Laboratory for Conservation and Utilization of Bio-Resources, and Key Laboratory for Microbial Resources of the Ministry of Education, Yunnan University, Kunming, 650091, People's Republic of China.
Appl Microbiol Biotechnol. 2014 Mar;98(6):2555-63. doi: 10.1007/s00253-013-5432-6. Epub 2013 Dec 10.
Malate synthase (Mls), a key enzyme in the glyoxylate cycle, is required for virulence in microbial pathogens. In this study, we identified the AoMls gene from the nematode-trapping fungus Arthobotrys oligospora. The gene contains 4 introns and encodes a polypeptide of 540 amino acids. To characterize the function of AoMls in A. oligospora, we disrupted it by homologous recombination, and the ΔAoMls mutants were confirmed by PCR and Southern blot analyses. The growth rate and colony morphology of the ΔAoMls mutants showed no obvious difference from the wild-type strains on potato dextrose agar (PDA) plate. However, the disruption of gene AoMls led to a significant reduction in conidiation, failure to utilize fatty acids and sodium acetate for growth, and its conidia were unable to germinate on minimal medium supplemented with sodium oleate. In addition, the trap formation was retarded in the ΔAoMls mutants, which only produced immature traps containing one or two rings. Moreover, the nematicidal activity of the ΔAoMls mutants was significantly decreased. Our results suggest that the gene AoMls plays an important role in conidiation, trap formation and pathogenicity of A. oligospora.
苹果酸合酶(Mls)是乙醛酸循环中的关键酶,对于微生物病原体的毒力是必需的。在本研究中,我们从食线虫真菌节丛孢菌中鉴定出 AoMls 基因。该基因包含 4 个内含子,编码 540 个氨基酸的多肽。为了研究 AoMls 在节丛孢菌中的功能,我们通过同源重组对其进行了敲除,并通过 PCR 和 Southern blot 分析对 ΔAoMls 突变体进行了验证。ΔAoMls 突变体在马铃薯葡萄糖琼脂(PDA)平板上的生长速率和菌落形态与野生型菌株没有明显差异。然而,基因 AoMls 的敲除导致产孢量显著减少,无法利用脂肪酸和醋酸钠进行生长,其分生孢子在补充油酸钠的基本培养基上无法发芽。此外,ΔAoMls 突变体中的陷阱形成受到抑制,仅产生含有一个或两个环的不成熟陷阱。此外,ΔAoMls 突变体的杀线虫活性显著降低。我们的结果表明,基因 AoMls 在节丛孢菌的产孢、陷阱形成和致病性中发挥重要作用。