Abstract
The great web-spinning sawfly Acantholyda posticalis is notorious for damaging Pinus forests across the Palearctic region. At present, uncertainties persist regarding its intraspecies variation and presumed subspecies. To use as tools for future studies, herein we developed genome-wide microsatellite markers for A. posticalis. Through searching, rigorous manual screening, and amplification trial, 56 microsatellite markers were obtained from the genome sequences. We characterized these markers across two populations from Shandong province (SD) and Heilongjiang province (HLJ) in China, and carried out cross-amplification in three related species. Out of the 56 markers tested, 10, 31, and 15 were categorized into high, moderate, and low polymorphic levels, respectively, based on their polymorphic information content (PIC) values. Meanwhile, 28, 19, and 4 microsatellite loci were successfully cross-amplified in Cephalcia yanqingensis, C. chuxiongica, and C. infumata, respectively, which could serve as potential molecular markers for their further studies. STRUCTURE and PCoA analyses revealed two distinct clusters corresponding to SD and HLJ, respectively, indicating a high resolution of these markers. Therefore，the 56 microsatellite markers identified here have the potential to serve as efficient tools for unraveling intraspecies variation and evolutionary history of A. posticalis.
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