麦冬Ophiopogon japonicus (L.f.)野生居群细胞学与遗传多样性研究
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摘要
麦冬(Ophiopogon japonicus (L.f.) Ker-Gawl)属于百合科沿阶草属,是多年生草本植物,作为沿阶草属的模式种,也是该属内分布最为广泛的种。前人研究表明重庆金佛山地区麦冬存在丰富的染色体数目变异。本研究对该处投影面积约为0.5平方公里,海拔高度在750-900米的区域内分布的麦冬进行穷尽式的采样。通过对金佛山生境的调查,麦冬染色体水平的探究、基因组水平的审视分析麦冬野生居群内变异和同域物种形成的可能性。实验结果以及论证如下：
(1)细胞学研究表明麦冬的体细胞染色体数目存在丰富变异,出现X=17和X=18两类基数变异,以及二倍体、四倍体、五倍体、六倍体的倍性变异。
(2)原位杂交结果显示,四倍体麦冬(2n=68)具有两对杂交信号。
(3)通过近缘属铃兰的引物及自行设计的EST-SSR引物扩增表明居群内不同基数的个体间存在变异。
染色体基数的变异可以使得生殖隔离得到快速建立,从而使X=17和X=18分别形成新的物种。在金佛山地区发生的这一染色体重组带来的瞬间成种模式,还为同域物种形成提供了新例证。最后,我们对麦冬野生群体遗传多样性的研究,为我们保护麦冬的遗传资源,制定相应的保护策略提供了重要的指导。
As a perennial herb, Ophiopogon japonicus belongs to Ophiopogon in liliaceae family. It is a model and most widely distributed species of Ophiopogon. Previous researches indicated that O. japonicus from Mt. Jinfoshan in Chongqin had profuse variation in chromosome number. Hence, we exhaustively sampled in Mt. Jinfoshan, the projection area of which is 0.5 square meter and altitude is 700-900 meter. Through investigating the habitat niche in Mt. Jinfoshan, studying O.japonicus both in the choromosome level and genomic level, this research showed that:
1, Abundant variation of somatic chromosome number existed in O. japonicus. Chromosome counting exposes two basic chromosome numbers(x=17&18 respectively) and ploidy variation like diploid, tetraploid, pentaploid, hexaploid frequently happended in this population
2, The result of FISH(Fluorescent in situ hybridization) showed that tetraploid has 2 pairs of signals.
3, The genomic data from amplification by primers based on convallariae that is a closely related genera of O. japonicus and self-designed primers according to EST-SSR revealed the distinct divergence between these individuals with different chromosome basic number.
Variation in chromosome basic number resulted in the reproductive isolation and contributes the species of x=17 and x=18 divergent into two species. Here we provide a new support of sympatric speciation in a case study of instantaneous speciation model in sibling species of O. japonicus. At last, protections for plant germplasm resources and genetic diversities are proposed basing on our research.
(1)细胞学研究表明麦冬的体细胞染色体数目存在丰富变异,出现X=17和X=18两类基数变异,以及二倍体、四倍体、五倍体、六倍体的倍性变异。
(2)原位杂交结果显示,四倍体麦冬(2n=68)具有两对杂交信号。
(3)通过近缘属铃兰的引物及自行设计的EST-SSR引物扩增表明居群内不同基数的个体间存在变异。
染色体基数的变异可以使得生殖隔离得到快速建立,从而使X=17和X=18分别形成新的物种。在金佛山地区发生的这一染色体重组带来的瞬间成种模式,还为同域物种形成提供了新例证。最后,我们对麦冬野生群体遗传多样性的研究,为我们保护麦冬的遗传资源,制定相应的保护策略提供了重要的指导。
As a perennial herb, Ophiopogon japonicus belongs to Ophiopogon in liliaceae family. It is a model and most widely distributed species of Ophiopogon. Previous researches indicated that O. japonicus from Mt. Jinfoshan in Chongqin had profuse variation in chromosome number. Hence, we exhaustively sampled in Mt. Jinfoshan, the projection area of which is 0.5 square meter and altitude is 700-900 meter. Through investigating the habitat niche in Mt. Jinfoshan, studying O.japonicus both in the choromosome level and genomic level, this research showed that:
1, Abundant variation of somatic chromosome number existed in O. japonicus. Chromosome counting exposes two basic chromosome numbers(x=17&18 respectively) and ploidy variation like diploid, tetraploid, pentaploid, hexaploid frequently happended in this population
2, The result of FISH(Fluorescent in situ hybridization) showed that tetraploid has 2 pairs of signals.
