西南牡丹品种起源的ISSR研究

doi:10.3864/j.issn.0578-1752.2015.05.11

摘要/Abstract

摘要： 【目的】通过对西南牡丹21个品种、中原牡丹18个品种、江南牡丹1个品种和1个野生种之间的亲缘关系进行研究，了解西南牡丹品种群的遗传背景，为品种选育奠定基础。【方法】通过改良的CTAB法对41份材料抽提总基因组DNA，采用ISSR分子标记的方法，通过优化的ISSR-PCR反应体系，从60个哥伦比亚大学（UBC）开发的通用引物中筛选27个进行PCR扩增差异比较，利用POPGENE软件计算多态位点百分率（%）；依据Nei’s遗传距离，运用NTSYSPC进行UPGMA聚类构建不同品种群和品种间亲缘关系。【结果】利用27个ISSR引物对41个牡丹样本扩增，共获得的317个条带，其中304个为多态性条带，多态性条带比率为95.41%，平均每个引物扩增条带为11.74；41个样本的遗传相似性系数为0.483—0.811，云南牡丹‘丽江紫5’和‘大关粉4’的遗传相似性系数最大，为0.811；云南牡丹‘大关粉4’和中原牡丹‘彩绘’的遗传相似性系数最小，为0.483；UPGMA聚类结果显示41个样本在阈值为0.625时，聚为四支：第一支由19份样本组成，其中包括5个中原牡丹品种和14个西南牡丹品种，其中，云南牡丹‘丽江紫5’与‘大关粉4’亲缘关系最近，而云南牡丹‘丽江紫5’与中原牡丹‘首案红’的亲缘关系最远，其遗传相似性系数仅为0.609；第二支包括10份样本：有‘乌龙捧盛’、‘豆绿’等5个中原品种和‘四川粉紫’、‘昭通粉’、‘丽江粉1’等5个西南牡丹品种，其中天彭牡丹‘紫金荷2’与‘四川粉紫’的亲缘关系最近，遗传相似性系数最大为0.770；天彭牡丹‘紫金荷2’与中原牡丹‘罗婺现瑞’亲缘关系最远，其遗传相似性系数为0.625；第三支包括11个样本：1个江南品种，8个中原牡丹品种；3个西南牡丹品种。亲缘关系最近的是中原牡丹‘菱花湛露’与‘朱砂垒’，其遗传相似性系数为0.779；亲缘关系最远的是‘朱砂垒’和‘凤丹白’；第四支为黄牡丹单独聚为一支。多数花色相同的供试中原品种表现出近缘关系，红色系的天彭牡丹‘胭脂楼’与紫红色系的中原牡丹品种有着较近的亲缘关系，而供试的紫红色系的天彭牡丹分别紫红色、粉色、紫色和红色的中原品种有近缘关系；云南紫牡丹品种与紫色、紫红色系的中原品种有一定关系，粉牡丹则与紫红、浅红、浅紫红、浅紫色等不同色系的中原牡丹和天彭牡丹都有一定的亲缘关系。天彭牡丹总是先与中原牡丹品种相聚，再与云南牡丹相聚；云南牡丹品种除‘狮山皇冠’、‘香玉板’外，不同产地、株型相似和花色相同的云南牡丹品种间遗传相似性较高，总是先聚为一分支后，才与其他中原品种相聚。【结论】西南牡丹品种栽培起源较复杂，天彭牡丹比云南牡丹与中原牡丹有着较近缘关系，云南牡丹不可能是天彭牡丹直接引种驯化产物，推测云南牡丹品种可能是由几个祖先品种演化的产物，但本地黄牡丹参与起源的可能性较小。

