Genetic Dessection And Mapping Of Bvp In Barley,Tilling-population Analysis Of Edr1 In Barley Cultivar "Xiyin2”

Basic vegetative period(BVP)is an important trait for determining flowering time and adaptation to variable environments.It respond to the vernalization,photoperiod and eps.Traditional genetic analysis using 557 F2 individuals derived from Mona×Turk revealed that the short BVP is governed by a single recessive gene(BVP-1)and was further validated in 2090 F3 individuals.The BVP-1 gene was first mapped to barley chromosome 1H using SSR markers.Comparative genomic analysis demonstrated that the chromosome region of BVP-1 is syntenic to rice chromosome 5 and Brachypodium chromosome 2.SSCP analysis was also implemented for mining polymorphic markers in the F2 population.Development of an EMS mutagenized population of the barley varieties"XiYin-2" and discovery of EDR1 mutations by TILLING technology.The results are as follows:1.We developed 563 F2 and between Mona X Turk to reveal that the short BVP is governed by a single recessive gene(BVP-1).2.Studying the genomes alignment between the barley annotation of BVP gene regions of Bin 14 with rice and Brachypodium special regions using the bioinformatics methods.Bioinformatics methods analysis showed that the target gene is located in the Bin 14 region on chromosome 1H,which displayed syntenic to a segment of rice chromosome 5 and a segment of Brachypodium distachyon chromosome 2.A BLASTN search using sequences of the BVP gene region identified a syntenic region of 443 kb on rice chromosome 5(Os05)from gene Os05g50980 to gene Os05g51754 and a syntenic region of 216 kb on Brachypodium chromosome 2(Bd2)from gene Bradi2g14190 to Bradi2g 14450.Twenty-five barley EST sequences were identified by using all the rice and Brachypodium putative genes in the syntenic region.3.Developed F2 population by crossing Mona x Turk obtained 14 polymorphic markers:SSR marker:WMC1E8,GBM1434,BMAG 579,GMS149,C1-CA608558;EST-SSR marker:GBM1461,SCSSR2748,SCSSR4163;RFLP marker:ABG373;SSCP marker:H1-AK250075,H4-AK252360,M9-Mot1;In/Del marker:F1-Ftsh4,W2-WG241.In which H1-AK250075,H4-AK252360,M9-Mot1,F1-Ftsh4,C1-CA608558 and W2-WG241 are new developed-markers,the others from public databases.4.By using the 14 obtained markers exploit the 563 F2 population,the BVP gene was mapped to the distal of chromosome 1HL which flanked by the gene-specific markers AK252360(0.2 cM)and CA608558(0.5 cM).5.Further analyses of the predicted Mot1 protein across the barley,wheat,rice,Brachypodium and Aegilops tauschii revealed higher amino acid identity between barley and wheat(96%identity)than between barley and rice(84%identity)or barley and Brachypodium(89%identity)6.A TILLING population of "Xiyin2" developed by treating with 30 mM EMS,has 17.35%variation and the mutant frequency is about 1 per 661 kb of EDR1 gene which is a key negative regulator of plant-powdery mildew interaction.7.Among 2012 M2 lines,we obtained three EDR1 mutants by the CEL1 based screening systems,it should be noted that a Ser to Pro amino acid change in the exons of an EDR1 mutant.