表达产物含有一段线粒体靶肽（mitochondrial枯吧eting peptide），推
测该基因编码产物可能在干扰线粒体的功能方面起作用。如果该推测成立，暗示三系和两系
水稻之间可能存在某种联系。Southern杂交分析表明在Tb7S基因组中SZ基因为单拷贝基因。
对获得的新基因有必要进行生物学功能的鉴定，而RNAi技术是一种高效的阻抑基因表
达的新技术。本研究成功构建了SZ C DNDNA 片段的RN＊1 植物表达载体
pCAMBIA130lAP人石，Sallnyll和Pmll／BstEll酶切鉴定表明正向和反向特异片段正确
插入表达载体，EcoR I／Sma酶切鉴定表明 ubiquitiin启动子也正确插入。构建的载体含有潮
霉素筛选标记基因，利用农杆菌介导法导入Tb7S愈伤组织，获得了抗性愈伤组织，后续工
作正在进行。
OBJECTIVE: The aim of the research is to isolate the genes associated with fertility transformation of thermo-sensitive genie male sterile rice. METHODS: Two-directional (forward and backward) Suppression Subtractive Hybridization (SSH) was performed on cDNAs of fertile young panicle and sterile young panicle of thermo-sensitive genie male sterile rice (Tb7S), and two Subtracted cDNA libraries were established with the forward and the backward subtracted cDNAs, positive clones from each subtracted cDNA libraries were selected for sequencing and BLAST analysis, the differential expression of 10 clones between fertile young panicle and sterile young panicle of Tb7S were analysed by RT-PCR; RACE technique was used to amplify the 5' cDNA fragment and 3' cDNA fragment of S2; The plant expression vector of pCAMBIA1301-P-A-S was constructed. In pCAMBIA1301-P-A-S, gene-specific sequences in the antisense (A) and sense (s) orientations were linked with a 1300-bp of the GUS gene and controlled by ubiquitin promoter, t
he 400-bp gene-specific sequences in the antisense (A) and sense (s) orientations were inserted in the Sal IIIBgl II and Pml l/Bst Ell sites, the ubiquitin promoter was inserted in the EcoR HSma I site, the callus from tissue cultured young embryo were used as material in agrobacterium-mediated genetic transformation.
RESULT: The experiment of subtractive efficiency indicated that the subtractive efficiency is high, and the plasmid digestion by EcoR I indicated that all the positive clones picked randomly contained cDNA fragments of 200-600bp. The sequence
analysis demonstrated that picked randomly 74 clones represented 61 different genes, which showed the repeat copies of subtracted population of cDNA fragments were few and the abundance between different genes was normalized. Sequencing and BLAST homology search revealed that 17 clones contain sequences of known gene fragments and 44 possibe novel genes show no sequence homologies with any know sequences in the database. 4 clones of 10 clones picked randomly were identified as differentially expressed genes by RT-PCR, among which 2 clones were expressed specifically in the fertile young panicle of Tb7S and the other 2 clones were expressed specifically in the sterile young panicle of Tb7S. A novel full-length of S2 cDNA containing 2976bp was obtained by RACE technique, which contain the ORF of 2367bp, 5'-UTR of 75bp and 3'-UTR of 534bp. The results of 3DPSSM Protein prediction and protein domain prediction indicated that it might be a regulatory factor involved in signal transduction;. The result of PSORT (P
rediction of Protein Localization Sites) indicates that the protein encoded by S2 has a mitochondrial targeting peptide. Southern Blotting showed that the S2 gene is a single cope gene.The plasmid digestion showed that the 400-bp gene-specific sequences in the antisense (A) and sense (s) orientations were inserted in the Sal 11/Bgl II and Pml l/Bst Ell sites, the ubiquitiin promoter was inserted in the EcoR l/Sma I site of the improved pCAMBIA1301. CONCLUSION: (1) All results confirmed the effectiveness and sensitivity of SSH;
(2) S2 gene is a single copy gene; (3) S2 gene is expressed specifically in the sterile young panicle of Tb7S ,which suggest that S2 gene might be a novel gene associated with fertility transformation of thermo-sensitive genie male sterile rice;
(3) The fact that the protein encoded by S2 cDNA has a mitochondrial targeting peptide suggests that there might be relationship between the two-line rice and three-line rice; (4) The RNAi plant expression vector(pCAMBIA1301A-P-A-S) was constructed successfully. In the pCAMBIA1301A-P-A-S, the gene-specific sequences in the antisense (A) and sense (s) orientations were linked with a fragment of the GUS gene and controlled by the ubiquitin promoter.
