Studies On Differential Proteomics Of Induced Resistance In Eucalyptus To Calonectria Pseudoreteaudii With Jasmonic Acid

Eucalyptus spp.is one of the three fast-growing forestation species in the world.It has important economic value in our country.Cylindrocladium leaf blight is a highly destructive disease for eucalyptus seedlings produced.Currently,chemical control is the main prevention to this disaster,but it has limitation as following:unsatisfactory control effect,pesticide residues,environmentally unfriendly and drug resistance.Therefore,it is an important and feasible way to combat this disease by enhances the disease resistance.JAs is one of the important chemical signal molecules that can start antiviral defense mechanism of host plants,besides we had found an up-regulated expression gene,JA mRNA domain,in the subtracted cDNA library of Eucalyptus grandis×E.urophylla 9224 resistant to Calonectria pseudoreteaudii.In this paper,we used proteomic technology to explore the impact of Calonectria pseudoreteaudii on Eucalyptus grandis 3 and E.grandis 3×E.urohylla guang 9.Meanwhile,we used the same way to investigate the mechanism of the host plants resistant to pathogen while being induced with the exogenous application of MeJA.The preliminary results were showed as the follow:1.TCA/acetone/phenol extraction was the most suitable extraction method for the eucalyptus leaves.2.When the lysis buffer was consisted of 7mol.L-1 Urea,2mol.L-1 Thiourea,4%CHAPS,60 mmol.L-1DTT and 0.5%IPG Buffer,the protein sample dissolved quite well.It is applicable to dissolve eucalyptus leaf protein.3.A higher-quality 2-DE map can get when use a 18cm,pH4-7 linear strip with 50?g sample volume and silver stain the gel.4.Differential expression proteins of the tested two resistant and susceptible material that under stress of Calonectria pseudoreteaudii after 24h were identified by doing MALDI-TOF-TOF/MS analysis.Ultimately,20 proteins were identified successfully.Function analysis of these proteins showed that:in addition to three EST corresponding proteins,three proteins of unknown function and two predicted proteins,these proteins were mainly involved in a plurality of physiological processes as antiviral defense,signal transduction and transcription,protein biosynthesis and modification,material and energy metabolism etc.The analysis revealed that:different eucalyptus species under the stress of Calonectria pseudoreteaudii their own defense ways there may be different,for disease resistance protein(TIR-NBS-LRR class)significantly upregulated in susceptible specie Eucalyptus grandis 3,while did no see significant expression in resistant species E.grandis 3×E.urohylla guang 9.5.Differential expression proteins of the tested two resistant and susceptible material that resistant to pathogen while being induced with the exogenous application of MeJA were identified by doing MALDI-TOF-TOF/MS analysis.Ultimately 15 proteins were identified successfully.Function analysis of these proteins showed that:In addition to three proteins of unknown function and two predicted proteins,these proteins were involved in several physiological processes as disease defense reaction,matter and energy metabolism,protein biosynthesis and other biochemical processes,such as tRNA modification and DNA repair.Analysis found:in the process of inducing resistance in eucalyptus to Calonectria pseudoreteaudii with exogenous MeJA,the upward trends of most up-regulated express proteins such as ADR 1-L1 protein,RP,tRNA-IPT,GPAT1 and DUF protein were clearly greater in Eucalyptus grandis 3 than E.grandis 3×E.urohylla guang 9,indicating that the value of exogenous MeJA in inducing eucalyptus disease resistance was more obvious in susceptible species.In addition,the respective differentially expressed proteins of MeJA-mediated induced disease resistance and Calonectria pseudoreteaudii stressed disease resistance were quite different,indicating that the defense ways of both may be different.