水稻根表固氮螺菌的鉴定

作者从水稻根表分离获得一批固氮细菌,对其中固氮活性较高的菌株R₃₈(固氮活性为429.02n·mol·C₂H₄/ml·hr.)进行形态特征和生理生化等特征的鉴定,确定R₃₈属于巴西固氮螺菌(Azospirillum brasilense)     

少动鞘脂单胞菌S1胞外多糖发酵工艺条件研究

研究了摇瓶培养条件下少动鞘脂单胞菌引的胞外多糖的发酵工艺。Ｓ１菌的发酵产胶可采用二步发酵法：第一阶段,培养基成分为：蔗糖１０ｇ,ＮＨ_４ＮＯ_３　０．５ｇ,ＫＨ_２ＯＰ_４　０．５ｇ,ＭｎＳＯ_４　０．３ｇ,ＭｇＳＯ_４　０．１ｇ,吐温８０ ０．０２ｇ溶于蒸馏水并定容至１０００ｍＬ,ｐＨ７．２±０．１,温度３３℃～３５℃,高溶氧。第二阶段,发酵 １０～１２ｈ后,补加蔗糖４０ｇ／Ｌ,温度     

土壤杆菌固氮生理特性研究

对根癌土壤杆菌C58/pGV3850菌株的固氮生理特性研究结果表明,该菌具有自生固氮活性,其固氮活性的最适pH为8.0,温度为30℃.固氮活性在对数生长后期(14h)最高,延缓期和静止期乙炔还原活性较低.该菌株好氧,通过氧呼吸和吸氢酶的作用达到避氧固氮.当通入氧气超过呼吸耗氧时,对固氮活性产生抑制作用.在培养过程中增加NH_4~+浓度,固氮活性会降低,当达到15mmol/L时完全丧失固氮活性,表现NH_4~+阻遏固氮酶蛋白合成.在乙炔还原测定系统中加入NH_4~+并不影响乙炔还原反应,说明没有NH_4~+关闭现象.培养过程中加入MSX(2.5mg/ml)能解除10mmol/L NH_4~+对固氮酶合成的阻遏作用.固定的游离氮不能以NH_4~+形式分泌于胞外.固氮过程中放出大量氢气,培养16小时产氢量达65μmol H_2/mg蛋白.在限碳条件下(0.2%蔗糖)其吸氢酶活力可达520nmol H_2·ml~(-1)·h~(-1).     

河西走廊豆科植物结瘤固氮特性初步研究

对甘肃河西走产野生豆科植物结瘤及固氮状况进行了调查。栽培豆科植物均能自然结瘤,野生豆科植物结瘤率仅５０％。该地区豆科植物根瘤大多数圆形、棒状或指状,形状较为规则。栽培植物根瘤多为粉红色,而野生揿为白色、黄色或棕色。ＡＲＡ测定表明,２５．６％根瘤为无效根瘤。不同种不同地区根瘤固氮活性相差较大,一般活力都较低,乙炔还原活性小１μｍｏｌＣ２Ｈ４／ｇＦＷ．ｈ的占检测样品数的２９．９％,最高者信为２８．１９     

液化沙雷氏菌胞外脂肪酶产生条件的研究

新分离的一株液化沙雷氏菌（Ｓｅｒｒａｔｉａｌｉｑｕｅｆａｃｉｅｎｓ）能产生大量胞外碱性脂肪酶,本文在摇瓶发酵水平上考察了培养基组成、培养条件等因素对其生成脂肪酶的影响。在优选条件下,即培养基组成为（％）玉米油１．２５,豆饼粉２．０,蛋白胨１．０,酵母膏０．２,Ｋ_２ＨＰＯ_４０．２,ＭｇＳＯ_４·７Ｈ_２Ｏ０．１,初始ｐＨ为７．２５时,２８℃,１５０ｒ／ｍｉｎ旋转摇床振荡培养４０ｈ,该菌株的发酵液酶活达到４３ｕ／     

八株芽孢杆菌菌株的分类及固氮活性的测定

从8个省市的不同土壤分离了200个菌株,筛选出8株（JR₁,JR₂,JR₅,ZZ₃,ZZ₁₂,ZZ₁₆,M₁,K₄）能在无氮培养基上生长良好的典型的芽抱杆菌,其中6株（JR₁,JR₂,JR₄和ZZ₃,ZZ₁₂,ZZ     

固氮螺菌耐高铵突变株的生物学特性

本文报道9株固氮螺菌耐铵突变株,在培养基中有30—120m mol/L NH₄⁺存在时,具有强弱不等的固氮酶活性。而出发菌株在有微量NH_4~+存在时,固氮酶活性完全被抑制。耐铵突变株的其它特性与出发菌株相似。     