引文
鲍文奎．1990．机会与风险—40 年育种研的思考．植物杂志，4：4-5
陈建南，傅鸿仪，路子显等．1997．科学通报，42(18)：1993-1997
陈良碧，周广洽，黄花玉祥．1994．温光条件对水稻安农S-1／衡农S-1的育性及生理的影响，植物学报,36(S)：119-1234
陈良碧，周广洽．1992．光温敏核不育水稻结实率与温度相关性研究．杂交水稻，6(2)：35-39
程式华，孙宗修．1993．国外光温敏核不育水稻的研究进展．种子世界，3：35
程式华，孙宗修，斯华敏．等，1994．农垦58S／农垦58F-2群体育性分离模式的年度间差异．中国水稻科学，8(2)：97-101
邓华凤，舒福北，袁定阳．1999．安农S-1的研究及其利用．杂交水稻，14(3)：1-3
傅荣昭，曹光诚，马江生等．1997．用基因枪法将人工雄性不育基因导入小麦的研究初报．遗传学报，24(4)：358-361
傅荣昭，孙勇如，贾士荣主编．1994．植物遗传转化手册．北京：中国科学技术出版社，134-143
高恒广．1993．三系不育系龙特浦A具有光温敏特性．杂交水稻，(1)：44-45
高一枝．1991．水稻短光敏雄性不育材料的发现初报．宜春农专学报．7(1)：1-5
何觉民，戴君惕，邹应斌，等．1994。生态遗传雄性不育理论与两系杂交植物Ⅰ：生态遗传雄性不育理论．湖南农业大学学报，20(1)：1-5
金冬雁，黎孟枫等译．1996．J．萨姆布鲁克，E．F．弗里奇，T．曼尼阿蒂斯等著．分子克隆实验指南，第二版．北京：科学出版社
康健，陈凡，吴乃虎．1998．光敏核不育水稻育性相关基因片段的分离．科学通报，43(19)：2078-2082
李传友，伏建民，金德敏等．1998．AFLP分析中多态性扩增产物的回收、克隆及鉴定．遗传,20(3)：1-4
李仕贵，周开达，朱立煌．1999，水稻温敏核不育基因的遗传分析和分子标记定位．科学通报，44(9)：955-958
李泽炳，肖翊华，朱英国等．1982．杂交水稻的研究与实践．上海：上海科学技术出版社
李子银，林兴华，谢岳峰等．1999．利用分子标记定位农垦58S的光敏核不育基因．植物学报，41(7)：731-735
凌定厚，陶利珍，马镇荣等．1998．以基因枪介导获转psl-Barnase基因的工程雄性不育水稻植株．遗传学报,25(5)：433-442
刘爱民，李必湖．1996．水稻温敏不育系育性与温度的相关性分析．杂交水稻，10(3)：25-28
李发生，谢戎，邵启明，等．1996．水稻低温敏两用系363S选育研究．西南农业大学学报，18(4)：338-342
刘军，袁自强，刘建东等．2000．应用抑制差减杂交分离水稻幼发育早期特异表达的基因．科学通报，45(13)：1392-1397
卢兴桂，顾铭洪。李成荃等．2001．两系杂交水稻理论与技术．北京：科学出版社．
罗玉英，刘玉乐，李胜国等．1997．TA29-Barnase嵌合基因导入对烟草花药绒毡层及花粉发育的影响．植物学报．39(10)：894-898
罗越华，周庭波，袁潜华，陈玉新．2001．水稻雄性不育研究的现状与展望．琼海：海南省生物技术研讨会论文汇编，91-97
梅明华，李泽炳．1995．水稻光温敏不育系与核质互作不育系的遗传关系剖析．遗传，17(1)：22-25
米志勇，王树生，吴乃．2000，水稻低分子量GTP结合蛋白基因OsRACD的分离．科学通报，45(19)：2047-2055
牛景，牛芝霞．2001．光敏核不育水稻杂种F2花药培养中获得了细胞质不育突变体．杂交水稻，16(2)：40
潘润森，李维明，闵绍楷，等．1994．光周期和温度对水稻雄性不育系龙特浦A育性恢复的影响．福建农业大学学报，23(3)：257-261
秦泰辰．1980．杂种优势利用原理和方法．南京：江苏科学技术出版社
邱芳，金德敏，伏健民等．1999．温敏核不育水稻5460S的细菌人工染色体文库的构建和鉴定．中国科学(C)，29(5)：518-524
曲雪萍，伏健民，李传友等．1999．水稻5460F的细菌人工染色体文库的构建和鉴定．科学通报,44)14)：1532-1536
石明松．1981．晚粳自然两用系的选育及其应用初报．湖北农业科学，7：44-48
孙宗修，程式华．1994．杂交水稻育种—从三系两系到一系．北京：中国农业科学出版社
童哲．1998．光敏核不育水稻的发育生物学研究评述．植物学报，40(3)：189-199
王斌，伏建民，邱方等．1998．TGMS水稻BAC文库的构建及与TGMS基因连锁的BAC克隆的筛选．遗传，20(增刊)：115
王培田．1976．水稻雄性不育发生的原因何在?遗传与育种，5：9-10
杨金水．1996，杂种优势机理探讨：作物雄性不育及杂种优势研究进展(Ⅰ)．北京：中国农业出版社．1-12
张宏，王波，薛爱群等．1998．雄性不育嵌合基因的构建及番茄转化研究．遗传，20(3)：5-7
杨仁崔，王乃元，梁康迳等．1993．籼稻温核不5460S的研究．福建农学院学报，22(2)：135-140
袁隆平．1987．杂交水稻的育种战略设想．杂交水稻，(1)：1-3
袁隆平．2000．两系法杂交水稻技术研究与中试开发．国家八六三计划项目验收报告
元生朝，张自国．1987．光照诱导湖光怪陆离核不育水稻育性转换的研究．武汉大学学报，HPGMR专刊：17-28
张自国，曾汉来，元生朝等．1992．再论光敏型核不育水稻育性转换的光温模式．华中农业大学学报，11(1)：1-6
中国农业科学院，湖南省农业科学院主编．1991．中国杂交水稻的发展．北京：中国农业出版社
曾汉来，张自国，元生朝等．1993．光敏型核不育水稻育性转换的温度敏感期研究．华中农业大学学报，12(5)：401-406
周国峰，龚光明，尹楚球等．1991．籼型两用不育系衡农S-1育性转换期及其遗传行为的初步观察．湖南农业科学，4：10-12
周坤炉．1994．籼型杂交水稻三系不育系选育．杂交水稻，1994(3-4)：22-26
周雪荣，彭仁旺，方荣祥等．表达核糖核酸酶基因的雄性不育油菜的获得．遗传学报，1997，24(6)：531-536
朱国，扬代常．1992．光周期敏感核不育水稻研与利用．武汉：武汉大学出版社
庄楚雄，徐是雄，卢永根等．1999．运用cDNA缩减杂交法克隆水稻花粉发育有关的cDNA．科学通报，44(17)：1842-1846
Aarts MG, Dirkse Wg, Stiekema W J, et al. 1993. Transposon tagging of a male sterility gene in Arabidopsis. Nature,363(6431): 715-717
Aarts MG, Hodge R, Kalantidis K, et al. 1997. The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes. Plant J. 12(3):615-623
Akagi H. Sakamoto M, Shinjyo C. et al. 1994. A unique sequence located downstream from the rice mitochondrial atp6 may cause male sterility. Curr Genet. 25(1):52-58
Bahn SC, Bae MS, Park YB, et al.2001.Molecular cloning and characterization of a novel low temperature-induced gene, blti2, from barley (Hordeum vulgare L.). Biochim Biophys Acta, 1522(2): 134-137
Cao GC,Sun YR, Chen ZK,et al. 1996.The cloning and construction study of artificial sterile gene and restore gene. Scentia Agricultural Sinica, 29(3):20-26
Carmeci C, Thompson DA, Kuang WW, et al. 1998. Moesin expression is associated with the estrogen receptor-negative breast cancer phenotype. Surgery, 124(2):211-217
Cho HJ, Kim S. Kim M, et al. 2001. Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells, 11(3):326-333
Chuang CF and Meyerowitz EM. 2000.Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana Proc. Natl. Acad. Sci. USA, 97(9), 4985-4990
Cui X, Wise RP, Schnable PS. 1996. The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize. Science,272(5266): 1334-1336
David J, Ursula HA, Michael J, et al. 1995. An analysis of differential display shows a strong bias towards high copy number mRNAs. Nucleic Acids Research, 23(21):4520-4523
Dessens JT, Margos G, Rodriguez MC, et al. 2000.Identification of differentially regulated genes of Plasmodium by suppression subtractive hybridization. Parasitol Today, 16(8):354-356
Diatchenko L, Lau Y, Campbell A, et al. 1996. Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc Natl Acad Sci USA, 93(12): 6025-6030
Dong NV, Subudhi PK ,Luong PN,et al.2000. Molecular mapping of a rice thermosenstive genetic male sterile gene (TGMS) by AFLP, RFLP, and SSR techniques. Theor Appl Genet, 100(5):727-734
Dudley NR, Labbe JC, Goldstein B.2002. Using RNA interference to identify genes required for RNA interference. Proc Natl Acad Sci U S A, 99(7):4191-4196
Fire A, Xu S, Montgomery MK, et al. 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391 (6669):806-811
