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1.
In order to investigate central metabolic changes inBeijerinckia indica, cells were grown on different carbon sources and intracellular flux distributions were studied under varying concentrations of nitrogen. Metabolic fluxes were estimated by combining material balances with extracellular substrate uptake rate, biomass formation rate, and exopolysaccharide (EPS) accumulation rate. Thirty-one metabolic reactions and 30 intracellular metabolites were considered for the flux analysis. The results revealed that most of the carbon source was directed into the Entner-Doudoroff pathway, followed by the recycling of triose-3-phosphate back to Hexose-6-phosphate. The pentose phosphate pathway was operated at a minimal level to supply the precursors for biomass formation. The different metabolic behaviors under varying nitrogen concentrations were observed with flux analysis.  相似文献   

2.
Abstract A total of 285 strains of Azospirillum were isolated from soils from seven geographic regions in New South Wales, Australia, using an immunomagnetic separation procedure which does not select strains according to their nitrogen-fixing ability. By combining amplification and restriction analysis of 16S rDNA (ARDRA) patterns with serological, morphological and biochemical results, we found that almost all isolates were A. brasilense and A. lipoferum. There was wide variation in the nitrogenase (acetylene reduction) activity of isolates grown in nitrogen-free, semisolid medium, with differences in average activities between regions. Isolates with zero or negligible nitrogenase activity were found in samples from only two regions, one of which had two out of 26 strains with no activity. Representative isolates, having the highest, the lowest, and intermediate nitrogen fixation rates for each site, were used to inoculate the roots of wheat plants in a model system. Most of the isolates, in association with wheat roots, reduced between 1 and 5 nmol C2H4· mg dry root−1· day−1, but certain strains gave considerably higher activities. The rank order of nitrogen fixation activity on wheat roots did not correlate well with that of nitrogen fixation in pure culture; some strains that fixed nitrogen vigorously in pure culture had low rates of fixation on roots, and vice versa. This inconsistency could not be explained by variations in the root colonizing ability of different strains. However, isolates of A. lipoferum had a higher average nitrogenase activity than A. brasilense, both in Nfb medium and in association with wheat roots. The majority of the most active nitrogen fixers were A. lipoferum. When wheat plants were inoculated with mixtures of two or four strains, nitrogen fixation rates were generally between the rates for the component strains when inoculated individually. There was no benefit from using mixtures of different strains. Received: 23 July 1997; Accepted: 4 December 1997  相似文献   

