Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Sciences, Nanjing Normal University, Nanjing 210023 Jiangsu, China.
Wuhan Sunhy Biology Co., Ltd.,Wuhan, 430000, Hubei, China.
Fungal Genet Biol. 2024 Aug;173:103899. doi: 10.1016/j.fgb.2024.103899. Epub 2024 May 25.
Fusarium head blight is a devastating disease that causes severe yield loses and mycotoxin contamination in wheat grain. Additionally, balancing the trade-off between wheat production and disease resistance has proved challenging. This study aimed to expand the genetic tools of the endophyte Phomopsis liquidambaris against Fusarium graminearum. Specifically, we engineered a UDP-glucosyltransferase-expressing P. liquidambaris strain (PL-UGT) using ADE1 as a selection marker and obtained a deletion mutant using an inducible promoter that drives Cas9 expression. Our PL-UGT strain converted deoxynivalenol (DON) into DON-3-G in vitro at a rate of 71.4 % after 36 h. DON inactivation can be used to confer tolerance in planta. Wheat seedlings inoculated with endophytic strain PL-UGT showed improved growth compared with those inoculated with wildtype P. liquidambaris. Strain PL-UGT inhibited the growth of Fusarium graminearum and reduced infection rate to 15.7 %. Consistent with this finding, DON levels in wheat grains decreased from 14.25 to 0.56 μg/g when the flowers were pre-inoculated with PL-UGT and then infected with F. graminearum. The expression of UGT in P. liquidambaris was nontoxic and did not inhibit plant growth. Endophytes do not enter the seeds nor induce plant disease, thereby representing a novel approach to fungal disease control.
镰刀菌穗腐病是一种毁灭性疾病,可导致小麦严重减产和真菌毒素污染。此外,平衡小麦生产和抗病性之间的权衡一直具有挑战性。本研究旨在扩展内生真菌拟茎点霉对禾谷镰刀菌的遗传工具。具体来说,我们使用 ADE1 作为选择标记,使用诱导型启动子驱动 Cas9 表达,对表达 UDP-葡萄糖基转移酶的拟茎点霉菌株 (PL-UGT) 进行了工程改造。我们的 PL-UGT 菌株在 36 小时后,将脱氧雪腐镰刀菌烯醇 (DON) 转化为 DON-3-G 的转化率为 71.4%。DON 失活可用于赋予植物体内的耐受性。与接种野生型拟茎点霉的小麦幼苗相比,接种内生菌株 PL-UGT 的小麦幼苗生长情况得到改善。菌株 PL-UGT 抑制了禾谷镰刀菌的生长,感染率降低到 15.7%。与此一致的是,当用 PL-UGT 预先接种花朵然后感染禾谷镰刀菌时,小麦籽粒中的 DON 水平从 14.25μg/g 降低到 0.56μg/g。PL-UGT 中的 UGT 表达无毒,不会抑制植物生长。内生菌不会进入种子,也不会引起植物病害,因此代表了一种控制真菌病害的新方法。