Puroindoline genes and proteins in tetraploid and hexaploid species of Triticum

Six tetraploid (5 Triticum turgidum and 1 Triticum timopheevii) and four hexaploid (three Triticum aestivum and one Triticum kiharae) taxa of Triticum were studied in order to identify novel variation in Pin genes and proteins which can be exploited in the improvement of cultivated wheat. Western blotting with a highly specific antibody showed that puroindoline proteins were present in all of the hexaploid lines but were absent from the tetraploids. The immunoreactive bands differed slightly in their relative mobilities and their relative amounts, which could have resulted from variation in the allelic forms of Pin a and Pin b. This was supported by HPLC analyses which showed differences in the retention times and peaks heights of the putative puroindoline components in T. kiharae and T. timopheevii. Sequence analyses of cDNAs also showed variation in the sequences of expressed puroindoline genes. In particular, a sequence encoding a new form of Pin b was present in T. aestivum ssp. macha.