3, The genomic data from amplification by primers based on convallariae that is a closely related genera of O. japonicus and self-designed primers according to EST-SSR revealed the distinct divergence between these individuals with different chromosome basic number.
Variation in chromosome basic number resulted in the reproductive isolation and contributes the species of x=17 and x=18 divergent into two species. Here we provide a new support of sympatric speciation in a case study of instantaneous speciation model in sibling species of O. japonicus. At last, protections for plant germplasm resources and genetic diversities are proposed basing on our research.
引文
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[12]Tetsuo Denda K N, Masatsugu Yokota.Karyotype of Ophiopogon reversus (Convallariaceae) from Taiwan
and the Southern Ryukyus[J].Taiwania,2006,51(2):117-122.
[13]Yamashita T.Karyotype Analysis of Six Species of the Genus Ophiopogon (Convallariaceae-Ophiopogoneae)[J].Journal of Japanese Botany,2001,76(2):100.
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[30]陈良兵,朱美霞,胡炳义.SSR在琼海和三亚两普通野生稻居群中的遗传变异[J].植纺学通 报，2006,23(2).
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[2]杨永平,李恒.沿阶草属的分类系统.云南植物研究,1990,12(S3).
[3]张大明.沿阶草族的染色体研究及系统学问题的探讨.中国科学院植物研究所博士学位论文,1998.
[4]Anonymous,四川植物志.四川人民出版社,第七卷.
[5]刘榴,张白嘉.麦冬及有效部位药理作用研究进展[J].四川中医，2010,28(12).
[6]郭海林,刘建秀,杭悦宇.麦冬研究进展[J].中国野生植物资源，2003,22(3).
[7]Sen S.Cytotaxonomy of Liliales[J].Feddes Repertorium,1975,86(5):255-305.
[8]Cutler D F.Vegetative anatomy of Ophiopogoneae (Convallariaceae)[J].Botanical Journal of the Linnean Society,1992,110(4):385-419.
[9]戴伦凯.沿阶草亚科植物的叶表皮特征及其在分类学上的意义[J].植物分类学报，1991,29(4)：335-346.
[10]刘峰群,贺承山.三种麦冬的显微图像分析与鉴定[J].解放军药学学报，2000,16(4)：221-223.
[11]Zhou Q,Zhou J,Chen J,Wang X.Karyotype analysis of medicinal plant <i>Liriope spicata</i> var.<i>prolifera</i> (Liliaceae)[J].Biologia,2009,64(4):680-683.
[12]Tetsuo Denda K N, Masatsugu Yokota.Karyotype of Ophiopogon reversus (Convallariaceae) from Taiwan
and the Southern Ryukyus[J].Taiwania,2006,51(2):117-122.
[13]Yamashita T.Karyotype Analysis of Six Species of the Genus Ophiopogon (Convallariaceae-Ophiopogoneae)[J].Journal of Japanese Botany,2001,76(2):100.
[14]杨永平,李恒,刘宪章,近藤胜彦.云南沿阶草属植物的核型研究[J].云南植物研究,1990,12(S3).
[15]梁国鲁,杨美全.川麦冬核型分析[J].西南农业大学学报,1998,20(4)：307-310.
[16]葛传吉,李岩坤,周月,徐炳声.山东地区药用拉物染色体数目观察(V)[J].山东中医学院学报,1988,12(33)：55-57.
[17]丁卓娅,同域物种形成—来自于麦冬(Ophiopogon japonicus)染色体异基数研究的例子,中国科学院研究生院硕士学位论文,2008.
[18]杨美全,李品明,曾维群,张兴翠,周斌,李晓林.川麦冬品种同工酶分析研究[J].中国中药杂志，1998,23(5).
[19]徐柯,郑鸣,曹毅,蒋彦,乔代蓉.四川麦冬自然居群间RAPD分析[J].中草药，2002,33(7)：648-652.
