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目的对蛭类转录组进行SSR分析,是开发功能分子标记、开展对药用蛭类资源的开发利用及进行遗传背景监测的必要手段。方法利用与医学密切相关的医蛭科棒纹牛蛭Poecilobdella javanica和日本医蛭Hirudo nipponia、黄蛭科宽体金线蛭Whitmania pigra和山蛭科洞穴山蛭Heamadipsa cavatuses sp.nov.共14个样本,通过RNA测序及转录组分析,筛选吸血差异unigene,并比较4个种的SSR标记特征和分布,用于挖掘抗凝血相关功能EST-SSR。结果通过SSR密度分析发现,SSR在棒纹牛蛭、日本医蛭、宽体金线蛭和洞穴山蛭转录组中的间隔长度为4 723、6 026、8 059、8 144 bp;SSR组成分析发现三碱基重复的基序占蛭类SSR类型组成中绝大多数;另外,洞穴山蛭的SSR组成与其他3种存在明显区别。比较4种吸血蛭吸血前后转录组中表达丰度显著差异的unigene群,发现4种蛭中包含SSR且抗凝血相关的重叠unigene类群在转录组中呈较高的片段多态性。结论 unigene片段长度、SSR的密度、组成和分布特点能反映物种之间的分类关系;4种蛭重叠的抗凝血相关分子功能对应的EST-SSR,可作为抗凝血相关的功能分子标记侯选EST-SSR。
OBJECTIVE: To perform SSR analysis on the transcriptome of leech, it is necessary to develop functional molecular markers for the development and utilization of medicinal leech resources and genetic background monitoring. Methods A total of 14 samples were obtained from Poecilobdella javanica and Hirudo nipponia, Whitmania pigra and Heamadipsa cavatuses sp.nov, closely related to medicine. RNA sequencing and transcriptome analysis to screen for differences in blood-sucking unigene, and to compare the characteristics and distribution of SSR markers in 4 species for mining anticoagulant-related EST-SSR. Results The results of SSR analysis showed that the length of SSR was 4 723, 6 026, 859, 8 144 bp in the Leech, Leech, Hirudo, Found that the three base repeat motifs accounted for the vast majority of the composition of SSR types of leeches. In addition, the SSR composition of the mountain Leech was significantly different from the other three. Four unigene groups with significant differences in expression abundance before and after blood-sucking blood-sucking were compared. Four leechs were found to contain SSR and the anticoagulant-associated overlapping unigene groups showed higher fragment polymorphisms in the transcriptome. Conclusion The unigene fragment length, SSR density, composition and distribution characteristics can reflect the taxonomic relationship among species. The EST-SSR corresponding to the anti-coagulation-related molecular function of 4 leptospirosis can be used as anticoagulant-related functional molecular marker Select EST-SSR.