3.
Beijerinckia indica subsp. indica is an aerobic, acidophilic, exopolysaccharide-producing, N2-fixing soil bacterium. It is a generalist chemoorganotroph that is phylogenetically closely related to facultative and obligate methanotrophs of the genera Methylocella and Methylocapsa. Here we report the full genome sequence of this bacterium.Beijerinckia indica subsp. indica ATCC 9039 is the type strain of the genus Beijerinckia (6), a member of the Rhizobiales order of the Alphaproteobacteria. Beijerinckia spp. are commonly found as free-living bacteria in acidic soils and also in plant rhizosphere and phyllosphere environments (4). Research on Beijerinckia has suffered from chronic taxonomic confusion, with some strains of Sphingomonas and Azotobacter being misidentified in the literature: e.g., a “Beijerinckia” reported to degrade PAH has been reclassified (3). However, some Beijerinckia spp. have received research attention due to their plant growth-promoting properties (7) and for their abundant production of exoheteropolysaccharide with potential biotechnological uses (5).Genomic DNA from Beijerinckia indica subsp. indica was used to create 3-kb, 8-kb, and 40-kb DNA libraries. Sequencing, assembly, and automated annotation were performed at the Joint Genome Institute using standard procedures (U.S. Department of Energy; http://www.jgi.doe.gov/sequencing/strategy.html). The total number of paired-end shotgun Sanger reads in the assembly was 33,870. In addition, Roche 454 sequence data were included into the final assembly. Large Newbler contigs of 454 reads were chopped into 4,975 overlapping fragments of 1,000 bp and entered into the assembly as pseudoreads.The genome of B. indica subsp. indica was 4,170,153 bp. In addition, two plasmids of 181,736 and 66,727 bp were present. There are a total of 3,982 open reading frames (ORFs) predicted using Glimmer, of which 3,784 are predicted protein-coding genes and 2,695 (70%) have been assigned a predicted function. There are 134 pseudogenes, 52 tRNA genes, and three operons each containing 16S, 23S, and 5S rRNA genes. The G+C content is 57.0% (56% and 54% in the plasmids).The bacterium lacks phosphofructokinase, the key enzyme of the Embden-Meyerhof pathway. Instead, it uses the Entner-Doudoroff or pentose phosphate pathway to catabolize sugars, which is typical of free-living Rhizobiales. The majority of the genes involved in N2 fixation are clustered in two genomic islands (10 kb and 51 kb), with the notable exception of the nifS gene encoding cysteine desulfurase.Beijerinckia indica is a metabolically versatile bacterium capable of growth on a variety of organic acids, sugars, and alcohols (4). In contrast, its close phylogenetic cousins Methylocella and Methylocapsa are highly specialized methanotrophs capable of growth on very few substrates (2). However, the genome size of Beijerinckia indica compared to that of Methylocella silvestris (4.17 versus 4.30 Mbp) and the numbers of predicted protein-encoding genes (3,788 versus 3,917) are remarkably similar. A BLAST analysis indicated that the 57% of the genes in the genome of B. indica have homologues in M. silvestris (stringency threshold expectation value [E] of 1e−50). Some key pathways of one-carbon metabolism (such as the tetrahydromethanopterin and serine pathways of formaldehyde metabolism) that are present in M. silvestris appear to be absent or incomplete in B. indica, which confirms previous experiments showing that the organism is incapable of methylotrophic growth (1). However, an operon encoding a putative propane monooxygenase homologous to soluble propane/methane monooxygenases of Methylocella silvestris BL2 was identified. More in-depth comparison of these genomes will help elucidate what defines their distinct lifestyles.  相似文献   

4.
Summary The nitrogen fixation rate in a 4-year-old stand of the woody legumeLeucaena leucocephala (Lam.) de Wit. was estimated in the field at a rather dry site in Tanzania by use of an acetylene reduction technique. The diurnal mean value during April–May was 35 nmol C2H4 mg–1 (dry weight) nodules h–1, with a variation between 22±8 and 48±12 nmol C2H4 mg–1 (dry weight) nodules h–1 in early morning and at midday, respectively. The nodule biomass was determined by auger sampling to be 51±16 kg (dry weight) ha–1. Most of the nodules were found at the 10–30 cm soil depth level. A rough calculation of the amount of nitrogen fixed annually arrived at 110±30 kg ha–1. The results give strong support for the use ofL. leucocephala for soil enrichment in less humid areas of tropical Africa.  相似文献   

5.
To investigate the role of ammonium-assimilating enzyme in heterocyst differentiation, pattern formation and nitrogen fixation, MSX-resistant and GS-impaired mutants of Anabaena 7120 were isolated using transposon (Tn5-1063) mutagenesis. Mutant Gs1 and Gs2 (impaired in GS activity) exhibited a similar rate of nitrogenase activity compared to that of the wild type under dinitrogen aerobic conditions in the presence and absence of MSX. Filaments of Gs1 and Gs2 produced heterocysts with an evenly spaced pattern in N2-grown conditions, while addition of MSX altered the interheterocyst spacing pattern in wild type as well as in mutant strains. The wild type showed complete repression of heterocyst development and nitrogen fixation in the presence of NO3 or NH4 +, whereas the mutants Gs1 and Gs2 formed heterocysts and fixed nitrogen in the presence of NO3 and NH4 +. Addition of MSX caused complete inhibition of glutamine synthetase activity in wild type but Gs1 and Gs2 remained unaffected. These results suggest that glutamine but not ammonium is directly involved in regulation of heterocyst differentiation, interheterocyst spacing pattern and nitrogen fixation in Anabaena.  相似文献   

