Dneasy animal tissue kit

Larval M. vitrata were collected from cultivated cowpea (Vigna unguiculata), and three alternative host plants – P. phaseoloides (dry season host), T. candida (short rainy season host), and L. sericeus (main rainy season host), in three divisions representing 6 departments in Southern Benin in 2012 (Figure 1). The divisions were Mono-Couffo, Zou-Collines and Ouémé-Plateau. Within each division, we collected from different locations to lessen the possibility that the same female individual laid larvae collected. Forty-nine, 50 and 49 individual M. vitrata samples were collected from V. unguiculata in Ouémé-Plateau, Zou-Collines, and Mono-Couffo, respectively. Forty-seven and 45 individual M. vitrata samples were collected from L. sericeus in Ouémé-Plateau, and Zou-Collines, respectively. Fifty-two, 52 and 58 individual M. vitrata samples were collected from T. candida in Ouémé-Plateau, Zou-Collines, and Mono-Couffo, respectively, and 49, 49 and 48 individual M. vitrata samples were collected from P. phaseoloides in Ouémé-Plateau, Zou-Collines, and Mono-Couffo respectively. Genomic DNA was extracted from the insect samples using DNeasy animal tissue kit and following manufacturer instructions (Qiagen, Valencia, CA). The DNA concentrations were adjusted to 10 ng/μl and used for genotyping.

To test the purity of each sample, the 5′ partial mtCOI gene region for species identification was amplified from B. tabaci samples using the new design primer pairs FHL (5′-TGRTTYTTTGGTCATCCVGAAGT-3′) and RHL(5′-TTACTGCACTTTCTGCCACATTAG-3′). DNA was extracted from 10 adult females separately from each sample (whole insect extractions) using the DNeasy animal tissue kit (Qiagen, Germany) following the manufacturer’s protocol and pooled in equal proportions.

Genomic DNA was extracted from individual whiteflies collected from the four established pure colonies using the DNeasy animal tissue kit (Qiagen, Germany) following the manufacturer's protocol. Obtained DNA from either male or female whiteflies was used for PCR amplifications followed by Sanger sequencing. For the purpose of Illumina sequencing, genomic DNA was extracted from only individual B. tabaci females collected from SSA1‐SG1_Kayingo, SSA1‐SG2_Kayingo, SSA1‐SG3_Dar es Salaam, and SSA2_Kiboga colonies.