Zhu Xiao-Guan, Chu Zhen-Jian, Ying Sheng-Hua, Feng Ming-Guang
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.
Fungal Biol. 2017 Nov;121(11):956-965. doi: 10.1016/j.funbio.2017.08.003. Epub 2017 Aug 21.
Krs is a class II lysyl-tRNA synthetase (KRS) that is involved in cytosolic protein synthesis in budding yeast but functionally has not been explored in filamentous fungi. Previous transcriptomic analysis has revealed that a Krs-coding gene is likely involved in pathogenesis of Beauveria bassiana, a classic insect pathogen as a global source of fungal insecticides. Here, we show that Krs is localized in the cytoplasm of hyphal cells and acts as a substantial virulence factor in B. bassiana. Deletion of krs resulted in 10-h delayed germination, decreased (15 %) thermotolerance, and lowered (46 %) UV-B resistance of aerial conidia despite limited impact on conidiation capacity and slight or inconspicuous influence on radial growth on rich and minimal media with different carbon (10 sugars/polyols) and nitrogen (17 amino acids) sources. The deletion mutant suffered 58 % reduction in submerged blastospore yield (an index of in vitro dimorphic transition rate) in a minimal medium, and the reduction increased to 71 % in another trehalose-based medium mimic to insect haemolymph. In standardized bioassays, median lethal actions of Δkrs against Galleria mellonella larvae through the infections passing and bypassing the insect cuticle were prolonged to 192 and 153 h from wild-type median lethal time (LT) estimates of 119 and 109 h, respectively. Microscopic examination revealed 2-d delayed presence of in vivo formed hyphal bodies in the haemolymph of the larvae infected by Δkrs in either mode. These findings unveil a vital role of Krs in conidial germination and dimorphic transition and its contribution to the fungal potential against arthropod pests.
Krs是一种II类赖氨酰-tRNA合成酶(KRS),参与芽殖酵母的胞质蛋白质合成,但在丝状真菌中的功能尚未得到探索。先前的转录组分析表明,一个编码Krs的基因可能参与球孢白僵菌的致病过程,球孢白僵菌是一种经典的昆虫病原体,也是全球真菌杀虫剂的来源。在这里,我们表明Krs定位于菌丝细胞的细胞质中,并在球孢白僵菌中作为一个重要的毒力因子发挥作用。krs的缺失导致萌发延迟10小时,耐热性降低(15%),气生分生孢子的抗UV-B能力降低(46%),尽管对分生孢子形成能力的影响有限,并且对在含有不同碳源(10种糖/多元醇)和氮源(17种氨基酸)的丰富和基本培养基上的径向生长有轻微或不明显的影响。缺失突变体在基本培养基中的浸没芽生孢子产量(体外双态转变率的指标)降低了58%,在另一种模拟昆虫血淋巴的海藻糖培养基中,降低率增加到71%。在标准化生物测定中,通过感染穿过和绕过昆虫角质层,Δkrs对大蜡螟幼虫的半数致死作用分别从野生型的半数致死时间(LT)估计值119小时和109小时延长至192小时和153小时。显微镜检查显示,在两种模式下,被Δkrs感染的幼虫血淋巴中体内形成的菌丝体的出现延迟了2天。这些发现揭示了Krs在分生孢子萌发和双态转变中的重要作用及其对真菌对抗节肢动物害虫潜力的贡献。