Interaction of maize roots and rhizosphere microorganisms

The interaction between the roots of Zea mays L. and its rhizosphere microflora was studied in nutrient solution with respect to two exemplary aspects - water soluble vitamins and the four most frequent carbohydrates. The exudation of vitamins and sugars into the medium is increased in a solid substrate (glass-ballotini simulating soil structure) in comparison with nutrient solution only. The amounts of vitamins released (1 to 421 ng per plant) are high enough for a pronounced influence on the rhizosphere microflora to be expected. However, the microbial population feeding on the exudates has very simple nutritional demands. In spite of appreciable amounts of vitamins found in the inoculated nutrient solution, the growth of vitamin requiring species is not stimulated compared to the soil population. The roots are mostly inhabited by shorter rods and occasionally by actinomycete-like filaments. The percentage of the root area covered by its microflora, assuming a monolayer, is 4% in the root region where root hairs are just emerging, 7% in the root hair zone and 20% in the oldest part of the roots, 10 mm below the grain. In the presence of rhizosphere microorganisms, root dry weight is lower than that of axenically grown plants because the diameter of the primary root is diminished. In addition, the vitamin and sugar content of the root is affected by rhizosphere microorganisms. The results can be interpreted as an indication that phytohormones may participate in interactions between roots and bacteria.     

恶臭假单胞菌P861(Gus)在油菜根部定殖的生态研究

本研究采用Gus 基因标记技术和常规方法跟踪考察了恶臭假单胞菌P861(Gus) 在缩影系统油菜根圈的定殖情况,以及缩影系统内土壤类型、土壤含水量对根部定殖的影响。土壤含水量分别为60% FC和75% FC时,P861(Gus) 在砂姜黑土中的定殖水平高于50% FC的,不但能散布至种子下8cm 以内的根段部位,且定殖水平分别为7.5×10²和2.8×10³cfu·g^-1。在灰潮土缩影中,P861(Gus) 在油菜根圈的定殖动态表现为在油菜播种后3 ～6 天,定殖密度可达最高水平(5.5×10⁶cfu·g^-1) ,然后急速下降,最后保持在一个相对稳定的较低水平(7.6×10²cfu·g^-1) 。P861(Gus) 在不同根段部位的定殖密度并无从上到下逐渐递减的规律。     

Inoculating wheat seedlings with exopolysaccharide-producing bacteria restricts sodium uptake and stimulates plant growth under salt stress

A pot experiment was conducted to elucidate the effects of inoculating five exopolysaccharide- (EPS-) producing bacterial strains on the dry matter yield and the uptake of K⁺, Na⁺, and Ca²⁺ by wheat seedlings grown in a moderately saline soil. The bacteria were isolated from the rhizosphere soil (RS) of wheat grown in a salt-affected soil and included Aeromonas hydrophila/caviae (strain MAS-765), Bacillus insolitus (strain MAS17), and Bacillus sp. (strains MAS617, MAS620 and MAS820). The inoculation substantially increased the dry matter yield of roots (149–527% increase) and shoots (85–281% increase), and the mass of RS (176–790% increase). All the strains, except MAS617, also increased the RS mass/root mass ratio as well as the population density of EPS bacteria on the rhizoplane, and both these parameters were significantly correlated with the content of water-insoluble saccharides in the RS. Inoculation restricted Na⁺ uptake by roots, which was not attributable to the binding of Na⁺ by the RS, or to the ameliorative effects of Ca²⁺ under salinity. The decreased Na⁺ uptake by roots of inoculated than uninoculated plants was probably caused by a reduced passive (apoplasmic) flow of Na⁺ into the stele due to the higher proportion of the root zones covered with soil sheaths in inoculated treatments. Among the strains tested, MAS820 was the most efficient in all respects, whereas MAS617 was the least effective. Results suggested that inoculating selected EPS-producing bacteria could serve as a useful tool for alleviating salinity stress in salt-sensitive plants.     

