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Quantification of ABCG4 in aphids

WJ Wikum H. Jayasinghe
HK Hangil Kim
YN Yusuke Nakada
CM Chikara Masuta
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Aphids were put on CMV-infected N. tabacum, [CMV-O + Y-sat]-infected N. tabacum, transgenic N. benthamiana expressing Y-sat dsRNA and wild-type N. benthamiana. For aphid ABCG4 mRNA (XM_022308170.1), total RNA was extracted separately from alate aphids of the third instar and from alate adults and amplified using primer pair 5′-AACTGCCCTGTCCCATCTAT-3′/5′-GGTGTGTCATTGATGGCTAG-3′. The β-tubulin gene was used as the reference as described above. Real-time qPCR was performed and the data were processed as explained in Supplementary Note 3. To detect a difference in ABCG4 mRNA accumulation level in aphids, two-tailed t test was performed. The data sets are presented in Supplementary Methods 1.

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