盐胁迫下烘产碱菌在水稻根表的定殖和异源固氮基因的表达

异源ｎｉｆ－ＬａｎＺ融合基因在粪产碱菌Ａ１５６１中的表达活性随盐浓度增加而升高,然而逐渐降。ｎｉｆＨ－ＬａｅＺ融合基因可以正常表达的盐浓度在０．１％～０．５％之间,盐浓度与０．０５％时活必同Ａ１５６１在盐浓度为０．０６％时趋化能力最强,随着盐浓度的提高逐渐下降,当盐浓度为３．０％时完全人趋化能力。一定的盐浓度（０．５％）对固氮立碱菌的数远大于对照。３种ｎｉｆ－ＬａｃＺ融合基因在根内的表达部位有显著     

Frankia sp．Asll和EaI827菌株的固氮研究

由辽东桤木和沙枣的根瘤中分离出两株内生菌(Frankia sp.),分别编号为Asll和EaI827。可在丙酸作碳源的限定培养基中诱导出泡囊,并表现较高固氮活性。不同浓度的NHt和水解酪蛋白对菌体的生长有促进作用,而对固氮活性却表现出明显的抑制作用。当氧分压为5％时,Asll菌株固氮活性较高,而F~1827菌株最适氧分压为1％。Asll菌株的固氮周期为8一11天,Eai827菌株在11一14天表现出较高的固氮活性。     

海水养殖生境中硫酸盐还原菌活性的抑制机制

【目的】海水养殖生境中的硫化物(H₂S)严重损害养殖生物健康,控制该条件下硫酸盐还原菌(sulfate-reducing bacteria,SRB)的代谢活性是有效抑制H₂S产生的重要途径。【方法】本研究利用稀释涂布-叠皿夹法对海水养殖生境底泥中SRB进行富集筛选,获得SRB菌株,通过投加硝酸盐对菌株产H₂S的活性进行抑制。【结果】获得的2株SRB Desulfovibrio sp.NY-1和Clostridium sp.NH-1,能够在35℃、pH为7.0及盐度为20–30 mg/L条件下,分别积累高达435和150 mg/L H₂S。硝酸盐不能有效抑制NY-1产H₂S的活性,基因调控作用以及缺乏将硝酸盐作为电子受体的酶体系是其不能被抑制的主要原因。硝酸盐对NH-1 H₂S产生活性有可逆性抑制,其具有硝酸盐异化还原成铵(dissimilatory nitrate reduction to ammonium,DNRA)的能力,优先利用硝酸盐作为电子受体。DNRA作用下的中间代谢产物亚硝酸盐是有效抑制菌株NH-1产H₂S活性的主要原因,其抑制机理主要为抑制菌株的生长繁殖。【结论】硝酸盐对不同SRB菌株具有不同的抑制机制和效果,在进行硫化物污染控制前需要对产生硫化物的SRB菌群进行分析判别。     

Pleomorphism and acetylene-reducing activity of free-living rhizobia.

Cowpea-type Rhizobium sp. strain 32H1 and Rhizobium japonicum USDA 26 and 110 grown on a glutamate-mannitol-gluconate agar medium showed increases in the number of pleomorphic cells coincident with their acetylene-reducing activity. Pleomorphs appeared to be inhibited in growth nonuniformly, because acetylene-reducing cultures were mixtures of rod, branched (V, Y, and T), and other irregularly shaped cells. In contrast, strain USDA 10 consistently failed to reduce acetylene, even though it also could grow and yield pleomorphic cells under various conditions. With minimal inhibitory supplements (5 micrograms per ml of medium) of nalidixic acid and novobiocin as cell division inhibitors, an increase in pleomorphic cells was observed, but the inhibited cultures displayed lower acetylene-reducing activity. A study of pleomorphic cells derived in different ways indicated that not all pleomorphs reduce acetylene.     

Glutamate as a differential nitrogen source for the characterization of acetylene-reducing Rhizobium strains

A majority (36 of 44) of Rhizobium japonicum strains tested reduced acetylene asymbiotically when grown on an agar medium containing 0.1% (w/v) L-glutamate as a sole nitrogen (N) source. Glutamate as N source led to pinpoint colonies and uniform glutamine synthetase activity of three selected, slow-growing acetylene-reducing strains ( R. japonicum L-259 and 110 and a cowpea-type Rhizobium 32H1). The three test strains were characterized further by antibiotic resistance, colony type, cellular morphology, and differential growth on different N sources. The evidence suggests that, in an agar medium, glutamate creates a growth condition leading to acetylene reduction activity, pinpoint colonies and pleomorphism.     

