Analysis on EST-SSR in transcriptome of medicinal leeches and mining of anticoagulant related functional molecular markers / 中草药

ABSTRACT

Objective: To expand the exploitation and genetic diversity conservation of medicinal leech resources, the effective functional molecular markers for identification are needed to be developed by transcriptome-SSR analysis. Methods: Totally 14 samples including Poecilobdella javanica and Hirudo nipponia of Hirudinidae, Whitmania pigra Whitman of Haemopidae and Heamadipsa cavatuses sp. nov. of Haemadipsidae were used in transcriptomic analysis by RNA-seqencing, the simple sequence repeats (SSR) distribution characteristics of four species were compared, and transcriptomic comparation of hematophagia samples and hungry samples were carried out to investigate the anticoagulation involved unigene. Finally, the data in two groups were combined to mining functional EST-SSR involved in anticoagulation. Results: The SSR density analysis showed the gap lengths of SSR in transcriptome of Poecilobdella javanica, Hirudo nipponia, Whitmania pigra Whitman, and Heamadipsa cavatuses were 4 723, 6 026, 8 059, and 8 144 bp. In the four species, trinucleotide repeats occupied overwhelming majority in composition of EST-SSR; On the whole, distribution of SSR types can distinct from each other, and Heamadipsa cavatuses was particularly different from others. Moreover, we found that the anticoagulation involved unigene group containing SSR had high fragment-polymophism. Conclusion: For the four leech species, the characteristics of density, composition, and distribution of EST-SSR can reflect its classification relationship. Meanwhile, the EST-SSRs in unigenes which involved in anticoagulation and overlaped in four leech species are ideal fragment group for screening anticoagulation functional marker.