State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Key Laboratory of Biopesticide and Chemical Biology of Ministry of Education & Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Int J Mol Sci. 2023 Mar 18;24(6):5810. doi: 10.3390/ijms24065810.
(Bt) is a widely used microbial pesticide. However, its duration of effectiveness is greatly shortened due to the irradiation of ultraviolet rays, which seriously hinders the application of Bt preparations. Therefore, it is of great importance to study the resistance mechanism of Bt to UV at the molecular level to improve the UV-resistance of Bt strains. In order to know the functional genes in the UV resistance, the genome of UV-induced mutant Bt LLP29-M19 was re-sequenced and compared with the original strain Bt LLP29. It was shown that there were 1318 SNPs, 31 InDels, and 206 SV between the mutant strain and the original strain Bt LLP29 after UV irradiation, which were then analyzed for gene annotation. Additionally, a mutated gene named , a member of helicase superfamily II, was detected as an important candidate. Then, was expressed and purified successfully. Through the result of the enzymatic activity , yqhH was found to have ATP hydrolase and helicase activities. In order to further verify its function, the gene was knocked out and complemented by homologous recombinant gene knockout technology. The survival rate of the knockout mutant strain Bt LLP29-Δ was significantly lower than that of the original strain Bt LLP29 and the back-complemented strain Bt LLP29-Δ-R after treated with UV. Meanwhile, the total helicase activity was not significantly different on whether Bt carried or not. All of these greatly enrich important molecular mechanisms of Bt when it is in UV stress.
（Bt）是一种广泛使用的微生物农药。然而，由于紫外线的照射，其有效期大大缩短，严重阻碍了 Bt 制剂的应用。因此，研究 Bt 在分子水平上对 UV 的抗性机制，提高 Bt 菌株的抗 UV 能力具有重要意义。为了了解 Bt 抗 UV 的功能基因，对经 UV 诱导的突变株 Bt LLP29-M19 的基因组进行了重测序，并与原始菌株 Bt LLP29 进行了比较。结果表明，UV 照射后，突变株与原始菌株 Bt LLP29 之间存在 1318 个 SNPs、31 个 InDels 和 206 个 SV，随后对这些基因进行了注释分析。此外，检测到一个名为的突变基因，它是解旋酶超家族 II 的一个成员，是一个重要的候选基因。然后，成功表达和纯化了。通过酶活性分析，发现 yqhH 具有 ATP 水解酶和解旋酶活性。为了进一步验证其功能，采用同源重组基因敲除技术对基因进行敲除和互补。敲除突变株 Bt LLP29-Δ 在 UV 处理后的存活率明显低于原始菌株 Bt LLP29 和互补菌株 Bt LLP29-Δ-R。同时，Bt 是否携带基因对总解旋酶活性没有显著影响。这些都极大地丰富了 Bt 在 UV 胁迫下的重要分子机制。
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