Identificationof Spiksprouting Resistance Of Wheat And Cloning And Expression Analysis Of Genes Related To Phs Of AIP2

PHS is a serious natural disaster, and the quality and value of sprouting wheat are decreased. In this study, we selected 8 copies of American wheat varieties, 14 accessions of India wheat varieties, 51 domestic wheat cultivars as materials, we combined four molecular markers(Vp1B3 、 wmc104 、 Xgwm155 、 Xbarc170)with experimental identification of pre harvest sprouting to to screen the PHS tolerance materials, and at the same time verified the effectiveness of the 4 molecular markers and screened the high and efficiency molecular markers. At the same time, we used homologous cloning method to clone AIP2 gene from Huashan and Xiaoyan 22 and the gene sequence and structure domain and function domain were analyzed. Then we analyed the expression characteristics of different periods, find AIP2 gene Sprouting resistance, with a view to the adoption of modern molecular biology techniques improved other good agronomic characters but easy to sense of Preharvest Sprouting in Wheat Varieties and which can broaden resistance gene resources of preharvest sprouting of wheat. The main experiment results are as follows:1. Germination experiment results showed that 73 wheat varieties can be roughly divided into four types, namely high resistance and resistance to preharvest sprouting, in the sense of preharvest sprouting and high feeling of Preharvest Sprouting varieties.There are 13 high resistance sprouting varieties and their germination index is less than 10%, among them there are four American wheat cultivars, three Indian wheat varieties and six Chinese wheat varieties; the average value 4.718%, variation range 2.857% to 9.619%. There are 32 varieties resistance sprouting varieties; their germination index is 10%~30%.There are 20 varieties in the sense of preharvest sprouting; their germination index of 30%~60%.There are 8 high varieties susceptible to preharvest sprouting varieties and their germination index is60%~100%.2. The detection results of the molecular markers showed that the marker Vp1B3 is associated with PHS resistance, but three other markers were not effective, the effectiveness of Xgwm155, wmc104, Xbarc170 are needed to be verified in further.3. Combined with germination experiment and molecular marker detection results, there are 8 high resistance sprouting varieties were selected from 73 wheat cultivars, theirgermination index were less than 10% and amplified 569 bp resistance band type with marker vp1B3. Respectively, they are Yannong 23、 Shannong 33、 Lankao 181、 Xinong 585、 U.S.105、 U.S. 231、 India UP2338 and India PBW550.4. The result showed the AIP2 gene was successful cloned from Psathyrostachys huashanica and xiaoyan 22, respectly named HSAIP2 and XYAIP2, the HSAIP2 gene of 840 bp complete open reading frame was obtained, which encoded 279 amino acid residues,XYAIP2 gene of 963 bp complete open reading frame was obtained, which encoded 320 amino acid residues, HSAIP2 and XYAIP2 protein were thought to belong to the family of zinc finger ring, because the all containd a conserved Ring domain. HSAIP2 and XYAIP2 gDNA contains 5 exons and 4 introns.5. The results of HSAIP2 and XYAIP2 gene expression analysis of flowers in different periods showed that:HSAIP2 gene relative expression quantity gradually increased with the mature of embryos,the expression reached the highest in 25 days of anthesis and it is about20 times as much as that of 5 days of flowering. Xiaoyan 22 AIP2 gene relative expression quantity was increased first and then decreased with the mature of embryos, the expression reached the highest in 25 days of anthesis and it is about 30 times as much as that of 5 days of flowering, the gene relative expression decreased from 30 days of anthesis, in 35 days it decreased to the lowest, which showed that AIP2 gene may be associated with preharvest sprouting resistance.