Fire A. 1999. RNA-triggered gene silencing. Trends Genet, 15(9):358-363
Godt D E and Roitsch T. 1997. Regulation and tissue-specific distribution of mRNAs for three extracellular invertase isoenzymes of tomato suggests an important function in establishing and maintaining sink metabolism. Plant Physiol, 115(1): 273-282
Goetz M, Godt DE, Guivarc'h A, et al,2001. Induction of male sterility in plants by metabolic engineering of the carbohydrate supply. Proc Natl Acad Sci U S A, 98(11):6522-6527
Hammond SM, Bernstein E, Beach D, et al. 2000. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature ,404(6775):293-296
He S, Abad AR, Gelvin SB, et al. 1996. A cytoplasmic male sterility-associated mitochondrial protein causes pollen disruption in transgenic tobacco. Proc Natl Acad Sci U S A, 93(21): 11763- 11768
He SC, Lyznik A, Mackenzie S, et al. 1995. Pollen fertility restoration by nuclear gene Fr in CMS Bean: nuclear-direct alteration of a mitochondrial population. Genetics, 139(2): 955-962
Hernould M, Suharsono S, Litvak S, et al. 1993. A. Male-sterility induction in transgenic tobacco plants with an unedited atp9 mitochondrial gene from wheat. Proc Natl Acad Sci U S A,90(6):2370-2374
Hernould M, Suharsono, Zabaleta E,et al. 1998. Impairment of tapetum and mitochondria in engineered male-sterile tobacco plants. Plant Mol Biol, 36(4):499-508
Hinderhofer K, Zentgraf U. 2001.1dentification of a transcription factor specifically expressed at the onset of leaf senescence. Planta, 213(3):469-473
Howad W, Kempken F. 1997. Cell type-specific loss of atp6 RNA editing in cytoplasmic male sterile Sorghum bicolor. Proc Natl Acad Sci U S A,94(20): 11090-11095
Howad W, Tang HV, Pring DR, et al. 1999. Nuclear genes from Tx CMS maintainer lines are unable to maintain atp6 RNA editing in any anther cell-type in the sorghum bicolor A3 cytoplasm. Curt Genet, 36(1-2):62-68
Hufton SE, Moerkerk PT, Brandwijk R, et al. 1999. A profile of differentially expressed genes in primary colorectal cancer using suppression subtractive hybridization. FEBS Lett ,463(1-2):77-82
Jia JH, Li CY, Qu XP, et al. 2000.Construction of genetic linkage map and chromosome of tms5 gene inrice. In:Abstract Book of Plant Genomics in China I.Dalian
Jin H, Cheng X, Diatchenko L, et al. 1997. Differential screening ora subtracted cDNA library: a method to search for genes preferentially expressed in multiple tissues. Biotechniques, 23(6): 1084-1086
Kempken F, Howard W, Piing DR. 1998. Mutations at specific atp6 codons which cause human mitocbondrial diseases also lead to male sterility in a plant. FEBS Lett, 441 (2): 159-160
Ketting RF, Fischer SE, Bernstein E, et al. 2001. Dicer functions in RNA interference and in synthesis of small RNA involved in developmental timing in C. elegans. Genes Dev, 15(20):2654-2659
Kim JY, Chung YS, Paek KH, et al.1999a. Isolation and characterization of a cDNA encoding the cysteine proteinase inhibitor, induced upon flower maturation in carnation using suppression subtractive hybridization. Mol Cells ,9(4):392-397
Kim JY, Chung YS, Ok SH, et al. 1999b. Characterization of the full-length sequences of phospholipase A2 induced during flower development. Biochim Biophys Acta, 1489(2-3):389-392
Kriete G, Niehaus K, Perlick AM, et al. 1996. Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J,9(6):809-818
Kuang WW, Thompson DA, Hoch RV, et al. 1998. Differential screening and suppression subtractive hybridization identified genes differentially expressed in an estrogen receptor-positive breast carcinoma cell line. Nucleic Acids RES, 26(4):1116-1123
Lamar EE, Palmer E. 1984. Y-encoded, species-specific DNA in mice: evidence that the Y chromosome exists in two polymorphic forms in inbred strains. Cell. 37( 1 ): 171 - 177
Landgren M, Zetterstrand M, Sundberg E, et al. 1996. Alloplasmic male-sterile Brassica lines containing B. tournefortii mitochondria express an ORF 3' of the atp6 gene and a 32 kDa protein. Plant Mol Biol, 32(5):879-890
Lang NT, Subudhi PK, Virmani SS, et al. 1999. Development of PCR-based markers for thermosensitive genetic male sterility gene tms3(t) in rice (Oryza sativa L.). Hereditas, 131 (2): 121-127
Larose M, Bouchard C, Chagnon YC. 2001. A new gene related to human obesity identified by suppression subtractive hybridization. Int J Obes Relal Metab Disord. 25(6):770-776
Liang P, Pardee AB. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science, 257:967-970
Lisitsyn N, Wiglet M. 1993, Cloning the differences between two complex genomes. Science,259(5097):946-951
Liu F, Cui X, Horner HT, et al. 2001.Mitochondrial aldehyde dehydrogenase activity is required for male fertility in maize. Plant Cell, 13(5):1063-1078
Liu N, Shan Y, Wang FP, et al. 2001. Identification of an 85-kb DNA fragment containing pmsl, a locus for photoperiod-sensitive genic male sterility in rice. Mol Genet Genomics, 266(2):271-275