覆盖模式及小麦根系对土壤微生物区系的影响

采用平皿分离培养法研究了5种栽培模式和小麦根系对土壤细菌、真菌及放线菌数量的影响。连续2年的定位测定结果表明:覆膜有利于土壤微生物数量增加。5种栽培模式中,小麦根区、根外土壤细菌数量均以覆膜模式下最高,分别为116.8×106cfu·g-1和86.7×106cfu·g-1;土壤真菌和放线菌数量均以垄沟覆膜(垄上覆膜、垄沟播种)模式下最高,分别为3.0×103cfu·g-1、1.4×103cfu·g-1和18.9×105cfu·g-1、19.7×105cfu·g-1。不同模式下小麦根系对土壤细菌和真菌数量影响较大,表现为根区高于根外;而根系对放线菌影响较小,只有补灌和覆膜2种模式为根区高于根外。多重比较结果显示,覆膜与其他模式之间细菌数量差异极显著,根区土壤细菌和真菌数量与根外存在显著差异。覆盖和根系能大幅度增加根区细菌、真菌和放线菌的数量,强化小麦根区根外细菌和真菌的数量差异。     

柑桔根际土壤微生物种群动态及根际效应的研究

在柑桔根际土壤微生物的种群结构中，细菌占优势，最高达76871×10⁷个每克干土；真菌次之，达17549个每克干土；放线菌最少，达12462个每克干土。三种菌的根际效应非常显著，根际细菌最高约为对照的67.6倍，真菌约为12.4倍，放线菌约为26.9倍，柑桔中，温州密柑根际土壤细菌最多，年平均为6490×10⁷个每克干土；红桔的放线菌及真菌最多，年均分别为1326个每克干土及1728个每克干土。柑桔根际土壤微生物区系的种群结构和数量变动，受土壤理化性质的影响，并随树种、树龄、砧木及季节而变化。柑桔根部及根颈部对脚腐病、根腐病及线虫病的感病性和耐病性同柑桔同柑桔根际土壤微生物区系中，有益微生物与有害微生物的种类、数量及分布有密切的关系。柑桔根际土壤有益微生物有促进生长发育和保护的作用。     

巨大芽孢杆菌在油菜根部定殖和促生作用的研究

采用基因标记技术和常规方法跟踪巨大芽孢杆菌A6 (gusA)在缩影系统油菜根际的定殖情况。A6 (gusA)菌在油菜不同根段部位的定殖密度表现从上到下逐渐递减的现象。随着接种后时间的延长而逐渐下降。在根段 8cm以外的根区几乎检测不到接种菌。在油菜播种后 3d ,定殖密度可达最高水平 (8 7×10 5cfug-1根 ) ,然后急速下降 ,30d后保持相对稳定的较低水平 (2 2× 10 2 cfug-1根 )。在促生试验中 ,表现在不同程度上增加植株干重、全氮、全磷和全钾的含量     

Effect of Azospirillum brasilense inoculation on root morphology and respiration in tomato seedlings

Summary The level of Azospirillum brasilense strain Cd colonization in the rhizosphere of some vegetables was 10⁴–10⁵ colony-forming units (CFU) per root of one plant in 2-week-old plants inoculated with 5 × 10⁸ Azospirillum cells. Significant increases in root length (35%) and in top (90%) and root (50%) dry weight and total leaf area (90%) were observed in 18-day-old inoculated tomato plants compared with non-inoculated controls. An inoculum concentration of 1 × 10⁸ to 5 × 10⁸ CFU/ml stimulated the appearance of root hairs. Large numbers of bacteria (1 × 10⁹ CFU/ml) caused asymmetrical growth of the root tip. In a petri dish system, Azospirillum (1 × 10⁸ CFU/ml) increased root dry weight (150%), protein content (20%), respiration rate per root (70%) and the specific activity of malate dehydrogenase (45%–65%) over non-inoculated controls. The specific respiration rate, expressed as micromol of O₂ per minute per milligram of dry weight of roots, was significantly lower in inoculated roots, suggesting that less energy was spent for accumulation of more dry material.     

