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草业学报 ›› 2023, Vol. 32 ›› Issue (9): 181-197.DOI: 10.11686/cyxb2022441

• 研究论文 • 上一篇    

基于SSR分子标记的自然状态下蕨麻采样策略研究

田甜1,2,3(), 李军乔1,2,3(), 马斌4, 王鑫慈1,2,3, 曲俊儒1,2,3   

  1. 1.青海民族大学生态环境与资源学院,青海 西宁 810007
    2.青海省特色经济植物高值化利用重点实验室,青海 西宁 810007
    3.青藏高原蕨麻产业研究院,青海 西宁 810007
    4.青海省湟源县种子站,青海 西宁 811699
  • 收稿日期:2022-11-09 修回日期:2023-01-12 出版日期:2023-09-20 发布日期:2023-07-12
  • 通讯作者: 李军乔
  • 作者简介:E-mail: ljqlily2002@126.com
    田甜(1997-),女,苗族,贵州铜仁人,在读硕士。E-mail: 1138115730@qq.com
  • 基金资助:
    青海省中央引导地方科技发展-科技成果转移转化项目(2023ZY020);青海民族大学2021年研究生创新项目(54M2021005)

Study on sampling strategy of Potentilla anserina in the wild based on SSR molecular markers

Tian TIAN1,2,3(), Jun-qiao LI1,2,3(), Bin MA4, Xin-ci WANG1,2,3, Jun-ru QU1,2,3   

  1. 1.College of Ecology and Environment Qinghai Nationality University,Xining 810007,China
    2.Qinghai Provincial Key Laboratory of High Value Utilization of Characteristic Economic Plants,Xining 810007,China
    3.Tibetan Plateau Juema Industry Research Institute,Xining 810007,China
    4.Qinghai Huangyuan Seed Station,Xining 811699,China
  • Received:2022-11-09 Revised:2023-01-12 Online:2023-09-20 Published:2023-07-12
  • Contact: Jun-qiao LI

摘要:

利用SSR分子标记法,对青海、甘肃、四川和西藏自治区自然状态下的蕨麻进行了采样策略研究,旨在为后期蕨麻采样、育种等研究提供理论依据。试验样品采集设置为距离原点1、2、3、4、5、10、15、20、25、30、40、50、60、80、90、100、120、150、160、200、250、300、400、500、1000、1500、2000、2500、5000、10000、15000、20000、30000、40000、50000 m处分别采样,后使用20对SSR引物对蕨麻6个居群210份样品DNA进行PCR扩增,通过毛细管电泳检测扩增片段大小,使用POPGENE、NTSYS、GenAIEx 6.5等软件进行分析,结果表明:1)遗传变异主要来源于居群内(变异方差分量和占比分比为12.745和84%),居群间变异较低(变异方差分量和占比分比为2.477和16%);2)6个居群间遗传多样性较高,其中祁连县遗传多样性最高(平均杂合度H=0.2797,香农指数I=0.4287),河南县的遗传多样性最低(H=0.2273,I=0.3542),6个居群整体遗传水平较高,表示在这6个居群间遗传变异低,采样时需要扩大范围;3)同一居群内采集蕨麻样品个体时,最短采样距离为5 km,有小型山脉时,跨越山脉即可采样。居群间采样分两种情况:1)具有离地面高度为1000~1500 m山脉时,翻越大型山脉后可直接采集蕨麻样本。2)在平原地区,由于花粉的长距离传播,居群间有基因交流,建议采样距离不小于100 km。

关键词: 蕨麻, SSR分子标记, 毛细管电泳, 采样策略

Abstract:

In this study, a SSR molecular marker method was used to evaluate sampling strategies of Potentilla anserina in Qinghai, Gansu, Sichuan, and the Tibetan Autonomous Region. The research aim was to provide a theoretical basis for future research on P. anserina sampling and breeding. The P. anserina was randomly selected at the sampling site as the first sample, then take samples at 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 40, 50, 60, 80, 90, 100, 120, 150, 160, 200, 250, 300, 400, 500, 1000, 1500, 2000, 2500, 5000, 10000, 15000, 20000, 30000, 40000, 50000 m from the first sample. 20 SSR primer loci were used for PCR amplification of 210 wild individuals from 6 populations, and the amplified fragment length was determined using capillary electrophoresis. SSR data were analyzed using POPGENE, NTSYS and GenAIEx 6.5 software packages. The result showed that: 1) The genetic variation was mainly founded in the population [estat variance (Est.Var)=12.745, 84%], and the variation among populations is low (Est.Var=2.477, 16%). 2) The genetic diversity among the six populations is variable. Qilian County was found to have the highest genetic diversity [average heterozygosityH)=0.2797, Shannon-Weiner index (I)=0.4287], and Henan County had the lowest genetic diversity (H=0.2273, I=0.3542). This indicates that the genetic variation among the six populations is relatively low, so it is suggested to increase the interval during sampling. 3) When collecting individual P. anserina samples in the same population, the shortest sampling distance is 5 km. If there are small mountains, sampling can be carried out across the mountains. Sampling between different populations can be divided into two situations: 1) When there is a larger mountain range with a height of 1000-1500 m, samples of the P. anserina population can be collected on either side of the mountain range to establish population variation. 2) In the plains where there is no large mountain range for isolation, the sampling distance to establish P. anserina population variation is not less than 100 km.

Key words: Potentilla anserine, SSR molecular markers, capillary electrophoresis, sampling strategy