Nitrogen fixation and delayed leaf senescence in soybeans

Delayed leaf senescence has been found in a soybean population which maintains its chlorophyll and ribulosebisphosphate carboxylase activity in leaves and nitrogen fixation (acetylene reduction) activity in root nodules throughout seed maturation. Incorporation of delayed leaf senescence into an agronomically desirable genetic background may help to increase seed yield and symbiotic nitrogen fixation during seed development.     

Nitrogen Fixation by Azotobacter vinelandii Strains Having Deletions in Structural Genes for Nitrogenase

Phenotypic reversal of Nif(-) mutant strains to Nif(+) under molybdenum-deficient conditions has been cited as evidence that Azotobacter vinelandii possesses two nitrogen fixation systems: the conventional molybdenum-enzyme system and an alternative nitrogen-fixation system. Since explanations other than the existence of an alternative system were possible, deletion strains of A. vinelandii lacking the structural genes for conventional nitrogenase (nifHDK) were constructed. These strains were found to grow in molybdenum-deficient nitrogen-free media, reduce acetylene (at low rates), and incorporate molecular nitrogen labeled with nitrogen-15. Thus it can be concluded that the phenotypic reversal phenomenon cannot be due to altered phenotypic expression of nif mutations under molybdenum-deficient conditions, but is due to the existence of an alternative nitrogen-fixation system in A. vinelandii as originally proposed.     

常压室温等离子体诱变选育高固氮酶活褐球固氮菌

为增强固氮酶活性以提高现有固氮菌的固氮能力，采用常压室温等离子体(atmospheric and room temperature plasma，ARTP)诱变技术处理褐球固氮菌（Azotobacter chroococcum）CICC21686，用乙炔还原法筛选具有高固氮酶活性的突变株28s-20，并观察该突变株酶活的遗传稳定性；通过梯度温度和pH培养，探究高酶活突变株的生物学活性；采用盆栽试验，探究突变株对玉米植株干重及全氮量的影响。结果表明，复筛获得突变株28s-20的固氮酶活较出发菌株提高了101.72%；经5次传代，证实该菌株具有良好的遗传稳定性；突变株最适生长温度为28 ℃，培养基初始pH为7.5；与野生株相比，突变株显著增加玉米干重及全氮量，分别提高21.16%和34.36%。诱变高酶活菌株为褐球固氮菌的菌种优化及农业生物固氮提供了技术支持。     

生物复合菌肥“地霸”的菌群分离鉴定及生物学特性的初步研究

对生物复合菌肥进行菌群的分离鉴定、含菌量的测定和比较以及菌株生物学特性的初步研究.该生物复合菌肥中含有活性较高的固氮菌(维氏固氮菌)、钾细菌和无机磷细菌,每克菌肥中细菌总数达到2.0亿～2.5亿.固氮菌在修改的Dobereiner无氮培养基上生长良好,固氮活性较强,该菌生长最适温度在20～32℃之间,生长的pH值范围是5.5～8.5,最适pH值是7.5.     

Nitrogen fixation in termites

Nitrogen fixation, measured by the reduction of acetylene to ethylene, was found in workers of the dry-wood termite Kalotermes minor. The soldiers and reproductive castes fixed little or no nitrogen. The fixation rates ranged between 24 and 566 micrograms of nitrogen fixed per month per gram (wet weight) of termite. Nitrogen fixation can be a significant source of nitrogen for these termites.     

水稻根际固氮菌NX-2的分离鉴定及其培养条件优化

筛选高效固氮能力的植物根际促生菌是研发微生物肥料的关键。研究从水稻根际土壤中分离筛选得到一株固氮菌NX-2,利用乙炔还原法测定其固氮酶活可达16.18μmol/(m L·h)(以C2H4浓度计)。通过形态观察、生理生化特性和16S r DNA序列分析,菌株NX-2被鉴定为褐球固氮菌。进一步研究其生长特性,结果显示最佳培养条件为培养温度28℃、初始p H值6.5、接种量3%。     

Biological nitrogen fixation in Lake Erie

Biological nitrogen fixation, as determined by acetylene reduction, occurs in Lake Erie. Fixation potential by blue-green algae in situ in water and by bacteria in collected sediments was demonstrated. Nitrogen-fixing activity occurred from June through November suggesting that it is significant over the extremes of seasonal variation in light, temperature, and nutrients.     

Nitrogen fixation by gloeocapsa

The continuous growth in a medium free of combined nitrogen and the experimental production of ethylene via acetylene reduction indicate that nitrogen fixation by blue-green algae is not solely confined to filamentous genera with heterocysts. Axenic cultures of Gloeocapsa sp., adapted to nitrate-free medium, form ethylene at rates comparable to those of species known to fix nitrogen.     

