薄壳山核桃品种亲缘关系分析与指纹图谱构建  

Genetic Relationship Analysis and Fingerprint Construction of Carya illinoensis Varieties

作　　者：何旭东[1] 郑纪伟[1] 田雪瑶 教忠意[1] 窦全琴[1] HE Xu-dong;ZHENG Ji-wei;TIAN Xue-yao;JIAO Zhong-yi;DOU Quan-qin(Jiangsu Academy of Forestry,Nanjing 211153,Jiangsu,China)

机构地区：[1]江苏省林业科学研究院,江苏南京211153

出　　处：《林业科学研究》2021年第4期95-102,共8页Forest Research

基　　金：江苏省科技支撑项目(BE2019397)。

摘　　要：[目的]基于荧光SSR标记结合高通量毛细管电泳技术,建立一种快速、高效、稳定、准确的薄壳山核桃基因分型体系,用以分析各品种间亲缘关系并构建指纹图谱,旨在为薄壳山核桃品种鉴定与新品种保护提供理论依据,也为薄壳山核桃种质资源评价、品种选育与推广提供有益参考。[方法]选取薄壳山核桃及其近缘种54对SSR引物进行初筛,最终确定10对标记合成荧光引物用于后续分析。基于毛细管电泳技术对25个薄壳山核桃品种进行基因分型,利用软件统计位点数据并计算各个位点的等位基因数(A)和多态信息含量(PIC)。利用SSR标记间的相互组合构建薄壳山核桃不同品种的指纹图谱。通过等位片段的转化对薄壳山核桃品种进行聚类,并分析其亲缘关系。[结果]10对荧光SSR标记共扩增出68条等位片段,平均为6.8个;位点Cc19最多,有12个等位基因,位点BFU-Jr19最少,有3个等位基因。多态信息含量(PIC)变化范围为0.2910~0.8435(平均为0.5883)。优选的4对核心引物Cc19、PM-GA31、PM-CIN4和PM-GA41组合能完全区分25个薄壳山核桃品种。25个品种遗传相似系数在0.62~0.99之间,并聚类成2个大的类群,部分亲缘关系较近的品种聚类在一起,部分品种聚类不能与遗传背景完全对应。[结论]与传统的显性标记及聚丙烯酰胺凝胶电泳分型技术相比,荧光SSR标记结合毛细管电泳技术构建薄壳山核桃品种指纹图谱切实可行,通量大且速度快,结果稳定可靠,通过标记组合可有效的对不同品种进行区分。在品种亲缘关系分析中,建议增加杂交亲本数量,并在全基因组范围内选取一定数量效率更高的标记,以便最大程度地揭示薄壳山核桃品种间亲缘关系的真实水平。[Objective]Since the morphological appearances among pecan(Carya illinoensis)varieties are extremely similar due to the narrow genetic basis of the crossing parents,making it difficult to precisely identify by phenotypic characters only,this study aims at elucidating the genetic relationships and constructing fingerprint among pecan varieties,and establishing a rapid,efficient,stable,and accurate genotyping system for pecan based on fluorescent SSR markers combining high throughput capillary electrophoresis technology.[Method]A total of 54 SSR primers were selected from pecan and related species for preliminary screening and ten of them were labeled by fluorophore for further analysis.25 pecan varieties were genotyped by capillary electrophoresis,and the number of allele(A)and polymorphic information content(PIC)of each locus was scored and calculated by software.The fingerprint of pecan varieties was constructed using different combination of SSR markers and the genetic relationships among varieties were also investigated through cluster analysis.[Result]A total of 68 alleles were detected by ten pairs of SSR markers with an average of 6.8 alleles.The most(12 alleles)were screened at the Cc19 locus and the least(3 alleles)were obtained at the BFU-Jr locus.The polymorphic information content varied with locus from 0.2910 to 0.8435(mean 0.5883).With the four optimal pairs of core primers,all of the 25 pecan varieties could be completely distinguished by the primer combinations of Cc19,PM-GA31,PM-CIN4,and PM-GA41.The cluster analysis demonstrated that the similarity coefficient of the 25 pecan varieties varied from 0.62 to 0.99.Two main clades were formed,in which some related varieties could be clustered together and some of them could not be completely corresponding with genetic background.[Conclusion]Compared with traditional dominant markers and polyacrylamide gel electrophoresis,the genotyping technology consisting of fluorescence-labeled SSR primers and capillary electrophoresis is quite practical with high t

关 键 词：薄壳山核桃 荧光标记 SSR 亲缘关系 指纹图谱 

分 类 号：S718.46[农业科学—林学] S718.49