Wu Lijuan, Hu Xinkun, Yan Shen, Wu Zenglin, Tang Xuzhong, Xie Lei, Qiu Yujie, Li Rui, Chen Ji, Tian Mengliang
College of Agronomy, Sichuan Agricultural University, Chengdu 611130, China.
Institute of Ecology, China West Normal University, Nanchong 637009, China.
Curr Issues Mol Biol. 2024 Sep 22;46(9):10618-10632. doi: 10.3390/cimb46090629.
(Berk.) is a complex is formed by larvae and . Infestation by , interaction with host larvae, and fruiting body development are three crucial processes affecting the formation of . However, research on the molecular mechanism of formation has been hindered by the lack of effective genetic transformation protocols. Therefore, -mediated transformation (ATMT) was adopted to genetically transform two strains and optimize the transformation conditions. The results revealed that the most suitable strain for transformation was AGL1, and that the surfactant Triton X-100 could also induce ATMT, although less effectively than acetosyringone (AS). In addition, the endogenous promoters of genes had a stronger ability to drive the expression of the target gene than did the exogenous promoter. The optimal transformation conditions were as follows: AS and hygromycin B concentrations of 100 μM and 50 μg/mL, respectively; OD of 0.4; cocultivation at 18 °C for 24 h; and used within three passages. The results lay a foundation for the functional study of key regulatory genes involved in the formation of .
(伯克)是由幼虫和……形成的复合体。……的侵染、与宿主幼虫的相互作用以及子实体发育是影响……形成的三个关键过程。然而,由于缺乏有效的遗传转化方案,对……形成的分子机制的研究受到了阻碍。因此,采用农杆菌介导的转化(ATMT)对两种……菌株进行遗传转化并优化转化条件。结果表明,最适合……转化的菌株是AGL1,表面活性剂吐温X - 100也能诱导ATMT,尽管其效果不如乙酰丁香酮(AS)。此外,……基因的内源启动子驱动靶基因表达的能力比外源启动子更强。最佳转化条件如下:AS和潮霉素B的浓度分别为100 μM和50 μg/mL;……的OD为0.4;在18℃共培养24小时;并且在三代内使用。这些结果为参与……形成的关键调控基因的功能研究奠定了基础。