Nitrogen-fixation associated with the marine blue-green alga,Trichodesmium, as measured by the acetylene-reduction technique

Summary The marine blue-green alga, Trichodesmium, was collected from the Gulf Stream, near Miami, and occurred in two distinct colonial forms both of which reduced acetylene to ethylene. Trichodesmium was more abundant during the summer but its acetylene-reducing potential showed no obvious seasonal variation. Illuminated Trichodesmium reduced acetylene to ethylene equally well either anaerobically or aerobically (20% oxygen). Acetylene-reduction in the dark, however, was oxygen-dependent and was usually 25% or less of the activity recorded in the light. 0.3–1.0 nmoles of ethylene were produced per minute per mg of protein, by illuminated cultures, and these values compare favorably with those recorded for other nitrogen-fixing blue-green algae. However, the possibility that bacteria contributed to the acetylene-reducing activity associated with Trichodesmium was not completely eliminated.Contribution No. 1578 from the University of Miami, Rosenstiel School of Marine and Atmospheric Science, 10 Rickenbacker Causeway, Miami, Florida 33149, U.S.A.     

Nitrogen fixation and nitrate utilization by marine and freshwater Beggiatoa

Four newly isolated marine strains of Beggiatoa and five freshwater strains were tested for nitrogen fixation in slush agar medium. All strains reduced acetylene when grown microaerobically in media containing a reduced sulfur source and lacking added combined nitrogen. The addition of 2 mmol N, as nitrate or ammonium salts, completely inhibited this reduction. Although not optimized for temperature or cell density, acetylene reduction rates ranged from 3.2 to 12 nmol·mg prot^-1 min^-1. Two freshwater strains did not grow well or reduce acetylene in medium lacking combined nitrogen if sulfide was replaced by thiosulfate. Two other strains grew well in liquid media lacking both combined nitrogen and reduced sulfur compounds but only under lowered concentrations of air. All freshwater strains grew well in medium containing nitrate as the combined nitrogen source. Since they did not reduce acetylene under these conditions, we infer that they can assimilate nitrate.     

Mutant Strain of Bradyrhizobium japonicum with Increased Symbiotic N(2) Fixation Rates and Altered Mo Metabolism Properties

Mutant strains of Bradyrhizobium japonicum that required higher levels of molybdate than the wild-type strain for growth on NO(3)-containing medium were obtained after transposon Tn5 mutagenesis of the wild-type strain. The mutant strains expressed more than fivefold-greater nitrate reductase activities in the range of 0.1 to 1.0 mM added molybdate compared with activities expressed upon incubation in non-Mo-supplemented medium, whereas the nitrate reductase activity of the wild-type strain (JH) was not markedly influenced by Mo supplementation. In free-living culture, mutant strains JH310 and JH359 expressed substantial nitrogenase activity, even in medium treated to remove molybdate, and nitrogenase activity was influenced little by Mo supplementation, whereas the wild-type strain required 100 nM added Mo for highest nitrogenase activity. Double-reciprocal plots of Mo uptake rates versus Mo concentration showed that both bacteroids and free-living cells of mutant strain JH359 had about the same affinity for Mo as did the parent strain. Bacteroids of both the mutants and the wild type also exhibited similar Mo accumulation rates over a 9-min period under very-low-Mo (4 nM) conditions. Nitrogenase activities for strain JH359 and for the wild-type strain in free-living culture were both strongly inhibited by tungsten; thus, the nitrogenase activities of both strains are probably the result of a "conventional" Mo-containing nitrogenase. Soybeans inoculated with strain JH359 and grown under either Mo-supplemented or Mo-deficient conditions had greater specific acetylene reduction rates and significantly greater plant fresh weight than those inoculated with the wild-type strain. Under Mo-deficient conditions, the acetylene reduction rates and plant fresh weights were up to 35 and 58% greater, respectively, for mutant-nodulated plants compared with wild-type-strain-nodulated plants.     

Mutant Strain of Bradyrhizobium japonicum with Increased Symbiotic N2 Fixation Rates and Altered Mo Metabolism Properties

