A Phytochrome-Interacting Factor Gene CaPIF7a Positively Regulates the Defense Response against Phytophthora capsici Infection in Pepper (Capsicum annuum L.)

Agronomy Pub Date : 2024-09-06 DOI:10.3390/agronomy14092035
Yu Li, Dan Wu, Ting Yu, Bing Liu, Xuchun Gao, Huibin Han, Jinyin Chen, Yong Zhou, Youxin Yang
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Abstract

Phytochrome-interacting factor (PIF) is a subfamily of the basic helix–loop–helix (bHLH) transcription factors (TFs) and plays key roles in plant responses to diverse biotic and abiotic stresses. In this work, a PIF gene named CaPIF7a was cloned and its role in the regulation of pepper’s resistance to Phytophthora capsici infection (PCI) was studied. The cloned CaPIF7a gene has a CDS length of 1383 bp, encodes a hydrophilic protein containing bHLH and APB characteristic domains, and subcellular localization results showed that CaPIF7a was located in the nucleus. Expression analysis showed that CaPIF7a gene has the highest expression level in leaf, and its expression was regulated under PCI and salicylic acid (SA) treatment. Silencing of CaPIF7a in pepper plants by virus-induced gene silencing (VIGS) reduces the resistance of pepper to PCI, with decreased expression of SA-responsive and SA-biosynthesis genes and obviously decreased SA content. DNA affinity purification sequencing (DAP-seq) was employed to identify the potential targets of CaPIF7a, and yeast one-hybrid (Y1H) verified that CaPIF7a could regulate the expression of CaHY5 by binding its promoter. These findings indicated that CaPIF7a might be a key modulator in plant immune response and presented a possible regulatory network of CaPIF7a in PCI.
植物色素相互作用因子基因 CaPIF7a 积极调控辣椒(Capsicum annuum L.)对疫霉菌感染的防御反应
植物色素互作因子(Phytochrome-interacting factor,PIF)是碱性螺旋环螺旋转录因子(bHLH)的一个亚家族,在植物对各种生物和非生物胁迫的反应中起着关键作用。本研究克隆了一个名为 CaPIF7a 的 PIF 基因,并研究了该基因在调控辣椒抗疫霉菌感染(PCI)中的作用。克隆的CaPIF7a基因CDS长度为1383 bp,编码一种亲水蛋白,含有bHLH和APB特征结构域,亚细胞定位结果显示CaPIF7a位于细胞核内。表达分析表明,CaPIF7a基因在叶片中的表达水平最高,其表达在PCI和水杨酸(SA)处理下受到调控。通过病毒诱导基因沉默(VIGS)技术沉默辣椒植株中的CaPIF7a基因,可降低辣椒对PCI的抗性,同时降低SA响应基因和SA生物合成基因的表达,并明显降低SA含量。DNA亲和纯化测序(DAP-seq)鉴定了CaPIF7a的潜在靶标,酵母单杂交(Y1H)验证了CaPIF7a可通过结合CaHY5的启动子调控其表达。这些发现表明CaPIF7a可能是植物免疫反应中的一个关键调控因子,并提出了CaPIF7a在PCI中可能的调控网络。
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