Koh HJ, Son YH, Heu MH, et al. Molecular mapping of a new genic male sterile gene causingf chalking endosperm in rice (Oryza sativa L). Euphitica, 106:57-62
Janke B, Dobrindt U, Hacker J, et al. 2001. A subtractive hybridisation analysis of genomic differences between the uropathogenic E. coli strain 536 and the E. coli K-12 strain MG1655. FEMS Microbiol Lett, 199(1):61-66
Mariani C, De Beuckeleer M, Truettner M, et al. 1990 Intmction of male sterility in plants by a chimeric ribonuclease gene. Nature,347:737-741
Mariani C, Gossele V, De Beukeleer M, et al. 1992. A chimeric ribonuclease inhibitor gene restores fertility to male sterile plants. Nature, 357:384-387
Marugma K, Araki H,kato H,et al. 1991. Thermonsensitive genic male sterility induced by irradiation. In:Khush GS ed. Rice Genet Ⅱ.manila, Phlippines:IRRI.227-235
Mascarenthas JP. 1993. Molecular mechanisma of pollen tube growth and differertitation. Plant Cell, 5(10):1303-1314
Matsuda N, Tsuchiya T, Kishitani S, et al. 1996. Partial male sterility in transgenic tobacco with antisense and sense PAL cDNA regulated by a tapetum-specific promoter. Plant Cell Physiol, 37(2):215-222
Mei MH,Chen,Zhang ZH, et al. 1999. pms3is the locus cauing the original photoperiod-sensitive male sterilitymutationof "Nongken 58S". Science in China(series C),42(3):316-322
Misquitta L and Paterson BM. 1999. Targeted disruption of gene function in Drosophila by RNA interference (RNA-i): A role for nautilus in embryonic somatic muscle formation. 96(4): 1451-1456
Miyasaka Y, Enomoto N, Nagayama K, et al. 2001. Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization. Br J Cancer, 85(2):228-234
Nicholson RH, Nicholson AW. 2002. Molecular characterization of a mouse cDNA encoding Dicer, a ribonuclease Ⅲ ortholog involved in RNA interference. Mamm Genome, 13(2):67-73
Okumura N, Nishizawa NK, Umehara Y ,et al. 1994. A dioxygenase gene (Ids2) expressed under iron deficiency conditions in the roots of Hordeum vulgate. Plant Molecular Biology, 25(4):705-719
Paddison PJ, Caudy AA, Bernstein E, et al. 2002. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev, 16(8):948-958
Porkka KP, Visakorpi T. 2001. Detection of differentially expressed genes in prostate cancer by combining suppression subtractive hybridization and cDNA library array. J Pathol, 193(1):73-79
Reddy OUK, Siddiq EA, Sarma NP, et al.2000.Genetic analysis of temoerature-sensitive male sterility in rice. Theor Appl Genetic, 100(5):794-801
Rutger JN. 1990. Sex-swithching rice may be short-out to hybridization. Rice Journal.93(6):4-5
Sharp PA. 1999. RNAi and double-strand RNA .Gen Dev, 13( 2): 139-141
Sompayrac L, Jane S, Burn TC, et al. 1995. Overcoming limitations of the mRNA differential display technique. Nucleic Acids Research, 23(22):4738-4739
Subudhi PK, Borkakati RP, Virmani SS. et al. 1997. Molecular mapping of a thermosenstive genetic malesterility gene in rice using bulked segregant analysis. Genome, 40:188-194
Sun Y, Hegamyer G,CoLburn NH. 1994. Molecular cloning of five messenger RNAs differential expressed in preneoplasmic or neoplasmic JB6 mouse epidermal cell: one is homologous to human tissue inhibitor of metalloproteinases-3. Cancer Res, 54(5): 1139-1144
Stamper SE, Dewey RE, et al. 1987. Characterization of the gene urf13-T and an unidentified reading frame, ORF 25, in maize and tobacco mitochondria. Curr Genet, 12(6):457-463
Stassar MJ, Devitt G, Brosius M,et al. 2001. Identification of human renal cell carcinoma associated genes by suppression subtractive hybridization. Br J Cancer, 85(9):1372-1382
Tavernarakis N, Wang SL, Dorovkov M, Ryazanov A, Driscoll M. 2000. Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes. Nat Genet,24(2): 180-183
Tsuchiya T, Toriyama K, Yoshikawa M, et al. 1995.Tapetum-specific expression of the gene for an endo-beta-1,3-glucanase causes male sterility in transgenic tobacco. Plant Cell Physiol,36(3):487-494
Van der Krol AR, Lenting PE. Veenstra J, et al. 1988. Antisense chalone synthase gene intransgenic plants inhibits flowerpigmentation. Nature, 333:866-869
van der Meer IM, Stare ME, van Tunen AJ, et al. 1992.Antisense inhibition of fiavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell ,4(3):253-262
Voiblet C, Duplessis S, Encelot N, et al. 2001 Identification of symbiosis-regulated genes in Eucalyptus globulus-Pisolithus tinctorius ectomycorrhiza by differential hybridization of arrayed cDNAs. Plant, 25(2): 181-191
Von Stein OD, Thies WG, Hofmann MA. 1997. A high throughput screening for rarely transcribed differentially expressed genes. Nucleic Acids Research, 25(13): 2598-2602
Wang B, Xu WW, Wang JZ, et al. 1995. Tagging and mapping of rice thermosenstive genic male sterile gene using molecular markers. Theor Appl Genet, 91(8):1111-1114
Wang Q, Yang C. Zhou J, et al. 2001. Cloning and characterization of full-length human ribosomal protein L15 cDNA which was overexpressed in esophageal cancer. Gene.263(1-2):205-209