关键词: 西南牡丹品种, 起源, ISSR标记, 亲缘关系

Abstract: 【Objective】 Forty-one tree peony samples consisting of 21 from Xinan group, 18 from Zhongyuan group, 1 from Jiangnan group and 1 wild species were used to detect their genetic diversity and verify their phylogenetic relationship and discuss their genetic backgrounds for the new cultivars breeding. 【Method】Total genomic DNA was extracted from the fresh leaves of tree peony by a modified CTAB method. A total of 27 primers were selected from 60 ISSR universal primers designed by UBC on the basis of the establishment of an optimal ISSR-PCR reaction system. They were used in the PCR amplification to compare the genetic difference among different cultivars. Comparative analysis on DNA fragments amplified by ISSR-PCR technology was made by the relative genetic software. Genetic parameter such as percentage of polymorphic loci (PPB) was calculated by using POPGENE 32. Cluster analysis (UPGMA) and dendrogram based on Nei’s genetic distance were made to construct the relationship between cultivars and cultivars groups by using the Numerical Taxonomy Multivariate Analysis System (NTSYS-pc) ver. 2.1 statistical package.【Result】A total of 317 bands were obtained by amplifying 41 cultivars from 27 primers, among which 304 bands were polymorphic and percentage of polymorphic bands (PPB) attained to 95.41%. In average, 11.74 bands were produced by each primer. The genetic similarity coefficients among all the tested 41 samples ranged from 0.483 to 0.811, which ‘Daguanfen 4’ and ‘Lijiangzi 5’ had the highest similarity coefficient with 0.811, otherwise ‘Daguanfen 4’and ‘Caihui’ had the lowest coefficient with 0.483. The 41 cultivars were divided into four branches based on UPGMA cluster at the coefficiency of 0.625. The first branch was composed of 19 samples, which included 5 Zhongyuan peony cultivars and 14 Xinan cultivars, the genetic similarity coefficient of ‘Lijiangzi 5’ and ‘Daguanfen 4’ had the closest phylogenetic relationship, ‘Lijiangzi 5’ and ‘Shouanhong’ had the farthest phylogenetic relationship with a genetic similarity coefficient of 0.609; The second branch included 10 samples in which there were 5 Zhongyuan cultivars and 9 Xinan cultivars. The closest phylogenetic relationship existed between ‘Zijinhe 2’ and ‘Sichuanfenzi’ with a genetic similarity coefficient of 0.770. However, the farthest phylogenetic relationship was between ‘Zijinhe 2’ and ‘Luowuxianrui’,with the minimum genetic similarity coefficient at 0.625; The third branch contained 11 samples: one Jiangnan cultivar, 7 Zhongyuan cultivars and 3 Xinan cultivars. The closest phylogenetic relationship was between ‘Linghuazhanlu’ and ‘Zhushalei’ with the genetic similarity coefficient of 0.779. The farthest genetic relationship was between ‘Zhushalei’ and ‘Fendangbai’. The fourth branch involved only one wild species of Paeonia lutea. The result showed that most of the Zhongyuan cultivars with similar flower color had the closer relationship. ‘Yanzhilou’ belonged to Tianpeng cultivars with red flower had the much more closer genetic relationship with the Zhongyuan cultivars bloomed in dark red flower, while other Tianpeng cultivars with dark red flower had the closer genetic relationship with Zhongyuan cultivars bloomed in purplish red, pink, purple and red. Yunnan cultivar with purple flower had a certain genetic relationship with Zhongyuan cultivars bloomed in purple and purplish red flower. But other pink flower cultivars had some certain genetic relationships with Zhongyuan cultivars in purplish red, light red, light purplish red and light purple flower color. Tianpeng cultivars firstly clustered with Zhongyuan cultivars and then with Yunnan cultivars. While Yunnan cultivars except ‘Shishanhuanguan’ and ‘Xianyuban’ gathered together firstly and then they clustered with Zhongyuan cultivars, which Yunnan cultivars from different places with similar and same flower-color had the closer genetic relationship. 【Conclusion】The origin of Xinan cultivar group is complicated. Tianpeng cultivars have a closer genetic relationship with Zhongyuan group than Yunnan cultivars. However, Yunnan cultivars could not share the common ancestors with Tianpeng cultivars. Yunnan cultivars might be differentiated by several ancestors under different conditions. More evidences were supposed to make certain their origins. Based on the cluster results, it could be inferred that P. lutea is almost involved in the origin of Yunnan cultivars.

Key words: tree peony from Xinan group, origination, ISSR marker, phylogenetic relationship

李宗艳，秦艳玲，蒙进芳，唐岱，王锦. 西南牡丹品种起源的ISSR研究[J]. 中国农业科学, 2015, 48(5): 931-940.

LI Zong-yan1, QIN Yan-ling1, MENG Jin-fang2, TANG Dai1, WANG Jin1. Study on the Origin of Tree Peony Cultivars from Southwest China Based on ISSR Technology[J]. Scientia Agricultura Sinica, 2015, 48(5): 931-940.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2015.05.11

https://www.chinaagrisci.com/CN/Y2015/V48/I5/931

参考文献

[1] 李嘉珏. 中国牡丹的起源研究. 北京林业大学学报, 1998, 20(2): 22-26.

Li J J. Study on the origin of peonies in China. Journal of Beijing Forestry University-26. (in Chinese), 1998, 20(2): 22

[2] Zhou Z Q, Pan K Y, Hong D Y. Phylogenetic analyses of Paeonia section Moutan (tree peonies, Paeoniaceae) based on morphological data. Acta Phytotaxomica Sinica, 2003, 41(5): 436-446.