Mutant strains of Bradyrhizobium japonicum that required higher levels of molybdate than the wild-type strain for growth on NO₃-containing medium were obtained after transposon Tn5 mutagenesis of the wild-type strain. The mutant strains expressed more than fivefold-greater nitrate reductase activities in the range of 0.1 to 1.0 mM added molybdate compared with activities expressed upon incubation in non-Mo-supplemented medium, whereas the nitrate reductase activity of the wild-type strain (JH) was not markedly influenced by Mo supplementation. In free-living culture, mutant strains JH310 and JH359 expressed substantial nitrogenase activity, even in medium treated to remove molybdate, and nitrogenase activity was influenced little by Mo supplementation, whereas the wild-type strain required 100 nM added Mo for highest nitrogenase activity. Double-reciprocal plots of Mo uptake rates versus Mo concentration showed that both bacteroids and free-living cells of mutant strain JH359 had about the same affinity for Mo as did the parent strain. Bacteroids of both the mutants and the wild type also exhibited similar Mo accumulation rates over a 9-min period under very-low-Mo (4 nM) conditions. Nitrogenase activities for strain JH359 and for the wild-type strain in free-living culture were both strongly inhibited by tungsten; thus, the nitrogenase activities of both strains are probably the result of a “conventional” Mo-containing nitrogenase. Soybeans inoculated with strain JH359 and grown under either Mo-supplemented or Mo-deficient conditions had greater specific acetylene reduction rates and significantly greater plant fresh weight than those inoculated with the wild-type strain. Under Mo-deficient conditions, the acetylene reduction rates and plant fresh weights were up to 35 and 58% greater, respectively, for mutant-nodulated plants compared with wild-type-strain-nodulated plants.     

Rhizobium trifolii 0403 Is Capable of Growth in the Absence of Combined Nitrogen

Rhizobium trifolii 0403 was treated with 16.6 mM succinate and other nutrients and thereby induced to grow in nitrogen-free medium. The organism grew microaerophilically on either semisolid or liquid medium, fixing atmospheric nitrogen to meet metabolic needs. Nitrogen fixation was measured via ¹⁵N incorporation (18% ¹⁵N enrichment in 1.5 doublings) and acetylene reduction. Nitrogen-fixing cells had a K_m for acetylene of 0.07 atm (ca. 7.09 kPa), required about 3% oxygen for optimum growth in liquid medium, and showed a maximal specific activity of 5 nmol of acetylene reduced per min per mg of protein at 0.04 atm (ca. 4.05 kPa) of acetylene. The doubling time on N-free liquid medium ranged from 1 to 5 days, depending on oxygen tension, with an optimum temperature for growth of about 30°C. Nodulation of white clover by the cultures showing in vitro nitrogenase activity indicates that at least part of the population maintained identity with wild-type strain 0403.     

Drag-reducing properties of bacteria from the skin mucus of the cornetfish (Fistularia commersonii)

Scanning electron microscopy and fluorescence microscopy after scanning with DAPI indicated that the skin mucus of the cornetfish contained large numbers of bacteria, 4 × 10⁸ per cm3. However, viable counts yielded only 2 × 104 per cm³. Twelve bacterial strains were isolated directly from the mucus and another ten strains were obtained following enrichment on pasteurized mucus medium. Most of the isolates belonged to the genus Pseudomonas; a smaller number were classified as Micrococcaceae. Cultures of 13 of the isolates were active in reducing friction in a turbulent flow rheometer. The surface active and drag-reducing properties of three strains—JR5, JR8, and GB7—were studied further. The drag-reducing activities, which were extracellular, were concentrated by ultra-filtration. The chemical composition of the concentrated preparations consisted of 14–24% protein and 38–75% polysaccharide. The major components of the polysaccharide fraction were galacturonic acid, galactosamine, and glucosamine, with lesser amounts of glucose and galactose. The most active preparation, from strain JR8, had a specific drag-reducing activity of 77 units per mg. Strain JR5 was the most hydrophobic as measured by the DOS and BATH tests. JR8 gave intermediate values, and GB8 showed low hydrophobicity values in both tests. The hydrocarbon-in-water emulsifying ability of the concentrated polymer fractions from JR8, GB7, and JR5 were high, intermediate, and low, respectively. The emulsifying and drag-reducing activities of the polymer fraction from strain JR8 were separated from each other by extraction with hydrocarbons. The emulsifying activity was due to a carbohydrate-protein complex, whereas the drag-reducing activity was associated with a uronic acid-containing polysaccharide. Offprint requests to: E. Rosenberg.     

Effects of vanadium and tungsten on the nitrogen-fixing cyanobacteriumNostoc linckia

The effects of vanadium and tungsten on growth of the heterocystous cyanobacteriumNostoc linckia (Roth.) Born. and the activity of its nitrogenase and nitrate reductase were studied. While vanadate stimulated the growth of molybdate-limited cells and also their acetylene-reducing ability, it did not stimulate nitrate reductase activity. Tungstate inhibited growth in molybdate-limited cells and also their acetylene reducing and nitrate reductase activities. Acetylene reduction response of cells grown in a tungstate-containing medium indicated certain difference between the modes of action of vanadium and molybdenum. Tungstate caused a slight increase in heterocyst frequency both in nitrogen-fixing and nitrate-grown material. Cyanophycin granules and polyhedral bodies disappeared in molybdate-deficient cells in which thick deposition of polyglucan-like granules occurred throughout the breakdown of cytoplasm and the thylakoids.