Cloning Of Related Genes And Identification Of Pre-harvest Sprouting Resistance In Common Wheat

Wheat seeds sprout under cool rainy or moist condition before harvest, which is called Pre-harvest sprout (PHS) is one of the devastating disaster of common wheat. Several studies were previously conducted to determine the negative effect of PHS on the yield and the quality. The white-grained wheat is became the mainly cultivars in China, but the resistant ability to PHS of white-grained wheat is weak, so rain during the harvest time would make a terrible loss. With the humid, rainy all over the harvest time in Sichuan Province, the PHS is appeared frequently. For decreasing the PHS appearance frequency, screen and identify of PHS resistance genes should be one of the most important works in world wheat genetic improve program.In this study, we choose173Chinese local cultivars and17cultivars/lines developed by our lab. All materials were planted in Qionglai breeding base in2010-2011and2011-2012. All materials’ PHS resistant ability was detected.33SSR primers which are related to PHS resistance were used to analyze their phylogenetic relationships of PHS. The allelic of PHS resistant gene VP1B3in the lines which are developed by us were cloning. At last, the results we obtained are listed below:1. The whole spike and seed germination were hold to detect the PHS resistance of190materials. The whole spike germination rate (SGR) and germination index (GI) were calculated. The result also shows that the PHS resistances of local cultivars were obvious lower than the released cultivars. Different from the released cultivars, the local cultivars which spikes with awns’ resistance were higher than those without awns. The PHS resistance of the cultivars with red-grained, were higher than those with white-grained.2.33SSR primers were used to analyze their phylogenetic relationships. Phylogenetic bands were amplified from reactions using these33screening primers. Primer Xwmc11can only amplify one type of bands.14primers were2and11primers were3. Xgwm194and Xgwm469can amplify4bands, and Xgwm174can get7bands. The bands statistics and analysis of Xgwm174indicated that:there are7types of bands, the cultivars with the amplified bands pattern Type D and type C accompany with the high PHS resistance, but the cultivars with the amplified bands pattern type A or no amplified band accompany with the low resistance of PHS.3. The availability of Vp1B3and Dorm-1was detected by the materials in this study. The results indicated that the845bp allelic of Vp1B3accompany with high resistance of PHS. Cultivars which amplified652bp and569bp are no significantly differences of mean value of SGR. Only25materials got both468bp and606bp which are amplified by primer Dorm-1, and none amplified single606bp band. Because in this study, all materials’GI were much higher than the previous study of Dorm-1, It is presumed that the single606bp band only appeared in the cultivars which GI is lower than10%.4. We used16wheat lines which are developed by us as materials and clone Vp1B3allelic, and use DNAman did the sequences comparative analysis. Allelic of Vp1B3were mutant in some materials. Compared with the sequence of Vp1Ba, R315have25bp deletion from102to126position, and a A to G mutant on326. J3091has a lot of mutants compare with Vp1Bb:a A deletion on74, and a insertion of10bases ATCCGGGAGC from254to263. Compared with the sequence of Vp1Bc, CN12, H-6, R316, You-II, J2865, MY11, J2871and J1809were have a A to G SNP on288, G to A on447. CN12, H-6, R316, You-II also have a A deletion on101position.