[3] 袁涛, 王莲英. 几个牡丹野生种的花粉形态及其演化、分类的探讨. 北京林业大学学报, 1999, 21(1): 17-21.

Yuan T, Wang L Y. Pollen morphology of several tree peony wild species and discussion on its evolution and taxonomy. Journal of Beijing Forestry University, 1999, 21(1): 17-21. (in Chinese)

[4] 于玲, 何丽霞, 李嘉珏, 成仿云. 牡丹野生种间蛋白质谱带的比较研究. 园艺学报, 1998, 25(1): 99-101.

Yu L, He L X, Li J J, Cheng F Y. Comparative studies on protein zones of wild tree peony species. Acta Horticulturae Sinica, 1998, 25(1): 99-101. (in Chinese)

[5] 邹喻苹, 蔡美琳, 王子平. 芍药属牡丹组的系统学研究―基于RAPD分析. 植物分类学报, 1999, 37(3): 220-227.

Zou Y P, Cai M L, Wang Z P. Systematic studies on Paeonia sect. Moutan DC. based on RAPD analysis. Acta Phytotaxonomica Sinica, 1999, 37(3): 220-227. (in Chinese)

[6] Tank D C, Sang T. Phylogenetic utility of the glycerol-3-phosphate acyltransferase gene: evolution and implications in Paeonia (Paeoniaceae). Molecular Phylogenetics and Evolution, 2001, 19(3): 421-429.

[7] 赵宣, 周志钦, 林启冰, 潘开玉, 洪德元. 芍药属牡丹组(Paeonia sect. Moutan)种间关系的分子证据: GPAT基因的PCR-RFLP和序列分析. 植物分类学报, 2004, 42(3): 236-244.

Zhao X, Zhou Z Q, Lin Q B, Pan K Y, Hong D Y. Molecular evidence for the interspecific relationship in Paeonia sect. Moutan: PCR-RFLP and sequence analysis of glycerol-3-phosphate acyltransferase (GPAT) gene. Acta Phytotaxonomica Sinica, 2004, 42(3): 236-244. (in Chinese)

[8] Sang T, Donoghue M J, Zhang D M. Evolution of alcohol dehydrogenase genes in peonies (Paeonia): phylogenetic relationships of putative nonhybrid species. Molecular Biology Evolution, 1997, 14(10): 994-1007.

[9] 林启冰, 周志钦, 赵宣, 潘开玉, 洪德元. 基于Adh 基因家族序列的牡丹组(Sect. Moutan DC.)种间关系. 园艺学报, 2004, 31(5): 627-632.

Lin Q B, Zhou Z Q, Zhao X, Pan K Y, Hong D Y. Interspecific relationships among the wild species of Paeonia Sect. Moutan DC. based on DNA sequences of Adh gene family. Acta Horticulturae Sinica, 2004, 31(5): 627-632. (in Chinese)

[10] Sang T, Crawford D J, Stuessy T F. Chloroplast DNA phylogeny, reticulate evolution, and biogeography of Paeonia (Paeoniaceae). American Journal of Botany, 1997, 84(9): 1120-1136.

[11] Sang T, Crawford D J, Stuessy T F. Documentation of reticulate evolution in peonies (Paeonia) using internal transcribed spacer sequences of nuclear ribosomal DNA: Implications for biogeography and concerted evolution. Proceedings of the National Academy of Sciences of USA, 1995, 92: 6813-6817.

[12] Zhao X, Zhou Zh Q, Lin Q B, Pan K Y, Li M Y. Phylogenetic analysis of Paeonia sect. Moutan (Paeoniaceae) based on multiple DNA fragments and morphological data. Journal of Systematic and Evolution, 2008, 46(4): 563-572.

[13] 洪德元, 潘开玉. 芍药属牡丹组的分类历史和分类处理. 植物分类学报, 1999, 37(4): 351-368.

Hong D Y, Pan K Y. Taxonomical history and revision of Paeonia sect. Moutan (Paeoniaceae). Acta Phytotaxonomica Sinica, 1999, 37(4): 351-368. (in Chinese)

[14] 张益民, 张赞平, 王世端. 芍药属叶片解剖结构的研究. 河南农业大学学报, 1988, 22(1): 117-126.

Zhang Y M, Zhang Z P, Wang S D. Study on leaf anatomy of genus Paeonia. Journal of Henan Agriculture University(in Chinese), 1988, 22(1): 117-126.

