Ku70 or Ku80 deficiencies in the fungus Botrytis cinerea facilitate targeting of genes that are hard to knock out in a wild-type context
Ku70 or Ku80 deficiencies in the fungus Botrytis cinerea facilitate targeting of genes that are hard to knock out in a wild-type context
复制标题
DOI：
10.1111/j.1574-6968.2008.01388.x
复制
发表时间：
2008-12-01
影响因子：
2.1
通讯作者：
Viaud, Muriel
中科院分区：
文献类型：
--
作者：
Choquer, Mathias;Robin, Guillaume;Viaud, Muriel
丝状子囊菌灰霉病菌（Botrytis cinerea）是了解植物病原真菌坏死性行为的研究最多的模型之一。对两株灰孢杆菌（B05.10和T4）的基因组进行了测序，这可能有助于阐明灰孢杆菌毒力多态性。在这项研究中，这两种菌株都进行了基因改造，以构建旨在靶向难以敲除的基因的受体菌株。B05.10菌株的BcKu70基因缺失和T4菌株的BcKu80基因缺失均影响了非同源末端连接（NHEJ） DNA修复机制。NHEJ在真菌转化过程中负责基因替代盒的异位整合，并导致同源重组（HR）的频率较低。Ku缺乏对葡萄球菌的体外和植株生长没有影响，但明显提高了倍半萜环化酶编码基因Cnd15的HR效率，该基因在野生型菌株中很难敲除。
The filamentous ascomycete Botrytis cinerea is one of the most studied models for understanding the necrotrophic behaviour of phytopathogenic fungi. The genomes of two strains of B. cinerea have been sequenced (B05.10 and T4), which may contribute to elucidating the virulence polymorphism in this fungus. In this study, both strains were genetically modified in order to construct recipient strains designed to target genes that are hard to knock out. Deletions of BcKu70 gene in B05.10 strain and BcKu80 gene in T4 strain both affected the nonhomologous end-joining (NHEJ) DNA repair mechanism. NHEJ is responsible for the ectopic integration of gene replacement cassettes during fungal transformation and leads to a lower frequency of homologous recombination (HR). Ku deficiencies in B. cinerea did not disturb in vitro or in planta growth, but clearly improved HR efficiency for the putative sesquiterpene cyclase-encoding gene Cnd15, which was hard to knock out in a wild-type strain.