橡胶树胶孢炭疽菌P型ATP酶基因CgATPase 01的敲除及表型分析

Knockout and Phenotypic Analysis of the CgATPase 01 Gene of Colletotrichum gloeosporioides

为了了解橡胶树胶孢炭疽菌的一个钙转运P型ATP酶基因Cg ATPase01的功能,本研究利用Splitmarker技术构建敲除载体,通过PEG介导原生质体转化对橡胶树胶孢炭疽菌菌株HBCg01的Cg ATPase 01基因进行敲除,根据表型变化及致病性检测对其功能进行分析。结果表明:通过Split-marker方法获得了敲除突变体△Cg ATPase01,表型分析发现该突变体与野生菌株间存在明显差异,病菌气生菌丝的颜色发生了改变,生长速率降低,产孢量下降,对橡胶树叶片的致病性减弱,推测该基因可能在橡胶树胶孢炭疽菌的产孢、生长发育和致病性等方面具有重要作用。本研究结果为进一步研究胶孢炭疽菌P型ATP酶基因功能和病原菌的致病机理奠定了基础。

To understand the function of cadmium-translocating P-type ATPase gene CgA TPase 01 in Colletotrichum gloeosporioides from rubber tree, the Split-marker strategy was applied to build a knockout cassette. The transformants which knocked out CgATPase 01 gene were obtained through PEG-mediated protoplast transformation. The function of CgATPase 01 gene was analyzed according to the phenotype and pathogenicity detection of the mutant. Results： The mutant of ΔCgATPase 01 was obtained by Split-Marker method. The phenotypic analysis revealed that there was significant difference between A CgATPase 01 and wild type. The mutant had a slower growth rate on plates, and reduced conidiation compared to the wild-type strain. Additionally, the mutants showed decreased pathogenicity to rubber tree. Therefore the CgATPase 01 gene in C. gloeosporioides plays an important role on conidiation, growth and pathogenicity. This study would lay a foundation for further study on the function of P-type ATPase gene and the pathogenic mechanism in C. gloeosporioides.