Xie Wanfeng, Lai Xiaolin, Wu Yuxiao, Li Zheyu, Zhu Jingwen, Huang Yu, Zhang Feiping
Jinshan College, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Key Laboratory of Integrated Pest Management in Ecological Forests (Fujian Province University), Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Plants (Basel). 2024 Sep 24;13(19):2672. doi: 10.3390/plants13192672.
Pine wilt disease, caused by , is a highly destructive and contagious forest affliction. Often termed the "cancer" of pine trees, it severely impacts the growth of Masson pine (). Previous studies have demonstrated that ectopic expression of the gene from in notably enhances resistance to pine wilt nematode infection. To further elucidate the transcriptional regulation and protein interactions of the in in response to pine wilt nematode infection, we cloned a 1984 bp promoter fragment of the gene, a transient expression vector was constructed by fusing this promoter with the reporter gene, which successfully activated the expression. DNA pull-down assays identified PmMYB8 as a trans-acting factor regulating gene expression. Subsequently, we found that the PmACRE1 protein interacts with several proteins, including the ATP synthase CF1 α subunit, ATP synthase CF1 β subunit, extracellular calcium-sensing receptor (PmCAS), caffeoyl-CoA 3-O-methyltransferase (PmCCoAOMT), glutathione peroxidase, NAD+-dependent glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase 1, cinnamyl alcohol dehydrogenase, auxin response factor 16, and dehydrin 1 protein. Bimolecular fluorescence complementation (BiFC) assays confirmed the interactions between PmACRE1 and PmCCoAOMT, as well as PmCAS proteins in vitro. These findings provide preliminary insights into the regulatory role of PmACRE1 in 's defense against pine wilt nematode infection.
松材线虫病由[病原体名称未给出]引起，是一种极具破坏性和传染性的森林病害。它常被称为松树的“癌症”，严重影响马尾松（[马尾松学名未给出]）的生长。先前的研究表明，将来自[物种名称未给出]的[基因名称未给出]基因在[宿主名称未给出]中异位表达，能显著增强对松材线虫感染的抗性。为了进一步阐明[宿主名称未给出]中[基因名称未给出]在响应松材线虫感染时的转录调控和蛋白质相互作用，我们克隆了[基因名称未给出]基因的一个1984 bp的启动子片段，通过将该启动子与报告基因[报告基因名称未给出]融合构建了一个瞬时表达载体，该载体成功激活了[报告基因名称未给出]的表达。DNA下拉实验确定PmMYB8为调控[基因名称未给出]基因表达的反式作用因子。随后，我们发现PmACRE1蛋白与几种蛋白质相互作用，包括ATP合酶CF1α亚基、ATP合酶CF1β亚基、细胞外钙敏感受体（PmCAS）、咖啡酰辅酶A 3 - O -甲基转移酶（PmCCoAOMT）、谷胱甘肽过氧化物酶、NAD⁺依赖性甘油醛 - 3 -磷酸脱氢酶、磷酸甘油酸激酶1、肉桂醇脱氢酶、生长素响应因子16和脱水素1蛋白。双分子荧光互补（BiFC）实验在体外证实了PmACRE1与PmCCoAOMT以及PmCAS蛋白之间的相互作用。这些发现为PmACRE1在[宿主名称未给出]抵御松材线虫感染中的调控作用提供了初步见解。