6.
Summary Different methods were used to demonstrate the existence of Ca++-binding sites (Ca++-bs) at the plasma membrane ofAmoeba proteus. In pinocytoting animals the number (indicated by the average distanced in nm) and size (average longitudinal axiss in nm) of Ca++-bs at the cytoplasmic surface of the cell membrane were significantly increased (d=162±15;n=41 ands=93±5;n=47) in comparison to controls (d=208 ±21;n=37 ands=59±8;n=45). The ratio of P: Ca obtained by X-ray microanalysis was in the range of 1.5. The differences observed in the two experimental groups of amoebae are explained by conformational changes in the molecular structure and an increased Ca++-permeability of the plasma membrane during induced pinocytosis.Microplasmodia of the acellular slime moldPhysarum polycephalum investigated for comparison were found to have no Ca++-bs at the interior cell surface.  相似文献   

7.
Summary Snails and nematodes, the potential cyanobacterial grazers, differ in their choice for cyanobacterial diet. Snails prefer non-mucilaginous forms while nematodes prefer mucilaginous forms. Such differences in feeding choice between the cyanobacteria suggests that it may not be possible to select strains of diazotrophic cyanobacteria that are resistant to all grazers. The potential consumption of cyanobacteria at an average field density of 20,000 snails ha−1 was estimated to be about 50 kg (fresh weight) ha−1 day−1. Dorylamus sp. was most dominant nematode associated with cyanobacterial consumption. Phytoextracts of neem (Azadirachta indica), bel (Aegle marmelos) and tobacco (Nicotiana tabacum) were effective in controlling these cyanobacterial grazers. The minimum concentration of neem, bel and tobacco phytoextract in water for 100 % mortality of snails were 0.1, 2.0 and 0.05%, respectively. However, trepellent level was only 0.01% for neem and tobacco phytoextract. Complete mortality of nematode (Dorylamus spp.) required a higher concentration level (2%) even in the most effective tobacco phytoextract. Lower levels of phytoextract (0.1%) were found to stimulate growth and nitrogen fixation of cyanobacteria. Application of these plant biomasses resulted in significant increase in cyanobacterial acetylene-reducing activity (ARA) and rice yield and a significant decrease in snail and nematode population. Augmentation of cyanobacterial acetylene-reducing activity was two to three times higher in comparison to the control in both the years of experimentation. Rice yield also increased between 3.8 and 58.5% over the control, depending on the quantity and nature of plant biomass. Tobacco waste was significantly superior in comparison to neem and bel biomass as carrier of cyanobacterial culture.  相似文献   

8.
Summary Roots of sugar cane had considerable nitrogenase activity and produced up to 5 n moles ethylene/h/g root by the reduction of acetylene. The rhizosphere soil and soil mid-way between the cane rows also reduced acetylene.Beijerinckia indica was abundant on roots and in the soil. Nitrogenase activity was also associated with roots ofPanicum maximum,Pennisetum purpureum andCymbopogon citratus.  相似文献   

9.
The DROOPING LEAF and OsETTIN2 genes promote awn development in rice   总被引:1,自引:0,他引:1  
The awn is a long needle‐like appendage that, in some grass species, is formed on the lemma that encloses floral organs together with the palea. In rice, most wild species and most strains of Oryza sativa ssp. indica generate an awn, whereas most strains of O. sativa ssp. japonica do not. In japonica, the long‐awn characteristic appears to have been lost during domestication and breeding programs. Here, we found that the genes DROOPING LEAF (DL) and OsETTIN2 (OsETT2) are involved in awn development in the awned indica strain Kasalath. Genetic analyses and RNA‐silencing experiments indicate that DL and OsETT2 act independently in awn formation, and that either gene alone is not sufficient for awn development. Scanning electron microscopy revealed that the top region of the lemma (a putative awn primordium) is larger in an awned floret than in an awnless floret. OsETT2 is expressed in the awn primordium in the awned indica floret, but not in the awnless japonica floret except in the provascular bundle. DL is expressed underneath the primordium at similar levels in both indica and japonica florets, suggesting non‐cell‐autonomous action. We hypothesize that loss of expression of OsETT2 in the awn primordium is probably associated with the failure of awn formation in japonica strains.  相似文献   

