芒与五节芒种间自然杂交研究
|
摘要
自然杂交一直是分类学和进化生物学的热点。芒属植物是一类新型的能源植物,芒属植物的分类及进化研究一直有待完善,在野外收集野生种植资源时,常遇见表型特征介于芒与五节芒之间的植株,通过表型形态鉴定难以准确确定其分类地位。本研究拟通过分析芒与五节芒表型性状及Adh1基因序列,初步探讨芒与五节芒的自然杂交起源过程中基因及表型的渗透。具体结果如下：
1、表型性状分析
测定了芒与五节芒共13份材料17项表型性状,所有表型性状数据采用ZScores进行标准化转换,计算13个材料两两间的欧式距离,在MVSP3.2软件上计算做主成分分析并进行UPGMA聚类。形态性状聚类分析结果表明疑似杂交种AD431与五节芒类群(AA335和AD625)聚为一类,而疑似杂交种AA343、AD431、AD628、AD607、AD633和AD606则与芒类群(AD623、AD512、AD620、AD619和AD627)聚为一类。
2、Adhl基因序列分析
测定13份材料Adhl基因的全部序列,分析已测定序列的序列特征,并结合GeneBank上11份已知序列,采用最大简约法(Maximum parsimony, MP)和邻接法(Neighbor-joining, NJ)分别构建分子系统树。结果显示芒属植物的Adh1基因包括6个外显子和5个内含子,外显子片段大小为60-210bp,平均大小为114bp。在13分材料用来构树的19个序列中,序列大小范围为1413-1444bp,平均大小为1430bp,当用ClustalX对齐序列并排除空位和多重迭代后显示序列长度为1473bp。序列A,T,C,G碱基含量分别为24.7%,30.3%,20.5%,24.5%,序列有明显的T碱基偏好,AT碱基含量(55.0%)明显大于CG碱基含量(45%)。颠换值为16,转换值为27,转换(SV)明显大于颠换(SI),转换与颠换的比为1.7。采用MEGA4.0程序上分别构建最大简约树和邻接树,结果显示五节芒和芒的材料分别聚成两个明显分开的分支,而疑似杂交种中均检测到两种Adh1基因单倍型的存在,其中一种单倍型在系统树中与芒聚为一类,另一单倍型则与五节芒聚为一类。
本研究结果表明：采用基于形态学性状的聚类分析并不能有效的将杂交种区分开来,而采用Adh1基因序列聚类分析则成功将杂交种鉴定出来。导致这两种聚类分析方法出现差异的本质原因在于：自然杂交所产生的杂交种,往往会与亲本再发生回交,即所谓的渐渗杂交现象,从而导致一个种群的基因逐渐渗入到了另一个种群的基因库中。种间的基因渗透,则降低了种群间的遗传差异性,使得种群间的遗传背景和表型性状趋于同化,从而导致这两个种群在进化上表现出趋近的亲缘关系。从本研究的测序结果可以发现,杂交种Adh1基因的内含子和外显子碱基序列均发生了改变,外显子碱基序列的变异更为丰富；而能够反映到形态上出现差别性状,仅仅只是那些由外显子上的显性基因所控制的少数性状,大量的中性突变在形态学上是没有反映的。因此基因序列聚类分析与形态性状聚类分析的结果出现差异是常见的,而基因序列聚类分析结果所反映出的物种变化更本质、更全面、更准确,用它所鉴定出的杂交种也更可靠。
Penetration of natural hybrid has always been the hottest issue in the field of taxonomy and evolutionary biology, owing to be hindered by the traits of its wide range of origins, frequent occurrences, and difficulties in identifications. Miscanthus is a new type of energy plant, the study of whose classification and evolution need to be improved urgently. It's hard to accurately distinguish a certain plant and identify its taxa by morphological phenotype whose characteristics is similar to and Miscanthus during the collection of wild plants resources. Though research and analysis to different phenotypes and Adh1 gene sequences of, a primary study into genetic and phenotypic penetration between sinensis and miscanthus in the process of the natural hybrid origin. The major results were as follows:
1) morphological phenotype analyse.
