Genome-Wide Analysis Of Wheat RNA Helicase And Expression Analysis Of Two Drought Related Genes

Wheat plays a crucial role in the world and China’s food production,more than 35% of the world’s population uses wheat as their staple food;with the increase of population,people’s demand for wheat is also increasing.Drought is the main abiotic stress that affects crop growth and limit crop yield,and is also the most vulnerable to stress.Helicase is widely exists in the life a kind of regulatory proteins in the body,mainly involved in RNA synthesis,editing,copy,the output of the m RNA,stability,and other physiological processes of RNA degradation,and the studies have shown that it plays an important role in the process of plant resistance.In recent years,more and more research shows that it plays an important role in the process of plant resistance.In this paper,we performed a genome-wide analysis of the RNA helicase genes in wheat and combined with transcriptome analysis data,two genes that strongly respond to drought and high temperature were cloned and analyzed for expression patterns in wheat cultivars with different drought resistance,in the hope of providing good candidates for resistance breeding of wheat gene.The main contents are as follows:1.Using bioinformatics methods,we finally identified 256 wheat RNA helicase family members in the wheat genome;we constructed a system using the helicase gene and wheat RNA helicase gene already studied in Arabidopsis and other species evolutionary tree,according to its conserved domain,the wheat RNA helicase gene was divided into three subfamilies: DEAD,DEAH,and DEx D/H.At the same time,256 wheat RNA helicase genes were analyzed by chromosome mapping,gene structure analysis,protein structural analysis and cis-acting element analysis.Chromosomal localization analysis revealed that the wheat RNA helicase gene was concentrated in both ends of the chromosome,while the centromere centrosome was less distributed.Gene structural analysis revealed that the DEAD-box RNA helicase gene has a simpler structure and more conserved structural patterns than the other two subfamilies.Protein structure analysis showed that wheat RNA helicase protein contains multiple conservd motifs.Analysis of cis-acting elements in the promoter region of wheat RNA helicase gene show that the wheat helicase gene may participate in the response process of various stresses.2.Analysis of the evolution of wheat RNA helicase gene revealed that the Ka/Ks data of wheat RNA helicase gene was 0.00801-0.6058,indicating that the wheat RNA helicase gene had a purification and selection effect,indicating that the genes were conserved and could not produce new mutations.Analysis of expression patterns of all wheat RNA helicase genes in different tissues and under conditions of drought and high temperature stress found that the expression level of wheat RNA helicase gene was higher in leaves,stems and panicles,and was expressed in grains and roots less;analysis of expression under high temperature and drought stress indicated that most wheat RNA helicases were involved in the responses of high temperature and drought stress,among which DEAD-box genes strongly respond to drought and high temperature stress.3.Combined with bioinformatics analysis and drought transcriptome data of small white wheat,we screened two drought resistant genes,TaDEAD1-B and TaDEAD23-A,which were strongly induced by drought and high temperature,from 256 wheat RNA helicase genes.q RT-PCR was used to analyze TaDEAD1-B and TaDEAD23-A expression patterns under drought stress in drought-resistant cultivar Luohan 7 and Huaimai 20 and non-drought resistant cultivar Xinong 979 and Yannong 886.The results showed that the two genes were up-regulated after drought stress at the seedling and jointing stages of the four varieties,indicating that the two genes could be expressed continuously and efficiently after the wheat was subjected to drought stress.4.The CDs sequences of TaDEAD1-B and TaDEAD23-A genes were cloned from wheat c DNA and constructed into the prokaryotic expression vector p Cold TMTF.After induction by IPTG,the recombinant protein was obtained.the helicase functional verification of purified recombinant proteins revealed that both helicase genes are ATP-dependent helicase genes.