Kwenda Stanford, Gorshkov Vladimir, Ramesh Aadi Moolam, Naidoo Sanushka, Rubagotti Enrico, Birch Paul R J, Moleleki Lucy N
Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, South Africa.
Kazan Institute of Biochemistry and Biophysics, Kazan Scientific Center, Russian Academy of Sciences, Kazan, Russia.
BMC Genomics. 2016 Jan 12;17:47. doi: 10.1186/s12864-016-2376-0.
Small RNAs (sRNAs) have emerged as important regulatory molecules and have been studied in several bacteria. However, to date, there have been no whole-transcriptome studies on sRNAs in any of the Soft Rot Enterobacteriaceae (SRE) group of pathogens. Although the main ecological niches for these pathogens are plants, a significant part of their life cycle is undertaken outside their host within adverse soil environment. However, the mechanisms of SRE adaptation to this harsh nutrient-deficient environment are poorly understood.
In the study reported herein, by using strand-specific RNA-seq analysis and in silico sRNA predictions, we describe the sRNA pool of Pectobacterium atrosepticum and reveal numerous sRNA candidates, including those that are induced during starvation-activated stress responses. Consequently, strand-specific RNA-seq enabled detection of 137 sRNAs and sRNA candidates under starvation conditions; 25 of these sRNAs were predicted for this bacterium in silico. Functional annotations were computationally assigned to 68 sRNAs. The expression of sRNAs in P. atrosepticum was compared under growth-promoting and starvation conditions: 68 sRNAs were differentially expressed with 47 sRNAs up-regulated under nutrient-deficient conditions. Conservation analysis using BLAST showed that most of the identified sRNAs are conserved within the SRE. Subsequently, we identified 9 novel sRNAs within the P. atrosepticum genome.
Since many of the identified sRNAs are starvation-induced, the results of our study suggests that sRNAs play key roles in bacterial adaptive response. Finally, this work provides a basis for future experimental characterization and validation of sRNAs in plant pathogens.
小RNA(sRNA)已成为重要的调控分子,并已在多种细菌中得到研究。然而,迄今为止,尚未对任何软腐肠杆菌科(SRE)病原菌群体中的sRNA进行全转录组研究。尽管这些病原菌的主要生态位是植物,但其生命周期的很大一部分是在宿主外的恶劣土壤环境中度过的。然而,人们对SRE适应这种恶劣的营养缺乏环境的机制的机制了解甚少。
在本文报道的研究中,通过使用链特异性RNA测序分析和计算机sRNA预测,我们描述了黑胫果胶杆菌的sRNA库,并揭示了众多sRNA候选物,包括那些在饥饿激活的应激反应中被诱导的候选物。因此,链特异性RNA测序能够在饥饿条件下检测到137种sRNA和sRNA候选物;其中25种sRNA是通过计算机预测出的该细菌的sRNA。对68种sRNA进行了功能注释。比较了在促进生长和饥饿条件下黑胫果胶杆菌中sRNA的表达:68种sRNA存在差异表达,其中47种sRNA在营养缺乏条件下上调。使用BLAST进行的保守性分析表明,大多数已鉴定的sRNA在SRE内是保守的。随后,我们在黑胫果胶杆菌基因组中鉴定出9种新型sRNA。
由于许多已鉴定的sRNA是饥饿诱导的,我们的研究结果表明sRNA在细菌适应性反应中起关键作用。最后,这项工作为未来对植物病原菌中sRNA的实验表征和验证提供了基础。