Rapid development and characterization of EST-SSR markers for the honey locust seed beetle, Megabruchidius dorsalis (Coleoptera: Bruchidae), using de novo transcriptome analysis based on next-generation sequencing
Ohbayashi, Kako | Ishikawa, Naoko | Hodoki, Yoshikuni | Okada, Yasukazu | Nakano, Shin-ichi | Ito, Motomi | Shimada, Masakazu
We developed 10 novel simple sequence repeat markers from expressed sequence tags for the honey locust seed beetle, Megabruchidius dorsalis Fåhraeus 1839 (Coleoptera: Bruchidae, using de novo transcriptome analysis based on next-generation sequencing. In a M. dorsalis Harataima (Kanagawa-pref.) population, the number of alleles per locus ranged from 2 to 6, with an average of 4.0. Observed and expected heterozygosities ranged from 0.13 to 0.72 and 0.17 to 0.72, respectively. We initially developed 11 novel markers, but one was eliminated because it showed significant linkage disequilibrium with another locus after Bonferroni correction. To check the applicability of the remaining 10 markers, we used them to analyze two additional geographic populations (Yashima, Akita-pref., and Kameoka, Kyoto-pref.). Mean numbers of alleles per locus in the Yashima and Kameoka populations were 2.6 and 2.9, respectively, with corresponding mean observed heterozygosities of 0.36 and 0.52. These results based on the two additional populations confirm that our developed markers worked efficiently. The simple sequence repeat markers developed from expressed sequence tags in the present study should, therefore, be useful for explicating the population genetic structure of M. dorsalis and for future paternal analyses.
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