10.
3种水稻土中7株固氮蓝细菌的分离与特征   总被引:1,自引:0,他引:1  
【背景】蓝细菌是水生和陆地生态系统中生物固氮的主要贡献者。【目的】增加对稻田土壤固氮蓝细菌的了解,获得用于进一步研究的可培养固氮蓝细菌菌株。【方法】选择3种具有不同固氮能力的水稻土,采用BG11-N培养基分离培养固氮蓝细菌菌株,对新分离菌株进行形态特征观察,通过基因组DNA的nifH基因扩增明确其固氮潜力,进一步采用乙炔还原法和~(15)N_2示踪法定量测定其固氮能力,通过基因组DNA的16SrRNA基因序列比对进行鉴定。【结果】在光照培养条件下,采用BG11-N培养基共分离纯化得到自养菌株7株,细胞呈圆形或椭圆形、单列、无分枝、丝状和念珠状,在固体培养基上形成团垫状菌落。新分离菌株在BG11-N培养基中生长状况良好,以基因组DNA为模板可扩增出nifH基因,乙炔还原法和~(15)N_2示踪法测定结果显示具有较高固氮能力,同时具有铁载体生成能力。结合16S rRNA基因序列比对和形态特征,7株菌被初步鉴定隶属于念珠藻科(Nostocaceae)。【结论】从水稻土中分离到在稻田生物固氮中发挥重要作用的蓝细菌(念珠藻科)菌株,可培养固氮蓝细菌菌株固氮能力较高,兼具铁载体生成能力,可作为进一步深入研究的微生物资源,具有潜在的研究应用价值。  相似文献   

11.
1. We propose that the appearance and establishment of Nostocales (cyanobacteria) species of the genera Aphanizomenon and Cylindrospermopsis in the warm subtropical Lake Kinneret (Sea of Galilee, Israel) from 1994 was linked to changes in climate conditions and summer nitrogen (N) availability. 2. From 1994 to 2009, an increase in frequency of events of elevated water temperature (>29 °C) in summer, and to some extent a greater frequency of lower summer wind speed events, affected water turbulence and water column stratification, thus providing better physical conditions for the establishment of these populations. 3. In recent years, N‐depleted conditions in Lake Kinneret in early summer have promoted the development and domination of Nostocales that could gain an ecological advantage owing to their N2‐fixing capability. 4. Nitrogen fixation rates coincided both with heterocyst abundance and with Nostocales biomass. The N supplied to the lake via nitrogen fixation ranged from negligible quantities when Nostocales represented only a minor component of the phytoplankton community to 123 tonnes when Cylindrospermopsis bloomed in 2005. This high N2 fixation rate equals the average summer dissolved inorganic nitrogen load to the lake via the Jordan River.  相似文献   

12.
The adaptive strategy of Tetraselmis indica to survive darkness was evaluated over a period of 182 days. Resting cells formation and reduction in catabolic activity facilitated dark survival and their rejuvenation. The occurrence of thick-walled cells (type 1) and thin-walled cells with the condensed chloroplast (type 2) in light-dark and dark conditions respectively indicated differential resting cell formation. The dissolved organic carbon remained nearly constant throughout the dark period signifying T. indica did not favour heterotrophy. Upon re-illumination of dark-adapted cells, the lag phase lengthened from 2 to 28 days when darkness period increased from 8 to 182 days, respectively. However, this negative influence on growth was compensated by an increase in growth rate from 0.35 to 0.95 d?1. This resurgence capability of T. indica after six months of dark exposure and distinctive resting cell formation indicate a differential population sustenance mechanism. Further elucidation of dark adaptability and survival strategies in T. indica may have significance in ecological and evolutionary processes.  相似文献   

13.
Acetylene reduction and nitrogen fixation by strains of Beijerinckia indica and B. lacticogenes increased with increased partial pressures of acetylene and nitrogen up to 80 kPa. The optical emission spectrophotometric method was used for the determination of 14N:15N ratios. The molar ratios of acetylene to nitrogen varied greatly from the theoretical value.  相似文献   

