粪产碱菌（Alcaligenes faecalis）吸收氢和同化CO2的特性

应用同位素氚(T_2)和~13C(~13CO_2),证明了水稻联合固氮菌——粪产碱菌A—15是一种含有吸氢酶的兼性化能自养细菌,具有较强的吸氢能力,吸氨酶活性可达到13.11μmol H_2 ml~(-1) cultureh~(-1);同时,它还可利用H_2为能源同化CO_2营化能自养生活,其RuBPC活性为24.65 nmolCO_2 mg~(-1) protein min~(-1)。无论在自养还是异养条件下,H_2都支持、并促进固氮活性。粪产碱菌培养在N_2条件下比在NH_4~ 条件下能积累更多的多聚-β羟基丁酸(PHB)。     

N_2和NH_4~ 培养下粪产碱菌固氮酶铁蛋白的生物合成及特性比较

应用DEAE-纤维素和凝胶柱层析,分别将N_2和NU_1~ (30 mmol/L)培养的粪产碱菌固氮酶铁蛋白(Af2和Af~ 2)分离并提纯52倍,在SDS-PAGE上呈均一状态。Af2的比活性达1540 nmol C_2H_4mg~(-1)protein min~(-1),Af~*2无活性。Af2和Af~*2理化性质基本相同,分子量为64.5 kD;均由2个亚基组成,每个亚基分子量为32.5kD;氨基酸种类相同,总残基数分别为537和553,不含色氨酸;每分子Af2和Af~ 2均含有4个Fe原子和4个酸不稳定S~(2-)原子;UV-vis光谱吸收特征相同;荧光探剂测定结果为:每分于Af2和Af~*2均络合2个分子MgATP或2个分子MgADP。     

响应面法优化粪产碱菌Alcaligenes faecalis DL-08产纤维素酶培养工艺

利用粪产碱菌Alcaligenes faecalis DL-08,进行产纤维素酶培养,在Plackett-Burman设计基础上,通过Design-Expert中心组合实验,建立了培养条件优化的二次回归模型,并通过响应曲面分析,确定了最佳培养条件。由响应面分析结果可知,产纤维素酶条件:温度37.98℃,碳氮比1∶1,接种量2%,初始p H 7,时间24 h。在此条件下,预测值OD520为1.237。经3组平行试验,OD520平均值为1.229,与模型预测值基本一致,且与未优化条件相比,纤维素酶活力提高了5.6%,可达1 303.2 U/m L。     

粪产碱菌(Alcaligenes faecalis)A-15与水稻根的结合作用

近年来,联合共生固氮作用已逐渐为人们所重视,与共生、自生固氮生物不同,这些细菌多聚集在植物根表,在根部的粘胶层中,部分细菌可进入根表层内。粪产碱茵A—15是从水稻根分离到的固氮细菌,     

N2和NH4^＋培养下粪产碱菌固氮酶钼铁蛋白的合成及其特性

应用柱层析和制备电泳分别将N_2和 NH_4~ 培养的粪产碱菌固氮酶钼铁蛋白(Af 1和Af 1)分离并纯化,两者理化性质十分相似。分子量分别为226 kD和 222 kD;α亚基和β亚基分子量分别为57 kD和60 kD;氨基酸种类相同,总残基数分别为1790和1750;每分子Af 1和Af 1均含有2个原子Mo和32个原子 Fe;金属原子簇的氧化还原当量数为6。Af 1的比活性为 1477 nmolC_2H_4 mg~(-1)protein min~(-1),Af 1无活性;分子结构两者有差异。     

氨和谷氨酰胺对浑球红假单胞菌（Rhodobacter sphaeroides）固氮酶...

