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Genome-wide identification of microRNA and siRNA responsive to endophytic beneficial diazotrophic bacteria in maize

ABSTRACT: Several reports have described the involvement of miRNAs in abiotic stresses. However, their role in biotic stress or to beneficial microbes has not been fully explored. In order to understand on the epigenetic regulation in plant in response to nitrogen-fixing bacteria association, we analyzed the sRNA regulation in maize hybrids (Zea mays – UENF 506-8) inoculated with the beneficial diazotrophic bacteria (Herbaspirillum seropedicae). Deep sequencing analysis was carried out to identify the sRNAs regulated in maize during association with diazotrophic bacteria. For this analysis, maize plants were germinated in wet paper and put in hydroponic system with Hoagland’s solution and then inoculated with H. seropedicae for seven days. Mock and inoculated plants were collected and total RNA from a pool of samples was extracted with Trizol reagent. The two sRNA libraries were sequenced by Illumina. The sequences were filtered to remove adaptors and contaminants rRNA and tRNAs, and sequences with 18-28 nt in length were selected. To identify the miRNAs present in these libraries, we used two strategies using the same website (http://srna-tools.cmp.uea.ac.uk): one to identify novel miRNAs using the maize genome (verson 2) and miRCat pipeline; and other to identify conserved miRNAs using the miRBase database (release 13.0, http://microrna.sanger.ac.uk) and miRProf pipeline. We identified 17 novel putative miRNAs candidates and mapped the precursor of these miRNAs in the maize genome. Furthermore, we identified 25 conserved miRNAs families and the differential expressions were analyzed with miRProf pipeline. The bioinformatics analysis of four up-regulated miRNAs (miR397, miR398, miR408 and miR528) in inoculated plant was validated using stem–loop RT-PCR assay. Our findings contribute to increase the knowledge of the molecular relation between plants and endophytic bacteria.

ORGANISM(S): Zea mays

PROVIDER: GSE47886 | GEO | 2013/08/01

SECONDARY ACCESSION(S): PRJNA208240

REPOSITORIES: GEO

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Project description:Several reports have described the involvement of miRNAs in abiotic stresses. However, their role in biotic stress or to beneficial microbes has not been fully explored. In order to understand on the epigenetic regulation in plant in response to nitrogen-fixing bacteria association, we analyzed the sRNA regulation in maize hybrids (Zea mays M-bM-^@M-^S UENF 506-8) inoculated with the beneficial diazotrophic bacteria (Herbaspirillum seropedicae). Deep sequencing analysis was carried out to identify the sRNAs regulated in maize during association with diazotrophic bacteria. For this analysis, maize plants were germinated in wet paper and put in hydroponic system with HoaglandM-bM-^@M-^Ys solution and then inoculated with H. seropedicae for seven days. Mock and inoculated plants were collected and total RNA from a pool of samples was extracted with Trizol reagent. The two sRNA libraries were sequenced by Illumina. The sequences were filtered to remove adaptors and contaminants rRNA and tRNAs, and sequences with 18-28 nt in length were selected. To identify the miRNAs present in these libraries, we used two strategies using the same website (http://srna-tools.cmp.uea.ac.uk): one to identify novel miRNAs using the maize genome (verson 2) and miRCat pipeline; and other to identify conserved miRNAs using the miRBase database (release 13.0, http://microrna.sanger.ac.uk) and miRProf pipeline. We identified 17 novel putative miRNAs candidates and mapped the precursor of these miRNAs in the maize genome. Furthermore, we identified 25 conserved miRNAs families and the differential expressions were analyzed with miRProf pipeline. The bioinformatics analysis of four up-regulated miRNAs (miR397, miR398, miR408 and miR528) in inoculated plant was validated using stemM-bM-^@M-^Sloop RT-PCR assay. Our findings contribute to increase the knowledge of the molecular relation between plants and endophytic bacteria. Screenning of sRNA transcriptome of maize plants inoculated with Herbaspirillum seropedicae after seven days