All of 13 materials's 17morphological characters had been determined and analysed.All the morphological characters had been standardized by the ZScores and computed theirs distance. The date operating in the MVSP3.2 by the UPGMA method. The phylogenic analysis based on morphological characters indicated that one putative hybrid (AD431) clustered together with M. floridulus samples and other putative hybrids (AD623、AD512、AD620、AD619 and AD627) grouped with M. sinensis samples.
2) Adhl gene analyse
All of 13 materials's Adh1 gene sequence which building-up the phylogenetic trees using the MP method and the NJ method with the other 11 sequences registered in the GeneBank were mensurated and cluster analysed. The results showed that the Adhl gene of the Miscanthus Anderss include 6 exons and 5 introns, and the exon region is 60bp to 210bp,the average value is 114bp. The 19 sequences's region is 1413bp to 1444bp and the average value is 1430bp. The length of sequence is 1473bp when analyse by the ClustalX. The A,T,C,G of the sequence take up 24.7%,30.3%, 20.5%,24.5%,and the sequence has the prominent T predilection, AT(55.0%) greater than CG(45%), and the ratio of between the SI(16) is 1.7two diverged types of Adhl gene sequences presented in all samples of putative hybrids, one of which is clustered with M. sinensis while the other is clustered with M. floridulus on the Adh1 phylogeny, suggesting that they are the true hybrids between M. sinensis and M. floridulus.
The phylogenic analysis based on morphological characters can not verification the hybrids, and the Adhl gene sequence analyse can.The Introgressive hybridizationhas been always appears in the nature hybrids, this phenomenon bring about the divergence between two analyse. The heredity background and the morphological characters has been assimilated by the Introgressive of gene and the otherness of the species has been reduced. Acid base of those hybrids's exons and introns has been changed and the exons are more plentifully. The disparate phylogenic controlled by the exons which in the dominant gene is the minority and the neutral mutation can't been expressed in the phylogenic. The otherness of the gene sequence analyse bwtween the morphological characters analyse is familiar, and the sequence analyse is more essential, general, exacting and the results when identifying the hybrids is more exacting.
1、表型性状分析
测定了芒与五节芒共13份材料17项表型性状,所有表型性状数据采用ZScores进行标准化转换,计算13个材料两两间的欧式距离,在MVSP3.2软件上计算做主成分分析并进行UPGMA聚类。形态性状聚类分析结果表明疑似杂交种AD431与五节芒类群(AA335和AD625)聚为一类,而疑似杂交种AA343、AD431、AD628、AD607、AD633和AD606则与芒类群(AD623、AD512、AD620、AD619和AD627)聚为一类。
2、Adhl基因序列分析
测定13份材料Adhl基因的全部序列,分析已测定序列的序列特征,并结合GeneBank上11份已知序列,采用最大简约法(Maximum parsimony, MP)和邻接法(Neighbor-joining, NJ)分别构建分子系统树。结果显示芒属植物的Adh1基因包括6个外显子和5个内含子,外显子片段大小为60-210bp,平均大小为114bp。在13分材料用来构树的19个序列中,序列大小范围为1413-1444bp,平均大小为1430bp,当用ClustalX对齐序列并排除空位和多重迭代后显示序列长度为1473bp。序列A,T,C,G碱基含量分别为24.7%,30.3%,20.5%,24.5%,序列有明显的T碱基偏好,AT碱基含量(55.0%)明显大于CG碱基含量(45%)。颠换值为16,转换值为27,转换(SV)明显大于颠换(SI),转换与颠换的比为1.7。采用MEGA4.0程序上分别构建最大简约树和邻接树,结果显示五节芒和芒的材料分别聚成两个明显分开的分支,而疑似杂交种中均检测到两种Adh1基因单倍型的存在,其中一种单倍型在系统树中与芒聚为一类,另一单倍型则与五节芒聚为一类。
本研究结果表明：采用基于形态学性状的聚类分析并不能有效的将杂交种区分开来,而采用Adh1基因序列聚类分析则成功将杂交种鉴定出来。导致这两种聚类分析方法出现差异的本质原因在于：自然杂交所产生的杂交种,往往会与亲本再发生回交,即所谓的渐渗杂交现象,从而导致一个种群的基因逐渐渗入到了另一个种群的基因库中。种间的基因渗透,则降低了种群间的遗传差异性,使得种群间的遗传背景和表型性状趋于同化,从而导致这两个种群在进化上表现出趋近的亲缘关系。从本研究的测序结果可以发现,杂交种Adh1基因的内含子和外显子碱基序列均发生了改变,外显子碱基序列的变异更为丰富；而能够反映到形态上出现差别性状,仅仅只是那些由外显子上的显性基因所控制的少数性状,大量的中性突变在形态学上是没有反映的。因此基因序列聚类分析与形态性状聚类分析的结果出现差异是常见的,而基因序列聚类分析结果所反映出的物种变化更本质、更全面、更准确,用它所鉴定出的杂交种也更可靠。
