植物内生固氮菌的研究进展

植物内生固氮菌是指定殖于植物体内与宿主植物进行联合固氮的一类微生物,不但具有固氮作用,还有生物防治、促进植物生长的作用。对植物内生固氮菌的研究始于上个世纪80年代,至今已从甘蔗、水稻等多种作物中分离到多种内生固氮菌。综述了10多年来内生固氮菌的研究进展,对固氮醋酸杆菌(Acetobater diazotrophicus)等几种内生固氮菌的生理、生物学特性及其侵染方式、传播途径和固氮机理进行了较全面的概述,并对应用内生固氮菌的可能性和意义,以及需要注意的问题作了一些探讨。     

干热河谷4种固氮植物根瘤固氮潜力及其影响因素

氮是除水分之外影响干热河谷生物活性的关键因子,豆科植物生物固氮是该地区氮素的重要来源之一。采用乙炔还原法测定了干热河谷不同季节燥红土和变性土林地中大叶相思、新银合欢、木豆和山合欢根瘤固氮酶活性(NAs)。结果表明:新银合欢(16.25μmol.g-1.h-1)和大叶相思(15.85μmol.g-1.h-1)根瘤NAs显著(P<0.001)高于山合欢(9.60μmol.g-1.h-1)和木豆(9.42μmol.g-1.h-1)。雨季根瘤NAs显著高于旱季,约为旱季的2.3倍。燥红土样地上植物根瘤NAs是变性土样地的1.3 1.6倍。研究揭示:除植被类型外,干热河谷植物根瘤NAs主要受土壤类型、季节和土壤含水量的影响,而受土壤温度的影响较小。     

玉米内生固氮菌的分离鉴定及对小麦幼苗的促生效应

应用研磨法从健康玉米植株的根茎叶中分离到4株具有高固氮酶活性的内生菌,根据它们在植物组织来源和分离时稀释度的不同将其分别命名为L1A1、L1A4、L1B9和L2A2(L代表来源于叶片,A代表10倍稀释,B代表100倍稀释).经形态学、生理生化特性及16S rDNA序列分析,对4株菌鉴定结果表明,L1A1为约氏不动杆菌(Acinetobacter johnsonii)、L1A4为Sphingomonas panni、L1B9为Sphingomonas yabuuchiae、L2A2为巴氏葡萄球菌(Staphylococcus pasteuri).这4株菌均具有不同程度的固氮酶活性和产IAA(吲哚乙酸)能力,其中L1A1、L2A2具有降解有机磷能力,L1A4具有体外拮抗小麦赤霉病菌的能力.接种这4株菌的小麦苗高、根长、鲜质量、干质量和叶绿素含量均比未接菌小麦高(P＜0.05),说明其具有促进小麦幼苗生长的作用.     

3种旱生禾草内生固氮菌的分离及促生性能测定

以3种多年生典型旱生禾草新农一号狗牙根(Cynodon dactylon‘ Xinnong No.1’)、沙生冰草(Agropyron de-sertorum)、偃麦草(Elytrigria repens)为材料,从根、叶组织中分离内生固氮菌株,测定其固氮酶活性、溶磷性及分泌生长素的能力,为禾草多功能促生菌剂的菌种筛选...     

A bradyrhizobial-<Emphasis Type="Italic">Penicillium</Emphasis> spp. biofilm with nitrogenase activity improves N<Subscript>2</Subscript> fixing symbiosis of soybean

A bradyrhizobial-fungal biofilm (i.e. Bradyrhizobium elkanii SEMIA 5019-Penicillium spp.) developed in vitro was assayed for its nitrogenase activity and was evaluated for N₂-fixing symbiosis with soybean under greenhouse conditions. The biofilm showed nitrogenase activity, but the bradyrhizobial strain alone did not. Shoot and root growth, nodulation and N accumulation of soybean increased significantly with an inoculum developed from the biofilm. This study concludes that such biofilmed inoculants can improve N₂-fixing symbiosis in legumes, and can also directly contribute to soil N fertility in the long term. Further studies should be conducted to investigate the performance of these inoculants under field conditions.     

Infectivity and acetylene reduction of diazotrophic rhizosphere bacteria in wheat (Triticum aestivum) seedlings under gnotobiotic conditions

Summary Wheat seedlings were inoculated with rhizosphere nitrogen-fixing bacteria and grown gnotobiotically for 15 days. The growth medium consisted of semisolid agar with or without plant nutrients. The bacteria, isolated from roots of field-grown wheat, were three unidentified Gram-negative rods (A1, A2, E1), one Enterobacter agglomerans (C1) and two Bacillus polymyxa (B1, B2). A strain of Azospirillum brasilense (USA 10) was included for comparison.Nitrogenase activity (acetylene reduction activity, ARA) was tested on intact plants after 8 and 15 days of growth. In semisolid agar without plant nutrients, five isolates showed ARA of 0.01–0.9 nmol C₂H₄ plant^–1 h^–1, while the two strains of B. polymyxa had higher ARA of 3.3–10.6 nmol C₂H₄ plant^–1 h^–1.Plant development was not affected by inoculation with bacteria, except that inoculation with B. polymyxa resulted in shorter shoots and lower root weight.Transmission electronmicroscopy of roots revealed different degrees of infection. A. brasilense, A1 and A2, occurred mainly in the mucilage on the root surface and between outer epidermal cells (low infectivity). B. polymyxa strains and E1 were found in and between epidermal cells (intermediate infectivity) while E. agglomerans invaded the cortex and was occasionally found within the stele (high infectivity).     

