极耐盐碱固氮菌的分离鉴定及固氮特性研究

我国盐碱土面积约9913万hm²,其中pH值高于9、盐含量大于0.6%的重度盐碱地每年以1.4%的速率增长。利用固氮微生物改善植物根际环境,提高作物产量,是盐碱地改良的重要方法。[目的] 从来自海南省三沙市热带珊瑚岛礁的土壤中,分离鉴定自生固氮菌,为极端盐碱地改良提供候选菌株。[方法] 通过形态学观察、生理生化特征分析和16S rRNA序列测定等方法进行菌种鉴定,分析其固氮、耐盐碱和促生长特性,盆栽试验验证其对玉米主要农艺性状的影响。[结果] 获得1株极端耐盐碱的固氮细菌DJ-1,其菌落呈圆形,菌体杆状,大小（0.5-1.3）μm×（0.3-0.5）μm,革兰氏染色阴性,与根癌土壤杆菌（Agrobacterium tumefaciens）的16S rRNA序列高度同源,确定其为根癌土壤杆菌。DJ-1在pH 9、NaCl含量为1%-4%的培养基上可正常生长,能耐受pH 12、NaCl含量8%的环境。从中克隆到固氮酶基因nifH。盆栽试验结果表明,DJ-1可显著促进玉米生长。[结论] 菌株DJ-1能耐受极端盐碱条件,且具有较强的固氮和促生长能力,有可能作为贫瘠盐碱耕地改良功能菌剂的候选菌株。     

Dinitrogen fixation (C2H2 reduction) by bacterial strains at various temperatures

Acetylene-reducing activities (ARA) of strains ofEnterobacter agglomerans, Azospirillum brasilense, Azotobacter chroococcum, and Bacillus, isolated from temperate or tropical soils, were compared at different temperatures to study temperature adaptability. All Enterobacter strains and Bacillus strain C-11-25 reduced C₂H₂ at temperatures as low as 5°C. ARA by Enterobacter strains declined sharply above 30°C but ARA by Bacillus strain C-11-25 continued to increase with an increase in temperature.A. brasilense strain sp 245, isolated from wheat roots in Brazil, reduced more C₂H₂ at lower temperatures than strain Cd, isolated from a Californian soil. Similarly, the temperate strain ofA. chroococcum was a better N₂ fixer than the tropicalA. chroococcum strain at lower temperatures. Tropical strains ofA. brasilense andA. chroococcum reduced more C₂H₂ than temperate strains at higher temperatures. Therefore, it appears that temperate and tropical N₂-fixing organisms adapt themselves to their particular environment and should have more potential to benefit crops grown at the particular temperatures favorable to them. Only Bacillus strain C-11-25 has potential to benefit both temperate and tropical crops because it reduced significant acetylene over a wide temperature range.     

自生固氮菌活化土壤无机磷研究

以土壤为磷源,通过液体培养试验研究了5株自生固氮菌(Azotobacter sp.)对土壤无机磷的活化利用.结果表明,自生固氮菌能释放大量的氢离子,使液体培养基的pH大幅度降低,氢离子的浓度至少提高58倍以上.自生固氮菌分泌有机酸的种类与数量因菌株不同而异,这些有机酸包括甲酸、乙酸、草酸、丁二酸、柠檬酸、苹果酸和乳酸等,其中均能分泌草酸和苹果酸.在接种自生固氮菌的液体培养基中,全磷含量显著高于不接种的液体培养基,土壤无机磷总量则显著降低.由于土壤是培养基磷的唯一来源,故自生固氮菌促进了土壤无机磷的溶解释放.相关分析表明,培养基的pH值与土壤无机磷总量呈极显著正相关(r=0.959**,n=6),与液体培养基中的无机磷和全磷呈显著或极显著负相关(r =-0.850*;r=-0.918**,n=6),说明自生固氮菌分泌的氢离子可能是溶解土壤无机磷的原因之一.接种自生固氮菌显著降低土壤钙磷,土壤中的铁磷、铝磷和闭蓄态磷的降幅因菌株不同而异,其原因可能与有机酸分泌的数量和种类有关.     

九龙江口红树林鹧鸪菜藻体自生固氮细菌

初步研究了福建九龙江口秋茄(Kandelia candel)红树林红藻鹧鸪菜(Caloglossa leprieurii)藻体异养自生固氮菌数量的季节变化和微生物区系。结果表明:鹧鸪菜上异养自生固氮细菌数量以春季最多(1.033×104cfu.g-1),冬季最少(0.567×104cfu.g-1),固氮菌的季节变化模式表现为春季>秋季>夏季>冬季;鹧鸪菜藻体氮含量也以春季最高(22.08g.kg-1),冬季最低(16.63g.kg-1),二者差异显著(P<0.05),且藻体含氮量的季节变化模式与固氮细菌数量一致;这与鹧鸪菜的生长和物质积累密切相关,鹧鸪菜与其藻体上的自生固氮菌可能存在着互惠互利的关系,这种关系同时也受到环境温度和水分等因子的综合影响;对9株固氮菌的初步鉴定结果显示,它们分属于固氮菌属(Azotobacter)与拜叶林克氏菌属(Beijerinckia)。     

耐氨固氮菌接种水稻所需最低活菌数的测定

耐氨固氮菌在谷芽上能生长,每粒谷芽上的菌数可达１０４－１０５个。当７０％以上的秧苗带有耐氨固氮菌时,经根系交叉感染能使所有的秧苗根系带上耐氨固氮菌。菌液浸泡谷芽接种水稻,耐氨固氮菌数／谷芽数达２０时,可使７０％的谷芽接种上菌,有２１０以上时,可使１００％的谷芽接种上菌。菌液淋施秧苗接种水稻,耐氨固氮菌数／秧苗数为１６００以上时,１００％的秧苗带有耐氨菌。菌液浸泡秧根接种水稻,耐氨固氮菌数／秧苗数为７０个以上时,可使１００％的秧苗根部带有耐氨菌。