摘要
为获得稳定可靠的马氏珠母贝SSR分子标记,本文利用MISA软件对转录组测序数据进行了大规模的EST-SSR标记发掘,并分析了位点信息及其多态性。结果表明,马氏珠母贝转录组测序所获得的74007条Unigenes中检测出9872个EST-SSR位点,位点出现频率为13.34%,平均每5102 bp含有1个SSR位点。在转录组SSR中共有132种重复基元类型,其中单核苷酸重复基元为主要类型,占SSR总数的81.46%;单核苷酸重复以A/T基序为主,占SSR总数的71.27%。基于筛选的SSR序列,应用Primer3软件进行引物的批量设计,共为5922条EST-SSR序列成功设计出17766对引物。随机选择80对引物对EST-SSR多态性进行检测,共有62对引物成功扩增出稳定条带,占引物总数的77.5%;其中,17对EST-SSR引物表现出个体多态性,多态性比率为27.42%。以上研究为马氏珠母贝遗传多样性、遗传图谱构建及分子辅助育种研究提供了有效工具,对于马氏珠母贝种质资源保护、优良品种培育和珍珠养殖业的健康发展均具有重要意义。
To enrich the SSR markers from the pearl oyster Pinctada martensii, the EST-SSRs(expressed sequence tag-derived-simple sequence repeats) were developed on a large scale based on the transcriptome sequencing data using program MISA, and polymorphism of SSRs were analyzed and characterized. The results indicate that 9872 EST-SSRs were recognized from 74007 unigenes generated from the transcriptome datasets, accounting for 13.34% of the total unigenes(one EST-SSR per 5102 bp sequence on average). In addition, 132 types of repeat motifs were classified in all EST-SSRs, of which the type of mono-nucleotide repeat was dominant(81.46%), and A/T was the most abundant mono-nucleotide motif(71.27%). Based on the identified SSRs, 17766 pairs of primer sets were designed from 5922 unigenes using program Primer 3. Eighty pairs of primer were randomly selected for PCR amplification to analyze the polymorphism of EST-SSRs, and 62 primers were amplified successfully, among which 17 were polymorphic and the polymorphic rate was 27.42%. These results will provide a useful tool for studies in genetic diversity, genetic map construction and molecular assisted breeding of P. martensii, and be of great importance in genetic resource conservation, stock breeding of P. martensii and sound development of the pearls culture.
出处
《海洋与湖沼》
CAS
CSCD
北大核心
2015年第3期687-693,共7页
Oceanologia Et Limnologia Sinica
基金
国家自然科学基金项目
31202023号
广东海洋大学优秀青年骨干教师特别资助计划
2014001号
广西北部湾海洋生物多样性养护重点实验室(钦州学院)开放课题
2015KB05号
关键词
马氏珠母贝
简单重复序列
转录组
分子标记
Pinctada martensii
simple sequence repeat
transcriptome
molecular marker