24株剑麻根际联合固氮菌的16S rDNA序列分析

对海南、广西和广东剑麻主产区种植基地的剑麻根际联合固氮菌进行了分离、纯化及固氮活性测定,获得了24株具有固氮活性的菌株,其固氮酶活性为5.8742～1765.6590 nmol/(mL.h)。16S rDNA序列分析结果表明:参试菌株的16S rDNA扩增区约为1500 bp,无太大的长度变异,其系统发育树表明这24株菌株分别属于6个不同的属。     

Nitrogen fixation in some anoxic lacustrine environments

Low rates of acetylene reduction to ethylene in water samples from two dystrophic lakes indicate the presence of nitrogenase and in situ nitrogen fixation. Highest rates were found in anoxic water from the aphotic zone. Environmental conditions in these lakes suggest the agents of fixation were bacteria.     

北京地区稻田固氮蓝藻的研究:Ⅱ.表层土壤和水稻茎基的固氮活性变化及原因分析

在水稻栽培期间应用乙炔还原法,原位测定光照和黑暗培养条件下的北京水稻田表层土壤(5cm)的固氮活性(ARA 表示)表明:在7月初(分蘖期)和8月底9月初(抽穗期)有前、后期两次高峰,前期峰是固氮蓝藻和固氮菌共同固氮,而后期峰主要是蓝藻固氮;稻田与休闲水田的表层土壤,在全生育期内固氮能力的大小随不同施氮肥水平而异;接种固氮蓝藻能使稻田与休闲水田表层土壤的固氮活性提高,但其特点不同。茎基的固氮活性变化受施肥、接种蓝藻等因素影响。     

刀孢蜡蚧菌的鉴定及其对南方根结线虫不同生活阶段的定殖

【目的】明确卵寄生真菌（Lecanicillium psalliotae）刀孢蜡蚧菌对根结线虫的防治潜力。【方法】采用分离自山东青岛城阳丝瓜根结线虫的卵寄生真菌CGMCC5329,通过形态学特征观测及分子生物学分析,鉴定该菌株为刀孢蜡蚧菌。显微观察刀孢蜡蚧菌对南方根结线虫（Meloidogyne incognita）不同生活阶段的定殖。【结果】刀孢蜡蚧菌能够寄生南方根结线虫生活史中的卵、2龄幼虫和雌虫,同时对卵块内卵孵化具有明显的抑制作用。该菌菌丝在卵表面形成侵染钉,卵壳皱缩凹陷,卵胚胎发育停止和内容物凝集,卵内长满菌丝,有时可见到菌丝上形成瓶梗细胞和分生孢子。第6天卵寄生率为85.76%;15 d卵块相对孵化抑制率为91.41%。被真菌寄生的2龄幼虫体表形成致密菌网,穿透体壁,体内长满菌丝,5 d后寄生率为79.23%。寄生的雌虫体壁形成致密菌网和侵染钉,体壁皱缩变形,有时内容物外渗。【结论】刀孢蜡蚧菌CGMCC5329不仅能寄生根结线虫生活史中定居态的雌虫和卵,还能寄生侵染态2龄幼虫。该菌是具有生物防治应用潜力的根结线虫寄生真菌。     

棕色固氮菌分离鉴定与初步应用研究

棕色固氮菌分离鉴定与初步应用研究结果表明,通过分离培养和鉴定,分离出的6个棕色固氮菌贵州地方株,其中4株有一定固氮能力,另外2株固氮能力较强。应用棕色固氮菌制成菌肥,可以显著提高黑麦草产量。     

地毯草根际固氮菌的分离及鉴定

结合气相色谱仪,利用乙炔还原等方法对地毯草(Axonopus compressus)根际固氮菌进行分离和鉴定。结果表明,从地毯草根际土壤中分离获得6株固氮菌株,菌株的固氮酶活性相差较大(92.5 C2H4~295.2 C2H4nmol.mL-1.h-1),菌株hnN2及hnN6具有较高的固氮酶活性(>200 C2H4nmol.mL-1.h-1);菌株经鉴定分属Bacillus(2株)、Azotobacte(r 3株)和Pseudomona(s 1株)3个属。     