Penetration of natural hybrid has always been the hottest issue in the field of taxonomy and evolutionary biology, owing to be hindered by the traits of its wide range of origins, frequent occurrences, and difficulties in identifications. Miscanthus is a new type of energy plant, the study of whose classification and evolution need to be improved urgently. It's hard to accurately distinguish a certain plant and identify its taxa by morphological phenotype whose characteristics is similar to and Miscanthus during the collection of wild plants resources. Though research and analysis to different phenotypes and Adh1 gene sequences of, a primary study into genetic and phenotypic penetration between sinensis and miscanthus in the process of the natural hybrid origin. The major results were as follows:
1) morphological phenotype analyse.
All of 13 materials's 17morphological characters had been determined and analysed.All the morphological characters had been standardized by the ZScores and computed theirs distance. The date operating in the MVSP3.2 by the UPGMA method. The phylogenic analysis based on morphological characters indicated that one putative hybrid (AD431) clustered together with M. floridulus samples and other putative hybrids (AD623、AD512、AD620、AD619 and AD627) grouped with M. sinensis samples.
2) Adhl gene analyse
All of 13 materials's Adh1 gene sequence which building-up the phylogenetic trees using the MP method and the NJ method with the other 11 sequences registered in the GeneBank were mensurated and cluster analysed. The results showed that the Adhl gene of the Miscanthus Anderss include 6 exons and 5 introns, and the exon region is 60bp to 210bp,the average value is 114bp. The 19 sequences's region is 1413bp to 1444bp and the average value is 1430bp. The length of sequence is 1473bp when analyse by the ClustalX. The A,T,C,G of the sequence take up 24.7%,30.3%, 20.5%,24.5%,and the sequence has the prominent T predilection, AT(55.0%) greater than CG(45%), and the ratio of between the SI(16) is 1.7two diverged types of Adhl gene sequences presented in all samples of putative hybrids, one of which is clustered with M. sinensis while the other is clustered with M. floridulus on the Adh1 phylogeny, suggesting that they are the true hybrids between M. sinensis and M. floridulus.
The phylogenic analysis based on morphological characters can not verification the hybrids, and the Adhl gene sequence analyse can.The Introgressive hybridizationhas been always appears in the nature hybrids, this phenomenon bring about the divergence between two analyse. The heredity background and the morphological characters has been assimilated by the Introgressive of gene and the otherness of the species has been reduced. Acid base of those hybrids's exons and introns has been changed and the exons are more plentifully. The disparate phylogenic controlled by the exons which in the dominant gene is the minority and the neutral mutation can't been expressed in the phylogenic. The otherness of the gene sequence analyse bwtween the morphological characters analyse is familiar, and the sequence analyse is more essential, general, exacting and the results when identifying the hybrids is more exacting.
