Relationships Of High-Molecular-Weight Glutenin Subunits 1Bx13 And 1By16 With Yield Related Traits In Wheat

High-molecular-weight glutenin subunits were major parts of wheat storage proteins, and played a significant role in determing wheat processing quality. An early research based on introgression lines with varied HMW-GS alleles showed that the introgression line with 1Bx13+1By16 had the highest grain protein content in all of the investigated introgression lines. However, the mechanism of 1Bx13 and 1By16 correlated with high grain protein content was still unknown.Few of the wheat varieties in China contained those good quality alleles except Jimai20 and several other cultivars,therefore,it had a great potential to use 1Bx13 and 1By16 to improve wheat processing quality in China. In this research 1Bx13ORF and 1By16ORF were cloned from Jimai20,constructed into expressing vectors and transferred into bread wheat through agrobacterium-mediated wheat mature embryo calli transformation system to do functional identification.We also used recombination inbred lines derived from crossing of Jimai20 and Yanzhan No.1 to analysis correlations between yield related traits and alleles 1Bx13+1Byl6. The results were outlined as following.1.We sequenced the full length of 1Bx13ORF and 1By16ORF based on the deletion subclones and BLAST them with 1Bx13ORF and 1By16ORF from NCB1.The results showed that there are several single nucleotide diversities between the two sequences which came from this research and came from NCBI, and some single nucleotide changes leaded to alteration of protein primary structure.They were AA 646 Arg-Gln, AA 702 Leu-Pro,AA 714 Pro-Ser,AA 777 Ser-Gln,and AA 792 Thr-Ala changes in 1Bx130RF, in which AA 646 change was located on repetitive domain while others located on C-terminal domain. In terms of 1By16ORF only one amino acid changed on AA 247 in repetitive domain from Glu on NCBI to Gly in this research. These protein primary structure changes resulted in a higher Gln content in 1Bx13ORF, which were advantageous to the aggregation and stability of the glutenin macropolymer.2.Over expressing vectors pCUbi1390-1Bx13 and pCUbi1390-1By16 were constructed to do agrobacterium-mediated wheat transformation.Resistant plantlets were obtained by using Agrobacterium C58C1 through agrobacterium-mediated wheat mature embryo calli transformation system, and at this moment they were cultured on plantlets growth medium.PCR analysis for the positive TO transgenic plants would be performed when they were transplanted in fields and grain protein content would be measured in T3 or T4 generation according to experimental schedule. The lower regeneration rate and higher browning rate during tissue culture were still a block to improve the efficiency wheat transgenic system.3. The correlations between 1Bx13+1By16 and some yield related traits were analysed using a recombination inbred lines derived from crossing of Jimai20 and Yanzhan No.1, and the result showed that there were no significant difference in spikes per plant, plant height, spike height, number of fertile spikelet, grain number per spike, thousand-grain weight between lines contained 1Bx13+1By16 from Jimai20 and lines contained 1Bx14+1Bx15 from Yanzhan No.1.So we inferred that 1Bx13+1By16 from Jimai20 or the chromosomal segment contained 1Bx13+1By16 from Jimai20 could improve wheat grain protein content while were with little influence on yield related traits, therefore, it was with a great productive potentialities.