农业生物技术学报2025，Vol.33Issue(3)：523-535,13.DOI:10.3969/j.issn.1674-7968.2025.03.005
利用杜仲漆酶基因(EuLAC1)创制抗灰霉病番茄新种质
Creating New Tomato(Solanum lycopersicum)Germplasm Resistance to Gray Mold Using Eucommia ulmoides Laccase 1(EuLAC1)Gene
摘要
Abstract
Laccase(LAC)is a glycoprotein oxidase that influences lignin synthesis,regulates the levels of phenolic compounds in plants,and participates in plant defense mechanisms against pests and diseases.In order to obtain a new tomato(Solanum lycopersicum)germplasm with high resistance to Botrytis cinerea and in compliance with transgenic safety,in this study,based on the pGM626-Act1 vector constructed in the laboratory in the previous stage,a gene-deleter system with the fruit-specific E8 promoter driving the FLP gene and the Act1 promoter driving the Eucommia ulmoides laccase 1(EuLAC1)gene in a plant expression vector was designed and constructed.Agrobacterium-mediated transformation technology was used to genetically transform the tomato variety'Micro-Tom'and 5 positive tomato transgenic plants overexpressing EuLAC1 were successfully obtained.The results of resistance analysis showed that the onset time of EuLAC1 overexpressing tomato plants was significantly delayed after inoculation with B.cinerea,which was in sharp contrast with wild type and empty vector control plants.Additionally,the lesion diameter in tomato plants overexpressing the EuLAC1 gene was significantly smaller than that in the control group(P＜0.05),which comprised the wild type and empty vector tomato plants.The result showed that overexpressing the EuLAC1 gene in tomato plants markedly improved their resistance to B.cinerea.Overexpression of the EuLAC1 gene also increased the activity of protective enzymes and the expression level of disease process-related protein(PR)genes in tomato plants.Deletion analysis of exogenous genes in transgenic tomato fruits revealed that no exogenous genes were detected in 22 out of 40 tomato fruits,with an exogenous gene deletion efficiency of 55％.This study provides a new technical method for the development of new disease-resistant tomato germplasm that meets transgenic safety standards.关键词
杜仲漆酶(EuLAC1)/番茄/遗传转化/灰霉病/抗病性分析Key words
Eucommia ulmoides laccase 1(EuLAC1)/Tomato/Genetic transformation/Gray mold/Disease resistance analysis分类
植物保护学引用本文复制引用
杨金玉,王作日,赵德刚,赵懿琛..利用杜仲漆酶基因(EuLAC1)创制抗灰霉病番茄新种质[J].农业生物技术学报,2025,33(3):523-535,13.基金项目
中央引导地方科技发展资金(黔科合中引地[2023]009) （黔科合中引地[2023]009）
国家自然科学基金(31870285 （）
31660076) （）
贵大人基合字(2021)82号 （2021）