[15] 袁涛, 王莲英. 中国栽培牡丹起源的形态分析. 山东林业科技, 2004(6): 1-3.

Yuan T, Wang L Y. Study on the origination of Chinese tree peony cultivars based on morphological analysis. Journal of Shandong Forestry Technology, 2004(6): 1-3. (in Chinese)

[16] 袁涛, 王莲英. 根据花粉形态探讨中国栽培牡丹起源. 北京林业大学学报, 2002, 24(1): 5-11

Yuan T, Wang L Y. Discussion on the origination of Chinese tree peony cultivars according to pollen grain morphology. Journal of Beijing Forestry University, 2002, 24(1): 5-11. (in Chinese)

[17] 于玲, 李嘉珏, 何丽霞. 甘肃紫斑牡丹与中原牡丹类染色体的比较研究. 园艺学报, 1997, 24(1): 79-83.

Yu L, Li J J, He L X. Comparative studies on chromosome in varieties of Paeonia rockii and Paeonia suffruticosa. Acta Horticulturae Sinica, 1997, 24(1): 79-83. (in Chinese)

[18] 孟丽, 郑国生. 部分野生与栽培牡丹种质资源亲缘关系的RAPD研究. 林业科学, 2004, 40(5) : 110-117.

Meng L, Zheng G S. Phylogenetic relationship analysis among Chinese wild species and cultivars of Paeonia sect. Moutan using RAPD marker. Scientia Silvae Sinicae, 2004, 40(5): 110-117. (in Chinese)

[19] Yu H P, Cheng F Y, Zhong Y, Cai C F, Wu J, Cui H L. Development of simple sequence repeat (SSR) markers from Paeonia ostii to study the genetic relationships among tree peonies (Paeoniaceae). Scientia Horticulturae, 2013, 164: 58-64.

[20] Han X Y, Wang L S, Shu Q Y, Liu Z A, Xu S X, Tetsumura T. Molecular characterization of tree peony germplasms using sequence- related amplified polymorphism markers. Biochemical Genetics, 2008,46(3/4): 162-179.

[21] 索志立, 张会金, 张治明, 陈富飞, 陈富慧. 紫斑牡丹与牡丹种间杂交后代的DNA分子证据. 云南植物研究, 2005, 27(1): 42-48.

Suo Z L, Zhang H J, Zhang Z M, Chen F F, Chen F H. DNA molecular evidences of the hybrids between Paeonia rockii and P. suffruticosa based on ISSR marker. Acta Botanica Yunnancia, 2005, 27(1): 42-48. (in Chinese)

[22] Wang J X, Xiao T, Zhang J M, Zhou S L. Isolation and characterization of fourteen microsatellites from a tree peony (Paeonia suffruticosa). Conservation Genetics, 2009, 10(4): 1029-1031.

[23] Homolka A, Berenyi M, Burg K, Kopecky D, Fluch S. Microsatellite markers in the tree peony, Paeonia suffruticosa (Paeoniaceae). American Journal of Botany, 2010, 97(6): e42-e44.

[24] Gao Z M, Wu J, Liu Z G, Wang L Sh, Ren H X, Shu Q Y. Rapid microsatellite development for tree peony and its implications. BMC Genomics, 2013, 14(1): 886-896.

[25] Hou X G, Guo, D L, Wang J. Development and characterization of EST-SSR markers in Paeonia suffruticosa (Paeoniaceae). American Journal of Botany, 2011, 98(11): E303-E305.

[26] Hao Q,Liu Z A, Shu Q Y, Zhang R, De Rick J, Wang L S. Studies on paeonia cultivars and hybrids identification based on SRAP analysis. Hereditas (Lund), 2008, 145(1): 38-47.

[27] 陈向明, 郑国生, 张圣旺. 牡丹栽培品种的RAPD分析. 园艺学报, 2001, 28(4): 370-372.

Chen X M, Zheng G S, Zhang S W. RAPD analysis of tree peony cultivars. Acta Horticulturae Sinica. 2001, 28(4): 370-372. (in Chinese)

[28] 王娟, 郭大龙, 侯小改, 范丙友, 刘改秀, 李友军. 不同花型牡丹品种亲缘关系的SRAP分析. 中国农学通报, 2011, 27(28): 167-171.

Wang J, Guo D L, Hou X G, Fan B Y, Liu G X, Li Y J. SRAP analysis of genetic relationships of the different tree peony flower forms. Chinese Agricultural Science Bulletin, 2011, 27(28): 167-171. (in Chinese)

[29] 史倩倩, 王雁, 周琳, 黄国伟. 十大花色中原牡丹传统品种核型分化程度. 林业科学研究, 2012, 25(4): 470-476.

Shi Q Q, Wang Y, Zhou L, Huang G W. Karyotypes and chromosomal differentiation in cultivars of tree peony (Paeonia suffruticosa). Forest Research, 2012, 25(4): 470-476. (in Chinese)

[30] 史倩倩, 王雁, 周琳, 黄国伟. 中原牡丹传统品种的核型及进化关系. 东北林业大学学报, 2012, 40(11): 23-29.

SHI Q Q, Wang Y, Zhou L, Huang G W. Karyotypes of twenty one traditional cuitivars of Paeonia suffruticosa and their evolutional relationship. Journal of Northeast Forestry University, 2012, 40(11): 23-29. (in Chinese)

[31] Suo Z L, Zhang C H, Zheng Y Q, He L X , Jin X B, Hou B X, Li J J. Revealing genetic diversity of tree peonies at micro-evolution level with hyper-variable chloroplast markers and floral traits. Plant Cell Reports, 2012, 31(12): 2199-2213.

[32] 李莹莹, 郑成淑. 利用CDDP标记的菏泽牡丹资源的遗传多样性. 中国农业科学, 2013, 46(13): 2739-2750.

Li Y Y, Zheng C S. Genetic diversity of tree peony cultivars resources in Heze revealed by CDDP markers. Scientia Agricuture Sinica, 2013, 46(13): 2739-2750. (in Chinese)

[33] 陈向明, 郑国生, 孟丽. 不同花色牡丹品种亲缘关系的RAPD-PCR分析. 中国农业科学, 2002, 35(5): 546-551.

Chen X M, Zheng G S, Meng L. RAPD-PCR analysis of genetic diversity of different color varieties 35 tree paeonia cultivars. Scientia Agricultura Sinica, 2002, 35(5): 546-551. (in Chinese)

[34] 李嘉珏. 中国牡丹与芍药. 北京: 中国林业出版社, 1999: 60.

Li J J. Chinese Tree Peony and Paeonia lactiflora. Beijing: China Forestry Press, 1999: 60. (in Chinese)

[35] 李嘉珏. 中国牡丹品种图志（西北·西南·江南卷）. 北京: 中国林业出版社, 2006: 63-197.

Li J J. Pictorical Record of Chinese Tree Peony Varieties (Xibei, Xinan, Jiangnan Volume). Beijing: China Forestry Press, 2006: 63-197. (in Chinese)

[36] 李宗艳, 郭盘江, 唐岱, 张海燕. 丽江牡丹不同品种的生物生态学特性及耐水淹胁近能力. 东北林业大学学报, 2006, 34(5): 44-46.

Li Z Y, Guo P J, Tang D. Zhang H Y. Biological characteristics and moisture-stress tolerance of Paeonia suffruticosa in Lijiang, Yunnan province. Journal of Northeast Forestry University, 2006, 34(5): 44-46. (in Chinese)

[37] 索志立. 牡丹品种鉴定用ISSR引物的筛选与开发. 生物技术通报, 2006(增): 342-346.

Suo Z L. Screening and development of ISSR primers for identification of tree peony cultivars in Paeonia suffruticosa Andrews. Biotechnology Bulletin, 2006(suppl.): 342-346. (in Chinese)

[38] 石颜通, 周波, 张秀新, 江海东, 薛祺, 王顺利. 牡丹89个不同种源品种遗传多样性和亲缘关系分析. 园艺学报, 2012, 39(12): 2499-2506.

Shi Y T, Zhou B, Zhang X X, Jiang H D, Xue Q, Wang S L. Assessment of genetic diversity and relationship of 89 tree peony cultivars from different provenances. Acta Horticulturae Sinica, 2012, 39(12): 2499-2506. (in Chinese)

[39] 侯小改, 尹伟伦, 李嘉珏, 王华芳. 部分牡丹品种遗传多样性的AFLP分析. 中国农业科学, 2006, 39(8): 1709-1715.

Hou X G, Yin W L, Li J J, Wang H F. AFLP analysis of genetic diversity of 30 tree peony (Paeonia suffruticosa Andr.) cultivars. Scientia Agricultura Sinica, 2006, 39(8): 1709-1715. (in Chinese)

[40] Wang X W, Fan H M, Li Y Y, Sun X, Sun X Z, Wang W L, Zheng C S. Analysis of genetic relationships in tree peony of different colors using conserved DNA-derived polymorphism markers. Scientia Horticulturae, 2014, 175: 68-73.