Project description:Analysis of gene expression changes of Mesorhizobium alhagi CCNWXJ12-2 under high salt stress. Mesorhizobium alhagi CCNWXJ12-2 is isolated from Alhagi sparsifolia in northwest of China. Total RNA extracted from Mesorhizobium alhagi CCNWXJ12-2 growing in TY medium containing 0.4 M NaCl and 0 M NaCl.
Project description:BACKGROUND: Mesorhizobium alhagi CCNWXJ12-2 is a ?-proteobacterium which could be able to fix nitrogen in the nodules formed with Alhagi sparsifolia in northwest of China. Desiccation and high salinity are the two major environmental problems faced by M. alhagi CCNWXJ12-2. In order to identify genes involved in salt-stress adaption, a global transcriptional analysis of M. alhagi CCNWXJ12-2 growing under salt-free and high salt conditions was carried out. The next generation sequencing technology, RNA-Seq, was used to obtain the transcription profiles. RESULTS: We have compared the transcriptome of M. alhagi growing in TY medium under high salt conditions (0.4 M NaCl) with salt free conditions as a control. A total of 1,849 differentially expressed genes (fold change ? 2) were identified and 933 genes were downregulated while 916 genes were upregulated under high salt condition. Except for the upregulation of some genes proven to be involved in salt resistance, we found that the expression levels of protein secretion systems were changed under high salt condition and the expression levels of some heat shock proteins were reduced by salt stress. Notably, a gene encoding YadA domain-containing protein (yadA), a gene encoding trimethylamine methyltransferase (mttB) and a gene encoding formate--tetrahydrofolate ligase (fhs) were highly upregulated. Growth analysis of the three gene knockout mutants under salt stress demonstrated that yadA was involved in salt resistance while the other two were not. CONCLUSIONS: To our knowledge, this is the first report about transcriptome analysis of a rhizobia using RNA-Seq to elucidate the salt resistance mechanism. Our results showed the complex mechanism of bacterial adaption to salt stress and it was a systematic work for bacteria to cope with the high salinity environmental problems. Therefore, these results could be helpful for further investigation of the bacterial salt resistance mechanism.
Project description:Mesorhizobium alhagi CCNWXJ12-2T, isolated from root nodules of the desert plant Alhagi sparsifolia, contains two type III secretion systems (T3SSs). T3SSs are specialized machinery with wide distribution in bacteria that inject effector proteins into target cells. Our previous study showed that the expression of M. alhagi T3SS1 is upregulated in high-salt conditions. Here, phylogenetic analysis of T3SS1 using the core protein RhcU suggested that T3SS1 belongs to the ?-Rhc II subgroup of the Rhc T3SS family. To elaborate the function of M. alhagi CCNWXJ12-2T T3SS1 in stress adaption, two T3SS1 mutants (?rhcQ and ?MA29250) were constructed and analyzed. ?-galactosidase transcriptional fusion assays showed that activity of the promoter of T3SS1 was induced by salts. Mutant ?rhcQ was more sensitive to NaCl and LiCl than the wild-type, but ?MA29250 was not. Both mutants were more sensitive to KCl than the wild-type. The intracellular Na+ concentration in ?rhcQ in high-NaCl conditions (0.4 M) increased by 37% compared to that of the wild-type strain, while the Na+ concentration in ?MA29250 increased by 13%. The K+ concentration in both mutants increased by 16% compared to the wild-type in high-KCl conditions (0.3 M). Strain ?rhcQ showed decreased survival compared to the wild-type after treatment with H2O2, while the survival rate of ?MA29250 was almost the same as that of the wild-type. Antioxidant enzyme activities in ?rhcQ were lower than those in the wild-type strain, but this was not the case for ?MA29250. Our data elucidate the beneficial effects of T3SS1 in the adaption of M. alhagi CCNWXJ12-2T to stress.
Project description:Lotus species are forage legumes with potential as pastures in low-fertility and environmentally constrained soils, owing to their high persistence and yield under those conditions. The aim of this work was the characterization of phenetic and genetic diversity of salt-tolerant bacteria able to establish efficient symbiosis with Lotus spp. A total of 180 isolates able to nodulate Lotus corniculatus and Lotus tenuis from two locations in Granada, Spain, were characterized. Molecular identification of the isolates was performed by repetitive extragenic palindromic PCR (REP-PCR) and 16S rRNA, atpD, and recA gene sequence analyses, showing the presence of bacteria related to different species of the genus Mesorhizobium: Mesorhizobium tarimense/Mesorhizobium tianshanense, Mesorhizobium chacoense/Mesorhizobium albiziae, and the recently described species, Mesorhizobium alhagi. No Mesorhizobium loti-like bacteria were found, although most isolates carried nodC and nifH symbiotic genes closely related to those of M. loti, considered the type species of bacteria nodulating Lotus, and other Lotus rhizobia. A significant portion of the isolates showed both high salt tolerance and good symbiotic performance with L. corniculatus, and many behaved like salt-dependent bacteria, showing faster growth and better symbiotic performance when media were supplemented with Na or Ca salts.
Project description:We report the recovery of a 7068-nt viral sequence from the "viral fossils" embedded in the genome of Alhagi sparsifolia, a typical desert plant. Although the full viral genome remains to be completed, the putative genome structure, the deduced amino acids and phylogenetic analysis unambiguously demonstrate that this viral sequence represents a novel species of the genus Badnavirus. The putative virus is tentatively termed Alhagi bacilliform virus (ABV). Southern blotting and inverse polymerase chain reaction (PCR) data indicate that the ABV-related sequence is integrated into the A. sparsifolia genome, and probably does not give rise to functional episomal virus. Molecular evidence that the ABV sequence exists widely in A. sparsifolia is also presented. To our knowledge, this is the first endogenous badnavirus identified from plants in the Gobi desert, and may provide new clues on the evolution, geographical distribution as well as the host range of the badnaviruses.
Project description:Endophytes have the potential to enhance the ability of plants to resist salt stress, improving crop development and yield. Therefore, in this study, we isolated an endophyte that produced large amounts of exopolysaccharides (EPSs) from the roots of sea rice and examined its effects on the physiological responses of rice (Oryza sativa L. ssp. japonica "Nipponbare") seedlings to salt stress using hydroponic experiments. The endophyte was named Pantoea alhagi NX-11 based on its morphological characteristics and 16S ribosomal DNA (rDNA) sequence alignment. Rice seedlings that had been inoculated with P. alhagi NX-11 exhibited a 30.3% increase in fresh weight, a 28.6% increase in root length, a 51.6% increase in shoot length, and a 26.3% increase in chlorophyll content compared with control seedlings under normal conditions. In addition, inoculated rice seedlings had a 37.5% lower malondialdehyde content, a 133% higher K+/Na+ ratio, and a 52.8% higher proline content after 7 days under salt stress, as well as up-regulated expression of proline synthase, down-regulated expression of proline dehydrogenase, and enhanced antioxidant enzyme activities. Interestingly, rice seedlings that were inoculated with an EPS-deficient strain named NX-11eps- that was obtained by atmospheric and room temperature plasma (ARTP) mutagenesis were damaged by salt stress and had similar physiological and biochemical indicators to the control group. Therefore, we speculate that the ability of P. alhagi NX-11 to enhance the salt tolerance of rice seedlings is related to the EPSs it produces.