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园艺学报 ›› 2018, Vol. 45 ›› Issue (1): 126-138.doi: 10.16420/j.issn.0513-353x.2017-0317

• 研究报告 • 上一篇    下一篇

基于ITS序列的中国樱桃、欧洲甜樱桃和毛樱桃种内遗传多样性及种间关系分析

王 浩?1,黄智林1,陈 涛2,张 静2,王 燕2,陈 清1,汤浩茹1,2,王小蓉1,2,*   

  1. 1四川农业大学园艺学院,成都 611130;2四川农业大学果蔬研究所,成都 611130
  • 出版日期:2018-01-25 发布日期:2018-01-25

Genetic Diversity and Relationship Analysis Among Cerasus pseudocerasus,C. avium,and C. tomentosa Based on Internal Transcribed Spacer(ITS)Sequences

WANG Hao1,HUANG Zhilin1,CHEN Tao2,ZHANG Jing2,WANG Yan2,CHEN Qing1,TANG Haoru1,2,and WANG Xiaorong1,2,*   

  1. 1College of Horticulture,Sichuan Agricultural University,Chengdu 611130,China;2Institute of Pomology and Olericulture,Sichuan Agricultural University,Chengdu 611130,China
  • Online:2018-01-25 Published:2018-01-25

摘要:

对中国樱桃[Cerasus pseudocerasus(Lindl.)G. Don](地方种质35份,野生资源35份),欧洲甜樱桃[C. avium(L.)Moench](18份)和毛樱桃[C. tomentosa(Thunb.)Wall.](7份)共95份材料的核糖体内转录间隔区(ITS,internal transcribed spacer)序列进行了测定和分析,以期从ITS的DNA序列变异角度揭示种内遗传多样性及种间遗传关系。结果表明:(1)96条ITS序列比对后长度为712 bp,G + C含量为58.1%,检测到71个变异位点(9.97%),共定义了37个单倍型;中国樱桃、欧洲甜樱桃和毛樱桃的单倍型多样性和核酸多样性分别为0.840和0.00466、0.928和0.00396、0.905和0.00564,中国樱桃中栽培种质遗传多样性明显低于其野生资源;(2)种间遗传关系显示,中国樱桃与欧洲甜樱桃之间的遗传距离较近(0.019),而与毛樱桃遗传距离较远(0.048);(3)邻接聚类和单倍型网络分析显示3个种分别聚为3个分支,种间具有较大的遗传分化。同时,选择3个种的代表单倍型进行其ITS序列的二级结构预测分析,3个种之间ITS1区、ITS2区的二级结构差异较大,最小自由能差异显著,进一步揭示了3个种间的遗传关系较远。综合分析认为,3个樱桃种内均具有较高的遗传多样性,种间具有较大的遗传分化,遗传关系较远,其中毛樱桃与中国樱桃和欧洲甜樱桃的遗传关系均较远。

关键词: 中国樱桃, 欧洲甜樱桃, 毛樱桃, ITS序列, 单倍型, 遗传多样性, 种间关系

Abstract:

In this study,we sampled a total of 95 samples,including 35 landraces and 35 wild individuals of Chinese cherry from 6 populations across 35 counties of 8 provinces in China,18 cultivars of European sweet cherry from 10 countries in the world,as well as 7 semi-wild Nanking cherry resources from 7 counties of 5 provinces in China. Based on internal transcribed spacer(ITS)sequences,we tried to study the genetic diversity within species and interspecific relationship among them. The final ITS dataset contained 712 aligned nucleotides,with 58.1% of G + C content,of which 71(9.97%)were variable sites. The ITS sequences defined 37 haplotypes. The haplotype diversity(Hd)and nucleic acid diversity(π)of C. pseudocerasus,C. avium,and C. tomentosa were(0.840,0.00466),(0.928,0.00396)and(0.905,0.00564),respectively. Within C. pseudocerasus,the genetic diversity of germplasm(Hd = 0.726,π = 0.00224)was significantly lower than that of wild resource(Hd = 0.914,π = 0.00681). Interspecific genetic relationship analysis showed close relationship(0.019)between Chinese cherry and sweet cherry,while they revealed further relationship with Nanking cherry. Both Neighbor-Joining reconstruction and haplotype network showed three distinct branches with obvious genetic differentiation among the three species. In addition,there were significant differences in the secondary structure of the ITS1 and ITS2 regions and t-detection of the minimum free energy. Based on above results,there were high genetic diversity and obvious genetic differentiation among three cherry cultivated species. Close relationship was detected between Chinese cherry and European sweet cherry,while far distance between Nanking cherry with them.

Key words: Cerasus pseudocerasus, C. avium, C. tomentosa, ITS sequence, haplotype, genetic diversity, interspecific relationship