基于SSR荧光标记的白蜡核心种质构建  

Construction of core collection of Fraxinus based on SSR molecular markers

作　　者：燕丽萍[1,2] 吴德军 毛秀红[1,2] 姚俊修 任飞[1] 李善文 王开芳[1] 王因花 刘翠兰[1,2] YAN Liping;WU Dejun;MAO Xiuhong;YAO Junxiu;REN Fei;LI Shanwen;WANG Kaifang;WANG Yinhua;LIU Cuilan(Shandong Provincial Academy of Forestry, Jinan 250014, Shandong, China;Shandong Provincial Key Laboratory of Forest Tree Genetic Improvement, Jinan 250014, Shandong, China)

机构地区：[1]山东省林业科学研究院,山东济南250014 [2]山东省林木遗传改良重点实验室,山东济南250014

出　　处：《中南林业科技大学学报》2019年第7期1-9,共9页Journal of Central South University of Forestry & Technology

基　　金：国家“十二五”科技支撑计划专题:“白蜡种质资源发掘与创新利用”(2013BAD01B06-6);山东省科技发展计划项目:“白蜡种质资源分子评价与体胚发育技术研究”(2014GNC111006)

摘　　要：白蜡为中国北方重要的盐碱地造林树种,研究其遗传多样性,进一步构建核心种质,对白蜡优异种质的筛选及开发利用具有重要作用,为广泛开展白蜡种质资源的保存评价、挖掘优异基因资源提供核心材料,同时也为白蜡育种提供优良基因资源。本研究运用SSR标记的方法,选取田间表型性状差异较大的4个白蜡品种华雄、鲁蜡5号、金叶白蜡和金枝白蜡,采用聚丙烯酰胺凝胶电泳从500对SSR引物中筛选出扩增条带清晰、多态性好、扩增稳定的42对引物。从筛选出的每对引物5′端添加荧光标记后,采用毛细管法通过DNA分析仪检测202个样品不同等位变异的扩增片段,从42对SSR引物中筛选出17对扩增稳定的引物,建立基于高通量荧光SSR标记的白蜡种质鉴定体系。研究了来自全国22个地区的具有代表性的202份白蜡种质资源的遗传多样性,通过R语言包Genetic Subsetter构建出了40份白蜡核心种质。结果表明:选用的17对引物都表现出多态性,共检测出142个等位变异,平均每对引物等位基因数为8.353个;平均有效等位基因数为3.342;平均多态性信息含量为0.635。可见白蜡属植物种间变异较大,具有丰富的遗传多样性。构建了40份白蜡核心种质,占原始种质的20%,t检测表明,所构建的核心种质遗传多样性丰富,其代表的遗传多样性指标与原始种质资源差异不显著,说明获得的核心种质资源能够充分、最大程度地代表原始种质。研究结果为白蜡种质资源的收集、保护、评价及利用提供了宝贵的理论依据和物质基础。利用17对高效引物构建了白蜡属植物SSR高通量鉴定体系,构建的40份白蜡核心种质能够最大程度的代表其遗传多样性。Fraxinus are important tree species in saline alkali land in northern China. It has an important effect to study its genetic diversity and further constructs core germplasm in screening and exploitation Fraxinus germplasm. It provides core materials for the deep study and breeding of Fraxinus. In this study, four different Fraxinus varieties, such as Huaxiong, Lula 5, golden leaf and golden branch, were selected. 500 pairs of SSR primers were used and 42 pairs of, which showed clear strip and good polymorphism stably, were selected by polyacrylamide gel electrophoresis. The selected primerswere then labeled with fluorescent markers at the 5′ end and were applied in capillary electrophoresis to detect the allelic variation in 202 Fraxinus samples. 17 pairs of stable fluorescent primers were used to establish the identification system of Fraxinus germplasm based on high throughput fluorescent SSR markers. The genetic diversity of 202 representative germplasm resources from 22 regions was studied. 40 core germplasms of Fraxinus were constructed by R language package Genetic Subsetter. It was showed that the 17 pairs of primers drew polymorphism, and 142 allelic variations were detected, on average 8.353 variation, in which 3.342 were valid,of each pair of primers. The average polymorphism information content (PIC) was 0.635. Thus, the varieties of Fraxinus have great variation and abundant genetic diversity. 40 core germplasm of Fraxinus were collected and 20% of the samples were the original germplasm. It was showed by the t-test that there was no significant difference between the genetic diversity of the core germplasm and the original germplasm resources, indicating that the obtained core germplasm resources could fully and maximally represent the original germplasm. These results provide valuable theoretical basis and germplasm materials for the protection, screening and utilization of Fraxinus germplasm resources. 17 pairs of stable fluorescent primers were used to establish the identification system of Frax

关 键 词：白蜡属 种质资源 SSR标记 遗传多样性 核心种质 

分 类 号：S718.46[农业科学—林学]