YANG Lei,ZHU Tongtong,LIU Yingbao,et al.Functional Analysis of Phospholipase PcPLD8 Gene From Phytophthora capsici[J].Northern Horticulture,2023,(17):1-7.[doi:10.11937/bfyy.20230315]
辣椒疫霉菌磷脂酶PcPLD8基因功能研究
- Title:
- Functional Analysis of Phospholipase PcPLD8 Gene From Phytophthora capsici
- 文章编号:
- 1001-0009(2023)17-0001-07
- 分类号:
- S 436.418
- 文献标志码:
- A
- 摘要:
-
辣椒是我国目前栽培面积最大的蔬菜之一，但其在生长过程中易受到多种病虫的危害，造成产量和品质的下降［1］。由辣椒疫霉菌（Phytophthora capsici）引起的辣椒疫病是一种世界分布的毁灭性土传病害，可引起辣椒根茎部黑腐、叶片脱落和果实腐烂，严重时植株枯萎死亡，给辣椒生产造成巨大的经济损失［2-3］。辣椒疫霉菌是一种异宗配合的卵菌病原菌，易发生变异，适应能力强，寄主范围广，可严重危害辣椒和其它作物［3-5］。现主要依靠化学农药防治辣椒疫病，但造成了病原菌的抗性增强、农药残留和环境污染。为更加有效地防治辣椒疫病，寻找辣椒疫霉菌的关键致病基因及其致病机制成为近年来的研究热点。磷脂酶是一类广泛存在于各种生物体内且可以水解甘油磷脂的酶，包括磷脂酶A（PLA1和PLA2）、磷脂酶B（PLB）、磷脂酶C（PLC）和磷脂酶D（PLD）。其中，磷脂酶D是一类特殊的酯键水解酶，它能水解磷脂生成磷脂酸和羟基化合物，并能催化某些含羟基的化合物结合到磷脂的酰基上，形成新的磷脂。PLD主要参与细胞信号传导、脂质代谢、植物生长发育、胁迫响应、免疫调节及真菌的生长发育和致病性调控等生理功能［6-10］。研究发现，酿酒酵母 （Saccharomyces cerevisiae）中的SPO14（PLD1）在膜脂质合成、细胞分裂、膜泡融合、孢子萌发等过程中发挥重要作用［11-13］。淡紫拟青霉（Paecilomyces lilacinus）PLD增强了该菌对南方根结线虫的寄生和致病能力［14］。禾谷镰刀菌（Fusarium graminearum）敲除FgPLD1后生长缓慢、产孢量少，致病力明显减弱［15］。在致病疫霉（P.infestans）中发现磷脂酶D分为6个亚家族，其中sPLD-like-1、sPLD-like-12和PLD-like-1在该菌侵染马铃薯过程中发挥作用，增强毒力，引起病害发生［16］。目前，植物和哺乳动物中PLD的作用已经研究的比较清楚，但植物病原真菌中PLD研究还相对较少，尤其是病原卵菌PLD的研究报道更少。长江大学微生物实验室前期成功从辣椒疫霉菌LT1534中分离鉴定到1个PLD基因，命名为PcPLD8，并对其进行了生物信息学分析［17］。为验证磷脂酶PcPLD8基因在辣椒疫霉菌致病中的作用，该研究通过烟草瞬时过表达对其进行了亚细胞定位分析，借由基因沉默和过表达技术筛选获得转化子，进行了菌丝形态、生长速率及致病力等表型分析，旨在为探索辣椒疫霉菌PLD的生物学功能和辣椒疫病的有效防控提供参考依据。
- Abstract:
- Phytophthora capsici was used as test material,the function of phospholipase D gene was studied by gene silencing and gene overexpression,the effects of gene on the growth and pathogenicity in P.capsici were revealed,in order to provide reference for further revealing the mechanism of phospholipase D in the pathogenesis of P.capsici. The results showed that the silenced transformants S1 and S5 of gene had sparse hyphae,many and short lateral branches,small colony diameter and slow growth rate,the pathogenicity to Nicotiana benthamiana decreased obviously,compared with the wild-type strain WT and the empty vector transformant CK. And over-expressed transformant OE2 showed little change in hypha morphology,growth rate and pathogenicity to N.benthamiana.
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备注/Memo
第一作者简介：杨磊（1995-），男，博士研究生，研究方向为病原微生物。E-mail：2524422614@qq.com. 责任作者：孙文秀（1979-），女，博士，教授，现主要从事植物病原卵菌致病机理等研究工作。E-mail：wenxiusun@163.com. 基金项目：国家自然科学基金资助项目（31701573）；湖北省教育厅科学研究计划重点资助项目（D20181302）。收稿日期：2023-02-04