Wang X, Feuerstein GZ. 2000. Suppression subtractive hybridization: application in the discovery of novel pharmacological targets. Pharmacogenomics, 1 (1): 101-108
Wassenegger M, Pelissier T. 1998. A model for RNA-mediated gene silencing in higher plants. Plant Mol Biol 37(2):349-362
Welsh J, Chada K, Dalai SS, et al. 1992. Arbitrarily primed PCR fingerprinting of RNA. Nucleic Acids Research, 20(9):4965-4970
Waterhouse PM, Graham MW, Wang MB, et al. 1998.Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA,95( 23): 13959-13964
Wianny F, Zernicka-Goetz M. 2000. Specific interference with gene function by double-stranded RNA in early mouse development. Nat Cell Biol, 2(2):70-75
Wilson ZA, Morroll SM, Dawson J, ct al.2001.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, 28(1):27-39
Wise RP, Gobelman-Werner K, Pei D, et al. 1999. Mitochondrial transcript processing and restoration of male fertility in T-cytoplasm maize. J Hered ,90(3):380-385
Xiong L, Lee MW, Qi M, et al. 2001.Identification of defense-related rice genes by suppression subtractive hybridization and differential screening. Mol Plant Microbe Interact, 14(5):685-692
Zamore PD, Tuschl T, Sharp PA, et al. 2000. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell,101(1):25-33
Zabaleta E, Mouras A, Hernould M, et al. 1996. Transgenic male-sterile plant induced by an unedited atp9 gene is restored to fertility by inhibiting its expression with antisense RNA. Proc Natl Acad Sci USA, 93(20): 11259-11263
Zhang L, Cilley RE, Chinoy MR. 2000. Suppression subtractive hybridization to identify gene expressions in variant and classic small cell lung cancer cell lines. J Surg Res, 93(1): 108-119
Zhang QF, Shen BZ. Dai XK, et al. 1994. Using bulked extremes and recessive class to map genes for photoperiod-sensitive genic male sterility in rice. Proc Natl Acad Sci U S A, 91 (18):8675-8679
Zhang Y, Shewry PR, Jones H, et al. 2001. Expression of antisense SnRK1 protein kinase sequence causes abnormal pollen development and male sterility in transgenic barley. Plant J, 28(4):431-441
Zhang YL, Ong CT, Leung KY. 2000. Molecular analysis of genetic differences between virulent and aviruient strains of Aeromonas hydrophila isolated from diseased fish.Microbiol. 146(Pt4):999-1009
Zhao R, Dielen V, Kinet JM, et al. 2000. Cosuppression of a plasma membrane H(+)-ATPase isoform impairs sucrose translocation, stomatal opening, plant growth, and male fertility. Plant Cell, 12(4):535-546
陈建南，傅鸿仪，路子显等．1997．科学通报，42(18)：1993-1997
陈良碧，周广洽，黄花玉祥．1994．温光条件对水稻安农S-1／衡农S-1的育性及生理的影响，植物学报,36(S)：119-1234
陈良碧，周广洽．1992．光温敏核不育水稻结实率与温度相关性研究．杂交水稻，6(2)：35-39
程式华，孙宗修．1993．国外光温敏核不育水稻的研究进展．种子世界，3：35
程式华，孙宗修，斯华敏．等，1994．农垦58S／农垦58F-2群体育性分离模式的年度间差异．中国水稻科学，8(2)：97-101
邓华凤，舒福北，袁定阳．1999．安农S-1的研究及其利用．杂交水稻，14(3)：1-3
傅荣昭，曹光诚，马江生等．1997．用基因枪法将人工雄性不育基因导入小麦的研究初报．遗传学报，24(4)：358-361
傅荣昭，孙勇如，贾士荣主编．1994．植物遗传转化手册．北京：中国科学技术出版社，134-143
高恒广．1993．三系不育系龙特浦A具有光温敏特性．杂交水稻，(1)：44-45
高一枝．1991．水稻短光敏雄性不育材料的发现初报．宜春农专学报．7(1)：1-5
何觉民，戴君惕，邹应斌，等．1994。生态遗传雄性不育理论与两系杂交植物Ⅰ：生态遗传雄性不育理论．湖南农业大学学报，20(1)：1-5
金冬雁，黎孟枫等译．1996．J．萨姆布鲁克，E．F．弗里奇，T．曼尼阿蒂斯等著．分子克隆实验指南，第二版．北京：科学出版社
康健，陈凡，吴乃虎．1998．光敏核不育水稻育性相关基因片段的分离．科学通报，43(19)：2078-2082
李传友，伏建民，金德敏等．1998．AFLP分析中多态性扩增产物的回收、克隆及鉴定．遗传,20(3)：1-4
李仕贵，周开达，朱立煌．1999，水稻温敏核不育基因的遗传分析和分子标记定位．科学通报，44(9)：955-958
李泽炳，肖翊华，朱英国等．1982．杂交水稻的研究与实践．上海：上海科学技术出版社
李子银，林兴华，谢岳峰等．1999．利用分子标记定位农垦58S的光敏核不育基因．植物学报，41(7)：731-735
凌定厚，陶利珍，马镇荣等．1998．以基因枪介导获转psl-Barnase基因的工程雄性不育水稻植株．遗传学报,25(5)：433-442
刘爱民，李必湖．1996．水稻温敏不育系育性与温度的相关性分析．杂交水稻，10(3)：25-28
李发生，谢戎，邵启明，等．1996．水稻低温敏两用系363S选育研究．西南农业大学学报，18(4)：338-342
刘军，袁自强，刘建东等．2000．应用抑制差减杂交分离水稻幼发育早期特异表达的基因．科学通报，45(13)：1392-1397
卢兴桂，顾铭洪。李成荃等．2001．两系杂交水稻理论与技术．北京：科学出版社．
罗玉英，刘玉乐，李胜国等．1997．TA29-Barnase嵌合基因导入对烟草花药绒毡层及花粉发育的影响．植物学报．39(10)：894-898
罗越华，周庭波，袁潜华，陈玉新．2001．水稻雄性不育研究的现状与展望．琼海：海南省生物技术研讨会论文汇编，91-97
梅明华，李泽炳．1995．水稻光温敏不育系与核质互作不育系的遗传关系剖析．遗传，17(1)：22-25
米志勇，王树生，吴乃．2000，水稻低分子量GTP结合蛋白基因OsRACD的分离．科学通报，45(19)：2047-2055
牛景，牛芝霞．2001．光敏核不育水稻杂种F2花药培养中获得了细胞质不育突变体．杂交水稻，16(2)：40
潘润森，李维明，闵绍楷，等．1994．光周期和温度对水稻雄性不育系龙特浦A育性恢复的影响．福建农业大学学报，23(3)：257-261
秦泰辰．1980．杂种优势利用原理和方法．南京：江苏科学技术出版社
邱芳，金德敏，伏健民等．1999．温敏核不育水稻5460S的细菌人工染色体文库的构建和鉴定．中国科学(C)，29(5)：518-524
曲雪萍，伏健民，李传友等．1999．水稻5460F的细菌人工染色体文库的构建和鉴定．科学通报,44)14)：1532-1536
石明松．1981．晚粳自然两用系的选育及其应用初报．湖北农业科学，7：44-48
孙宗修，程式华．1994．杂交水稻育种—从三系两系到一系．北京：中国农业科学出版社
童哲．1998．光敏核不育水稻的发育生物学研究评述．植物学报，40(3)：189-199
王斌，伏建民，邱方等．1998．TGMS水稻BAC文库的构建及与TGMS基因连锁的BAC克隆的筛选．遗传，20(增刊)：115
王培田．1976．水稻雄性不育发生的原因何在?遗传与育种，5：9-10
杨金水．1996，杂种优势机理探讨：作物雄性不育及杂种优势研究进展(Ⅰ)．北京：中国农业出版社．1-12
张宏，王波，薛爱群等．1998．雄性不育嵌合基因的构建及番茄转化研究．遗传，20(3)：5-7
杨仁崔，王乃元，梁康迳等．1993．籼稻温核不5460S的研究．福建农学院学报，22(2)：135-140
袁隆平．1987．杂交水稻的育种战略设想．杂交水稻，(1)：1-3
袁隆平．2000．两系法杂交水稻技术研究与中试开发．国家八六三计划项目验收报告
元生朝，张自国．1987．光照诱导湖光怪陆离核不育水稻育性转换的研究．武汉大学学报，HPGMR专刊：17-28
张自国，曾汉来，元生朝等．1992．再论光敏型核不育水稻育性转换的光温模式．华中农业大学学报，11(1)：1-6
中国农业科学院，湖南省农业科学院主编．1991．中国杂交水稻的发展．北京：中国农业出版社
曾汉来，张自国，元生朝等．1993．光敏型核不育水稻育性转换的温度敏感期研究．华中农业大学学报，12(5)：401-406
周国峰，龚光明，尹楚球等．1991．籼型两用不育系衡农S-1育性转换期及其遗传行为的初步观察．湖南农业科学，4：10-12
