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脂肪酰辅酶A还原酶基因RcFAR1的功能表征及在月季愈伤组织中的抗虫性

Functional characterization of fatty acyl-CoA reductase gene RcFAR1 and its insect resistance in callus of Rosa chinensis

  • 摘要: 为了探究白粉菌侵染诱导月季对甜菜夜蛾产生抗性的分子机制,明确脂肪酰辅酶A还原酶基因RcFAR1在该过程中的功能。采用生物信息学方法对月季RcFARs基因家族进行鉴定与分析;通过RT-PCR技术克隆受白粉菌诱导上调表达的RcFAR1基因;利用农杆菌介导法将其转化月季愈伤组织,获得过表达RcFAR1的阳性愈伤;采用GC-MS检测阳性愈伤组织中脂肪醇类化合物的含量变化;通过产卵行为测试分析转基因愈伤组织对甜菜夜蛾雌虫产卵选择的影响。结果表明:月季RcFARs家族成员均具有典型的FAR保守结构域;过表达RcFAR1的阳性愈伤组织中1-十六醇和1-十八醇含量显著积累;甜菜夜蛾雌虫对空载体转化的对照愈伤组织具有显著产卵偏好,对转基因愈伤组织的产卵驱避率达45%。RcFAR1通过介导病原触发的脂肪醇生物合成,其产物作为嗅觉信号分子驱避甜菜夜蛾产卵。因此,建立了基于愈伤组织的植物-病原-昆虫三方互作模型,为解析植物诱导防御机制提供了简便有效的实验系统,RcFAR1可作为抗虫月季分子育种的重要候选靶点。

     

    Abstract: This study aims to investigate the molecular mechanism underlying the resistance of rose (Rosa chinensis) to the beet armyworm (Spodoptera exigua) induced by powdery mildew (Podosphaera pannosa) infection, and to clarify the function of the fatty acyl-CoA reductase gene RcFAR1 in this process.Bioinformatic methods were employed to identify and analyze the RcFARs gene family in rose. The RcFAR1 gene, which is up-regulated upon powdery mildew infection, was cloned using RT-PCR. It was then transformed into rose callus via Agrobacterium-mediated transformation to obtain callus overexpressing RcFAR1. Gas chromatography-mass spectrometry (GC-MS) was used to detect changes in fatty alcohol content in the positive callus. An oviposition bioassay was conducted to analyze the effect of the transgenic callus on the oviposition choice of female S. exigua moths.The results showed that all members of the rose RcFARs family possess the typical conserved FAR domain. The contents of 1-hexadecanol and 1-octadecanol were significantly accumulated in the RcFAR1-overexpressing callus. Female S. exigua moths showed a significant oviposition preference for control callus transformed with the empty vector, with an oviposition repellent rate of 45% against the transgenic callus.RcFAR1 mediates pathogen-triggered fatty alcohol biosynthesis, and its products act as olfactory signals to repel S. exigua oviposition. This study establishes a callus-based tripartite interaction model (plant-pathogen-insect), providing a simple and effective experimental system for deciphering plant-induced defense mechanisms. RcFAR1 represents a promising candidate target for the molecular breeding of insect-resistant rose.

     

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