[20]黄玉吉,陈菁瑛,苏海兰,黄颖桢,万学锋.麦冬遗传多样性的RAPD分析及分子指纹图谱构建[J].中国农学通报,2009,25(23).
[21]李懋学,张教方.植物染色体研究技术[J].东北林业大学出版社,1991.
[22]John H A,Birnstie.M1,Jones K W.RNA-DNA HYBRIDS AT CYTOLOGICAL LEVEL[J]. (Translated from English)Nature,1969,223(5206):582-& (in English).
[23]Jiang J M,Gill B S.Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research[J].Genome,2006,49(9):1057-1068.
[24]向素琼汪卫星,梁国鲁.45SrDNA在不同倍性沙田柚染色体上的荧光原位杂交分析.[J].园艺学报，2007,34(1)：75-80.
[25]Valarik M,Bartos J,Kovarova P,Kubalakova M,de Jong J H,Dolezel J.High-resolution FISH on super-stretched flow-sorted plant chromosomes[J]. (Translated from English)Plant Journal,2004,37(6):940-950 (in English).
[26]Koornneef M,Fransz P,de Jong H.Cytogenetic tools for Arabidopsis thaliana[J]. (Translated from English)Chromosome Research,2003,11(3):183-194 (in English).
[27]易能君,王明麻,施季森.林木群体遗传分化的多元分析[J].南京林业大学学报，1995,19(1).
[28]Yang J-F,Zhao L-L,Shao C-W,Ji X-S,Liao X-L,Chen S-L.Isolation and characterization of polymorphic microsatellite loci from black snapper Sebastodes fuscescens (Houttuyn)[J].Conservation Genetics,2009,10(5):1541-1543.
[29]Chen F,Chen D,Valles M-P,Gao Z,Chen X. Analysis of Diversity in Chinese Cultivated Barley with Simple Sequence Repeats:Differences Between Eco-Geographic Populations[J].Biochemical Genetics,2010,48(1-2):44-56.
[30]陈良兵,朱美霞,胡炳义.SSR在琼海和三亚两普通野生稻居群中的遗传变异[J].植纺学通 报，2006,23(2).
[31]Chen S-L,Xing S-C,Xu G-B,Liao X,Yang J-F.Isolation and characterization of 10 polymorphic microsatellite loci from small yellow croaker (Pseudosciaena polyactis)[J].Conservation Genetics,2008,10(5):1469-1471.
[32]Lu Z,Wang Y,Zhang X,Korpelainen H,Li C.Genetic variation of isolated Picea balfouriana populations from the southeast of the Qinghai-Tibet Plateau[J].Annals of Forest Science,2009,66(6):607.
[331 Peleg Z,Fahima T,Abbo S,Krugman T,Saranga Y.Genetic structure of wild emmer wheat populations as reflected by transcribed versus anonymous SSR markers[J].Genome/National Research Council Canada=Genome/Conseil National De Recherches Canada,2008,51(3):187-195.
[34]Raji A A J,Fawole I,Gedil M,Dixon A G O.Genetic differentiation analysis of African cassava (Manihot esculenta) landraces and elite germplasm using amplified fragment length polymorphism and simple sequence repeat markers[J].Annals of Applied Biology,2009,155(2):187-199.
[35]Wang X,Guo X,Zhang Y,Meng X,Qiu X,Liu S,Zhang T.Development of polymorphic EST-derived SSR markers for the shrimp, Fenneropenaeus chinensis[J].Conservation Genetics,2008,10(5):1455-1457.
[36]Sharifi Tehrani M,Mardi M,Sahebi J,Catalan P,Diaz-Perez A.Genetic diversity and structure among Iranian tall fescue populations based on genomic-SSR and EST-SSR marker analysis[J].Plant Systematics and Evolution,2009,282(1-2):57-70.
[37]Kanagarajan S,Muthusamy S,Chen E C-F,Tsay H-S.Development and characterization of microsatellite markers in the fungus Antrodia cinnamomea from dbEST database[J].Conservation Genetice,2008,10(5):1487-1490.
[38]Dong P,Wei Y-M,Chen G-Y,Li W,Wang J-R,Nevo E,Zheng Y-L.EST-SSR diversity correlated with ecological and genetic factors of wild emmer wheat in Israel[J].Hereditas,2009,146(1):1-10.
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