周坤炉．1994．籼型杂交水稻三系不育系选育．杂交水稻，1994(3-4)：22-26
周雪荣，彭仁旺，方荣祥等．表达核糖核酸酶基因的雄性不育油菜的获得．遗传学报，1997，24(6)：531-536
朱国，扬代常．1992．光周期敏感核不育水稻研与利用．武汉：武汉大学出版社
庄楚雄，徐是雄，卢永根等．1999．运用cDNA缩减杂交法克隆水稻花粉发育有关的cDNA．科学通报，44(17)：1842-1846
Aarts MG, Dirkse Wg, Stiekema W J, et al. 1993. Transposon tagging of a male sterility gene in Arabidopsis. Nature,363(6431): 715-717
Aarts MG, Hodge R, Kalantidis K, et al. 1997. The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes. Plant J. 12(3):615-623
Akagi H. Sakamoto M, Shinjyo C. et al. 1994. A unique sequence located downstream from the rice mitochondrial atp6 may cause male sterility. Curr Genet. 25(1):52-58
Bahn SC, Bae MS, Park YB, et al.2001.Molecular cloning and characterization of a novel low temperature-induced gene, blti2, from barley (Hordeum vulgare L.). Biochim Biophys Acta, 1522(2): 134-137
Cao GC,Sun YR, Chen ZK,et al. 1996.The cloning and construction study of artificial sterile gene and restore gene. Scentia Agricultural Sinica, 29(3):20-26
Carmeci C, Thompson DA, Kuang WW, et al. 1998. Moesin expression is associated with the estrogen receptor-negative breast cancer phenotype. Surgery, 124(2):211-217
Cho HJ, Kim S. Kim M, et al. 2001. Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells, 11(3):326-333
Chuang CF and Meyerowitz EM. 2000.Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana Proc. Natl. Acad. Sci. USA, 97(9), 4985-4990
Cui X, Wise RP, Schnable PS. 1996. The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize. Science,272(5266): 1334-1336
David J, Ursula HA, Michael J, et al. 1995. An analysis of differential display shows a strong bias towards high copy number mRNAs. Nucleic Acids Research, 23(21):4520-4523
Dessens JT, Margos G, Rodriguez MC, et al. 2000.Identification of differentially regulated genes of Plasmodium by suppression subtractive hybridization. Parasitol Today, 16(8):354-356
Diatchenko L, Lau Y, Campbell A, et al. 1996. Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc Natl Acad Sci USA, 93(12): 6025-6030
Dong NV, Subudhi PK ,Luong PN,et al.2000. Molecular mapping of a rice thermosenstive genetic male sterile gene (TGMS) by AFLP, RFLP, and SSR techniques. Theor Appl Genet, 100(5):727-734
Dudley NR, Labbe JC, Goldstein B.2002. Using RNA interference to identify genes required for RNA interference. Proc Natl Acad Sci U S A, 99(7):4191-4196
Fire A, Xu S, Montgomery MK, et al. 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391 (6669):806-811
Fire A. 1999. RNA-triggered gene silencing. Trends Genet, 15(9):358-363
Godt D E and Roitsch T. 1997. Regulation and tissue-specific distribution of mRNAs for three extracellular invertase isoenzymes of tomato suggests an important function in establishing and maintaining sink metabolism. Plant Physiol, 115(1): 273-282
Goetz M, Godt DE, Guivarc'h A, et al,2001. Induction of male sterility in plants by metabolic engineering of the carbohydrate supply. Proc Natl Acad Sci U S A, 98(11):6522-6527
Hammond SM, Bernstein E, Beach D, et al. 2000. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature ,404(6775):293-296
He S, Abad AR, Gelvin SB, et al. 1996. A cytoplasmic male sterility-associated mitochondrial protein causes pollen disruption in transgenic tobacco. Proc Natl Acad Sci U S A, 93(21): 11763- 11768
He SC, Lyznik A, Mackenzie S, et al. 1995. Pollen fertility restoration by nuclear gene Fr in CMS Bean: nuclear-direct alteration of a mitochondrial population. Genetics, 139(2): 955-962
Hernould M, Suharsono S, Litvak S, et al. 1993. A. Male-sterility induction in transgenic tobacco plants with an unedited atp9 mitochondrial gene from wheat. Proc Natl Acad Sci U S A,90(6):2370-2374
Hernould M, Suharsono, Zabaleta E,et al. 1998. Impairment of tapetum and mitochondria in engineered male-sterile tobacco plants. Plant Mol Biol, 36(4):499-508
Hinderhofer K, Zentgraf U. 2001.1dentification of a transcription factor specifically expressed at the onset of leaf senescence. Planta, 213(3):469-473
Howad W, Kempken F. 1997. Cell type-specific loss of atp6 RNA editing in cytoplasmic male sterile Sorghum bicolor. Proc Natl Acad Sci U S A,94(20): 11090-11095
Howad W, Tang HV, Pring DR, et al. 1999. Nuclear genes from Tx CMS maintainer lines are unable to maintain atp6 RNA editing in any anther cell-type in the sorghum bicolor A3 cytoplasm. Curt Genet, 36(1-2):62-68
Hufton SE, Moerkerk PT, Brandwijk R, et al. 1999. A profile of differentially expressed genes in primary colorectal cancer using suppression subtractive hybridization. FEBS Lett ,463(1-2):77-82
Jia JH, Li CY, Qu XP, et al. 2000.Construction of genetic linkage map and chromosome of tms5 gene inrice. In:Abstract Book of Plant Genomics in China I.Dalian
Jin H, Cheng X, Diatchenko L, et al. 1997. Differential screening ora subtracted cDNA library: a method to search for genes preferentially expressed in multiple tissues. Biotechniques, 23(6): 1084-1086