Reduced root water uptake after drying and rewetting

The ability of plants to extract water from soil is controlled by the water‐potential gradient between root and soil, by the hydraulic conductivity of roots, and, as the soil dries, by that of the soil near the roots (rhizosphere). Recent experiments showed that the rhizosphere turned hydrophobic after drying and it remained temporarily dry after rewetting. Our objective was to investigate whether rhizosphere hydrophobicity is associated with a reduction in root water uptake after drying and rewetting. We used neutron radiography to trace the transport of deuterated water (D₂O) in the roots of lupines growing in a sandy soil. The plants were grown in aluminum containers (28 × 28 × 1 cm³) filled with a sandy soil. The soil was initially partitioned into different compartments using a 1‐cm layer of coarse sand (three vertical × three horizontal compartments). We grew plants in relatively moist conditions (0.1 < θ < 0.2). Three weeks after planting, we let the upper left compartment of soil to dry for 2–3 d while we irrigated the rest of the soil. Then, we injected D₂O in this compartment and in the upper right compartment that was kept wet. We monitored D₂O transport in soil and roots with time‐series neutron radiography. From the changes of D₂O concentration inside roots, we estimated the root water uptake. We found that root water uptake in the soil region that was let dry and rewetted was 4–8 times smaller than that in the region that was kept moist. The reduced uptake persisted for > 1–0.5 h. We conclude that a reduction in hydraulic conductivity occurred during drying and persisted after rewetting. This reduction in conductivity could have occurred in roots, in the rhizosphere, or more likely in both of them.     

Soil microorganisms antagonistic towards Rhizobium japonicum

Success in introducing Rhizobium japonicum strains into soil is related to their interaction with native microorganisms including some that are antagonistic. Actinomycetes, bacteria, fungi and rhizobiophages antagonistic towards strains of R. japonicum were counted directly using soil samples from field plots under different crop and soil management systems. The antagonistic actinomycete population varied from 1.3 × 10³ to 4.5 × 10⁵ g^?1 dry soil and ranged up to 90% of total actinomycetes. Soybean rhizosphere soil samples included antagonistic actinomycetes ranging up to 70% of total actinomycetes. The antagonistic bacterial population was less than 10% of total bacteria and the proportion did not vary significantly with crop or soil management practices. Antagonistic fungi were observed for many of the soils examined but they could not be counted. There were few rhizobiophages and they were found most frequently in soybean rhizospheres. Occasional bacterial and actinomycete colonies that stimulated growth of R. japonicum were randomly observed among the soil samples tested.     

Responses by iron-efficient and inefficient oat cultivars to inoculation with siderophore-producing bacteria in a calcareous soil

Rhizosphere bacteria may enhance plant uptake of Fe by producing siderophores that chelate sparingly soluble Fe³⁺ in calcareous soils. To evaluate the extent to which plants benefit from colonization of the roots by prolific siderophore-producing bacteria, we inoculated two oat cultivars with six strains of bacteria that produced high concentrations of siderophores under Felimiting conditions in vitro. Oat cv Coker 227, an Fe-efficient cultivar, which produces the phytosiderophore avenic acid, and cv TAM 0-312, and Fe-inefficient cultivar, which does not produce the phytosiderophore, were grown in a calcareous soil (Weswood silt loam) on a light bench in the laboratory. Half of the plants were fertilized with a nutrient solution containing 5 mM Fe and half with a nutrient solution containing no Fe. After 6 weeks of growth, we compared colonization of the roots by the inoculant bacteria and the dry weight and Fe content of roots and shoots. Three species of Pseudomonas colonized the roots of both oat cultivars in high numbers (10⁶ cells g^-1 root dry weight), whereas the remaining bacteria colonized the roots in substantially lower numbers (10⁴ cells g^-1 root dry weight). Plants fertilized with 5 mM Fe were larger and supported greater numbers or rhizosphere bacteria per gram of root than plants not supplied with Fe. Comparisons of the Fe content and dry weight of roots and shoots revealed few significant differences between inoculated and uninoculated plants, or among the plants inoculated with the different strains of siderophore-producing bacteria. The differences that were observed revealed no consistent response to inoculation. We conclude that inoculation of the roots of the two oat cultivars with bacteria that produce high concentrations of siderophores in response to an Fe deficiency had little or no effect on Fe acquisition by the plants.     