14.
Nutrient conservation in plants and soil fertility may be intricately linked. We studied nitrogen conservation in small Scots pine (Pinus sylvestrisL.) trees growing in stands on organogenic Dystric Histosols and on mineral Podzols. Nitrogen-resorption efficiency (NRE) and proficiency (NRP) of senescent needles, and mean residence time of nitrogen (MRT) were studied in relation to needle surface area, needle longevity, and leaf mass per area (LMA). Trees on Podzols had higher nitrogen concentration in green needles than the trees on Dystric Histosols, but the nitrogen concentration of yellowing needles was similar for trees on both soil types. NRE averaged 65±3.5% (mean±SD) and 56±7.2% for the trees on Podzols and Dystric Histosols, respectively. Neither NRP (0.44±0.05% and 0.35±0.07%, respectively) nor MRT (8.4±2.3 and 6.1±1.2 years) differed significantly between the stands on the two soil types. Mean needle surface area was significantly smaller in trees on Dystric Histosols (76±29 mm2) than on Podzols (131±38 mm2), whereas needle longevity varied between 2 and 4 years independently of the soil type. Trees invested, on average, the same amount of dry matter per unit of needle area on both soil types. Growth of trees, measured as increment of shoot length, was more restricted on Dystric Histosols (55±18 mm yr–1) than on Podzols (184±44 mm yr–1). The results of the correlation analysis applied to pooled data were inconsistent with the relations between traits of stress resistance syndrome observed in inter-specific comparisons. The study indicated that Scots pine trees relocated nitrogen from senescent foliage more efficiently on mineral Podzols than on organogenic Dystric Histosols, but the minimum nitrogen concentration of needles appeared to be similar on both soil types.  相似文献   

15.
We have developed an efficient transformation system for Tylophora indica, an important medicinal plant in India, using Agrobacterium rhizogenes strains LBA9402 and A4 to infect excised leaf and stem explants and intact shoots at different sites. The induction of callus and transformed roots was dependent on the bacterial strain, explant type and inoculation site used. Transformed roots were induced only in explants infected with A. rhizogenes strain A4, while an optimal transformation frequency of up to 60% was obtained with intact shoots inoculated at the nodes. The presence of the left-hand transferred DNA (TL-DNA) in the genome of T. indica roots induced by A. rhizogenes was confirmed by PCR amplification of the rooting locus genes of A. rhizogenes. Root growth and the production of tylophorine, the major alkaloid of the plant, varied substantially among the nine root clones studied. Both parameters increased over time in liquid cultures, with maximum biomass and tylophorine accumulation occurring within 4–6 weeks of growth in fresh medium. Interestingly, in liquid culture, the culture medium also accumulated tylophorine up to concentrations of 9.78±0.21 mg l–1.  相似文献   

16.
Antipchuk  A. F.  Kosenko  L. V. 《Microbiology》2004,73(1):51-55
The influence of lipopolysaccharides (LPS), glucans, and their unseparated complexes on nodulation activity of rhizobia and efficiency of their symbioses with pea plants was studied in vegetation tests. Two Rhizobium leguminosarum bv. viciae strains which differed in their symbiotic properties were used: strain 31 (fix+, efficient, moderately virulent, and moderately competitive) and strain 248b (fix, inefficient, highly virulent, and highly competitive). Preparations of LPS–glucan complex and the respective LPS from the highly virulent strain 248b increased the nodulation activity of both strains by 10–26%. Analogous preparations from a less virulent strain 31 did not have this ability. Unseparated LPS–glucan complexes from these strains increased the productivity of plants infected with the efficient strain by 18–23% but did not change it in plants inoculated with the other, inefficient strain. No significant influence of LPS preparations on the symbiosis productivity was observed. Glucans from both strains enhanced the nodulation ability of the highly virulent strain by 36–56%. In addition, treatment of pea plants with glucan from strain 248b increased nitrogen fixation by root nodules by 27% in plants inoculated with strain 31. In general, the formation and efficiency of the symbiosis of R. leguminosarum bv. viciae with pea plants was more influenced by preparations from strain 248b, highly virulent but deficient in nitrogen fixation, than by preparations from the nitrogen fixation–proficient but less virulent strain 31.  相似文献   