Kempken F, Howard W, Piing DR. 1998. Mutations at specific atp6 codons which cause human mitocbondrial diseases also lead to male sterility in a plant. FEBS Lett, 441 (2): 159-160
Ketting RF, Fischer SE, Bernstein E, et al. 2001. Dicer functions in RNA interference and in synthesis of small RNA involved in developmental timing in C. elegans. Genes Dev, 15(20):2654-2659
Kim JY, Chung YS, Paek KH, et al.1999a. Isolation and characterization of a cDNA encoding the cysteine proteinase inhibitor, induced upon flower maturation in carnation using suppression subtractive hybridization. Mol Cells ,9(4):392-397
Kim JY, Chung YS, Ok SH, et al. 1999b. Characterization of the full-length sequences of phospholipase A2 induced during flower development. Biochim Biophys Acta, 1489(2-3):389-392
Kriete G, Niehaus K, Perlick AM, et al. 1996. Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J,9(6):809-818
Kuang WW, Thompson DA, Hoch RV, et al. 1998. Differential screening and suppression subtractive hybridization identified genes differentially expressed in an estrogen receptor-positive breast carcinoma cell line. Nucleic Acids RES, 26(4):1116-1123
Lamar EE, Palmer E. 1984. Y-encoded, species-specific DNA in mice: evidence that the Y chromosome exists in two polymorphic forms in inbred strains. Cell. 37( 1 ): 171 - 177
Landgren M, Zetterstrand M, Sundberg E, et al. 1996. Alloplasmic male-sterile Brassica lines containing B. tournefortii mitochondria express an ORF 3' of the atp6 gene and a 32 kDa protein. Plant Mol Biol, 32(5):879-890
Lang NT, Subudhi PK, Virmani SS, et al. 1999. Development of PCR-based markers for thermosensitive genetic male sterility gene tms3(t) in rice (Oryza sativa L.). Hereditas, 131 (2): 121-127
Larose M, Bouchard C, Chagnon YC. 2001. A new gene related to human obesity identified by suppression subtractive hybridization. Int J Obes Relal Metab Disord. 25(6):770-776
Liang P, Pardee AB. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science, 257:967-970
Lisitsyn N, Wiglet M. 1993, Cloning the differences between two complex genomes. Science,259(5097):946-951
Liu F, Cui X, Horner HT, et al. 2001.Mitochondrial aldehyde dehydrogenase activity is required for male fertility in maize. Plant Cell, 13(5):1063-1078
Liu N, Shan Y, Wang FP, et al. 2001. Identification of an 85-kb DNA fragment containing pmsl, a locus for photoperiod-sensitive genic male sterility in rice. Mol Genet Genomics, 266(2):271-275
Koh HJ, Son YH, Heu MH, et al. Molecular mapping of a new genic male sterile gene causingf chalking endosperm in rice (Oryza sativa L). Euphitica, 106:57-62
Janke B, Dobrindt U, Hacker J, et al. 2001. A subtractive hybridisation analysis of genomic differences between the uropathogenic E. coli strain 536 and the E. coli K-12 strain MG1655. FEMS Microbiol Lett, 199(1):61-66
Mariani C, De Beuckeleer M, Truettner M, et al. 1990 Intmction of male sterility in plants by a chimeric ribonuclease gene. Nature,347:737-741
Mariani C, Gossele V, De Beukeleer M, et al. 1992. A chimeric ribonuclease inhibitor gene restores fertility to male sterile plants. Nature, 357:384-387
Marugma K, Araki H,kato H,et al. 1991. Thermonsensitive genic male sterility induced by irradiation. In:Khush GS ed. Rice Genet Ⅱ.manila, Phlippines:IRRI.227-235
Mascarenthas JP. 1993. Molecular mechanisma of pollen tube growth and differertitation. Plant Cell, 5(10):1303-1314
Matsuda N, Tsuchiya T, Kishitani S, et al. 1996. Partial male sterility in transgenic tobacco with antisense and sense PAL cDNA regulated by a tapetum-specific promoter. Plant Cell Physiol, 37(2):215-222
Mei MH,Chen,Zhang ZH, et al. 1999. pms3is the locus cauing the original photoperiod-sensitive male sterilitymutationof "Nongken 58S". Science in China(series C),42(3):316-322
Misquitta L and Paterson BM. 1999. Targeted disruption of gene function in Drosophila by RNA interference (RNA-i): A role for nautilus in embryonic somatic muscle formation. 96(4): 1451-1456
Miyasaka Y, Enomoto N, Nagayama K, et al. 2001. Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization. Br J Cancer, 85(2):228-234
Nicholson RH, Nicholson AW. 2002. Molecular characterization of a mouse cDNA encoding Dicer, a ribonuclease Ⅲ ortholog involved in RNA interference. Mamm Genome, 13(2):67-73
Okumura N, Nishizawa NK, Umehara Y ,et al. 1994. A dioxygenase gene (Ids2) expressed under iron deficiency conditions in the roots of Hordeum vulgate. Plant Molecular Biology, 25(4):705-719
Paddison PJ, Caudy AA, Bernstein E, et al. 2002. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev, 16(8):948-958
Porkka KP, Visakorpi T. 2001. Detection of differentially expressed genes in prostate cancer by combining suppression subtractive hybridization and cDNA library array. J Pathol, 193(1):73-79
Reddy OUK, Siddiq EA, Sarma NP, et al.2000.Genetic analysis of temoerature-sensitive male sterility in rice. Theor Appl Genetic, 100(5):794-801
