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MaSnf1,一个蔗糖非发酵蛋白激酶基因,参与了绿僵菌的碳源利用、应激耐受和毒力。

MaSnf1, a sucrose non-fermenting protein kinase gene, is involved in carbon source utilization, stress tolerance, and virulence in Metarhizium acridum.

机构信息

Genetic Engineering Research Center, School of Life Sciences, Chongqing University, Chongqing, 400045, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2014 Dec;98(24):10153-64. doi: 10.1007/s00253-014-6066-z. Epub 2014 Sep 12.

Abstract

The protein kinase sucrose non-fermenting-1(Snf1) regulates the derepression of glucose-repressible genes and plays a major role in carbon source utilization. In this study, MaSnf1, a sucrose non-fermenting protein kinase gene, has been identified from the entomopathogenic fungus Metarhizium acridum, which has a great potential as a biocontrol agent. The functions of MaSnf1 were characterized using gene disruption and complementation strategies. Disruption of MaSnf1 reduced the conidial yield and delayed the conidial germination on potato dextrose agar (PDA) medium. MaSnf1 is also important for response to ultraviolet radiation and heat shock stress and carbon source utilization in M. acridum. Bioassays by topical inoculation and intrahemocoel injection showed that the MaSnf1 deletion mutant exhibited greatly reduced pathogenicity. The reduced expression level of chitinase gene (Chi) and protease gene (Pr1A) in MaSnf1-disruption transformant (ΔMaSnf1) most likely affects the initial penetration into its host. Additionally, the reduced expression level of acidic trehalase gene (ATM1) probably causes a decline in growth rate in insect hemolymph. Inactivation of MaSnf1 led to a significant decrease in virulence, probably owing to reduction in conidial germination, and appressorium formation, impairment in penetration, and decrease in growth rate in insect hemolymph.

摘要

蛋白激酶蔗糖非发酵 1(Snf1)调节葡萄糖抑制基因的去阻遏作用,并在碳源利用中起主要作用。在这项研究中,从昆虫病原真菌绿僵菌中鉴定出蔗糖非发酵蛋白激酶基因 MaSnf1,它作为生物防治剂具有巨大的潜力。使用基因敲除和互补策略来表征 MaSnf1 的功能。MaSnf1 的敲除减少了分生孢子的产量,并延迟了在土豆葡萄糖琼脂(PDA)培养基上的分生孢子萌发。MaSnf1 对于绿僵菌对紫外线辐射和热休克应激以及碳源利用的反应也很重要。通过表面接种和血腔注射的生物测定表明,MaSnf1 缺失突变体的致病性大大降低。MaSnf1 敲除转化体(ΔMaSnf1)中几丁质酶基因(Chi)和蛋白酶基因(Pr1A)的表达水平降低,很可能影响其宿主的初始穿透。此外,酸性海藻糖酶基因(ATM1)的表达水平降低可能导致在昆虫血淋巴中的生长速率下降。MaSnf1 的失活导致毒力显著降低,可能是由于分生孢子萌发和附着胞形成减少,穿透受损以及在昆虫血淋巴中的生长速率下降所致。

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