Development of a transformation system for Hirsutella spp. and visualization of the mode of nematode infection by GFP-labeled H. minnesotensis.
Development of a transformation system for Hirsutella spp. and visualization of the mode of nematode infection by GFP-labeled H. minnesotensis.
复制标题
被毛菌转化系统的开发。
DOI：
10.1038/srep10477
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发表时间：
2015-07-20
影响因子：
4.6
通讯作者：
Xiang M
中科院分区：
文献类型：
--
作者：
Sun J;Park SY;Kang S;Liu X;Qiu J;Xiang M
红丝毛孢和明尼苏达毛孢是自然界中大豆囊线虫（Heterodera glycines）第二阶段幼虫（J2）的内生真菌。它们也寄生于琼脂板上的甘氨酸H. J2和秀丽隐杆线虫。建立了农杆菌介导的Hirsutella spp转化条件，得到的转化子与相应的野生型菌株相似。显微镜下分析了表达ZsGreen的明尼苏达血吸虫感染甘氨酸H. J2和秀丽隐杆线虫第二幼虫期（L2）的过程。秀丽隐杆线虫L2和甘氨酸螺旋线虫J2的分生孢子分别在12 ~ 24 hpi和12 ~ 32 hpi之间形成感染钉，在接种后8 h内与经过的线虫粘附，在8 ~ 12 hpi之间形成感染钉。在线虫体腔内，秀丽隐杆线虫L2的菌丝增殖速度约为32 hpi，甘氨酸线虫J2的菌丝增殖速度约为40 hpi。对于秀丽隐杆线虫L2和甘氨酸线虫J2，真菌消耗整个身体并生长产生分生孢子，分别约为156和204 hpi。有效的转化方案和对感染过程的更好理解为研究线虫寄生的分子和细胞机制提供了坚实的基础。
Hirsutella rhossiliensis and H. minnesotensis are endoparasitic fungi of the second-stage juvenile (J2) of the soybean cyst nematode (Heterodera glycines) in nature. They also parasitize both H. glycines J2 and Caenorhabditis elegans on agar plates. Agrobacterium tumefaciens-mediated transformation conditions were established for these Hirsutella spp. The resulting transformants were similar to the corresponding wild-type strains. The infection processes of H. glycines J2 and C. elegans second larval stage (L2) by H. minnesotensis expressing ZsGreen were microscopically analyzed. Conidia of H. minnesotensis adhered to passing nematodes within 8 h post-inoculation (hpi), formed an infection peg between 8 and 12 hpi and penetrated the nematode cuticle between 12 and 24 hpi for C. elegans L2 and between 12 and 32 hpi for H. glycines J2. Hyphal proliferation inside of the nematode coelom was observed at approximately 32 hpi for C. elegans L2 and at approximately 40 hpi for H. glycines J2. The fungus consumed the whole body and grew out to produce conidia at approximately 156 and 204 hpi for C. elegans L2 and H. glycines J2, respectively. The efficient transformation protocol and a better understanding of infection process provide a solid foundation for studying the molecular and cellular mechanisms underlying fungal parasitism of nematodes.