| 熊翠玲*，张璐*，付中民，王鸿权，侯志贤，童新宇，李汶东，郑燕珍，陈大福**，郭睿**,2017,基于RNA-seq数据大规模开发中华蜜蜂幼虫的SSR分子标记[J].环境昆虫学报,39(1):68-74 |
| 基于RNA-seq数据大规模开发中华蜜蜂幼虫的SSR分子标记 |
| Large-scale development of SSR primers for Apis cerana ceranalarvae based on its RNA-seq datasets |
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| DOI： |
| 中文关键词: RNA-seq 中华蜜蜂 幼虫肠道 SSR分子标记 |
| 英文关键词:RNA-seq Apis cerana cerana larval gut SSR |
| 基金项目:现代农业产业技术体系建设专项资金（CARS-45-KXJ7）；福建农林大学科技发展资金（KF2015123）；国家自然科学基金（30800806） |
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| 中文摘要: |
| 中华蜜蜂是东方蜜蜂的一个亚种，也是我国的特有蜂种。本研究基于已获得的中华蜜蜂幼虫肠道转录组数据预测SSR分子标记，并进行SSR位点的信息分析和SSR引物的发掘。利用MISA软件对幼虫肠道转录组中数据组装得到的43557条unigenes进行搜索，共预测出13448个SSR位点，它们分布于7763条unigene中，其中最主要的重复类型为二核苷酸重复（58.03%），其次为三核苷酸重复（28.23%）和四核苷酸重复（9.72%）。二核苷酸重复中的基序主要是 AT/AT（占总量的30.4%)。对于所有的SSR位点，利用Primer Premier 5软件成功设计出21627对引物，随机选取48对引物对5个不同来源的中华蜜蜂幼虫肠道样品进行SSR位点扩增，共有15对成功扩增出符合预期的目的片段。研究结果表明利用转录组数据大规模开发中华蜜蜂幼虫的SSR引物是可行的，本研究开发出的SSR引物为研究中华蜜蜂分子遗传学奠定了基础。 |
| 英文摘要: |
| Apis cerana cerana is a subspecie of Apis cerana, and it's also an unique honeybee specie in China. In the present study, SSR molecular markers for A. c. cerana were predicted based on the transcriptome datasets of A. c. cerana larval gut previously obtained. Further, we performed infromation analysis of SSR loci and development of SSR primers. First, MISA software was used to search 43557 unigenes assembled from RNA-seq data of eastern honeybee larval gut, and a total of 13448 SSR loci were found, which distributed in 7763 unigenes. Among these SSR loci, the main repeat types were di-nucleotide repeats (58.03%), followed by tri-nucleotide repeats (28.23%)and finally tetra nucleotide repeats (9.72%). The AT/AT motif was the most abundant (30.4%)in di-nucleotide repeats. In total, 21627 pairs of primers were designed from all SSRs. 48 pairs of SSR primers were randomly selected and used to amplify the SSR loci inA. c. cerana larva gut samples from five different regions in China, and 15 SSR primer pairs could amplify the target fragments with expected size. Our findings suggested that large-scale development of A. c. cerana larvae's SSR primers on basis of transcriptome data is feasible, additionally, the 15 SSRs primers developed here can lay a foundation for study on molecular genetics of A. c. cerana. |
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