引文
[1]解新明,周峰,赵燕慧,卢小良.多年生能源禾草的产能和生态效益[J].生态学报,2008,28(5)：2329-2342.
[2]萧运峰,王锐,高洁.五节芒生态—生物学特性的研究[J].四川草原,1995,(01)：25-29.
[3]刘亮,朱明,朱太平.芒荻类植物资源的开发和利用[J].自然资源学报,2001,16(06)：562-563.
[4]刘亮.中国植物志(第十卷,第二分册)[M].北京：科学出版社,1997,4-55.
[5]Benth G. Notes on Gramineae [J].J Linn Soc Bot,1881,19:14～135.
[6]吴征镒,路安民,汤彦承,等.中国被子植物综论[M].北京：科学出版社,2003,342-343.
[7]Chen S L. Renvoize S A. A new species and a new combination of Miscanthus(poaceae) From China [J]. Kew Bull,2005,60(4):605～607.
[8]Hunter. A karyosystematic investigation in the Gramineae[J]. Canad. Jour. Res,1930(11):213-241.
[9]Darlington C D,Wylie A P. Chromosome atlas of flowering plants[M]. London, George Allen and Unwin Ltd,1956(6):422.
[10]Hirayoshi I, Nishikawa K, Kubono M. Cyto-genetical studies on forage plants on the chromosome number of Ogi(Miscanthus sacchariflorus)[J]. Res Bull Fac Agr Gifu Univ,1957(8)8-13.
[11]Adati S. Studies on the genus Miscanthus with special reference to the Japanese species suitable for breeding purpose as fodder crops(in Japanese with English summary)[J]. Bull Fac Agr Gifu Univ,1958(17):111-112.
[12]Wikberg S. The genus Miscanthus, a summary of available literature[M]. Project Agro-fibre. Thesis, University of Lund, Sweden,1990.
[13]Linde Laursin. Cytogenetic analysis of Miscanthus'Giganteus', an interspecific hybrid[J]. Hereditas,1993,119:297-300.
[14]何立珍,李爱华,刘选民,周仆华.南荻的染色体核型模式[J].湖南农业大学学报,1996,22(5)：461-465.
[15]蔡青,文建成,范源洪.甘蔗属及其近缘植物的染色体分析[J].西南农业学报,2002,15(2)：16-19.
[16]陈少风.荻属系统学研究[D].湖南农业大学,博士学位论文,2007.
[17]Von Wuhlisch G, Deuter M, Muhs H. Identifizierung verschiedener Miscanthus-Sorten mittels Isoenzymen[J]. Journal of Agronomy & Crop Science,1993,172:247-254.
[18]黄家雍,廖江雄,诸葛莹.甘蔗与河八王、五节芒、滇蔗茅属间交配性及杂种F1无性系的形态学和同工酶分析[J].西南农业报,1997,10(3)：92-98.
[19]Hernandez P, Dorado G, Laurie DA, Martin A, Snape JW. Microsatellites RFLP probes from maize are efficient sources of molecular markers for the biomass energy crop Miscanthus[J]. Theor Appl Genet,2001,102:616-622.
[20]Greef M J, Deuter M, Jung C, Schondelmaier J. Genetic diversity of European Miscanthus species revealed by AFLP fingerprinting[J]. Genet Res Crop Evol,1997,44:185-195.
[21]S G Atienza, Z Satovic, K K Petersen, O Dolstra. Preliminary genetic linkage map of Miscanthus sinensis with RAPD markers[J]. Theor Appl Genet,2002,105:946-952.
[22]Hodkinson, Chase, Renvoize. Characterization of a Genetic Resource Collection for Miscanthus (Saccharinae, Andropogoneae, Poaceae) using AFLP and ISSR PCR[J]. Annals of Botany,2002,89:627-636.
