SYMBIOTIC NITROGEN FIXATION IN CEANOTHUS ROOTS

Ceanothus greggii var. perplexans is a common shrub in the southern California chaparral. Clusters of nodules found under the canopy of this species are modified roots which contain a nitrogen-fixing endophyte, Frankia ceanothi (Actinomycetales), within the cortex. The nodule density per m² obtained from root system excavations is much lower than that reported for different Ceanothus species in northern California. Field observations indicate that soil moisture is an important factor in nodule formation. Anatomical studies with the scanning electron microscope and acetylene reduction assays support the hypothesis that the vesicles, spherical swellings of hyphal endings (1.2–3.0 μm in diam), are indeed the sites of N₂ fixation. No bacteria-like bodies were found. The acetylene reduction rates of C. greggii endophytes were of the same order of magnitude as those reported for other members of the genus Frankia. It is estimated that 100 grams of nitrogen are fixed per year per hectare for a specific area in the southern Californian chaparral where C. greggii comprises 1/3 of the ground cover. This amount appears to be large enough to replace the nitrogen that is lost annually by drainage and runoff from winter rain storms.     

MEASURING RATES OF SYMBIOTIC NITROGEN FIXATION BY ALNUS TENUIFOLIA

Rates of nitrogen fixation were determined for alder (Abuts tenuifolia Nutt.) with the C₂H₂ assay and verified by the ¹⁵N method. Rates of C₂H₂ reduction obtained varied between 0.0342 and 10.9 μmoles per gram of nodule tissue (fresh weight) per hour with a seasonal average of 2.70 μmoles g^-1 hr^-1. Expressed as nitrogen fixation rates these are equivalent to 0.0114, 3.63, and 0.9 μmoles g^-1 hr^-1 of elemental nitrogen, respectively. Nitrogen fixation (¹⁵N) rates averaged 2.20 μmoles N g^-1 hr^-1 with a standard deviation of 1.14. The influence of pC₂H₂, pO₂, temperature during incubation, length of incubation time, presence vs. absence of N₂ in the incubation atmosphere, and amount of nodule biomass per sample were investigated.     

羊奶果结瘤固氮特性研究

羊奶果根瘤多年生,初发生时呈2—3分又状,后经多次分叉生长,形成珊瑚状的根瘤簇,外形球状或扁球状,直径可达数厘米。根瘤固氮活性较高,全年平均固氮活性8.86微摩乙烯/克鲜瘤/小时。固氮活性秋夏较高,冬春较低。根瘤离体后固氮能力持续时间较长,可达9小时。固氮作用最适温度为30℃,低温使固氮活性显著下降,短时间高温(37℃)能提高根瘤固氮活性,但持续高温会抑制固氮活性。 羊奶果根瘤具有氢酶,表现出较高吸氢活性(9.52微摩氢/克鲜瘤/小时),且持续时间较长。外源氢能明显提高根瘤固氮活性,提高幅度达60％左右。 羊奶果各器官硝酸还原酶活力为根>根瘤>叶片>枝条,叶片NR活力呈季节性变化,秋夏较高,冬春较低。根瘤NR活力变化较大,且与根瘤固氮活性变化的趋势基本一致。     

磁场对大豆共生固氮的效应

恒定磁场处理慢生大豆根瘤菌“005”和接种后的大豆植株,发现磁场可以提高根瘤的固氮活性。在一定的磁场强度(70—100mT)下,固氮活性平均可以提高4—5倍,植株的结瘤数和根瘤重量平均提高2—3倍。从这样的根瘤中所分离出的根瘤菌,由慢生型转变成快生型,在100植株中有17株的根瘤分离出快生菌。生长世代时间和培养溶液中的pH值与慢生型不同,而与快生型相同。     

新疆干旱区豆科植物结瘤的固氮特性

调查了新疆干旱区72种豆科植物的结瘤固氮活性,其中33种尚未见报道。这些植物所结根瘤在外形上多数不规则,以皮层厚和白色、棕色者居多,与非干旱区的根瘤形态显著不同。根瘤固氮活力相差较大,但比一般豆科植物根瘤活性高,最高者可达当地大豆根瘤的42倍。根瘤活性与宿主的抗逆境能力有关。此外,从11种豆科植物根瘤观察到10种具有吸氢活性。对干旱区豆科共生固氮生理生态的特性进行了讨论。     

大豆-根瘤菌共生体系光合与固氮关系研究

水培大豆和田间生长的大豆,接种根瘤菌 Rhizobium B16-11C 后植株全氮含量、叶片叶绿素含量和净光合速率及种子产量都明显增加。比较 Clark 大豆的结瘤品系和不结瘤品系获类似结果。摘除根瘤后3天内叶片净光合速率无明显变化。大豆植株遮阴、去叶或切掉地上部导致根瘤活性明显下降。但去豆荚不能提高根瘤固氮的比活性。根瘤活性的日变化不能用根瘤蔗糖、淀粉含量或周围温度的变化来解释,其控制因子尚待深入研究。     

