Sinorhizobium meliloti nifA is important in fixing nitrogen during symbiosis. A nifA null mutant induces small white invalid nodules in the roots of host plant. The additional phenotypic alterations associated with th...Sinorhizobium meliloti nifA is important in fixing nitrogen during symbiosis. A nifA null mutant induces small white invalid nodules in the roots of host plant. The additional phenotypic alterations associated with the disruption of the nifA gene are reported in this study. Under a free-living state, S. meliloti nifA mutant reduces its ability to swarm on a half-solid plate. Interestingly, the AHL （Acylhomoserine lactones） contents in the nifA mutant are lower than that of the wild type during the lag phase, whereas it is reversed in the logarithmic and stationary phases. Quantitative spectrophotometric assays reveal that the total amount of extracellular proteins of the nifA mutant are lower than that of the wild type. In addition, the mutant abolishes its nodulation competitive ability during symbiosis. These findings indicate that NifA plays a regulatory role in multiple cellular processes in S. meliloti.展开更多
The inhibitory effect of gadolinium on Sinorhizobium fredii USDA 205 was studied on a global scale using twodimensional gel electrophoresis and MALDI-TOF MS. The results indicated that 22 proteins were significantly a...The inhibitory effect of gadolinium on Sinorhizobium fredii USDA 205 was studied on a global scale using twodimensional gel electrophoresis and MALDI-TOF MS. The results indicated that 22 proteins were significantly affected by 1 mmol · L^-1 Gd^3 ＋ treatment when compared with an untreated control. Among these proteins, nine were up-regulated and thirteen were down-regulated. The differently expressed proteins were classified into 8 functional categories based on their functions, including transporters, proteins for cellular defence, and proteins involved in metabolism.展开更多
Seventeen Sinorhizobium meliloti strains from seven provinces in China were used to screen highly effective strains for alfalfa cultivar in a greenhouse study and their symbiotic relationship and competitive ability w...Seventeen Sinorhizobium meliloti strains from seven provinces in China were used to screen highly effective strains for alfalfa cultivar in a greenhouse study and their symbiotic relationship and competitive ability were studied in the field. CCBAU30138 was the most effective strain, as evidenced by increase in dry weights. A field experiment showed that the inoculation of alfalfa with CCBAU30138 resulted in increases of 11.9% and 19.6% of dry matter production and crude protein production, respectively, in forage of monocultured plants. The total dry matter yields of alfalfa and tall fescue in binary culture were increased by 16.3% by inoculation of alfalfa with this strain. These results showed that S. rneliloti strain CCBAU30138 was an effective inoculant both in the greenhouse and in the field. The analysis of randomly amplified polymorphic DNA （RAPD） by polymerase chain reaction （PCR） from nodule extracts showed that the strain CCBAU30138 had high competitiveness in the field. It occupied 47.5% of nodules in alfalfa monoculture and 44.4% of nodules in alfalfa-tall fescue binary culture after 20 weeks of growth. In conclusion, a simple system to select highly effective and competitive symbiotic strains specific to alfalfa was established. Using this system, a s.train suitable for the alfalfa cultivar ‘Vector＇ grown in Wuqiao County of Hebei Province was obtained.展开更多
Several studies have demonstrated that the Rhizobium nifA gene is an activator of nitrogen fixation acting in nodule bacteria. To understand the effects of the Sinorhizobium meliloti nifA gene on Alfalfa, the cDNA-AFL...Several studies have demonstrated that the Rhizobium nifA gene is an activator of nitrogen fixation acting in nodule bacteria. To understand the effects of the Sinorhizobium meliloti nifA gene on Alfalfa, the cDNA-AFLP technique was employed to study the changes in gene expression in nifA mutant nodules. Among the approximately 3,000 transcriptderived fragments, 37 had differential expression levels. These expression levels were subsequently confirmed by reverse Northern blot and RT-polymerase chain reaction. Sequence analyses revealed that 21 cDNA fragments corresponded to genes involved in signal communication, protein degradation, nutrient metabolism, cell growth and development.展开更多
