41The rice weevil, Sitophilus oryzae is one of the primary pests of stored grains worldwide. To 42 develop and implement an effective integrated pest management strategy, an understanding of the 43 population structuring of this destructive pest is vital. In this study we used Illumina paired-end 44 sequencing to develop S. oryzae species-specific microsatellite markers, and used these markers to 45 conduct a preliminary assessment of population structuring in four populations of S. oryzae from three 46 countries (Australia, China, and USA). 7,635,996 raw sequencing reads were produced, with 11,794 47 microsatellites detected and 214,257 primer options designed. 48 microsatellite markers were selected 48 for further validation, with 10 markers amplifying consistently across the four S. oryzae populations.
49These markers displayed a high level of polymorphism overall (6.67 alleles/locus), though this was 50 slightly lower within populations (3.10 -4.88 alleles/locus). We used the markers to conduct a 51 preliminary assessment of genetic structuring among the four S. oryzae populations: three laboratory 52 cultures (New South Wales, Queensland, and Santai) and a field collected population from Kansas. 53 Analyses suggest high levels of genetic differentiation between the sample locations, with a global F ST 54 of 0.239, and pairwise F ST values ranging from 0.100 to 0.395. Bayesian clustering analyses suggest 55 these four populations formed four distinct clusters, with a similar pattern identified by Principal 56 Coordinate Analysis. These microsatellite markers, together with our preliminary population genetic 57 analyses, will provide a valuable resource for population genetic research, and contribute to effective 58 integrated pest management strategies in the future. 59 60 61 62 63 4
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