Isolation And Preliminary Functional Analysis Of A CaRH36 Gene Of Capsicum Annuum

Hot pepper(Capsicum annuum L.)is a kind of higher important economic vegetables of our country,and is a a soil-borne,easy onset typical solanaceae plants but also by other different abiotic stress influence.Revealing the molecular mechanism of disease resistance and stress resistance in pepper,can promote the breeding and application disease-resistant or stress resistanc varieties,improve the pepper production efficiency.More attention were payyed on the function of the gene of Hot pepper by people.The DEAD-box RNA participated in RNA metabolism,played important role in biotic and abiotic stress,and related with the growth and development of plant.A novel gene CaRH36 was isolated from the hot pepper and its initial function was analyzed in this study.The results are as follow:1?With the arabidopsis DEAD-box RNA helicase homologous EST to design and synthesize specificity primer.Using the primer based on PCR cDNA library screening method from chili cDNA library to separate a gene encoded a DEAD-box RNA helicase cloning.Its length was 1815bp,and it contained 493 amino acid residue open reading frame.The sequence of CaRH36 amino acids containing ten conservative domain,we find it homologous with thaliana DEAD-box RNA helicase AtRH36 and Medicago truncatula DEAD-box RNA helicse MtRH1.The CaRH36 genes was infered as DEAD-box RNA an enzyme family member,named for CaRH36.2?Prediction of protein localization,we found that the gene may also be in its endoplasmic reticulum(membrane)and nucleus.Through real-time quantitative fluorescence qRT-PCR experiment,the conclusion is drawn that this gene were down-regulated in high temperature and low temperature,mechanical damage and bacteria.The gene were expressed in stems,leaves,the flowers of the pepper,and its expression in them were different.This showed that the gene played role in the growth and development of the hot pepper.3?By constructing the overexpression vector and using of agrobacterium mediated tobacco dordrecht transformation,got 34 transgenic plants and corresponded T1 generation seeds.Consistent with the use of growing the T1 generation seedlings,CaRH36 overexpression tobacco T1 plants showed resistance to high temperature and under high temperature transgenic tobacco show a lighter injuries and smaller conductivity compared with the control.