Alfalfa （Medicago sativa L.） is being grown in harsh environment in Iraq and is mostly subjected to abiotic stresses such as drought, salinity, pH and temperature. Both alfalfa and its nitrogen fixing symbiotic bact...Alfalfa （Medicago sativa L.） is being grown in harsh environment in Iraq and is mostly subjected to abiotic stresses such as drought, salinity, pH and temperature. Both alfalfa and its nitrogen fixing symbiotic bacteria Sinorhizobium meliloti are affected by these environmental stresses. Enhancing nitrogen fixation biologically could be achieved through selection of tolerant strains of S. meliloti to these environmental stresses and inoculating them to the crop, also growing tolerant cultivars. This study examines phenotypic diversity for tolerance to drought, salinity, temperature and pH. Sixty isolates sampled from different areas of Iraq. The results revealed high degree of phenotypic diversity in Sinorhizobium populations. Furthermore, the isolates which showed tolerance to drought stress also showed tolerance to salinity and high degree of temperature, indicating direct relationship between three physiological path ways. Also 58.3% of drought tolerant isolates were alkaline tolerant they tolerated up to pH 9, point to say almost all drought tolerant isolates in this study illustrated strong ＋ positive reaction to catalase enzyme. And 91.6% themes were negative for Gelatinase enzyme test. While only 50% of drought sensitive isolates were negative for drought sensitive isolates could grow at high temperature （42 ℃）.展开更多
Though the majority of bacteria can form structured communities known as biofilms, mutations can cause bacterial strains to vary in their ability to form a biofilm. In this study, the apparent diffusion coefficient of...Though the majority of bacteria can form structured communities known as biofilms, mutations can cause bacterial strains to vary in their ability to form a biofilm. In this study, the apparent diffusion coefficient of polystyrene microspheres 0.29 μm in diameter, which were executing Brownian motion inside bacterial colonies, was used as a quantitative parameter of the ability of a strain to form a biofilm and of the biofilm development. The study was performed using five Sinorhizobium meliloti strains, the biofilm-forming strains Rm8530 expR+, Rm8530 exoY, and Rm9034 expG, and the non-biofilm forming strains Rm1021 and Rm9030-2 expA1. The green fluorescent beads were placed with each strain in a separate channel of a microfluidic device. Thus, as the bacterial colonies grew under identical conditions over a 4-day period, the motion of the fluorescent microspheres was recorded and the diffusion coefficients were measured every 24 hours via particle tracking algorithms. It was found that each strain displayed a unique pattern of change in diffusion coefficient over time. Also, for a given biofilm-forming strain, there was a clear correlation between the value of the diffusion coefficient and the appearance and motility of the bacterial community. Thus, the diffusion coefficient can be used to identify different S. meliloti strains, and for the biofilm-forming strains, it is also a quantitative indicator of the stage of biofilm development.展开更多
Soil bacteria Sinorhizobium meliloti had enormous agricultural value, due to their ability in fixing nitrogen symbiotically with an important forage crop legume--alfalfa. The aims of the present study were （1） to is...Soil bacteria Sinorhizobium meliloti had enormous agricultural value, due to their ability in fixing nitrogen symbiotically with an important forage crop legume--alfalfa. The aims of the present study were （1） to isolate indigenous S. meliloti from different field sites in lraq; （2） to evaluate the isolates tolerance to induced drought using polyethylene glycol-6000; （3） assessing genetic diversity and genetic relationships among isolates of natural population with drought tolerant abilities. Drought tolerance study revealed vast variations between Sinorhizobium isolates, the highest tolerant isolates to drought were 12 from total thirty （40%）, tolerated from -3 up to -4 MPa, while the drought sensitive isolates tolerated up to -1.5 MPa, except isolate Bs 58 which tolerated up to -1 Mpa water potential. The growth declined with the increase of drought stress. Cluster analysis based on Polymerase Chain Reaction-Restriction Fragment Length Polymorphism （PCR-RFLP） of the 16S rDNA showed two divergent groups with 41% similarity, the first group included three drought sensitive isolates （Bs 44, Bs 54 and Bs 58）, the second group comprised the rest nine isolates （moderate and high drought tolerant）, except for Bs 55 which was drought sensitive isolate, all isolates in the two groups showed no differences between them. The PCR-RFLP of 16S rDNA method revealed a genetic variance between the drought sensitive and tolerant isolates.展开更多
基金This work was supported by the National Grand Fundamental Research 973 Project (No. 2001CB108901)National Science Foundation of China (No. 30400267).
