Idnurm Alexander, Howlett Barbara J
School of Botany, The University of Melbourne, Victoria 3010, Australia.
Eukaryot Cell. 2002 Oct;1(5):719-24. doi: 10.1128/EC.1.5.719-724.2002.
A pathogenicity gene has been identified in Leptosphaeria maculans, the ascomycetous fungus that causes blackleg disease of canola (Brassica napus). This gene encodes isocitrate lyase, a component of the glyoxylate cycle, and is essential for the successful colonization of B. napus. It was identified by a reverse genetics approach whereby a plasmid conferring hygromycin resistance was inserted randomly into the L. maculans genome. Twelve of 516 transformants tested had reduced pathogenicity on cotyledons of B. juncea and B. napus, and 1 of these 12 had a deletion of the isocitrate lyase gene, as well as an insertion of the hygromycin resistance gene. This mutant was unable to grow on fatty acids, including monolaurate, and the isocitrate lyase transcript was not detected. When the wild-type gene was reintroduced into the mutant, growth on monolaurate was restored and pathogenicity was partially restored. L. maculans isocitrate lyase is produced during infection of B. napus cotyledons, while the plant homologue is not. When 2.5% glucose was added to the inoculum of the isocitrate lyase mutant, lesions of sizes similar to those caused by wild-type isolate M1 developed on B. napus cotyledons. These findings suggest that the glyoxylate pathway is essential for disease development by this plant-pathogenic fungus, as has been shown recently for a fungal and bacterial pathogen of animals and a bacterial pathogen of plants. Involvement of the glyoxylate pathway in pathogenesis in animals and plants presents potential drug targets for control of diseases.
在引起油菜(甘蓝型油菜)黑胫病的子囊菌——大斑壳针孢菌中,已鉴定出一个致病基因。该基因编码异柠檬酸裂解酶,这是乙醛酸循环的一个组成部分,对甘蓝型油菜的成功定殖至关重要。它是通过反向遗传学方法鉴定出来的,即将一个赋予潮霉素抗性的质粒随机插入大斑壳针孢菌基因组中。在测试的516个转化体中,有12个在芥菜型油菜和甘蓝型油菜子叶上的致病性降低,其中1个缺失了异柠檬酸裂解酶基因,同时插入了潮霉素抗性基因。该突变体无法在包括月桂酸单酯在内的脂肪酸上生长,且未检测到异柠檬酸裂解酶转录本。当将野生型基因重新导入该突变体时,其在月桂酸单酯上的生长得以恢复,致病性也部分恢复。在感染甘蓝型油菜子叶期间会产生大斑壳针孢菌异柠檬酸裂解酶,而植物同源物则不会。当向异柠檬酸裂解酶突变体的接种物中添加2.5%的葡萄糖时,甘蓝型油菜子叶上会出现与野生型分离株M1引起的病斑大小相似的病斑。这些发现表明,乙醛酸途径对于这种植物病原真菌的病害发展至关重要,最近对动物的一种真菌和细菌病原体以及植物的一种细菌病原体的研究也表明了这一点。乙醛酸途径参与动植物发病机制为疾病控制提供了潜在的药物靶点。