[23J陈少风,董穗穗,吴伟.基于ITS序列探讨荻属及其近缘植物的系统发 育关系[J].武汉植物学研究,2007,25(3)：239-244.
[24]Rieseberg LH,Wendel J. Introgression and is consequences in plants [A]. Oxford University Press, New York,1997,70-103.
[25]Pigliucci M, Murren CJ, Schlichting CD. Phenotypic plasticity and evolution by genetic assimilation [J] Exp Biol,2006,209:362-367.
[26]李自超,张洪亮,曹永生.中国地方稻种初级核心种质取样策略研究[J].作物学报,2003,29(1)：20-24.
[27]Kilsung. Origion of Cultivated Rice[M]. Tokyo:J apan Seieniifie Soeieties Press,1988,12:170-175
[28]张洪亮.稻种资源核心种质研究及其信息处理系统骨架的建立[D].北京：中国农业大学,2004.
[29]N Bapa Rao, T. Sri Valli and N Lakshmi. Cytogenetic studies on the interspecific hybrid Capsicum baccatum L. x C. frutescens L. and its progeny[J].Euphytica,1992 (59):134-140.
[30]杜玉娟.中国沙棘×肋果沙棘自然杂交带的形态学分析[J],西北师范大学学报,2008(13),144.
[31]王瑞江.国产海桑属植物的两个杂交种[J].广西植物,1999(3)199-204.
[32]Linde laursen. Cytogenetic analysis of Miscanthus'Giganteus', an interspecific hybrid [J]. Hereditas,1993(119):297-300.
[33]Aparicio RG, Albaladejo M, Porras and G. Ceballoce. Isozyme Evidence for Natural Hybridization in Phlomis (Lamiaceae):Hybrid Origin of the Rare P. margaritae[J]. Annals of Botany,2000(85):7-12.
[34]Renchao Zhou. Molecular criteria for determining new hybrid species-Anapplication to the Sonneratia hybrids Molecular[J]. Phylogenetics and Evolution,2005(35) 595-601.
[35]Rieseberg LH. Hybrid origins of plant species[J]. Annual Review of ecology and Systematics,1997(18):359-389.
[36]Ertugrul Yuzbas loglu. Natural hybridization between Phlomis lycia D. DonP. Bourgaei Boiss, (Lamiaceae) revealed by RAPD markers[J]. Genetica,2008,133:13-20.
[37]Weiping Li. Natural hybridization between Aster ageratoides var. scaberulus and Kalimeris indica (Asteraceae):evidence from morphology, karyotype, and ITS sequences[J]. Botanical Studies,2006,47:191-197.
[38]A Franzke, K Mummenho. Recent hybrid speciation in Cardamine (Brassicaceae) conversion of nuclear ribosomal ITS sequences in statu nascendi[J]. Theor Appl Genet,1999,98:831-834.
[39]Brunner S. Evolution of DNA sequence nonhomologies among maize inbreds[J]. Plant Cell,2005,17(2):343-360.
[40]Hie K,San Miguel PJ, Bennetzen JL. A complex history of rearrangement in an orthologous region of the maize, sorghum, and rice Genomes[J]. Proc natl Acad,2003,100(21):12265-12270.
[41]廖芳.基于Adh1基因分析高粱属的系统进化[J].遗传,2009.31：523-530.
[42]袁长春,何雪宝,袁秋梅.绿绒蒿自然杂交种及其亲本cpDNA trn Ltrn F基因的遗传学分析[J].植物研究,2007,29(1)：103-108.
[43]李小娟,王留阳.麻花艽和管花秦艽(龙胆科)之间自然杂交类型的分子验证[J].云南植物研究,2007,29(1)：91-97.