文摘Sinorhizobium meliloti nifA is important in fixing nitrogen during symbiosis. A nifA null mutant induces small white invalid nodules in the roots of host plant. The additional phenotypic alterations associated with the disruption of the nifA gene are reported in this study. Under a free-living state, S. meliloti nifA mutant reduces its ability to swarm on a half-solid plate. Interestingly, the AHL （Acylhomoserine lactones） contents in the nifA mutant are lower than that of the wild type during the lag phase, whereas it is reversed in the logarithmic and stationary phases. Quantitative spectrophotometric assays reveal that the total amount of extracellular proteins of the nifA mutant are lower than that of the wild type. In addition, the mutant abolishes its nodulation competitive ability during symbiosis. These findings indicate that NifA plays a regulatory role in multiple cellular processes in S. meliloti.
基金Project supported by the National Natural Science Foundation of China (G2001CB108902 ,2004CB418506)
文摘The inhibitory effect of gadolinium on Sinorhizobium fredii USDA 205 was studied on a global scale using twodimensional gel electrophoresis and MALDI-TOF MS. The results indicated that 22 proteins were significantly affected by 1 mmol · L^-1 Gd^3 ＋ treatment when compared with an untreated control. Among these proteins, nine were up-regulated and thirteen were down-regulated. The differently expressed proteins were classified into 8 functional categories based on their functions, including transporters, proteins for cellular defence, and proteins involved in metabolism.
基金Project supported by the National "Eleventh Five Years Plan" Key Project on Science and Technology (No. 2006BAD02A15)the National Natural Science Foundation of China (No. 30671222).
文摘Seventeen Sinorhizobium meliloti strains from seven provinces in China were used to screen highly effective strains for alfalfa cultivar in a greenhouse study and their symbiotic relationship and competitive ability were studied in the field. CCBAU30138 was the most effective strain, as evidenced by increase in dry weights. A field experiment showed that the inoculation of alfalfa with CCBAU30138 resulted in increases of 11.9% and 19.6% of dry matter production and crude protein production, respectively, in forage of monocultured plants. The total dry matter yields of alfalfa and tall fescue in binary culture were increased by 16.3% by inoculation of alfalfa with this strain. These results showed that S. rneliloti strain CCBAU30138 was an effective inoculant both in the greenhouse and in the field. The analysis of randomly amplified polymorphic DNA （RAPD） by polymerase chain reaction （PCR） from nodule extracts showed that the strain CCBAU30138 had high competitiveness in the field. It occupied 47.5% of nodules in alfalfa monoculture and 44.4% of nodules in alfalfa-tall fescue binary culture after 20 weeks of growth. In conclusion, a simple system to select highly effective and competitive symbiotic strains specific to alfalfa was established. Using this system, a s.train suitable for the alfalfa cultivar ‘Vector＇ grown in Wuqiao County of Hebei Province was obtained.
文摘Several studies have demonstrated that the Rhizobium nifA gene is an activator of nitrogen fixation acting in nodule bacteria. To understand the effects of the Sinorhizobium meliloti nifA gene on Alfalfa, the cDNA-AFLP technique was employed to study the changes in gene expression in nifA mutant nodules. Among the approximately 3,000 transcriptderived fragments, 37 had differential expression levels. These expression levels were subsequently confirmed by reverse Northern blot and RT-polymerase chain reaction. Sequence analyses revealed that 21 cDNA fragments corresponded to genes involved in signal communication, protein degradation, nutrient metabolism, cell growth and development.