17.
Summary Nitrogenase activity was measured by acetylene reduction in excised Myrica gale nodules collected throughout the growing season at two sites associated with a small lake in central Massachusetts. One site was in an open, weakly minerotrophic peatland dominated by M. gale and the other was on the lakeshore. Nitrogenase activity appeared in late May when the leaves unfolded, reached a peak in July when the maximum number of leaves was present, and declined until it was no longer measurable in late October several days after all leaves had fallen. Summer activities were substantially higher at the peatland site than the lakeshore. Maximum activities were 19.8±1.9 and 8.1±0.7 mol/h x g dry weight (x±SE; N=20) at the peatland and lakeshore sites respectively.Nitrogenase activities were very low at 5° C and increased linearly from 10 to 30° C, the highest temperature examined. The maximum soil temperature measured was 20° C, and no significant diurnal fluctuation in activity was detected.Annual nitrogen fixation calculated from the seasonal nitrogenase activity curve was 34 kg N/ha x yr at the peatland site with mean dry weight nodule biomass of 104 kg/ha, and 24 kg N/ha x yr at the lakeshore with 111 kg/ha nodule biomass. These rates of nitrogen fixation are equivalent to 4–5x the amount of nitrogen contained in bulk precipitation and are major components in the nitrogen budgets of the M. gale plants and wetlands in which they grow.  相似文献   

18.
Summary Azotobacter associated withEichhornia crassipes were found to be concentrated in the slime formed around petiole bases. Massive colonies formed where raw slime was applied to nitrogen-free medium inoculated with Azotobacter. Slime in which Azotobacter had been killed did not stimulate the formation of massive colonies unless live Azotobacter cells were mixed into it. Acetylene reduction tests indicated that Azotobacter associated with Eichhornia did not fix nitrogen unless the plant was suffering extreme nitrogen deficiency, and that the maximum rate of fixation was inadequate for rapid plant growth.  相似文献   

19.
In this study, a process of removing a half volume of culture broth and replacing it with an equal volume of substituted solution was developed to enhance the production of heteropolysaccharide-7 (PS-7) by Beijerinckia indica HS-2001. The optimal substitution time and volume of the substituted solution were found to be 48 h after cultivation and 50% of the initial volume of the culture broth. The optimal composition of the substituted solution was determined to be 20.0 g/L glucose, 10.0 g/L soybean pomace, 0.1 g/L MgSO4·7H2O, 0.9 g/L NH4NO3, and 5.0 g/L potassium phosphate, which was the same composition as the medium developed in a previous study for the production of PS-7 by B. indica HS-2001. The total amount and productivity of PS-7 by B. indica HS-2001 with a substitution under optimized conditions in a 7 L bioreactor for 96 h were 49.28 g and 0.51 g/h, respectively, which were 1.76 and 1.31-foldgreater values than those without a substitution for 72 h.  相似文献   

20.
Summary The Warburg's manometric technique was used to measure the rate of oxygen consumption of the second generation of laboratory-reared snails, Biomphalaria alexandrina and Bulinus truncatus at two temperatures of 25° and 30°C. The individual weight of the experimental snails ranged between 40 and 78 mg for B. alexandrina, between 60 and 90 mg for B. truncatus.At 25°C, the uninfected snails B. alexandrina consumed oxygen at an average rate of 0.096 ± 0.020 ml/g wet wt/hr. The rate of oxygen consumption increased to an average of 0.147 ± 0.008 ml/g wet wt/hr for uninfected snails maintained at 30°C (about 53 per cent increase). The average RW value for uninfected snails maintained at 25°C was 0.80.The snail Bulinus truncatus showed higher oxygen requirements than the snail Biomphalaria alexandrina. At 25°C, it consumed oxygen at an average rate of 0.124 ± 0.016 ml/g wet wt/hr. At 30°C, the rate of oxygen consumption reached a value of 0.220 + 0.006 ml/g wet wt/hr. The average RQ for Bulinus truncatus maintained at 25°C was 0.87.The rate of oxygen consumption of the schistosome — infected Biomphalaria alexandrina snails, maintained at 25°C decreased to an average rate of 0.059 ± 0.010 ml/g wet wt/hr, (an average of 39 per cent decrease). The respiratory quotient (RQ) also decreased to an average value of 0.58. Further research is suggested to clarify the metabolism of both schistosome-infected and uninfected snails.Read at the Ist African Symposium on Bilharziasis, Cairo Egypt, U.A.R., February, 1969.From the Laboratory of Bilharziasis Research, National Research Centre, Dokki, Egypt, U.A.R.  相似文献   

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