闽楠基因组SSR的开发及其通用性分析  

Development of Genome SSR in Phoebe bournei and Generality Analysis

作　　者：熊远芳 江淑珍 连辉 刘丹 李煜 叶宝鉴 陈世品[1,2] Xiong Yuanfang;Jiang Shuzhen;Lian Hui;Liu Dan;Li Yu;Ye Baojian;Chen Shipin(College of Forestry,Fujian Agriculture and Forestry University,Fuzhou,350002;Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization,College of Landscape Architecture,Fujian Agriculture and Forestry University,Fuzhou,350002;College of Architecture and Urban Planning,Fujian University of Technology,Fuzhou,350108)

机构地区：[1]福建农林大学林学院,福州350002 [2]福建农林大学园林学院,兰科植物保护与利用国家林业和草原局重点实验室,福州350002 [3]福建工程学院建筑与城乡规划学院,福州350108

出　　处：《分子植物育种》2024年第23期7764-7773,共10页Molecular Plant Breeding

基　　金：福建省林木种苗科技攻关六期项目(KLB19003A)资助。

摘　　要：本研究以闽楠优良基因型为供试材料,利用IlluminaMiSeq测序平台对SSR富集文库进行高通量测序,共获得5178284条序列,数据过滤后得到4758764个高质量reads。通过MISA软件搜索到1590126个SSR位点,SSR特征分析表明闽楠SSR的优势重复类型为单核苷酸、二核苷酸和三核苷酸,SSR位点数分别为130952(8.24%)、1229255(77.31%)、200055(12.58%)。通过Prel程序初步对获得的SSR位点进行了多态性评估,并利用Primer3 v2.3.6成功设计了引物。用6个闽楠基因型对136对基因组SSR引物进行筛选验证,最终得到23对条带清晰、特异性好、稳定性好且多态性高的引物。用筛选出的23对多态性引物对8个闽楠近缘种进行通用性检测,结果表明,23对引物在楠属中通用率为100%;在刨花润楠、红楠、黄绒润楠、绒毛润楠中通用率分别为95.65%、86.96%、82.61%、86.96%;在香樟和阴香中通用率分别为78.26%和69.57%。开发的基因组SSR为闽楠遗传多样性研究及分子标记辅助育种提供科学依据。The superior genotypes of Phoebe bournei were used as test materials in this study,the SSR-enriched library was high-throughput sequenced using Illumina MiSeq sequencing platform,and a total of 5178284 sequences were obtained.After data filtering,4758764 high-quality reads were obtained.MISA software was used to search 1590126 SSR loci.SSR characteristic analysis showed that the dominant repeat types of P.bournei SSR were mononucleotide,dinucleotide and trinucleotide.The SSR number was 130952(8.24%),1229255(77.31%)and 200055(12.58%),respectively.The polymorphism of the obtained SSR loci was evaluated preliminarily by Prel program,and primers were successfully designed using Primer3 v2.3.6.136 pairs of genomic SSR primers were screened and verified by 6 P.bournei genotypes,and 23 pairs of primers with clear bands,good specificity,good stability and high polymorphism were obtained.They were used to detect the transferability of 8 sibling species of P.bournei.The results showed that the universal rate of 23 primers in Phoebe was 100%.In Machilus pauhoi,M.thunbergii,M.velutina,M.grijsiithe,the common rates were 95.65%,86.96%,82.61%and 86.96%,respectively.The primer universal rates of Cinnamomum camphora and C.burmanni were were 78.26%and 69.57%.The development of genomic SSRs provided scientific basis for genetic diversity research and molecular mark-er-assisted breeding.

关 键 词：闽楠(Phoebe bournei) 高通量测序 SSR 通用性 

分 类 号：S792.24[农业科学—林木遗传育种]