State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.
Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN 47907, USA.
Mol Plant Pathol. 2018 Apr;19(4):909-921. doi: 10.1111/mpp.12576. Epub 2017 Sep 20.
Fusarium graminearum is a causal agent of wheat scab disease and a producer of deoxynivalenol (DON) mycotoxins. Treatment with exogenous cyclic adenosine monophosphate (cAMP) increases its DON production. In this study, to better understand the role of the cAMP-protein kinase A (PKA) pathway in F. graminearum, we functionally characterized the PKR gene encoding the regulatory subunit of PKA. Mutants deleted of PKR were viable, but showed severe defects in growth, conidiation and plant infection. The pkr mutant produced compact colonies with shorter aerial hyphae with an increased number of nuclei in hyphal compartments. Mutant conidia were morphologically abnormal and appeared to undergo rapid autophagy-related cell death. The pkr mutant showed blocked perithecium development, but increased DON production. It had a disease index of less than unity and failed to spread to neighbouring spikelets. The mutant was unstable and spontaneous suppressors with a faster growth rate were often produced on older cultures. A total of 67 suppressor strains that grew faster than the original mutant were isolated. Three showed a similar growth rate and colony morphology to the wild-type, but were still defective in conidiation. Sequencing analysis with 18 candidate PKA-related genes in three representative suppressor strains identified mutations only in the CPK1 catalytic subunit gene. Further characterization showed that 10 of the other 64 suppressor strains also had mutations in CPK1. Overall, these results showed that PKR is important for the regulation of hyphal growth, reproduction, pathogenesis and DON production, and mutations in CPK1 are partially suppressive to the deletion of PKR in F. graminearum.
镰刀菌禾谷孢是小麦赤霉病的病原体,也是脱氧雪腐镰刀菌烯醇(DON)真菌毒素的产生者。用外源环腺苷酸(cAMP)处理会增加其 DON 产量。在这项研究中,为了更好地理解 cAMP-蛋白激酶 A(PKA)途径在禾谷镰刀菌中的作用,我们对编码 PKA 调节亚基的 PKR 基因进行了功能表征。缺失 PKR 的突变体是有活力的,但在生长、产孢和植物感染方面表现出严重缺陷。pkr 突变体产生的菌落紧凑,气生菌丝较短,菌丝隔室中的细胞核数量增加。突变体分生孢子形态异常,似乎经历了快速的自噬相关细胞死亡。pkr 突变体表现出产囊体发育受阻,但 DON 产量增加。它的疾病指数小于 1,无法传播到相邻的小穗。该突变体不稳定,在较老的培养物上经常产生生长速度更快的自发抑制子。总共分离出 67 株生长速度快于原始突变体的抑制子菌株。其中 3 株的生长速度和菌落形态与野生型相似,但产孢仍有缺陷。对 3 个代表性抑制子菌株中的 18 个候选 PKA 相关基因进行测序分析,仅发现 CPK1 催化亚基基因发生突变。进一步的表征表明,其他 64 株抑制子菌株中的 10 株也存在 CPK1 突变。总的来说,这些结果表明 PKR 对菌丝生长、繁殖、致病性和 DON 产生的调节很重要,CPK1 中的突变对禾谷镰刀菌中 PKR 的缺失有部分抑制作用。