Institute of Biotechnology, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China.
Appl Microbiol Biotechnol. 2013 Jun;97(12):5433-45. doi: 10.1007/s00253-012-4593-z. Epub 2013 Jan 8.
The sucrose non-fermenting protein kinase 1 gene (SNF1) regulates the derepression of glucose-repressible genes in microorganisms. In this study, we cloned an ortholog of SNF1 from Penicillium digitatum and characterized its functions through a gene knock-out strategy. Growth of the PdSNF1 mutant (ΔPdSNF1) on the synthetic medium (SM) supplemented with pectin or polygalacturonic acid was severely disturbed. The appearance of disease symptoms on the ΔPdSNF1 mutant-inoculated citrus fruits was significantly delayed as well. The expression levels of the cell wall-degrading enzyme (CWDE) genes (e.g., XY1, PL1, PNL1, and EXPG2) after pectin induction were up-regulated in wild type, but unchanged or less up-regulated in the ΔPdSNF1 mutant. During infection in citrus fruit, the up-regulation of XY1 was delayed in the ΔPdSNF1 mutant. Disruption of PdSNF1 also resulted in impaired conidiation and caused malformation of the conidiophore structures. In addition, the expression of BrlA, a gene that regulates conidiophore development, was significantly impaired in the ΔPdSNF1 mutant. However, the expression of FadA, encoding the α-subunit of a heterotrimeric G protein, was up-regulated in this mutant. Collectively, our results demonstrate that the PdSNF1 plays a role in adapting P. digitatum to alternative carbon sources. Its involvements in the virulence of P. digitatum is probably via regulation of the expression of CWDE genes; and it is also involved in conidiation, probably through activation of the conidiation signaling pathway while inactivating the mycelial growth-signaling pathway.
蔗糖非发酵蛋白激酶 1 基因(SNF1)调节微生物中葡萄糖抑制基因的去阻遏。在这项研究中,我们从桔青霉中克隆了 SNF1 的同源物,并通过基因敲除策略对其功能进行了表征。PdSNF1 突变体(ΔPdSNF1)在补充果胶或多聚半乳糖醛酸的合成培养基(SM)上的生长受到严重干扰。在柑橘果实上接种ΔPdSNF1 突变体时,发病症状的出现也明显延迟。在果胶诱导后,细胞壁降解酶(CWDE)基因(如 XY1、PL1、PNL1 和 EXPG2)的表达水平在野生型中上调,但在ΔPdSNF1 突变体中不变或上调较少。在柑橘果实感染过程中,ΔPdSNF1 突变体中 XY1 的上调被延迟。PdSNF1 的破坏也导致分生孢子形成受损,并导致分生孢子结构畸形。此外,调控分生孢子发育的 BrlA 基因的表达在ΔPdSNF1 突变体中显著受损。然而,编码异三聚体 G 蛋白α亚基的 FadA 基因的表达在该突变体中上调。总之,我们的结果表明,PdSNF1 在桔青霉适应替代碳源中起作用。其在桔青霉毒力中的作用可能是通过调节 CWDE 基因的表达;它还参与分生孢子形成,可能通过激活分生孢子信号通路而失活菌丝生长信号通路。