Molecular Cloning And Sequence Analysis Of Glutenin Subunit Genes From Xinjiang Rice Wheat(Triticum Petropavlovskyi Udacz. Et Migusch)

Xinjiang rice wheat (T. petropavlovskyi Udacz. et Migush, 2n=6x=42, AABBDD) is one of the endemic Chinese wheats, only distributing in Xinjiang and Xizang (Tibet), China. Comparison with hexaploid bread wheat, there were one or two different chromosomes in Xinjiang rice wheat. In our previous work, we found that one Xinjiang rice wheat accession, Daomai2, has novel subunits 1Dx2.1+1Dy10.1, which are very rare in cultivated wheat. In order to study this HMW glutenin subunit combination, the gene coding 1Dx2.1 was cloned and sequenced completely. And one LMW glutenin gene has also been cloned. The results were list as follows:1. In genomic PCR amplification using primers P1 and P2, one DNA band of approximately 900bp was amplified for Xinjiang rice wheat accession Daomai2. After the fragment was recovered and cloned into the pMD18-T vector, one positive clone LMWXJ-1 (Genbank accession number: AY695380) with proper insert size was screened and sequenced. LMWXJ-1 has a typical glutenin structure similar to those of published LMW-GS genes. By the deduced amino acid sequence comparison, it was found that LMWXJ-1 has higher similarity to the genes at Glu-A3 and Glu-D3 locus (with the highest similarity 82% and 80% respectively), while it has a lower similarity to the genes at Glu-B3 locus (68%).2. HMW glutenin subunits from Xinjiang rice wheat accession Daomai2 were separated by SDS-PAGE. Daomai2 has four expressed HMW glutenin subunits. Subunit 1Dx2.1 migrated faster than 1Ax1, whereas slower than 1Dx2. Subunit 1Dy10.1 migrated slower than1Dy12, while slightly faster than 1Dy10. This glutenin...