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大别山不同龄级映山红种群遗传多样性的SSR分析

Genetic Diversity of Rhododendron simsii Populations on Dabieshan at Different Life Stages Based on SSR Markers

  • 摘要:
    目的 利用SSR标记比较大别山黄狮寨不同年龄级映山红(Rhododendron simsii Planch.)种群的遗传多样性及遗传结构,探究映山红不同世代间遗传多样性的变化规律,为大别山野生映山红资源的合理利用和高效保护提供科学依据。
    方法 按照基径大小和丛枝多少将大别山黄狮寨典型映山红种群划分为老树、成树、小树、幼苗4个年龄级,筛选出12对多态性强的SSR引物用于PCR扩增,扩增产物经6%变性聚丙烯酰胺凝胶电泳检测并银染。构建"0/1"矩阵,利用POPGENE 32.0软件分析种群遗传多样性。基于Nei's遗传距离,采用软件MEGA5.0进行UPGMA聚类。
    结果 不同龄级的映山红遗传多样性差别较大,幼苗和老树种群多样性最差,小树种群多样性最丰富。12个微卫星标记观测等位基因数为3~9个,平均5.08;有效等位基因数为2.254 9~6.129 7,平均3.460 5;观察杂合度HO和期望杂合度HE分别为0.676 4~0.881 2和0.607 7~0.690 7。Shannon信息指数(I)以小树群体最高,成树次之,幼苗最低。近交系数Fis为-0.638 3~0.174 4,平均为-0.294 6;总近交系数Fit为-0.615 1~0.270 6,平均为-0.162 1,表明各龄级种群内主要繁殖方式为杂交。分子方差分析(AMOVA)表明89.76%的遗传变异存在于年龄级内,仅10.24%存在于年龄级间。基因流水平高,仅1个位点Nm < 1。遗传一致度最高的为小树和成树种群。
    结论 大别山黄狮寨映山红种群遗传多样性丰富,种群间中度分化,遗传变异主要存在于年龄级内。

     

    Abstract:
    Objective Microsatellite markers were used to compare the genetic diversity and genetic structure of Rhododendron simsii populations at four different life stages sampled from Huangshizhai of Dabieshan region to clarify the mechanism of genetic diversity among different age groups.
    Method  The populations of R. simsii were divided into four age groups (old trees, adult, juvenile, and seedlings) according to their sizes of basal diameter and numbers of branches. Twelve SSR markers were used for PCR amplification, and the products were subjected to 6% polyacrylamide gel electrophoresis, which were further visualized through silver staining. The "0/1" matrix was constructed and POPGENE 32.0 software was used to analyze the genetic diversity. Based on Nei's genetic distance, UPGMA dendrogram was constructed with MEGA 5.0 software.
    Result  Different genetic diversities were observed in the four R. simsii populations, the lowest genetic diversity was observed in seedlings population, followed by old trees population, while the highest genetic diversity was observed in juvenile population. The observed and effective number of alleles per locus ranged from 3 to 9 and 2.254 9-6.129 7, with the average of 5.08 and 3.460 5, respectively. Moreover, HO and HE were in the ranges of 0.676 4-0.881 2 and 0.607 7-0.690 7, respectively. The Shannon's information index (I) was the highest in juvenile population, followed by adult population, and lowest in seedling population. Fis and Fit ranged within-0.638 3-0.174 4 and-0.615 1-0.270 6, with the mean of-0.294 6 and-0.162 1, indicating the outcross took place frequently. In particular, the AMOVA analysis showed that 89.76% of genetic variation was maintained within age groups, while only 10.24% presented among age groups. Frequent gene flow was observed, as only one locus showed Nm less than 1.0.The genetic identify between adult and juvenile groups was the highest.
    Conclusion High genetic diversity existed among R. simsii populations located on Huangshizhai. Moderate differentiation was observed, which was maintained between age groups.

     

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