基于转录组数据的意大利蝗微卫星位点分析与分子标记开发
nalysis of microsatellite loci from Calliptamus italicus (Orthopera: Acrididae) based on a transcriptome dataset
桑 迪；徐 叶；王伟亮；向 敏；季 荣；王 晗
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DOI：10.7679/j.issn.2095-1353.2020.066
作者单位：中亚区域跨境有害生物联合控制国际研究中心，新疆师范大学生命科学学院，乌鲁木齐 830054； 新疆玛纳斯县蝗虫鼠害预测预报防治站，玛纳斯 832200
中文关键词：意大利蝗；微卫星；转录组；分子标记
英文关键词：Calliptamus italicus; microsatellite; transcriptome; molecular marker
中文摘要：
【目的】 意大利蝗Calliptamus italicus (L.)是新疆荒漠半荒漠草原重要害虫。本研究利用已获得的意大利蝗转录组数据，鉴定其微卫星位点。【方法】 使用MISA筛选SSR位点，利用Primer Premier 5设计引物，通过PCR扩增对引物进行验证。【结果】 在意大利蝗转录组数据库中，共检测出156 500个SSR位点，分布在126 369条unigene中。其中，单核苷酸重复为60.88%，二核苷酸重复为23.58%，三核苷酸重复和四核苷酸重复分别为12.99%和2.04%。单核苷酸重复主要为A/T（44.38%），二核苷酸重复主要为AC/GT（12.99%）和AG/CT（8.05%）。基于筛选的SSR位点设计引物，随机挑选24对引物，对10个不同地理种群意大利蝗成虫DNA样品进行PCR扩增，共有6对引物扩增成功。【结论】 本研究利用转录组数据发掘意大利蝗SSR位点，为意大利蝗分子标记、种群遗传及功能基因等研究奠定基础。
英文摘要：
[Objectives] To
investigate genetic variation in Calliptamus italicus（L.）, an important pest of desert and semi-desert grasslands in
Xinjiang. [Methods] Microsatellite loci were analyzed from the transcriptome
dataset of C. italicus, and primers
designed for PCR amplification. SSR loci were
identified using MISA, and primers were designed using Primer 5 for
experimental validation. [Results] In total, 156 500 SSR loci distributed in 126 369 unigenes were detected. The
majority of these were mono-nucleotides (60.88%), followed by di-nucleotides
(23.58%), tri-nucleotides (12.99%), and tetra-nucleotides (2.04%). The most abundant mono-nucleotide
repeat motif was A/T (44.38%). The dominant motif among the di-nucleotide repeat units was AC/GT (12.99%), followed by AG/CT (8.05%). 24 primer pairs were
randomly selected to validate SSR loci detected with genomic DNA from multiple
individuals from 10 different geographic populations. 6 primer pairs amplified the
expected products. [Conclusion] Based on transcriptome
data, 156 500 SSR loci were identified which can
provide a foundation for research on molecular markers, population genetics and functional genes of C. italicus.