印度东部农田枯草芽孢杆菌和巨芽孢杆菌分离株的形态生化和分子特征。

IF 2.8 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Sujaan Aehsas, Jyoti Prakash Sahoo, Neelanjana Choudhury, Shaikh Nausad Hossain, Anjali Lata Sahoo, Sunil Kumar Sunani
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引用次数: 0

摘要

背景:本研究旨在利用营养琼脂连续稀释和涂布平板技术,研究不同环境(城市公园(SJ1)和农业研究场地(SJ2)土壤的微生物多样性,以鉴定具有农业或生物技术应用潜力的区域特异性细菌菌株。方法和结果:分离细菌通过选择性传代培养纯化,通过革兰氏染色和生化试验(MR-VP,过氧化氢酶,氧化酶)进行鉴定,并在4°C保存。提取基因组DNA,针对16S rRNA基因进行pcr扩增,测序后进行blast鉴定。系统发育分析(MEGA 12,邻居连接,1000次引导)和进化速率估计(最大似然,γ分布位点变异)来评估遗传关系和差异。形态学和生化分析均显示革兰氏阳性杆状菌,过氧化氢酶阳性,氧化酶阴性,甲基红阴性,Voges-Proskauer阳性。提取基因组DNA,扩增16S rRNA基因(~ 1500 bp),测序,BLASTN分析。SJ1与芽孢杆菌的同源性为99.84 ~ 99.92%,SJ2与巨型Priestia megaterium的同源性为99.92%。系统发育分析表明,SJ1在枯草芽孢杆菌菌株SBIS1 (GenBank: PV523294.1)中,SJ2在巨芽孢杆菌菌株PIIS1 (GenBank: PV523528.1)中,表明不同的进化谱系。最大似然估计显示出不同的进化速率,即SJ1表现出高速率异质性,而SJ2表现出均匀速率。SJ1的5个离散γ类别的平均相对进化率在0.08到2.75之间,而SJ2的平均相对进化率在0.90到1.10之间。SJ1显示(25.23%),20.09% (T), 23.43% (C)和31.26% (G);而SJ2 (24.87%), 21.26% (T), 24.20% (C), 29.68% (G)。SJ1(1268个对齐位点)和SJ2(750个对齐位点)的最大对数似然值分别为-3315.740和-1034.491,证实了稳健的系统发育推断。结论:该研究成功鉴定出具有独特进化速度、遗传组成和分离株之间强大差异的不同菌株,支持其进一步农业或生物技术探索的适用性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Morpho-biochemical and molecular characterization of Bacillus subtilis and Priestia megaterium isolates from eastern Indian farmlands.

Background: The study aimed to investigate the microbial diversity of soils from distinct environments, an urban park (SJ1) and an agricultural research field (SJ2) using serial dilution and spread plate techniques on nutrient agar, to identify region-specific bacterial strains with potential agricultural or biotechnological applications.

Methods and results: Bacterial isolates were purified through selective sub-culturing, characterized via Gram staining and biochemical tests (MR-VP, catalase, oxidase), and preserved at 4 °C. Genomic DNA was extracted, PCR-amplified targeting the 16S rRNA gene, and sequenced for BLAST-based identification. Phylogenetic analysis (MEGA 12, neighbor-joining, 1000 bootstraps) and evolutionary rate estimation (maximum likelihood, gamma-distributed site variation) were performed to assess genetic relationships and divergence. Morphological and biochemical analyses revealed Gram-positive rods in both isolates, exhibiting catalase positive, oxidase negative, methyl red negetive, and Voges-Proskauer positive activity. Genomic DNA was extracted, and the 16S rRNA gene was amplified (~ 1500 bp), sequenced, and analyzed via BLASTN. SJ1 showed 99.84-99.92% identity with Bacillus spp., while SJ2 matched Priestia megaterium (99.92%). Phylogenetic analysis placed SJ1 within Bacillus subtilis strain SBIS1 (GenBank: PV523294.1), and SJ2 within P. megaterium strain PIIS1 (GenBank: PV523528.1), indicating distinct evolutionary lineages. Maximum likelihood estimation revealed differing evolutionary rates, i.e., SJ1 exhibited high rate heterogeneity, while SJ2 showed uniform rates. Mean relative evolutionary rates across five discrete gamma categories for SJ1 ranged from 0.08 to 2.75, while for SJ2 they ranged from 0.90 to 1.10. SJ1 showed 25.23% (A), 20.09% (T), 23.43% (C), and 31.26% (G); while SJ2 had 24.87% (A), 21.26% (T), 24.20% (C), and 29.68% (G). Maximum log-likelihood values were -3315.740 for SJ1 (1268 aligned sites) and -1034.491 for SJ2 (750 aligned sites), confirming robust phylogenetic inference.

Conclusion: The study successfully identified distinct bacterial strains with unique evolutionary rates, genetic compositions, and robust divergence between the isolates, supporting their suitability for further agricultural or biotechnological exploration.

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来源期刊
Molecular Biology Reports
Molecular Biology Reports 生物-生化与分子生物学
CiteScore
5.00
自引率
0.00%
发文量
1048
审稿时长
5.6 months
期刊介绍: Molecular Biology Reports publishes original research papers and review articles that demonstrate novel molecular and cellular findings in both eukaryotes (animals, plants, algae, funghi) and prokaryotes (bacteria and archaea).The journal publishes results of both fundamental and translational research as well as new techniques that advance experimental progress in the field and presents original research papers, short communications and (mini-) reviews.
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