[44]Suo Qiu. Molecular evidence for natural hybridization between Sonneratia alba and S.griffithii[J]. Journal of Systematics and Evolution,2008(10): 391-395.
[45]Carolyn J, Ferguson, Donald A, Levin, and Robert K. Jansen. Natural hybridization between an outcrossing and a selfing Phlox (Polemoniaceae): the maternal species of F1 hybrids[J]. Plant Syst.1999(218)153-158.
[46]Thompson JD, Gibson TJ. plewnia F. The Cluctal_X windows interface: flexible strategies formultiple sequences alignment aidcd by quality analysis tools[J]. Nucleic Acids Res.1997 (24):4876-4882.
[47]Tamura K, Dudley J, Nei M, Kumar S. MEGA4:Molecular Evolutionary Genetics Analysis(MEGA)software version 4.0[J]. MolBiol Evol, 2007,24(8):1596-1599.
[48]Rieseberg LH. Wendel J. Introgression and is consequences in plants [A]. Oxford University Press,New York:70-103.
[49]Mason-Gamer RJ. Allohexaploidy, introgression, and the complex phyloge-netic history of Elymus repens(Poaceae) [J]. Molecular Phylogenetics and Evolution,2008.47:598-611.
[50]Koch MA, Dobes C, Kiefer C, Schmickl R,klimes L, Lysak MA. Supernetwork identifies multiple events of plastid trnF(GAA) pseudogene evolutiong in the Braddicaceae[J]. Molecular Biology and Evolution, 2007:24,63-73
[51]Wortley AH, Rudall PJ, Harris DJ, Scotland RW. How much date are needed to resolve a difficult phylogeny? Case study in Lamiales [J]. Sysematic Bioligy,2005:54,697-709.
[52]Bergthorsson U, Richardsom AO, Young GJ, Goertzen LR. Massive horizontal transfer of mitochondrial genes from diverse land plant donors to the basal angionsperm Amborella[J]. Proceedings of the National Academy of Sciences USA; 2004:101,177747-177752.
[53]赵辉,李启寨,李俊.相邻碱基组分与产生SNP的转换或颠换在植物基因组中的研究[J].中国科学C辑生命科学,2006,36(1)：1-8.
[2]萧运峰,王锐,高洁.五节芒生态—生物学特性的研究[J].四川草原,1995,(01)：25-29.
[3]刘亮,朱明,朱太平.芒荻类植物资源的开发和利用[J].自然资源学报,2001,16(06)：562-563.
[4]刘亮.中国植物志(第十卷,第二分册)[M].北京：科学出版社,1997,4-55.
[5]Benth G. Notes on Gramineae [J].J Linn Soc Bot,1881,19:14～135.
[6]吴征镒,路安民,汤彦承,等.中国被子植物综论[M].北京：科学出版社,2003,342-343.
[7]Chen S L. Renvoize S A. A new species and a new combination of Miscanthus(poaceae) From China [J]. Kew Bull,2005,60(4):605～607.
[8]Hunter. A karyosystematic investigation in the Gramineae[J]. Canad. Jour. Res,1930(11):213-241.
[9]Darlington C D,Wylie A P. Chromosome atlas of flowering plants[M]. London, George Allen and Unwin Ltd,1956(6):422.
[10]Hirayoshi I, Nishikawa K, Kubono M. Cyto-genetical studies on forage plants on the chromosome number of Ogi(Miscanthus sacchariflorus)[J]. Res Bull Fac Agr Gifu Univ,1957(8)8-13.
[11]Adati S. Studies on the genus Miscanthus with special reference to the Japanese species suitable for breeding purpose as fodder crops(in Japanese with English summary)[J]. Bull Fac Agr Gifu Univ,1958(17):111-112.
[12]Wikberg S. The genus Miscanthus, a summary of available literature[M]. Project Agro-fibre. Thesis, University of Lund, Sweden,1990.
