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一个含有三种等离子体的CRISPR-Cas9平台,用于设计Bacillus velezensis 916作为有效的生物控制剂
Lian Li1, Kecheng Luo1, Shuangyu Zhang1
1Jiangsu Provincial Key Construction Laboratory of Probiotics Preparation, Huaiyin Institute of Technology, Huaian, China.
Applied and environmental microbiology
|September 2, 2025
概括
一个新的CRISPR-Cas9基因编辑平台增强了Bacillus velezensis的生物控制. 这种系统有助于促进循环脂的产生和抗击疾病的能力,
科学领域:
- 微生物学
- 生物技术
- 植物病理学
背景情况:
- 由于其通过非核糖体合成酶 (NRPS) 产生循环脂 (CLPs),Bacillus velezensis (Bv) 是一种重要的植物疾病生物控制剂.
- 已知产生四种CLP的Bv916菌株对米病和角叶斑具有有效性.
- 提高Bv916的有效性和安全性需要食品级的基因编辑系统.
研究的目的:
- 为生物控制剂Bacillus velezensis Bv916开发一个食品级的CRISPR-Cas9基因编辑平台.
- 通过修改Bv916中的NRPS基因集群来优化CLP的产生.
- 增强Bv916对植物病原体的生物控制功效.
主要方法:
- 在Bv916中使用热敏源,构成性促进物 (P43) 和特定促进物 (Psrf) 构建一个三等离子CRISPR-Cas9系统.
- 通过ComX和Reca的促进物替代进行基因编辑,实现高单基因和双基因编辑效率.
- 通过用强大的构成性促进剂 (PB,PA,P43,PrepU) 替换四个NRPS基因集群 (loc,srf,bl,fen) 来产生BvLSBF.
主要成果:
- 在5天内,CRISPR-Cas9平台在Bv916中显示出高的单基因 (96%) 和双基因 (61%) 编辑效率.
- 衍生品BvLSBF的产量显著增加了四种CLP:洛基洛米 (6. 8倍),表面素 (5. 9倍),巴基洛米L (10. 9倍) 和风 (6. 2倍).
- 与野生型Bv916相比,BvLSBF对植物病原体具有明显增强的对抗活性.
结论:
- 为Bv916成功建立了强大的食品级CRISPR-Cas9平台,使得有效的基因编辑成为可能.
- 开发的平台通过NRPS基因集群的促进器工程来促进CLP生产.
- 这项研究为开发Bv916为可持续农业的先进细胞工厂提供了基础,
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