Rutger JN. 1990. Sex-swithching rice may be short-out to hybridization. Rice Journal.93(6):4-5
Sharp PA. 1999. RNAi and double-strand RNA .Gen Dev, 13( 2): 139-141
Sompayrac L, Jane S, Burn TC, et al. 1995. Overcoming limitations of the mRNA differential display technique. Nucleic Acids Research, 23(22):4738-4739
Subudhi PK, Borkakati RP, Virmani SS. et al. 1997. Molecular mapping of a thermosenstive genetic malesterility gene in rice using bulked segregant analysis. Genome, 40:188-194
Sun Y, Hegamyer G,CoLburn NH. 1994. Molecular cloning of five messenger RNAs differential expressed in preneoplasmic or neoplasmic JB6 mouse epidermal cell: one is homologous to human tissue inhibitor of metalloproteinases-3. Cancer Res, 54(5): 1139-1144
Stamper SE, Dewey RE, et al. 1987. Characterization of the gene urf13-T and an unidentified reading frame, ORF 25, in maize and tobacco mitochondria. Curr Genet, 12(6):457-463
Stassar MJ, Devitt G, Brosius M,et al. 2001. Identification of human renal cell carcinoma associated genes by suppression subtractive hybridization. Br J Cancer, 85(9):1372-1382
Tavernarakis N, Wang SL, Dorovkov M, Ryazanov A, Driscoll M. 2000. Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes. Nat Genet,24(2): 180-183
Tsuchiya T, Toriyama K, Yoshikawa M, et al. 1995.Tapetum-specific expression of the gene for an endo-beta-1,3-glucanase causes male sterility in transgenic tobacco. Plant Cell Physiol,36(3):487-494
Van der Krol AR, Lenting PE. Veenstra J, et al. 1988. Antisense chalone synthase gene intransgenic plants inhibits flowerpigmentation. Nature, 333:866-869
van der Meer IM, Stare ME, van Tunen AJ, et al. 1992.Antisense inhibition of fiavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell ,4(3):253-262
Voiblet C, Duplessis S, Encelot N, et al. 2001 Identification of symbiosis-regulated genes in Eucalyptus globulus-Pisolithus tinctorius ectomycorrhiza by differential hybridization of arrayed cDNAs. Plant, 25(2): 181-191
Von Stein OD, Thies WG, Hofmann MA. 1997. A high throughput screening for rarely transcribed differentially expressed genes. Nucleic Acids Research, 25(13): 2598-2602
Wang B, Xu WW, Wang JZ, et al. 1995. Tagging and mapping of rice thermosenstive genic male sterile gene using molecular markers. Theor Appl Genet, 91(8):1111-1114
Wang Q, Yang C. Zhou J, et al. 2001. Cloning and characterization of full-length human ribosomal protein L15 cDNA which was overexpressed in esophageal cancer. Gene.263(1-2):205-209
Wang X, Feuerstein GZ. 2000. Suppression subtractive hybridization: application in the discovery of novel pharmacological targets. Pharmacogenomics, 1 (1): 101-108
Wassenegger M, Pelissier T. 1998. A model for RNA-mediated gene silencing in higher plants. Plant Mol Biol 37(2):349-362
Welsh J, Chada K, Dalai SS, et al. 1992. Arbitrarily primed PCR fingerprinting of RNA. Nucleic Acids Research, 20(9):4965-4970
Waterhouse PM, Graham MW, Wang MB, et al. 1998.Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA,95( 23): 13959-13964
Wianny F, Zernicka-Goetz M. 2000. Specific interference with gene function by double-stranded RNA in early mouse development. Nat Cell Biol, 2(2):70-75
Wilson ZA, Morroll SM, Dawson J, ct al.2001.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, 28(1):27-39
Wise RP, Gobelman-Werner K, Pei D, et al. 1999. Mitochondrial transcript processing and restoration of male fertility in T-cytoplasm maize. J Hered ,90(3):380-385
Xiong L, Lee MW, Qi M, et al. 2001.Identification of defense-related rice genes by suppression subtractive hybridization and differential screening. Mol Plant Microbe Interact, 14(5):685-692
Zamore PD, Tuschl T, Sharp PA, et al. 2000. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell,101(1):25-33
Zabaleta E, Mouras A, Hernould M, et al. 1996. Transgenic male-sterile plant induced by an unedited atp9 gene is restored to fertility by inhibiting its expression with antisense RNA. Proc Natl Acad Sci USA, 93(20): 11259-11263
Zhang L, Cilley RE, Chinoy MR. 2000. Suppression subtractive hybridization to identify gene expressions in variant and classic small cell lung cancer cell lines. J Surg Res, 93(1): 108-119
Zhang QF, Shen BZ. Dai XK, et al. 1994. Using bulked extremes and recessive class to map genes for photoperiod-sensitive genic male sterility in rice. Proc Natl Acad Sci U S A, 91 (18):8675-8679
Zhang Y, Shewry PR, Jones H, et al. 2001. Expression of antisense SnRK1 protein kinase sequence causes abnormal pollen development and male sterility in transgenic barley. Plant J, 28(4):431-441
Zhang YL, Ong CT, Leung KY. 2000. Molecular analysis of genetic differences between virulent and aviruient strains of Aeromonas hydrophila isolated from diseased fish.Microbiol. 146(Pt4):999-1009
Zhao R, Dielen V, Kinet JM, et al. 2000. Cosuppression of a plasma membrane H(+)-ATPase isoform impairs sucrose translocation, stomatal opening, plant growth, and male fertility. Plant Cell, 12(4):535-546