Development of low-Gly m Bd 30K(P34) allergen breeding lines using molecular marker in soybean

Soybean (Glycine max (L.) Merr.) is an important source of vegetable oil and high protein. Use of soybean meal by the food industry is increasing, but severely limiting dietary choices and the quality of life of food-allergic individuals. Gly m Bd 30K (P34) is known as the main seed allergens in soybean-sensitive patients. The objective of this work was to determine the molecular basis of the low mutation of soybean P34 and to design molecular marker for the selection of the causative mutations for wild homozygous, heterozygous and mutant homozygous. We developed a co-dominant marker based on the sequence of Glyma08g12270 containing a four-base pair insertion at the P34 start codon. Also, we made a polyclonal antibody for investigation of P34 protein levels. Using a co-dominant marker and a polyclonal antibody, polymorphism and amount of protein for Glyma08g12270 were analyzed in F2 and F3 generation crossing PI 567476 and Hwanggumkong, Korean cultivar. To investigate the association of the P34 genotype with the P34 protein phenotype, segregating populations in F2 were developed from crossed Hwanggum and PI567476. For the 258 samples analyzed, the ratio of homozygous wild-type: heterozygous: homozygous mutant P34 genotypes was 34:94:30=1:2:1(test for goodness of fit by X2 analysis). As results, the polymorphism analysis was accustomed to a difference of protein level of wild homozygous, heterozygous and mutant homozygous.

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