文冠果转录组SSR特征分析及EST-SSR标记开发  

Characterization of SSRs and development of EST-SSR markers in transcriptome of Xanthoceras sorbifolia Bunge

作　　者：刘玉林[1,2] 李伟[2] 董树斌[3] 沐先运[3] 张志翔[3] LIU Yulin LI Wei DONG Shubin MU Xianyun ZHANG Zhixiang(College of Forestry, Northwest A ＆ F University, Tangling, Shaanaci 712100, China College of Biological Science and Biotechnology College of Nature Conservation, Beijing Forestry University, Beijing 100083, China)

机构地区：[1]西北农林科技大学林学院,陕西杨凌712100 [2]北京林业大学生物科学与技术学院,北京100083 [3]北京林业大学自然保护区学院,北京100083

出　　处：《西北农林科技大学学报（自然科学版）》2017年第3期89-95,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"十二五"科技支撑计划项目(2011BAD22B08);国家林业公益性行业科研专项(201004001);西北农林科技大学博士科研启动基金项目(Z109021508)

摘　　要：【目的】利用高通量测序技术分析文冠果转录组中SSR位点分布类型与特征,并设计开发EST-SSR标记,为文冠果的遗传多样性分析提供标记资源。【方法】利用MISA软件筛选文冠果51 867条unigenes中的SSR(2~6bp)位点,统计分析SSR位点的分布特征、发生频率和平均分布距离。利用Prime 3在线软件随机设计合成60对EST-SSR标记,用我国7个省份的16份文冠果材料对所设计的标记进行多态性筛选;用Popgene 32软件分析标记的等位基因数(NA)、观测杂合度(HO)、期望杂合度(HE)和多态性信息含量(PIC)等。【结果】从文冠果unigenes中筛选出6 707个SSR位点,其平均发生频率为12.93%,平均分布距离5.38kb,平均长度17.30bp,其中2、3核苷酸重复单元的发生频率分别为6.24%和4.93%。设计的60对标记中有47对能扩增出与目的片段长短相符的产物,其中14对多态性较高,共扩增到52个等位基因,HO和HE分别为0~0.813和0.353~0.762,PIC为0.283~0.701。【结论】文冠果转录组SSR重复单元以2、3核苷酸重复为主;新开发的14个EST-SSR标记可作为文冠果种质资源多样性分析的标记资源。【Objective】High-throughput sequencing technology was used to analyze the distribution types and characteristics of SSRs in transcriptome of Xanthoceras sorbifolia and EST-SSR markers were developed to provide basis for genetic diversity and evolutionary origin analysis of X.sorbifolia.【Method】The number,distribution characteristics,frequency of occurrence and mean distribution distance of SSRs screened by MISA from 51 867 unigenes of X.sorbifolia were counted and analyzed.Sixty EST-SSR primers were designed by on line software Prime 3and 16 germplasms of X.sorbifoliafrom seven provinces of China combined with 6% modified polyacrylamide gel electrophoresis were used for polymorphism screening of these markers.The number of alleles,observed heterozygosity and expected heterozygosity of these markers were analyzed using Popgene 32.【Result】A total of 6 707 SSRs were identified from the X.sorbifoliaunigenes.The average frequency of occurrence,distribution distance and average SSR length of these SSRs were 12.93%,5.38 kb and 17.30 bp.The frequencies of occurrence of dinucleotide and trinucleotiderepeats were 6.24% and 4.93%,respectively.Among the 60 markers,47amplified the targeted fragments and 14 markers appeared to be polymorphic with a total of 52 alleles,the observed and expected heterozygosity ranged from 0to 0.813 and 0.353 to 0.762,respectively.The PIC values varied from 0.283to0.701.【Conclusion】The SSR repeat units in transcriptome of X.sorbifolia were mainly dinucleotide and trinucleotide repeats and the 14EST-SSR markers developed in this work are useful for genetic diversity analysis in X.sorbifolia.

关 键 词：文冠果 转录组 EST-SSR标记 多态性分析 

分 类 号：S565.9[农业科学—作物学]