苏云金芽孢杆菌 ATCC10792 的全基因组序列和基于新型生物标志物提高对苏云金芽孢杆菌和蜡样芽孢杆菌群的区分度。
Whole genome sequence of Bacillus thuringiensis ATCC 10792 and improved discrimination of Bacillus thuringiensis from Bacillus cereus group based on novel biomarkers.
机构信息
Department of Food Science and Biotechnology, College of Agriculture & Life Science, Kangwon National University, Chuncheon, Gangwon, 200-701, South Korea.
National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, 430070, PR China.
出版信息
Microb Pathog. 2019 Apr;129:284-297. doi: 10.1016/j.micpath.2019.02.014. Epub 2019 Feb 10.
BACKGROUND
Among the Bacillus cereus group, B. thuringiensis, is one of the most extensively used biological control agent. The present study reports the complete genome and four novel plasmid analysis of the type strain B. thuringiensis ATCC 10792.
METHODS
Complete genome sequencing of Bacillus thuringiensis ATCC 10792, assembled using de-novo (v.3.2.0, assembly name MIRA3), Pac-Bio sequencers and Hierarchical Genome Assembly Process software (version 4.1) and real-time polymerase chain reaction (qPCR) is a consistent technique for quantifying gene expression based on specific biomarkers, in addition the efficiency of the primers were analysed based on artificially spiked food samples on lettuce, kimbab and spinach with B. thuringiensis ATCC 10792.
RESULTS
Complete genome annotation was performed, and a total of 6269 proteins with 5427594 bps were identified and four novel plasmid (poh2, poh3, poh4, poh5) a total of 134, 131, 96, 21 proteins with 113294; 92,949; 86488; 11332 bps were identified. Six selective genes (lipoprotein-lipo, methyltransferase-MT, S-layer homology domain protein-BC, flagellar motor protein-motB, transcriptional regulator-XRE, crystal protein-cry2) and associated four novel plasmids were investigated along with the characteristics and expression profiles of two housekeeping genes (chaperonin protein-GroEL and topoisomerase enzyme-gyrB). Although from the assessment of 120 strains, both GroEL and gyrB showed 100% specificity towards detection of both B. thuringiensis in artificially spiked vegetable samples. All the eight genes revealed no specificity towards any of the 9 non- Bacillus strains.
CONCLUSION
In our study based on the complete genome and plasmid sequence of B. thuringiensis ATCC 10792, among the six discriminating genes, specifically GroEL, gyrB and XRE showed promising results in identifying B. thuringiensis ATCC 10792, and there detection limit was 3.0-9.6 log CFU/g in the food samples respectfully. The key role in control of the predatory biological agent.
背景
在蜡状芽孢杆菌组中，苏云金芽孢杆菌是应用最广泛的生物防治剂之一。本研究报告了苏云金芽孢杆菌 ATCC 10792 的全基因组和四个新型质粒分析。
方法
使用 de-novo（v.3.2.0，组装名称 MIRA3）、Pac-Bio 测序仪和层次基因组组装过程软件（版本 4.1）对苏云金芽孢杆菌 ATCC 10792 进行全基因组测序，并通过实时聚合酶链反应（qPCR）进行定量基因表达的一致技术，该技术基于特定生物标志物，此外，还基于在生菜、紫菜卷和菠菜上添加苏云金芽孢杆菌 ATCC 10792 的人工污染食品样本分析了引物的效率。
结果
完成了全基因组注释，共鉴定出 6269 种蛋白，总长度为 5427594bp，共鉴定出 4 个新型质粒（poh2、poh3、poh4、poh5），总长度分别为 134131、9698、86488、11332bp。对 6 个选择基因（脂蛋白-脂、甲基转移酶-MT、S-层同源结构域蛋白-BC、鞭毛运动蛋白-motB、转录调节因子-XRE、晶体蛋白-cry2）和相关的 4 个新型质粒进行了研究，并对 2 个管家基因（伴侣蛋白-GroEL 和拓扑异构酶酶-gyrB）的特征和表达谱进行了研究。尽管从对 120 株菌的评估来看，GroEL 和 gyrB 对人工污染蔬菜样本中的苏云金芽孢杆菌均具有 100%的特异性。所有 8 个基因对 9 株非芽孢杆菌均无特异性。
结论
在本研究中，基于苏云金芽孢杆菌 ATCC 10792 的全基因组和质粒序列，在 6 个鉴别基因中，特异性基因 GroEL、gyrB 和 XRE 显示出在鉴定苏云金芽孢杆菌 ATCC 10792 方面有很好的效果，在食品样本中的检测限分别为 3.0-9.6 log CFU/g。在控制捕食性生物制剂方面发挥着关键作用。
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