Nesher Iris, Minz Anna, Kokkelink Leonie, Tudzynski Paul, Sharon Amir
Department of Molecular Biology and Ecology of Plants, Tel Aviv University, Tel Aviv 69978, Israel.
Eukaryot Cell. 2011 Aug;10(8):1122-30. doi: 10.1128/EC.00321-10. Epub 2011 Apr 1.
Colletotrichum gloeosporioides is a facultative plant pathogen: it can live as a saprophyte on dead organic matter or as a pathogen on a host plant. Different patterns of conidial germination have been recognized under saprophytic and pathogenic conditions, which also determine later development. Here we describe the role of CgRac1 in regulating pathogenic germination. The hallmark of pathogenic germination is unilateral formation of a single germ tube following the first cell division. However, transgenic strains expressing a constitutively active CgRac1 (CA-CgRac1) displayed simultaneous formation of two germ tubes, with nuclei continuing to divide in both cells after the first cell division. CA-CgRac1 also caused various other abnormalities, including difficulties in establishing and maintaining cell polarity, reduced conidial and hyphal adhesion, and formation of immature appressoria. Consequently, CA-CgRac1 isolates were completely nonpathogenic. Localization studies with cyan fluorescent protein (CFP)-CgRac1 fusion protein showed that the CgRac1 protein is abundant in conidia and in hyphal tips. Although the CFP signal was equally distributed in both cells of a germinating conidium, reactive oxygen species accumulated only in the cell that produced a germ tube, indicating that CgRac1 was active only in the germinating cell. Collectively, our results show that CgRac1 is a major regulator of asymmetric development and that it is involved in the regulation of both morphogenesis and nuclear division. Modification of CgRac1 activity disrupts the morphogenetic program and prevents fungal infection.
它既可以在死的有机物上作为腐生菌生存,也可以在寄主植物上作为病原菌生存。在腐生和致病条件下,已经识别出不同的分生孢子萌发模式,这些模式也决定了后续的发育。在这里,我们描述了CgRac1在调节致病萌发中的作用。致病萌发的标志是在第一次细胞分裂后单侧形成单个芽管。然而,表达组成型活性CgRac1(CA-CgRac1)的转基因菌株显示同时形成两个芽管,在第一次细胞分裂后两个细胞中的细胞核都继续分裂。CA-CgRac1还导致了各种其他异常,包括建立和维持细胞极性困难、分生孢子和菌丝附着力降低以及不成熟附着胞的形成。因此,CA-CgRac1分离株完全无致病性。用青色荧光蛋白(CFP)-CgRac1融合蛋白进行的定位研究表明,CgRac1蛋白在分生孢子和菌丝尖端丰富。尽管CFP信号在萌发的分生孢子的两个细胞中均匀分布,但活性氧仅在产生芽管的细胞中积累,表明CgRac1仅在萌发细胞中活跃。总体而言,我们的结果表明CgRac1是不对称发育的主要调节因子,并且它参与形态发生和核分裂的调节。CgRac1活性的改变破坏了形态发生程序并阻止真菌感染。