【摘要】 【目的】研究刺激植物响应蛋白Epl1对木本植物山新杨的刺激响应功能,为开发新一代提高植物免疫的新型诱导剂奠定基础。【方法】从棘孢木霉ACCC30153菌株中克隆获得一个刺激植物响应蛋白基因Epl1,并进行序列分析和原核表达,将获得的原核重组蛋白rEpl1与山新杨组培苗进行互作,初步探讨rEpl1蛋白对山新杨生长、生理指标、以及生长和防御相关基因转录水平的影响。【结果】刺激植物响应蛋白基因Epl1的cDNA序列全长为417bp,预测蛋白分子量12.6 kDa,属于Cerato-platanin家族;通过构建原核表达载体,成功获得重组蛋白rEpl1;在1μg/mL rEpl1诱导下,山新杨组培苗生长较快、根系旺盛,生物量明显增加,CAT活性在诱导第1 d时为对照的3.51倍,可溶性糖含量和脯氨酸含量在诱导初期急剧积累;山新杨生长素基因LAX2/AUX和水杨酸防御基因JAR2的转录水平分别在蛋白诱导2 d和12 h时达到峰值,分别为对照的873.10和388.02倍。【结论】Epl1基因的克隆、序列分析和原核表达为进一步研究Epl1蛋白诱导植物系统抗病性提供了理论基础;重组蛋白rEpl1与山新杨组培苗互作实验,初步Epl1蛋白能够刺激木本植物在生理及分子水平上的响应,从而促进生长,提高抗性。更多还原

【Abstract】 [Objective]To develop a new inducer of enhancing plant immunity,the function of the eliciting plant response protein Epl1 to the woody plant Populus davidiana × P.alba var.pyramidlis(Pd Pap) was studied.[Method]An eliciting plant response protein gene Epl1 was cloned and analyzed from Trichoderma asperellum ACCC30536.The recombinant protein rEpl1 was obtained by prokaryotic expression,and the effect of rEpl1 was discussed on the growth,physical signs,and the transcription level of gene related to growth and defence of Pd Pap during the interaction between rEpl1 and Pd Paptissue culture seedlings.[Result]The results showed that the c DAN sequence of the gene Epl1 was417 bp long with a predicted protein of 12.6 k Da molecular weight,which belonged to the cerato-platanin family.The prokaryotic expression plasmid was constructed and the recombinant protein rEpl1 was successfully expressed.Under 1 μg/mL rEpl1 induction,the Pd Pap seedlings had an fast growth,developed root and the increasing biomass;the CAT activity of Pd Pap seedlings was 3.51 folds of the control at 1 st day,and the soluble sugar content and proline content of Pd Pap seedlings were sharply accumulated;the transcription levels of auxin gene(LAX2/AUX) and defence gene(JAR2) of Pd Pap seedlings reached the peak at 2 nd day and 12 hours,and the peak was 873.10 and 388.02 folds of the control,respectively.[Conclusion]So,the cloning,sequence analysis and prokaryotic expression of the gene Epl1 provided fundamental basis for studying Epl1 protein eliciting plant systemic resistance;through the interaction between the recombinant protein rEpl1 and Pd Pap seedlings,it was confirmed that Epl1 protein could stimulate the response of the wood plant on the level of physiology and molecular,which could promote plant growing and improve plant resistance.更多还原