浑球红假单胞菌菌株601具有迅速对外源氨作出“关闭”固氮酶活性的反应。氨对固氮酶的抑制作用,可被谷氨酰胺合成酶(GS)抑制剂MSX所解除。反之,加入Glu代谢抑制剂DON,可延长氨抑制的持续时间。Gln对固氮酶也有抑制作用。在脱腺苷化GS的透性细胞中,加入Gln可抑制固氮酶活性,同时,GS腺苷化状态提高。然而,氨则对透性细胞的固氮酶活性和GS腺苷化状态没有影响。     

光与氨对Rhodopseudomonas capsulata固氮活性的调节

光强是调节氨瞬间抑制Rps.capsulata光合固氮活性的一个因子。与弱光(500 lx)比较,强光(30000 lx)对固氮活性氨抑的启动推迟。被氨抑制了的固氮活性在强光下较在弱光下提前解抑。经光合作用解联剂处理的菌体,强光拮抗固氮活性氨抑的现象消失。菌体ATP库水平分析表明:在氨关闭固氮活性时,库量升高。氨的同化被阻抑时,氨对光合固氮的瞬间抑制消失,菌体ATP库保持恒定。对光强与氨抑制固氮活性之间可能涉及的机制进行了探讨。     

Nitrogenase activity in composting horse bedding and leaves

To date little or no nitrogen fixation has been found in composting materials. However, some materials have high C:N ratios and thus might be expected to support nitrogen fixation. We examined windrows of composting horse bedding and leaves for nitrogenase activity via acetylene reduction and used carbon monoxide controls to differentiate ethylene evolved by nitrogenase from that evolved by endogenous processes. In both piles temperatures were substantially elevated and partial pressures of oxygen greatly reduced thus indicating vigorous microbial activity. In the horse bedding, there were only low rates of ethylene evolution and little of this was due to nitrogen fixation. In contrast, much higher rates of ethylene evolution were measured in the leaf pile and 94% of this was due to nitrogenase activity. We estimate that the leaf pile fixed approximately 0.062 mg N/g leaves during a 5-week period.     

沙坡头地区生物土壤结皮的固氮活性及其对水热因子的响应

氮是除水分之外影响干旱区生态系统生物活性的关键因子。生物土壤结皮是干旱半干旱荒漠地表景观的重要组成部分, 也是荒漠生态系统氮素的主要贡献者。通过野外调查采样, 利用开顶式生长室, 模拟不同降水梯度, 采用乙炔还原法连续测定了沙坡头地区典型生物土壤结皮(藻类结皮、地衣结皮和藓类结皮)在其主要固氮活跃期(6-10月, 湿润期)的固氮活性, 及其对水热因子的响应特征。结果表明, 试验期三类生物土壤结皮的固氮活性介于2.5 × 10³-6.2 × 10⁴ nmol C₂H₄·m^-2·h^-1之间, 其中藻类结皮的最高(平均达2.8 × 10⁴ nmol C₂H₄·m^-2·h^-1), 地衣结皮的次之(2.4 × 10⁴ nmol C₂H₄ ·m^-2·h^-1), 藓类结皮的最低(1.4 × 10⁴ nmol C₂H₄·m^-2·h^-1), 差异显著(p < 0.001)。在模拟降水3 mm时, 三类结皮均可达到最大固氮速率, 当发生> 3 mm的降水事件时, 它们的固氮速率无显著增加; 不同结皮的固氮活性与温度均呈显著的负相关关系(r_藻类结皮 = -0.711, r_地衣结皮 = -0.732, r_藓类结皮 = -0.755, p < 0.001), 藻类和藓类结皮的固氮活性的最适温度区间为25-30 ℃, 地衣结皮为20-30 ℃。三类结皮之间的这种固氮差异主要归因于结皮组成生物体即隐花植物的差异, 藻类结皮主要成分为大量的蓝细菌和一些绿藻, 地衣结皮也由大量的固氮藻和真菌共生形成, 而藓类结皮的主要组成部分苔藓植物并不具有固氮作用, 其微弱的固氮量是结皮中混生的少量蓝细菌或地衣所致。     

