Dataset Information

Proteomic and mutant analysis of hydrogenase gene hypE in symbiotic nitrogen fixation in Mesorhizobium huakuii

ABSTRACT: Hydrogenases catalyze the simple yet importantredox reaction between protons and electrons and H2, thus, mediate interactions between microorganisms and their chemical environment. A mutation in M. huakuii hypE gene was generated by homologous recombination. The hypE mutant grew a little faster than its parental 7653R. Real-time quantitative PCR showed that the hypE gene expression is significantly upregulated in all the detected stages of nodule development. The wild type contained normal spherical shape of the nodules, while the hypE mutant elicited small-size nodules. Plants inoculated with mutant HKhypE showed a significantly more than 47% decrease in acetylene reduction activity compared to the wild-type 7653R. Proteomics analysis showed seven proteins were up-regulated and eighty-two proteins were down-regulated in hypE triple mutant bacteroids.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Mesorhizobium Huakuii

TISSUE(S): Root Nodule

SUBMITTER: Guojun Cheng

LAB HEAD: Guojun Cheng

PROVIDER: PXD026564 | Pride | 2023-10-24

REPOSITORIES: Pride

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Proteomic and Mutant Analysis of Hydrogenase Maturation Protein Gene hypE in Symbiotic Nitrogen Fixation of Mesorhizobium huakuii.

Long Songhua S Su Min M Chen Xiaohong X Hu Aiqi A Yu Fuyan F Zou Qian Q Cheng Guojun G

International journal of molecular sciences 20230808 16

Hydrogenases catalyze the simple yet important redox reaction between protons and electrons and H2, thus mediating symbiotic interactions. The contribution of hydrogenase to this symbiosis and anti-oxidative damage was investigated using the M. huakuii hypE (encoding hydrogenase maturation protein) mutant. The hypE mutant grew a little faster than its parental 7653R and displayed decreased antioxidative capacity under H2O2-induc ...[more]

PMID: 37628715

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Project description:Transcriptional profiling of hop samples throughout in vitro culture on medium with sucrose and hormones IAA and BAP Study of gene expression involved in stress response during in vitro culture and in organogenic nodule formation Hop (Humulus lupulus L.) is an economically important plant forming organogenic nodules which can be used for genetic transformation and micropropagation. We are interested in the mechanisms underlying reprogramming of cells through stress and hormone treatments. To investigate the spatio-temporal sequence of events that underlies competence acquisition for organogenesis three main stages were selected: (i) internodes at the time of excision from the parent plant (T0); (ii) internodes grown for 15 days on culture medium in which several prenodular structures are formed inside the calli (T15d); and (iii) nodule forming tissue after 28 days of culture (T28d). Additionally a control was included corresponding to 28 days of culture without hormones, lacking nodule formation and plantlet regeneration abilities, in order to identify genes specifically involved in morphogenesis (T28dWH). Organogenic nodule formation is a morphogenic process that shares features with somatic embryogenesis though in the former no shoot/root pole is established and plantlet regeneration can occur from different peripheral regions of nodules. Previous studies have shown that the organogenesis-determining period (when cells are determined to form nodules) occurs in between 15 and 25 days of culture corresponding to prenodular and first nodular stages. When nodules are fully developed they are surrounded by layers of elongated, highly vacuolated cells that may degenerate when nodules start differentiating plantlets. Plantlet regeneration from organogenic nodules can be observed after 45 days of culture. However, organogenic nodules after 28 days of culture are already determined to undergo plantlet regeneration, since they no longer require exogenous supply of hormones. Explants cultured in medium without hormones can develop incipient prenodular structures with vascular tissue but not nodules, thus lacking morphogenic potential. Keywords: Plant development, Time course 3 time points: 3 Biological replicates and 1 technical replicate (1 dye-swap). 1 control at beginning of culture and 1 control wihout hormones. One replicate per array. Each clone printed twice in the same sub-grid

Project description:PhyR is an unusual type of response regulator consisting of a receiver domain and an extracytoplasmic function (ECF) sigma factor-like domain. It was recently described as a master regulator of general stress response in Methylobacterium extorquens. Orthologues of this regulator are present in essentially all free-living Alphaproteobacteria. In most of them, phyR is genetically closely linked to a gene encoding an ECF sigma factor. Here, we investigate the role of these two regulators in the soybean symbiont Bradyrhizobium japonicum USDA110. Using deletion mutants and phenotypic assays, we showed that PhyR and the ECF sigma factor sigma(EcfG) are involved in heat shock and desiccation resistance upon carbon starvation. Both mutants had symbiotic defects on the plant hosts Glycine max (soybean) and Vigna radiata (mungbean). They induced fewer nodules than the wild type and these nodules were smaller, less pigmented, and their specific nitrogenase activity was drastically reduced 2 or 3 weeks after inoculation. Four weeks after infection, soybean nodule development caught up to a large extent whereas most mungbean nodules remained defective even 5 weeks after infection. Remarkably, both mutants triggered aberrant nodules on the different host plants with ectopically emerging roots. Microarray analysis revealed that PhyR and sigma(EcfG) control congruent regulons suggesting both regulators are part of the same signalling cascade. This finding was further substantiated by in vitro protein-protein interaction studies which are in line with a partner-switching mechanism controlling gene regulation triggered by phosphorylation of PhyR. The large number of genes of unknown function present in the PhyR/sigma(EcfG) regulon and the conspicuous symbiotic phenotype suggest that these regulators are involved in the Bradyrhizobium-legume interaction via yet undisclosed mechanisms. Comparative analysis of the B. japonicum phyR mutant 8402, ecfG mutant 8404 mutant and the wild type during exponential growth in rich medium (PSY) and after 24 hours starvation in carbon source-free minimal medium

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Dataset Information

Proteomic and mutant analysis of hydrogenase gene hypE in symbiotic nitrogen fixation in Mesorhizobium huakuii

Dataset's files

Publications

Proteomic and Mutant Analysis of Hydrogenase Maturation Protein Gene <i>hypE</i> in Symbiotic Nitrogen Fixation of <i>Mesorhizobium huakuii</i>.

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