Placement effects of organic sources on nitrogenase activity and nitrogen-fixing bacteria in flooded rice soils

The placement effects of organic sources (cellulose and rice straw) at different depths on nitrogenase activity (acetylene reduction assay, ARA) associated with four tropical rice soils were investigated. Results showed that placement of organic sources in the top profile (1–2 cm) produced a higher nitrogenase activity; while placement in the 2–4 cm and 4–6cm profiles significantly decreased the activity irrespective of soil type. The effect of organic sources on nitrogenase varied depending on the soil type. Cellulose and rice straw caused differential stimulation in response to the placement. Evidence was provided for the creation of favourable redox potential supporting higher nitrogenase activity in series receiving organic amendments in the top profile. Higher nitrogenase activity in the top profile was associated with greater populations of nitrogen-fixing Azospirillum sp., anaerobic nitrogen fixers and Azotobacter sp. Moreover, the lower amounts of reducing sugars and higher carbon dioxide evolution gave evidence of higher microbial activity in the top profile. These results clearly indicate the positive relationship between nitrogenase activity, nitrogen-fixing populations and redox status in the top layers of flooded tropical rice soils. Received: 5 June 1996     

Effects of root-associated N2-fixing cyanobacteria on the growth and nitrogen content of wheat (Triticum vulgare L.) seedlings

Summary Following co-cultivation of wheat with N₂-fixing cyanobacterial isolates capable of forming associations, Nostoc 2S6B, 2S9B or Anabaena C5, for 15 days in the presence or absence of combined N a large stimulation of root length was observed without any increase in root dry weight. Increases in the N concentrations of both roots and shoots occurred following co-cultivation with most cyanobacteria tested. The increase in plant N concentrations appeared to be dependent on the wheat cultivar and the cyanobacterial isolate used. Nostoc isolates had similar nitrogenase activities when associated with roots and when grown in shake-flask cultures. The nitrogenase activity of roots colonized by Anabaena C5 or Nostoc 2S6B was higher following removal of loosely associated cyanobacteria.     

Perennial winged bean yield and nitrogen fixation improvement with soil fertility treatments of a typic eutrustox

The perennial legume, Winged Bean (Psophocarpus tetragonolobus (L) DC), has potential as a high protein food crop for the humid, tropical regions of the world. Edible seed pods, oil seed grain, leaves, flowers, and unique high protein tubers provide abundant nutritious components desirable for improved human diets. However, soil characteristics and fertility levels influence plant growth, yields and nitrogen fixation capability of this legume. Objectives of this study were to determine soil‐plant nutrient influences on vigorous regrowth, pod and tuber yields, nitrogenase (C₂H₂ red.) activity levels, and nodule cytosol components of the perennial Siempre cultivar grown on a Typic Eutrustox during three years, 1978–1980.

Available soil phosphorus was a first limiting plant nutrient during all three years of plant age. Effects of combined 100 mg P with 200 mg K/kg soil were highly significant for every parameter and plant age. Pod and seed yields more than doubled with PK addition compared to the check. Tuber growth, nodule mass and nitrogenase activity levels more than trebled with PK treatments as compared to the check. Both elemental P and K were significantly increased within the nodule cytosol of fertilized plants. Cytosol Na was significantly decreased with soil K additions. The best fit multiple regression was: nitrogenase = 1.99 nodule wt. + 6.34 tuber wt. + 0.39 tuber % N + 5.08 cytosol P + 1.55 cytosol K ‐ 0.45 cytosol Na, R² = 95.5, C.V. = 11.2%. The dominant nodule cytosol enzymes, aspartate aminotransferase (AST) and glutamine synthetase (GS), significantly increased with soil K additions regardless of P treatments. Glutamate dehydrogenase (GDH) and glutamate synthase (GOGAT) also contributed significantly with multiple regression for nitrogenase = 1.07 GS + 2.1 AST + 1.74 GOGAT ‐ 1.76 GDH + 12.89 Ureide, R² = .89, C.V. = 17.3%. Highly significant increases in nodule cytosol ureide composition with K soil additions has interest because of the role as a nitrogenous nonprotein component for many legumes. Increases in growth, nodulation and nitrogenase activity levels resulted with increased K levels of 0, 100, 200 and 300 mg K/kg soil when soil P and Ca were not limiting.     

联合固氮菌与根瘤菌协同作用对小麦幼苗的影响

小麦种子分别接种不同组合的联合固N菌和根瘤菌,播种到灭菌蛭石上培养28d后分别测定小麦幼苗F_v/F_m值、鲜物质量、干物质量、含N量及根系固N酶活性结果表明,接种联合固N菌植株均测到鲜根系固N酶活性,最高达386.6(±3.5)nmol/g,且联合固N菌和根瘤菌协同作用时植物干物质量、F_v/F_m值和含N量均略高于其单独作用,尤其是来自干旱半干旱草原羊草根际的联合固N菌——催娩克雷伯氏菌al(Klebsiella oxytoca)和内蒙古草原草木犀根瘤的中华苜蓿根瘤菌(Sinorhizobium,melilot)菌株Ⅱ的组合固N能力更强,对农业生产有一定应用潜力。