Laboratorio de Biotecnología Molecular, Instituto de Biotecnología Misiones "Dra. María Ebe Reca" (InBioMis), CONICET, Facultad de Ciencias Exactas, Químicas y Naturales/FCEQyN, Universidad Nacional de Misiones/UNaM, Posadas, Misiones, Argentina.
Cátedra de Bacteriología, Dpto. de Microbiología, Facultad de Ciencias Exactas, Químicas y Naturales/FCEQyN, Universidad Nacional de Misiones/UNaM, Posadas, Misiones, Argentina.
PLoS One. 2021 Mar 11;16(3):e0248274. doi: 10.1371/journal.pone.0248274. eCollection 2021.
Plant growth-promoting bacteria (PGPB) are a heterogeneous group of bacteria that can exert beneficial effects on plant growth directly or indirectly by different mechanisms. PGPB-based inoculant formulation has been used to replace chemical fertilizers and pesticides. In our previous studies, two endophytic endospore-forming bacteria identified as Bacillus altitudinis were isolated from roots of Ilex paraguariensis St. Hil. seedlings and selected for their plant growth-promoting (PGP) properties shown in vitro and in vivo. The purposes of this work were to assemble the genomes of B. altitudinis 19RS3 and T5S-T4, using different assemblers available for Windows and Linux and to select the best assembly for each strain. Both genomes were also automatically annotated to detect PGP genes and compare sequences with other genomes reported. Library construction and draft genome sequencing were performed by Macrogen services. Raw reads were filtered using the Trimmomatic tool. Genomes were assembled using SPAdes, ABySS, Velvet, and SOAPdenovo2 assemblers for Linux, and Geneious and CLC Genomics Workbench assemblers for Windows. Assembly evaluation was done by the QUAST tool. The parameters evaluated were the number of contigs ≥ 500 bp and ≥ 1000 bp, the length of the longest contig, and the N50 value. For genome annotation PROKKA, RAST, and KAAS tools were used. The best assembly for both genomes was obtained using Velvet. The B. altitudinis 19RS3 genome was assembled into 15 contigs with an N50 value of 1,943,801 bp. The B. altitudinis T5S-T4 genome was assembled into 24 contigs with an N50 of 344,151 bp. Both genomes comprise several genes related to PGP mechanisms, such as those for nitrogen fixation, iron metabolism, phosphate metabolism, and auxin biosynthesis. The results obtained offer the basis for a better understanding of B. altitudinis 19RS3 and T5S-T4 and make them promissory for bioinoculant development.
植物促生细菌(PGPB)是一组异质细菌,通过不同的机制直接或间接地对植物生长产生有益影响。基于 PGPB 的接种剂配方已被用于替代化肥和农药。在我们之前的研究中,从巴拉圭冬青幼苗的根部分离出两种内生芽孢形成细菌,鉴定为高山芽孢杆菌,并选择了它们在体外和体内表现出的植物促生(PGP)特性。本工作的目的是使用适用于 Windows 和 Linux 的不同组装器组装高山芽孢杆菌 19RS3 和 T5S-T4 的基因组,并为每种菌株选择最佳组装。还对两个基因组进行了自动注释,以检测 PGP 基因并与其他报告的基因组序列进行比较。文库构建和草图基因组测序由 Macrogen 服务公司完成。使用 Trimmomatic 工具过滤原始读取。使用 SPAdes、AByss、Velvet 和 SOAPdenovo2 组装器在 Linux 下,以及 Geneious 和 CLC Genomics Workbench 组装器在 Windows 下对基因组进行组装。使用 QUAST 工具评估组装。评估的参数包括≥500bp 和≥1000bp 的 contig 数量、最长 contig 的长度和 N50 值。使用 PROKKA、RAST 和 KAAS 工具进行基因组注释。两个基因组的最佳组装均使用 Velvet 获得。高山芽孢杆菌 19RS3 基因组组装成 15 个 contig,N50 值为 1,943,801bp。高山芽孢杆菌 T5S-T4 基因组组装成 24 个 contig,N50 值为 344,151bp。两个基因组都包含几个与 PGP 机制相关的基因,例如固氮、铁代谢、磷代谢和生长素生物合成。获得的结果为更好地理解高山芽孢杆菌 19RS3 和 T5S-T4 提供了基础,并为生物接种剂的开发提供了有希望的方向。