Project description:Plant defense responses to biotic stresses are complex biological processes, all governed by sophisticated molecular regulations. Induced systemic resistance (ISR) is one of these defense mechanisms where beneficial bacteria or fungi prime plants to resist pathogen or pest attacks. In ISR the defense arsenal in plants remains dormant and it is only triggered by the infection, allowing a rapid response and a better allocation of plant resources. We recently described that the well-known beneficial bacterium Paraburkholderia phytofirmans PsJN is able to reduce Arabidopsis thaliana susceptibility to Pseudomonas syringae Pst DC3000 through ISR. Nevertheless, the molecular networks governing this beneficial interaction remain unknown. To tackle this issue we analyzed the temporal changes in the transcriptome of PsJN-inoculated plants before and after the infection with Pst DC3000 strain. These data were used to perform a gene network analysis to identify highly connected transcription factors that may be acting as hubs in controlling this ISR response. Before the pathogen challenge, strain PsJN regulated 405 genes (corresponding to 1.5% of the analyzed genome). PsJN-inoculated plants presented a faster and stronger transcriptional response 1-hour post infection (hpi) compared with the non-inoculated plants, which presented the highest transcriptional changes 24 hpi. A principal component analysis showed that PsJN-induced plant responses to the pathogen could be differentiated from those induced by the sole pathogen. Forty-eight transcription factors were regulated by PsJN 1 hpi, and a system biology analysis revealed a network with 4 modules where LHY, WRKY28, MYB31 and RRTF1 are highly connected transcription factors, that can be acting as hub regulators in this interaction. These modules are related to Jasmonate, Ethylene, Salicylic acid and ROS pathways. Additionally, the down- and up-regulation of ANAC32 and ORA59, respectively, support an important role of PDF1.2. These results indicate that a rapid and specific response of PsJN-inoculated plants to this virulent pathogen could be the pivotal element in the protection mechanism. A role for specific transcriptional regulators in the orchestration of this complex molecular response is also proposed

Project description:BACKGROUND: The coevolution and interaction between plants and microorganisms have long been a subject of significant research interest. Dark septate endophytes (DSE) have garnered great attention in contemporary research due to their functional diversity, in vitro cultivation ability, and ability to establish symbiotic associations with host plants. In the present study, three DSE strains, namely Acrocalymma vagum, Zopfiella marina, and Phoma herbarum, which were obtained from the roots of Astragalus membranaceus, were introduced into maize plants through inoculation. We evaluated the effects of DSE inoculation on maize growth and root secretion activity through a multi omics methods, and proposed mechanisms for 'internal pathways' and 'external pathways'.RESULTS: The findings indicated that A. vagum exhibited superior growth-promoting ability on maize compared to Z. marina and P. herbarum.GO and KEGG enrichment analysis found that A. vagum inoculation resulted in significant enrichment of differentially expressed genes in annotation functions related to hormone regulation and lipid metabolism. A. vagum inoculation revealed that the gene pathways involved in plant hormone signaling and plant pathogen interactions play a crucial role in promoting host growth, and A. vagum inoculation group exhibited the highest number of differentially expressed genes, the most intricate protein-protein interaction (PPI) model, and the most pronounced relationship between differentially expressed genes. After the inoculation of A.vagum, the levels of salicylic acid, zeatin, and IAA in maize plants significantly increased. Additionally, the diversity and abundance of endophytic fungi, as well as the proportion of harmful bacteria and beneficial fungi, had significantly increased. Compared with Z. marina and P. herbarum, the net photosynthetic rate (Pn) and stomatal conductance (Gs) of A.vagum inoculated plants significantly increased. Inoculation with A.vagum could enhance the ability of corn roots to secrete lipids, sugars, and amino acids, resulted in a notable augmentation of beneficial bacteria and fungi, accompanied by a significant reduction in the proportion of harmful bacteria in the rhizosphere soil, such as Fusarium solani and Fusarium lacertarum, exhibited significant inhibition, whereas Bacillus niabensis and Bacillus nealsonii demonstrated enrichment trends. Soil pH, organic matter, available potassium content, acid phosphatase, alkaline phosphatase and urease activity exhibited significant increases following the inoculation of A. vagum. Variance decomposition and structural equation modeling (SEM) analysis indicated that the 'internal pathway', maize growth is mainly influenced by the interaction of endogenous hormones, endophytic microorganisms, and photosynthetic parameters, whereas within the 'external pathway', the interaction between soil microorganisms and soil physicochemical properties exerted a dominant influence. Compared with the Z. marina and P. herbarum inoculation, A. vagum inoculation showed a more significant impact on maize growth, both in terms of 'internal pathway' and 'external pathway', in terms of pathway level and quantity.CONCLUSIONS: These findings provide a new perspective for understanding the potential mechanisms of 'microbe-plant' interactions and also contribute to the exploration of targeted functional microorganisms that promote growth and stress resistance.

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Genome-wide identification of microRNA and siRNA responsive to endophytic beneficial diazotrophic bacteria in maize

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