[13]Linde Laursin. Cytogenetic analysis of Miscanthus'Giganteus', an interspecific hybrid[J]. Hereditas,1993,119:297-300.
[14]何立珍,李爱华,刘选民,周仆华.南荻的染色体核型模式[J].湖南农业大学学报,1996,22(5)：461-465.
[15]蔡青,文建成,范源洪.甘蔗属及其近缘植物的染色体分析[J].西南农业学报,2002,15(2)：16-19.
[16]陈少风.荻属系统学研究[D].湖南农业大学,博士学位论文,2007.
[17]Von Wuhlisch G, Deuter M, Muhs H. Identifizierung verschiedener Miscanthus-Sorten mittels Isoenzymen[J]. Journal of Agronomy & Crop Science,1993,172:247-254.
[18]黄家雍,廖江雄,诸葛莹.甘蔗与河八王、五节芒、滇蔗茅属间交配性及杂种F1无性系的形态学和同工酶分析[J].西南农业报,1997,10(3)：92-98.
[19]Hernandez P, Dorado G, Laurie DA, Martin A, Snape JW. Microsatellites RFLP probes from maize are efficient sources of molecular markers for the biomass energy crop Miscanthus[J]. Theor Appl Genet,2001,102:616-622.
[20]Greef M J, Deuter M, Jung C, Schondelmaier J. Genetic diversity of European Miscanthus species revealed by AFLP fingerprinting[J]. Genet Res Crop Evol,1997,44:185-195.
[21]S G Atienza, Z Satovic, K K Petersen, O Dolstra. Preliminary genetic linkage map of Miscanthus sinensis with RAPD markers[J]. Theor Appl Genet,2002,105:946-952.
[22]Hodkinson, Chase, Renvoize. Characterization of a Genetic Resource Collection for Miscanthus (Saccharinae, Andropogoneae, Poaceae) using AFLP and ISSR PCR[J]. Annals of Botany,2002,89:627-636.
[23J陈少风,董穗穗,吴伟.基于ITS序列探讨荻属及其近缘植物的系统发 育关系[J].武汉植物学研究,2007,25(3)：239-244.
[24]Rieseberg LH,Wendel J. Introgression and is consequences in plants [A]. Oxford University Press, New York,1997,70-103.
[25]Pigliucci M, Murren CJ, Schlichting CD. Phenotypic plasticity and evolution by genetic assimilation [J] Exp Biol,2006,209:362-367.
[26]李自超,张洪亮,曹永生.中国地方稻种初级核心种质取样策略研究[J].作物学报,2003,29(1)：20-24.
[27]Kilsung. Origion of Cultivated Rice[M]. Tokyo:J apan Seieniifie Soeieties Press,1988,12:170-175
[28]张洪亮.稻种资源核心种质研究及其信息处理系统骨架的建立[D].北京：中国农业大学,2004.
[29]N Bapa Rao, T. Sri Valli and N Lakshmi. Cytogenetic studies on the interspecific hybrid Capsicum baccatum L. x C. frutescens L. and its progeny[J].Euphytica,1992 (59):134-140.
[30]杜玉娟.中国沙棘×肋果沙棘自然杂交带的形态学分析[J],西北师范大学学报,2008(13),144.
[31]王瑞江.国产海桑属植物的两个杂交种[J].广西植物,1999(3)199-204.
[32]Linde laursen. Cytogenetic analysis of Miscanthus'Giganteus', an interspecific hybrid [J]. Hereditas,1993(119):297-300.
[33]Aparicio RG, Albaladejo M, Porras and G. Ceballoce. Isozyme Evidence for Natural Hybridization in Phlomis (Lamiaceae):Hybrid Origin of the Rare P. margaritae[J]. Annals of Botany,2000(85):7-12.
[34]Renchao Zhou. Molecular criteria for determining new hybrid species-Anapplication to the Sonneratia hybrids Molecular[J]. Phylogenetics and Evolution,2005(35) 595-601.
