欢迎访问林业科学,今天是

林业科学 ›› 2017, Vol. 53 ›› Issue (6): 37-46.doi: 10.11707/j.1001-7488.20170605

• 论文与研究报告 • 上一篇    下一篇

基于SSR标记的木荷核心种质构建

杨汉波1, 张蕊1, 王帮顺2, 徐肇友2, 周志春1   

  1. 1. 中国林业科学研究院亚热带林业研究所 浙江省林木育种技术研究重点实验室 杭州 311400;
    2. 浙江省龙泉市林业科学研究院 龙泉 323700
  • 收稿日期:2016-06-29 修回日期:2016-08-24 出版日期:2017-06-25 发布日期:2017-07-14
  • 通讯作者: 张蕊
  • 基金资助:
    "十二五"国家科技支撑计划课题(2012BAD01B04);浙江省竹木农业新品种选育重大科技专项竹木育种协作组项目(2012C12908-6);福建省林木种苗科技攻关五期项目木荷课题;江西省林业厅林业科技创新专项项目(201503)

Construction of Core Collection of Schima superba Based on SSR Molecular Markers

Yang Hanbo1, Zhang Rui1, Wang Bangshun2, Xu Zhaoyou2, Zhou Zhichun1   

  1. 1. Zhejiang Provincial Key Laboratory of Tree Breeding Research Institute of Subtropical Forestry Chinese Academy of Forestry Hangzhou 311400;
    2. Longquan Forestry Research Institute, Zhejiang Province Longquan 323700
  • Received:2016-06-29 Revised:2016-08-24 Online:2017-06-25 Published:2017-07-14

摘要: [目的] 通过对比分析与评价以确定木荷核心种质构建的最适取样策略和比例,并构建木荷核心种质;在此基础上,进一步建立核心种质分子身份信息,为木荷种质资源的深入研究和加强利用、发掘优异基因资源提供理论依据和核心材料,同时也可为其他多年生木本植物核心种质的构建提供参考。[方法] 利用13对SSR引物,以来自7个省(市)29个地区的754份木荷种质资源为材料,利用M策略(M)、随机取样法(R)、遗传多样性最大化法(SAGD)和等位基因最大化法(SANA)分别构建核心种质。采用等位基因数(Na)、有效等位基因数(Ne)和Shannon's信息指数(I)等遗传多样性指标进行比较分析来确定最适合的构建方法。[结果] 13对SSR引物共检测到128个等位基因(Na),平均有效等位基因数(Ne)为3.47,Shannon's信息指数(I)为1.39,表明木荷种质资源具有丰富的遗传多样性。SANA、SAGD和M策略构建的核心种质均优于R策略。SANA和SAGD法抽取的核心种质对原有种质均具有较好的代表性,但等位基因保留率较低。M策略构建的核心种质等位基因(Na)保留比例明显高于其他3种策略所构建的核心种质。根据遗传多样性参数综合考虑,同时考虑抽样数量,M策略构建的核心种质能以最小的取样量、最大程度地保留原有种质的遗传多样性,为最优的取样策略。采用主坐标分析法显示,M策略构建的核心种质能够较全面地代表木荷种质资源的遗传多样性,利用该策略得到的115份木荷核心种质,保留了原有种质15.3%的种质材料,等位基因数(Na)、有效等位基因数(Ne)和Shannon's信息指数(I)的保留率分别达到93.8%,115.6%和109.9%。依据13对SSR引物的扩增数据,经过多态性谱带的有序编码转换,构建了115份木荷核心种质的特异分子身份信息,置信概率达到99.99%,具有有效性和唯一性。[结论] M策略是较适宜的构建木荷核心种质的方法,构建的115份核心种质能最大程度代表木荷种质资源的遗传多样性,同时,本研究所采用的方法对其他多年生木本植物核心种质的构建具有重要的参考价值。

关键词: 木荷, SSR标记, 核心种质, 分子身份信息

Abstract: [Objective] The most suitable sampling strategy and proportion for core collection of Schima superba were defined through a comparative analysis and evaluation, and a core collection of S. superba was constructed. Molecular identity of the core collection was further studied, providing theoretical basis and core materials for further research and utilization of S. superba germplasms. It also provides a basis for the core collection construction of other woody plants.[Method] Taking 754 accessions of S. superba as materials that were come from seven provinces of China. The methods of M strategy (maximization strategy, M), random sampling (R), simulated annealing algorithm maximizing the genetic diversity (SAGD) and simulated annealing algorithm maximizing the number of alleles (SANA) were used to construct the core collections using 13 SSR primer pairs. The parameters of genetic diversity, such as number of alleles (Na), effective number of alleles (Ne) and Shannon's information index (I), were used to determine the optimal method.[Result] 128 alleles (Na) were detected in 13 SSR primer pairs, the average of Ne was 3.47. High genetic diversity was revealed in the germplasm resources of S. superba (I=1.39). Comparative analysis showed that the core collection constructed by SANA, SAGD and M strategies were all better than R strategy. The retained of Na in the core collection was low, although the core collection with SANA and SAGD are better representation of the original collection. The retained of Na in the core collection that was constructed by M strategy was apparently higher than other core collections that were constructed by other three strategies. According to the parameters of genetic diversity, and the number of sampling, the core collection constructed by M strategy could preserve the greatest level of genetic diversity of germplasm resources with the minimum sample size, indicating that M strategy was optimal in terms of establishing the most representative core collection, although SANA and SAGD methods all have good representation. The principal coordinate analysis (PCoA) showed that the core collection constructed by M strategy could more comprehensively represent at the genetic diversity level of S. superba. The 115 accessions of S. superba includes 15.3% samples of the germplasms, the retention ratio of Na, Ne and I were 93.8%, 115.6% and 109.9%, and the specific molecular identity for 115 core collections were established with 99.99% probability of confidence. These results demonstrated that the 115 accessions could stand for the initial collection, at the same time a set of unique molecular identity was established for 115 accessions based on 13 SSR loci.[Conclusion] M strategy is a suitable method for constructing S. superba core collection. These results demonstrated that the 115 accessions as core collection could represent the original germplasms, at the same time this research method of the construction of core collection would provide an example for other tree species.

Key words: Schima superba, SSR marker, core collection, molecular identity

中图分类号: