Characterization of a Double Mutation of eIF(iso)4E.a and eIF(iso)4E.c with a Diverse Genetic Background and TuMV Resistance in Chinese Cabbage (Brassica rapa ssp. pekinensis)
Abstract：Eukaryotic translatoin initiation factor 4E (eIF4E) and its iso form (eIF(iso)4E) have been demonstrated to play central roles in plant-vorus interactions. Broad-spectrum turnip mosaic virus (TuMV) resistance in Chinese cabbage (Brassica rapa ssp. pekinensis) has been demonstrated to be related to BraA.eIF(iso)4E.a and BraA.eIF(iso)4E.c. By now only two double allelic mutant lines (RLR22 and BP8407), were reported, and they shared the same pattern of mutation in eIF(iso)4E.c. To explore diversiform allelic variation in BraA.eIF(iso)4E.a and BraA.eIF(iso)4E.c, germlines with confirmed TuMV resistance or susceptibility were analyzed in this study using candidate gene re-sequencing approach. One line (He102) with new double mutation of eIF(iso)4E.a and eIF(iso)4E.c resistant to TuMV was identified. Polymorphisms between wild and mutant sequences showed that in line He102 eIF(iso)4E.a was a pseudogene with exons 4 and 5 deleted, while eIF(iso)4E.c contained a frame-shift mutation which resulted in early termination. These mutations were responsible for the loss of TuMV resistance as wild-type. Chinese cabbage genes could recover virus susceptibility in Arabidosis lsp1 mutant while mutated genes could not. Codominant functional markers related to the mutation on both alleles were developed. The categorized double allelic mutated line was a more valuable resistant resource with a diverse background, and the developed markers could be used directly for germplasm screening and broad-spectrum TuMV-resistant variety breeding through marker-assisted selection. The study provides a representative exaple exemple for allelic-variation identification by candidate gene re-sequencing with mini-core germplasm collection as materials, which were carefully selected based on target traits.
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