[35]Rieseberg LH. Hybrid origins of plant species[J]. Annual Review of ecology and Systematics,1997(18):359-389.
[36]Ertugrul Yuzbas loglu. Natural hybridization between Phlomis lycia D. DonP. Bourgaei Boiss, (Lamiaceae) revealed by RAPD markers[J]. Genetica,2008,133:13-20.
[37]Weiping Li. Natural hybridization between Aster ageratoides var. scaberulus and Kalimeris indica (Asteraceae):evidence from morphology, karyotype, and ITS sequences[J]. Botanical Studies,2006,47:191-197.
[38]A Franzke, K Mummenho. Recent hybrid speciation in Cardamine (Brassicaceae) conversion of nuclear ribosomal ITS sequences in statu nascendi[J]. Theor Appl Genet,1999,98:831-834.
[39]Brunner S. Evolution of DNA sequence nonhomologies among maize inbreds[J]. Plant Cell,2005,17(2):343-360.
[40]Hie K,San Miguel PJ, Bennetzen JL. A complex history of rearrangement in an orthologous region of the maize, sorghum, and rice Genomes[J]. Proc natl Acad,2003,100(21):12265-12270.
[41]廖芳.基于Adh1基因分析高粱属的系统进化[J].遗传,2009.31：523-530.
[42]袁长春,何雪宝,袁秋梅.绿绒蒿自然杂交种及其亲本cpDNA trn Ltrn F基因的遗传学分析[J].植物研究,2007,29(1)：103-108.
[43]李小娟,王留阳.麻花艽和管花秦艽(龙胆科)之间自然杂交类型的分子验证[J].云南植物研究,2007,29(1)：91-97.
[44]Suo Qiu. Molecular evidence for natural hybridization between Sonneratia alba and S.griffithii[J]. Journal of Systematics and Evolution,2008(10): 391-395.
[45]Carolyn J, Ferguson, Donald A, Levin, and Robert K. Jansen. Natural hybridization between an outcrossing and a selfing Phlox (Polemoniaceae): the maternal species of F1 hybrids[J]. Plant Syst.1999(218)153-158.
[46]Thompson JD, Gibson TJ. plewnia F. The Cluctal_X windows interface: flexible strategies formultiple sequences alignment aidcd by quality analysis tools[J]. Nucleic Acids Res.1997 (24):4876-4882.
[47]Tamura K, Dudley J, Nei M, Kumar S. MEGA4:Molecular Evolutionary Genetics Analysis(MEGA)software version 4.0[J]. MolBiol Evol, 2007,24(8):1596-1599.
[48]Rieseberg LH. Wendel J. Introgression and is consequences in plants [A]. Oxford University Press,New York:70-103.
[49]Mason-Gamer RJ. Allohexaploidy, introgression, and the complex phyloge-netic history of Elymus repens(Poaceae) [J]. Molecular Phylogenetics and Evolution,2008.47:598-611.
[50]Koch MA, Dobes C, Kiefer C, Schmickl R,klimes L, Lysak MA. Supernetwork identifies multiple events of plastid trnF(GAA) pseudogene evolutiong in the Braddicaceae[J]. Molecular Biology and Evolution, 2007:24,63-73
[51]Wortley AH, Rudall PJ, Harris DJ, Scotland RW. How much date are needed to resolve a difficult phylogeny? Case study in Lamiales [J]. Sysematic Bioligy,2005:54,697-709.
[52]Bergthorsson U, Richardsom AO, Young GJ, Goertzen LR. Massive horizontal transfer of mitochondrial genes from diverse land plant donors to the basal angionsperm Amborella[J]. Proceedings of the National Academy of Sciences USA; 2004:101,177747-177752.
[53]赵辉,李启寨,李俊.相邻碱基组分与产生SNP的转换或颠换在植物基因组中的研究[J].中国科学C辑生命科学,2006,36(1)：1-8.