Development and characterization of microsatellite markers in the earthworm Drawida gisti Michaelsen, 1931 and cross-amp

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Development and characterization of microsatellite markers in the earthworm Drawida gisti Michaelsen, 1931 and cross‑amplification in two other congeners Hongyi Liu1   · Yufeng Zhang2   · Guobing Wang1   · Jie Chen3 · Qingzheng Zhang1   · Honghua Ruan1  Received: 5 June 2020 / Accepted: 29 August 2020 © Springer Nature B.V. 2020

Abstract The earthworm (Drawida gisti) is an ecologically important sentinel species for soils that is widely distributed throughout Eastern Asia; however, the molecular tools required for genetic diversity studies of this earthworm are still rare. The aim of the study was to develop and characterize microsatellite markers in D. gisti and to evaluate their transferability to other Drawida species. We employed a RAD-seq approach to develop 12 microsatellite markers for D. gisti. The characterization and analysis of loci was achieved using 24 individuals of D. gisti from a natural population. The number of alleles per locus ranged from four to eleven, with an average of 6.5. Observed and expected heterozygosities varied from 0.708 to 0.958, and from 0.568 to 0.883, respectively. No loci presented significant deviations from the Hardy–Weinberg equilibrium, while linkage disequilibrium was detected between three loci. Cross-species amplification tests suggested that the transferability of ten loci was positive for the two congeners D. japonica and D. ghilarovi. This set of microsatellite markers may be used to evaluate the genetic diversity and population structures of D. gisti and related species in the future. Keywords  Drawida gisti · Earthworm · Microsatellite · Cross-species amplification · RAD-seq

Introduction Earthworms are considered to be “ecosystem engineers”, as they play critical roles in soil ecosystems by making direct contributions to their physical and chemical properties, as well as nutrient circulation [1, 2]. There are upwards of 6000 species of earthworms globally [1]. The major factors attributable to this hyper-diversity include their variable living environments and poor dispersal ability [3–5]. Recent studies suggest that extant earthworms consist primarily of Crassiclitellata and Moniligastridae [1, 2]. In * Honghua Ruan [email protected] 1



College of Biology and the Environment, Nanjing Forestry University, No.159 Lonpan Road, Nanjing 210037, Jiangsu, China

2



Hebei Key Laboratory of Animal Diversity, Langfang Normal University, Langfang 065000, China

3

Key Laboratory for Ecology and Pollution Control of Coastal Wetlands (Environmental Protection, Department of Jiangsu), School of Environmental Science and Engineering, Yancheng Institute of Technology, Yancheng 224051, China



contrast to Crassiclitellata, the number of Moniligastridae species is much smaller. Consequently, few studies on the genetic diversity and molecular phylogeny of species in the Moniligastridae family have been conducted, with the genus Drawida Michaelsen, 1900 being the largest group [6]. Drawida gisti Michaelsen, 1931 is a common species of genus Drawida, whi

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