Chen Yixiong, Feng Peng, Shang Yanfang, Xu Yong-Jiang, Wang Chengshu
Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Fungal Genet Biol. 2015 Aug;81:142-9. doi: 10.1016/j.fgb.2014.10.018. Epub 2014 Nov 18.
Fungal polyketide synthases (PKSs) and their related gene clusters are highly diversified at both inter- and intra-specific levels. The most well characterized PKS enzymes include those responsible for the biosynthesis of polyketide pigments such as melanins. The genome of the insect pathogenic fungus Metarhizium robertsii contains 20 type I PKSs but none has been functionally characterized. In this study, two PKS genes (designated as MrPks1 and MrPKs2) showing homologies to those counterparts for the biosynthesis of heptaketide pigments and dihydroxynaphthalene (DHN)-melanins, respectively, were deleted in two different strains of M. robertsii. The results indicated that disruption of MrPks1 but not MrPks2 impaired fungal culture pigmentation and cell wall structure. In addition to the negative effect of the DHN-melanin pathway inhibitor, it was postulated that DHN-melanin would not be produced by M. robertsii. Various assays revealed that the stress resistance abilities against ultraviolet radiation, heat shock and oxidants, as well as virulence against insects were not impaired in ΔMrPks1 and ΔMrPks2 isolates when compared with the wild-type strain. Thus, the non-melanin pigment(s) produced by the fungus do not contribute to cell damage protection and pathogenicity in M. robertsii. Physiological differences were evident in the two examined wild-type strains. The results from this study advance the understanding of functional divergence of fungal PKSs.
真菌聚酮合酶(PKSs)及其相关基因簇在种间和种内水平上高度多样化。最具特征的PKS酶包括那些负责聚酮色素(如黑色素)生物合成的酶。昆虫病原真菌罗伯茨绿僵菌的基因组包含20种I型PKSs,但尚未对其进行功能表征。在本研究中,分别在两种不同的罗伯茨绿僵菌菌株中缺失了两个与七酮色素和二羟基萘(DHN)-黑色素生物合成对应基因同源的PKS基因(分别命名为MrPks1和MrPKs2)。结果表明,MrPks1的缺失而非MrPks2的缺失会损害真菌培养物的色素沉着和细胞壁结构。除了DHN-黑色素途径抑制剂的负面影响外,推测罗伯茨绿僵菌不会产生DHN-黑色素。各种分析表明,与野生型菌株相比,ΔMrPks1和ΔMrPks2分离株对紫外线辐射、热休克和氧化剂的抗逆能力以及对昆虫的毒力均未受损。因此,该真菌产生的非黑色素对罗伯茨绿僵菌的细胞损伤保护和致病性没有贡献。在所检测的两种野生型菌株中生理差异明显。本研究结果推进了对真菌PKSs功能差异的理解。