Comparison of the amino acid sequences of prolamins and protein allergens from rice seeds. (A) Phylogenetic analysis of Cys-rich prolamins, Cys-depleted prolamins, and protein allergens of RA16/17. The phylogenetic tree was constructed by the neighbor-joining method using MEGA version 458 (http://www.megasoftware.net). The reliability of different phylogenetic groupings was evaluated by the bootstrap test (1000 replicates). Magenta, blue, and green letters indicate Cys-rich prolamins, Cys-depleted prolamins, and RA16/17 proteins, respectively. Numbers in parentheses indicate the numbers of Cys residues in the predicted mature-sized proteins. The NCBI accession numbers are Os03 g0766100, NP_001051380; Os03 g0766250, NP_001173654; Os05 g0328800, NP_001055211; Os05 g0329100, NP_001055213; Os05 g0329300, NP_001055214; Os05 g0329400, NP_001055215; Os05 g0329700, NP_001055216; Os05 g0330600, NP_001055218; Os05 g0331550, NP_001174359; Os05 g0331800, NP_001055221; Os05 g0332000, NP_001055222; Os06 g0507200, NP_001057724; Os07 g0206400, NP_001059151; Os07 g0206500, NP_001059152; Os07 g0219300, NP_001059197; Os07 g0219400, NP_001059198; Os07 g0220050, NP_001175103; Os11 g0535100, NP_001068024; Os12 g0269101, NP_001176891; Os12 g0269200, NP_001066544. (B) The amino acid sequences of Cys-rich and Cys-depleted prolamins were aligned with the CLUSTALW program. The genes of Os03 g0766100 and Os05 g0329100 encode the Cys-rich 10-kDa prolamin (crP10) and Cys-depleted 13-kDa prolamin (cpP13), respectively, analyzed in our recent paper.10 Magenta-on-black letters indicate Cys residues in the predicted mature-sized proteins. White-on-black letters indicate amino acid residues conserved in more than 10 of the sequences analyzed.