甘蔗器官固氮酶活性及其对接种固氮菌的响应

以巴西固氮甘蔗品种B1、B8和广西主栽甘蔗品种ROC22和ROC16为材料,通过桶栽砂培方法,研究不同品种甘蔗及接种固氮菌R16SH后不同生育期和不同器官(根、茎、叶片、叶鞘)的固氮酶活性.结果显示,4个品种甘蔗及接菌后各器官的固氮酶活性均在伸长初期或伸长盛期达到高峰,且以根和茎较高,叶、叶鞘次之;B1、B8根的接菌效果最佳,其固氮酶活性分别比对照增加13.22%和12.42%,而B1和ROC16茎的固氮酶活性分别比对照提高12.19%和12.02%,叶片提高了8%~9%,叶鞘的固氮酶活性变化最小;4个品种甘蔗固氮酶活性表现为B8>B1>ROC22>ROC16,且品种B8与ROC22和ROC16存在显著或极显著差异;相对于品种ROC22和ROC16,固氮菌R16SH对B1与B8的株高以及B1的茎径有更好的生长诱导效应,且在伸长期表现最明显.研究表明,甘蔗的固氮酶活性在品种、器官及生育期间均存在明显差异,接种固氮菌能不同程度提高各品种器官的固氮酶活性,且各品种根、茎以及B1和B8的叶片固氮酶活性显著或极显著差异增加;巴西固氮甘蔗品种表现出比广西主栽甘蔗品种更强的固氮酶活性,且对接种固氮菌反应更敏感.     

Relation between nitrogenase synthesis and activity in a marine unicellular diazotrophic strain, Gloeothece sp. 68DGA (Cyanophyte), grown under different light/dark regimens

The relationship between the abundance of nitrogenase and its activity was studied in the marine unicellular cyanobacterium Gloeothece sp. 68DGA cultured under different light/dark regimens. The Fe‐ and MoFe‐protein of nitrogenase and nitrogen (N₂)‐fixing (acetylene reduction) activity were detected only during the dark phase when the cells were grown under a 12 h light/12 h dark cycle (12L/12D). Nitrogenase activity appeared about 4 h after entering the dark phase. Maximum nitrogenase activity occurred at around the middle of the dark phase, and the activity rapidly decreased to zero before the start of the light phase. The rapid decrease of nitrogenase activity and the Fe‐protein of nitrogenase near the end of the dark phase in 12L/12D were partly recovered by the addition of l ‐methionine‐sulfoximine, an inhibitor of glutamine synthetase. Diurnal oscillation of the abundance of nitrogenase was maintained in the first subjective dark phase (i.e. the period corresponding to the dark phase) after the cells were transferred from 12L/12D to continuous illumination. However, enzyme activity was detected only when photosynthetic oxygen (O₂) evolution was completely suppressed by reducing the light intensity or by the addition of 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea. Nitrogenase always appeared in the cells about 16 h after starting the light phase, even when the 12L/12D cycle was modified by the addition or subtraction of a single 6 h period of light or dark. These results suggest the following: (i) N₂‐fixation by Gloeothece sp. 68DGA is primarily regulated by an endogenous circadian oscillator at the level of nitrogenase synthesis. (ii) The endogenous circadian rhythm resets on a shift of the timing of the light phase. (iii) Nitrogenase activity is not always reflected in the presence of nitrogenase. (iv) The activity of nitrogenase is negatively regulated by fixed nitrogen and the concentration of ambient O₂.     

生物固氮在我国农业生产中的研究现状及发展对策

简述了生物固氦在我国农业生产中3个热点的研究现状:提高现有固氦生物的固氦能力、化学模拟固氦酶、固氦微生物的生物工程,同时提出了生物固氦在令后生态农业中的发展对策。     

广西甘蔗根际高效联合固氮菌的筛选及鉴定

对广西主要甘蔗产区的根际联合固氮细菌进行了收集和评价,拟筛选获得对甘蔗具有潜在促生性能的联合固氮菌,为甘蔗生产节肥减耗提供依据。结合nifH基因扩增和固氮酶活性分析方法筛选获得36个固氮细菌菌株;进一步对所获得固氮菌株的固氮能力、溶磷性、分泌植物生长素IAA的特性等促进植物生长潜能进行评价,获得了5个同时具有较强固氮能力、降解无机磷和分泌植物生长激素IAA的功能菌株;通过Biolog鉴定系统和16S rRNA序列分析对5个具有较好应用潜力的固氮菌进行分类鉴定。结果表明这5个菌株分别属于Klebsiella sp.、Bacillus megaterium、Pseudomonas sp.、Pantoea sp.和Burkholderia sp.。本研究结果表明广西甘蔗根际联合固氮菌具有较大的开发利用潜力。