文摘Alfalfa （Medicago sativa L.） is being grown in harsh environment in Iraq and is mostly subjected to abiotic stresses such as drought, salinity, pH and temperature. Both alfalfa and its nitrogen fixing symbiotic bacteria Sinorhizobium meliloti are affected by these environmental stresses. Enhancing nitrogen fixation biologically could be achieved through selection of tolerant strains of S. meliloti to these environmental stresses and inoculating them to the crop, also growing tolerant cultivars. This study examines phenotypic diversity for tolerance to drought, salinity, temperature and pH. Sixty isolates sampled from different areas of Iraq. The results revealed high degree of phenotypic diversity in Sinorhizobium populations. Furthermore, the isolates which showed tolerance to drought stress also showed tolerance to salinity and high degree of temperature, indicating direct relationship between three physiological path ways. Also 58.3% of drought tolerant isolates were alkaline tolerant they tolerated up to pH 9, point to say almost all drought tolerant isolates in this study illustrated strong ＋ positive reaction to catalase enzyme. And 91.6% themes were negative for Gelatinase enzyme test. While only 50% of drought sensitive isolates were negative for drought sensitive isolates could grow at high temperature （42 ℃）.
文摘Though the majority of bacteria can form structured communities known as biofilms, mutations can cause bacterial strains to vary in their ability to form a biofilm. In this study, the apparent diffusion coefficient of polystyrene microspheres 0.29 μm in diameter, which were executing Brownian motion inside bacterial colonies, was used as a quantitative parameter of the ability of a strain to form a biofilm and of the biofilm development. The study was performed using five Sinorhizobium meliloti strains, the biofilm-forming strains Rm8530 expR+, Rm8530 exoY, and Rm9034 expG, and the non-biofilm forming strains Rm1021 and Rm9030-2 expA1. The green fluorescent beads were placed with each strain in a separate channel of a microfluidic device. Thus, as the bacterial colonies grew under identical conditions over a 4-day period, the motion of the fluorescent microspheres was recorded and the diffusion coefficients were measured every 24 hours via particle tracking algorithms. It was found that each strain displayed a unique pattern of change in diffusion coefficient over time. Also, for a given biofilm-forming strain, there was a clear correlation between the value of the diffusion coefficient and the appearance and motility of the bacterial community. Thus, the diffusion coefficient can be used to identify different S. meliloti strains, and for the biofilm-forming strains, it is also a quantitative indicator of the stage of biofilm development.
文摘Soil bacteria Sinorhizobium meliloti had enormous agricultural value, due to their ability in fixing nitrogen symbiotically with an important forage crop legume--alfalfa. The aims of the present study were （1） to isolate indigenous S. meliloti from different field sites in lraq; （2） to evaluate the isolates tolerance to induced drought using polyethylene glycol-6000; （3） assessing genetic diversity and genetic relationships among isolates of natural population with drought tolerant abilities. Drought tolerance study revealed vast variations between Sinorhizobium isolates, the highest tolerant isolates to drought were 12 from total thirty （40%）, tolerated from -3 up to -4 MPa, while the drought sensitive isolates tolerated up to -1.5 MPa, except isolate Bs 58 which tolerated up to -1 Mpa water potential. The growth declined with the increase of drought stress. Cluster analysis based on Polymerase Chain Reaction-Restriction Fragment Length Polymorphism （PCR-RFLP） of the 16S rDNA showed two divergent groups with 41% similarity, the first group included three drought sensitive isolates （Bs 44, Bs 54 and Bs 58）, the second group comprised the rest nine isolates （moderate and high drought tolerant）, except for Bs 55 which was drought sensitive isolate, all isolates in the two groups showed no differences between them. The PCR-RFLP of 16S rDNA method revealed a genetic variance between the drought sensitive and tolerant isolates.