农田环境中固氮菌的促生潜能与分布特点

【目的】了解小麦、水稻、玉米、蔬菜等作物根际固氮菌的优势种群、固氮菌菌株的固氮、抗病、促生潜能以及菌株在系统发育地位和来源作物种类上的分布特点。【方法】采用无氮培养基培养固氮菌,乙炔还原法测定菌株固氮酶活性,平板对峙法测定菌株拮抗病原真菌性能,ACC（1-氨基环丙烷-1-羧酸）唯一氮源法定性测定菌株产ACC脱氨酶特性,比色法定量测定ACC脱氨酶活性,通过16S rDNA序列测定和相似性分析研究菌株的分类地位。【结果】94株供试菌株的固氮酶活性在0.99-180.59 nmol C2H4/h•mg蛋白,其中大于10 nmol C2H4/h•mg蛋白的菌株有42株,占全部供试菌株的44.7%;类芽孢杆菌属（Paenibacillus)和芽孢杆菌属（Bacillus）是主要类群,分别占供试菌株总数的33.0%和26.6%,且不具有寄主专一性。供试菌株中有6株分别对核盘菌（Sclerotinia sclerotiorum）、玉米赤霉菌（Gibberella zeae）和大丽轮枝菌（Verticillium dahliae）3种植物病原真菌表现出拮抗作用,占菌株总数的6.4%,抑菌率为23.9%-65.9%。有20株固氮菌能够产生ACC脱氨酶,占全部供试菌株的21.3%,活性在0.33-21.98 µmol α-丁酮酸/h•mg蛋白,主要分布在芽孢杆菌属、类芽孢杆菌属和根瘤菌属（Rhizobium）。【结论】小麦、水稻、玉米、白菜、芹菜等作物根际以及农田环境中固氮菌的优势种群为类芽孢杆菌属和芽孢杆菌属,多数固氮菌菌株具有较高的固氮潜能,部分菌株具有ACC脱氨酶活性和促生潜能,少数菌株具有抗病潜能;固氮、抗病、促生潜力菌株主要分布在类芽孢杆菌属、芽孢杆菌属和根瘤菌属,随作物分布广泛,无专一性。     

红豆草在旱农地区固氮肥田作用研究(Ⅰ)

本文研究了红豆草(Onobrychis Viciaeffolia Scop)在旱农地区的结瘤固氮作用。研究表明,在该地区红豆草结瘤状况良好,单株根瘤量多达1.9—3.0克,但多属无效根瘤。用乙炔还原法测定根瘤的固氮活性为0.048毫微克乙烯分子/毫克根瘤/分钟。此外,研究了红豆草结瘤固氮的环境因素(包括土壤有效养分、微量元素及水分、光照等)。研究认为,在该地区肥力较低的土壤,需要施用一定氢肥,这对提高红豆草的经济产量是重要的。     

北方稻田异养固氮活性变化

用乙炔还原法研究吉林延边地区与北京两个不同生态区稻田异养固氮活性变化,结果表明,两地稻田根际土乙炔还原活性(ARA)均在分蘖期及抽穗期出现高峰,并前期高峰比后期高峰高。北京稻田根系 ARA 也与根际土同步出现两次高峰,但前期高峰低于后期高峰。延边稻苗根系和两地茎基部 ARA 只有抽穗的一次高峰。延边稻田茎叶 ARA 有明显的单峰变化,而北京稻田则极微,这与两地空气湿度差异一致。在 ARA 高峰期两地相比,根系 ARA 较一致,根际土及茎部 ARA 以北京远高于延边,此现象又与两地气温差异一致。延边稻田土移至北京种稻时,各部位 ARA 变化趋势接近延边原地,而变化量则接近北京稻田。     

禾谷镰刀菌侵染拟南芥的研究

本研究采用禾谷镰刀菌侵染拟南芥,结果表明,在拟南芥开花初期,采用分生孢子悬浮液喷雾接种,能够引起花组织和角果产生病变,菌丝进一步扩展到整个植株中,最终引起植株坏死。用组织染色及特异引物进行PCR扩增,证实接种菌已侵入植物组织。说明拟南芥可用来进行禾谷镰刀菌侵染过程的相关分析,也可为筛选抗赤霉病的基因提供新思路。     

Quantitative imaging of nitrogen fixation by individual bacteria within animal cells

Biological nitrogen fixation, the conversion of atmospheric nitrogen to ammonia for biosynthesis, is exclusively performed by a few bacteria and archaea. Despite the essential importance of biological nitrogen fixation, it has been impossible to quantify the incorporation of nitrogen by individual bacteria or to map the fate of fixed nitrogen in host cells. In this study, with multi-isotope imaging mass spectrometry we directly imaged and measured nitrogen fixation by individual bacteria within eukaryotic host cells and demonstrated that fixed nitrogen is used for host metabolism. This approach introduces a powerful way to study microbes and global nutrient cycles.     

萨克拉普奇尼亚菌串孢变种对南方根结线虫卵和雌虫的致病性

根结线虫卵和雌虫的寄生真菌对根结线虫病害生物防治具有很大应用潜力。本项研究采用分离自北方根结线虫卵的萨克拉普奇尼亚菌串孢变种Pochonia suchlasporia var.catenata CFCC84965,测定其对南方根结线虫卵和雌虫的致病性。被寄生的卵内充满菌丝且胚胎破坏,胚前发育期的卵对菌丝的侵染敏感,即将孵化出2龄幼虫的卵很少被寄生。菌丝环绕于被寄生卵周围形成致密菌网和侵染钉,卵壳皱缩和凹陷,有时可见到菌丝上形成瓶梗细胞和分生孢子;卵胚胎发育停止和幼虫发育畸形,卵内容物聚集变态,卵壳破裂,内容物外渗。萨克拉普奇尼亚菌串孢变种CFCC84965对卵的寄生率为65.18%。该菌对南方根结线虫雌虫具有致病性,菌丝接触雌虫体壁形成侵染钉,穿透体壁,虫体皱缩变形。该菌对根结线虫雌虫的寄生致病是首次报道。