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     

导入额外拷贝nifA基因对费氏中华根瘤菌共生固氮作用的影响

以广宿主、稳定性质粒pTR102为载体构建重组质粒pHN306,其上克隆有来自肺炎克氏杆菌（Klebsiellapneumoniae）的nifA基因和来自pDB30所含的发光酶标记基因（luxAB）。经三亲本接合转移,将pHN306导入费氏中华根瘤菌（Sinorhizobiumfredii）HN01,GR3和YC4。与出发菌相比较的盆栽试验结果表明：HN01（pHN306）和GR3（pHN306）分别在大豆渝豆一号和黑龙33上能显著提高瘤数,瘤重,植株地上部分干重和总氮量,YC4（pHN306）在大豆渝豆一号上也能显著提高瘤数,癌重和总氮量,对植株地上部分干重表现出一定的促进作用。结果表明：nifA基因对固氮效率和结瘤能力的促进作用与受体根瘤菌和大豆品种等因素有关。以luxAB为报告基因进行的菌落和根瘤发光检测结果表明：pHN306可在供试根瘤菌中稳定遗传。     

COMPARATIVE STUDIES OF NITROGEN FIXATION AND PHOTOSYNTHESIS IN ANABAENA CYLINDRICA

Manometric methods were developed to allow short-time measurements on relatively small amounts of cell material. The methods provided measurement of the nitrogen uptake and oxygen evolution attributable to nitrogen fixation either in the presence or absence of carbon dioxide. The methods were applied to observe effects of cell history, light intensity, temperature, pH, and ammonia on nitrogen fixation and photosynthesis with 2 new and significant findings. Nitrogen fixation is markedly accelerated by a preceding period of cellular nitrogen depletion. Nitrogen fixation is depressed by light intensities greater than those required to saturate photosynthesis.     

NITROGEN FIXATION BY LEGUMES AND BLUE-GREEN ALGAL-LICHEN CRUSTS IN A COLORADO DESERT ENVIRONMENT

Potential sources of fixed nitrogen in a Colorado desert environment were examined by the acetylene reduction method at the Deep Canyon Desert Research Center, near Palm Desert, California. In field and greenhouse studies all members of the genera Astragalus, Dalea, Lotus, Lupinus, Melilotus, and Prosopis examined formed active nodules (acetylene reduction) with indigenous soil bacteria. No evidence of nodulation was found for Acacia greggii, Cercidium floridum, or Hoffmannseggia microphylla. Lotus tomentellus was estimated to fix 0.1 kg N ha⁻¹ by the time of flowering under field conditions. Several members of the genus Dalea showed substantial rates of acetylene reduction in the greenhouse: D. emoryi, 16.1 + 3.5, D. mollissima, 11.4 + 3.7, D. schottii 2.9 + 1.7, D. spinosa 2.5 + 0.4 μmoles ethylene plant⁻¹ hr⁻¹. In greenhouse assays where water was supplied continuously, blue-green algal-lichen crusts reduced acetylene at an average rate of 11.0 + 5.7 nmoles ethylene cm⁻² hr⁻¹ with a maximum of 57.1. But when in situ assays were done following irrigation of a field plot with 2.3 cm of water, much lower activities were observed with a maximum activity of only 6.4 nmoles cm⁻² hr⁻¹.     

固氮鱼腥藻的蛋白水解酶及其部分性质的研究

应用溶菌酶处理,超声破碎细胞,通过差异离心和解离剂处理,将细胞的不同部分分开。以水解α-酪蛋白反应检测蛋白水解酶活性,结果表明,在细胞的可溶部分、内细胞质膜、外膜及胞浆周围区均存在蛋白水解酶活性。异形胞可溶部分的蛋白酶活比营养细胞可溶的蛋白酶活高4—5倍。在固氮条件下,营养细胞可溶性蛋白酶反应最适温度为50—55℃,65℃时,酶活迅速下降。有Ca^2 5mmol／l时,蛋白酶在60℃稳定,无Ca^2 存在下,酶液在60℃预处理10分钟,酶活性丧失80％以上。酶反应的最适pH为8—10。而且氮饥饿之后,培养基的pH值对细胞蛋白酶活水平有明显影响,氮饥饿24小时内,蛋白酶活迅速增加,但在碱性条件下,酶活水平比在中性条件下要高得多。邻菲哕啉对蛋白酶活性有严重的抑制,EDTA仅有轻微抑制,而PMSF对细胞蛋白酶活的抑制作用,与细胞氮饥饿时间有关。     

用毛地黄皂苷分离异形胞及其对光合和固氮的某些特性的影响

本文叙述一种从多变鱼腥藻(Anabaena variabilis)中分离异形胞的简易方法。这种新方法是用毛地黄皂苷和甘露醇的 TES 缓冲液处理藻丝,破碎营养细胞,并结合分级离心的方法获得异形胞。所分离异形胞的纯度,在显微镜下观察达到90%左右。当提供 ATP 和Na_2S_2O_4时,能够测到所分离异形胞的固氮活性,其最大速率是5.31毫微克分子 C_2H_2/10~6异形胞/小时,为整体藻丝活性的10%。这种比活性,在4小时内,甚至更长的时间内,保持不变。但是,在氢气下和照光条件下,分离的异形胞缺乏受光促进的固氮活性。分离的异形胞在77°K下波长为430nm 光激发的荧光光谱和完整藻丝的相比,它缺乏属于光合系统Ⅱ的685nm 和695nm 的荧光发射峰,而仅具有光合系统 I 的730nm 荧光发射峰。当提供 DCIP 和抗坏血酸时,被压碎的异形胞能够光还原甲基紫精,其活性为360消耗 O_2的微克分子/毫克叶绿素/小时。上述结果表明用毛地黄皂苷法分离的异形胞具有较完整的 DCIPH_2→MV 的 PSI 活性。