协同抑制番茄ACO1对果实成熟及病程相关蛋白基因表达的影响

中国农业科学 2007,40(3):558-565

Scientia Agricultura Sinica

协同抑制番茄ACO1对果实成熟及病程相关

蛋白基因表达的影响

胡宗利1，陈国平1，吕丽娟1，陈绪清1， GRIERSON Donald 2

（1重庆大学生物工程学院，重庆400030；2 Plant Sciences Division, School of Biosciences, University of Nottingham, Sutton Bonington Campus,

Loughborough, Leicestershire LE12 5RD, UK）

摘要：【目的】探讨协同抑制番茄ACO1基因对果实成熟和病程相关蛋白基因表达、内源乙烯生物合成及果实耐贮性的影响。【方法】采用PCR或RT-PCR方法克隆了番茄ACC氧化酶1、ACC氧化酶3、EBF1、PR1、PR5以及

NP24基因片段，并以此制备探针，以协同抑制ACO1的转基因番茄和野生型番茄为研究对象，进行Northern杂交，

同时测定了伤害叶片和果实的乙烯释放量，并进行了果实贮藏试验等。【结果】Northern杂交结果表明，番茄ACO1

基因表达被抑制后，与果实成熟相关基因LeACO3和LeEBF1，以及病程相关蛋白基因LePR1、LePR5 和LeNP24的

表达量急剧降低。乙烯释放量测定试验和果实贮藏试验结果表明，协同抑制LeACO1番茄完整和受伤叶片以及完整

果实内源乙烯释放量相对于野生型番茄大大减少，成熟果实贮藏时间延长。【结论】协同抑制番茄ACO1基因表达

的同时，与果实成熟相关基因和病程相关蛋白基因的表达也不同程度地受到抑制，而且其内源乙烯生物合成减少，

果实耐贮性增强。

关键词：协同抑制；番茄ACO1；果实成熟；病程相关蛋白基因

The Influence of Co-suppressing Tomato 1-Aminocyclopropane-1 -Carboxylic Acid Oxidase I on the Expression of Fruit

Ripening-Related and Pathogenesis-Related Protein Genes

HU Zong-li1, CHEN Guo-ping1, Lü Li-juan1, CHEN Xu-qing1, GRIERSON Donald 2

(1College of Bioengineering, Chongqing University, Chongqing 400030; 2Plant Sciences Division, School of Biosciences,

University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE12 5RD, UK)

Abstract: 【Objective】The purpose of this study is to explore the influence of co-suppressing tomato ACC oxidase I on the

expression of fruit ripening-related and pathogenesis-related protein genes and on the biosynthesis of endogenous ethylene and

storage ability of fruits. 【Method】Specific fragments of several fruit ripening-related and pathogenesis-related protein genes from

tomato (Lycopersicon esculentum) were cloned, such as the 1-aminocyclopropane-1-carboxylic acid oxidase I gene (LeACO1),

1-aminocyclopropane-1-carboxylic acid oxidase III gene (LeACO3), EIN3-binding F-box 1 gene (LeEBF1), pathogenesis-related

protein 1 gene (LePR1), pathogenesis-related protein 5 gene (LePR5) and pathogenesis-related protein osmotin precursor gene

(LeNP24) by PCR or RT-PCR. Then these specific DNA fragments were used as probes to hybridize with the total RNAs

extracted from the wild type tomato (AC++) and the LeACO1 co-suppression tomatoes (V1187 and T4B), respectively. At the same time,

ethylene production measurement and storage experiment of tomato fruits were carried out.【Result】The hybridization results

indicated that the expression of fruit ripening-related genes, such as LeACO3 and LeEBF1, and pathogenesis-related protein genes,

such as LePR1, LePR5 and LeNP24, were reduced sharply. The ethylene production in the fruits and wounded leaves decreased.

收稿日期：2006-06-01；接受日期：2006-11-07

基金项目：教育部“留学回国基金”项目（教外司留[2005]55号）；教育部“春晖计划”项目（教外司留[2003]589号）

作者简介：胡宗利（1971-），女，重庆江津人，讲师，博士，研究方向为分子生物学。E-mail：huzongli71@。通讯作者陈国平（1959-），男，湖北大冶人，教授，研究方向为分子生物学与基因工程。E-mail：chenguoping@