Fang Weiguo, Pei Yan, Bidochka Michael J
Department of Biological Sciences, Brock University, 500 Glenridge Avenue, St Catharines, ON L2S 3A1, Canada.
Biotechnology Research Center, Southwest University, Beibei Chongqing 400716, P. R. China.
Microbiology (Reading). 2007 Apr;153(Pt 4):1017-1025. doi: 10.1099/mic.0.2006/002105-0.
Regulators of the G protein signalling (RGS) pathway have been implicated in the control of a diverse array of cellular functions, including conidiation in filamentous fungi. However, the regulatory processes involved in conidiation in insect-pathogenic fungi are poorly understood. Since conidia are the infective propagules in these fungi, an understanding of the regulatory processes involved in conidiation is essential to the development of an effective biocontrol fungus. Here, the cloning and characterization of an RGS protein gene, cag8 (conidiation-associated gene), from the insect-pathogenic fungus Metarhizium anisopliae is reported. Phylogenetic analysis showed that CAG8 was orthologous to the RGS protein FlbA from Aspergillus nidulans. Complementation of A. nidulans DeltaflbA, which cannot conidiate, with M. anisopliae cag8 restored conidiation. Gene disruption of cag8 in M. anisopliae resulted in the lack of conidia on agar plates and on infected insects, reduced mycelial growth, decreased virulence, lysis during growth in liquid medium as well as lack of pigmentation and irregularly shaped blastospores. Transcript levels of ssgA (hydrophobin-encoding gene) were markedly reduced in a Deltacag8 strain, while pr1A (subtilisin-like protease) transcription was unaffected. These results suggest that cag8 is involved in the modulation of conidiation, virulence and hydrophobin synthesis in M. anisopliae.
G蛋白信号通路调节因子(RGS)已被证明参与控制多种细胞功能,包括丝状真菌的分生孢子形成。然而,人们对昆虫病原真菌分生孢子形成过程中的调控机制了解甚少。由于分生孢子是这些真菌的感染性繁殖体,因此了解分生孢子形成过程中的调控机制对于开发有效的生物防治真菌至关重要。本文报道了从昆虫病原真菌绿僵菌中克隆和鉴定一个RGS蛋白基因cag8(分生孢子形成相关基因)。系统发育分析表明,CAG8与构巢曲霉的RGS蛋白FlbA直系同源。用绿僵菌cag8对不能形成分生孢子的构巢曲霉ΔflbA进行互补,恢复了分生孢子形成能力。绿僵菌中cag8的基因破坏导致琼脂平板和被感染昆虫上分生孢子缺失、菌丝生长减少、毒力降低、在液体培养基中生长时裂解以及缺乏色素沉着和芽生孢子形状不规则。在Δcag8菌株中,疏水蛋白编码基因ssgA的转录水平显著降低,而枯草杆菌蛋白酶样蛋白酶pr1A的转录不受影响。这些结果表明,cag8参与了绿僵菌分生孢子形成、毒力和疏水蛋白合成的调控。