Microbial colonisation of roots as a function of plant species

Fifteen plants species were grown in the greenhouse on the same soil and sampled at flowering to obtain rhizosphere soil and root material. In both fractions, the data on fungal and bacterial tissue obtained by amino sugar analysis were compared with the total microbial biomass based on fumigation-extraction and ergosterol data. The available literature on glucosamine concentrations in fungi and on muramic acid concentrations in bacteria was reviewed to prove the possibility of generating conversion values for general use in root material. All microbial properties analysed revealed strong species-specific differences in microbial colonisation of plant roots. The root material contained considerable amounts of microbial biomass C and biomass N, reaching mean levels of 10.9 and 1.4 mg g⁻¹ dry weight, respectively. However, the majority of CHCl₃ labile C and N, i.e. 89 and 55% was root derived. The average amount of ergosterol was 13 μg g⁻¹ dry weight and varied between 0.0 for Phacelia roots and 45.5 μg g⁻¹ dry weight for Vicia roots. The ergosterol content in root material of mycorrhizal and non-mycorrhizal plant species did not differ significantly. Fungal glucosamine was converted to fungal C by multiplication by 9 giving a range of 7.1-25.9 mg g⁻¹ dry weight in the root material. Fungal C and ergosterol were significantly correlated. Bacterial C was calculated by multiplying muramic acid by 45 giving a range from 1.7 to 21.6 mg g⁻¹ dry weight in the root material. In the root material of the 15 plant species, the ratio of fungal C-to-bacterial C ranged from 1.0 in mycorrhizal Trifolium roots to 9.5 in non-mycorrhizal Lupinus roots and it was on average 3.1. These figures mean that the microbial tissue in the root material consists on average of 76% fungal C and 24% bacterial C. The differences in microbial colonisation of the roots were reflected by differences in microbial indices found in the rhizosphere soil, most strongly for microbial biomass C and ergosterol, but to some extent also for glucosamine and muramic acid.     

RHIZOSPHERE MICROBIAL POPULATIONS IN CONTAMINATED SOILS

Rhizosphere microbial populations may increase bioremediation of soil contaminated with organic chemicals. A growth chamber study was conducted to evaluate rhizosphere microbial populations in contaminated and non-contaminated soil. Alfalfa (Medicago sativa L.) and alpine bluegrass (Poa alpina L.) were grown in soil containing a mixture of organic chemicals for 14 weeks. The equal millimolar mixture of hexadecane, (2,2-dimethylpropyl)benzene, cis-decahydronaphthalene (decalin), benzoic acid, phenanthrene, and pyrene was added at levels of 0 and 2000 mg/kg. Organic chemical degrader (OCD) populations were assessed by a Most-Probable-Number technique, and bacteria and fungi were enumerated by plate count methods. Different methods for expressing OCD rhizosphere populations were investigated to determine the effect it had on interpretation of the results. At 9 weeks, the OCD numbers were significantly higher in rhizosphere and contaminated soils than in bulk and non-contaminated soils, respectively. Alfalfa rhizosphere OCD levels were 4 × 10⁷/g for contaminated and 6 × 10⁶/g for non-contaminated soils. Bluegrass rhizosphere OCD levels were 1 × 10⁷/g and 1 × 10⁶/g in contaminated and non-contaminated soils, respectively. Selective enrichment of OCD populations was observed in contaminated rhizosphere soil. Higher numbers of OCD in contaminated rhizospheres suggest potential stimulation of bioremediation around plant roots.     

Rhizobacteria: Influence of cultivar and soil type on plant growth and yield of potato

The influence of potato cultivar and soil type on effectiveness of plant growth-promoting rhizobacteria (PGPR) was examined. Rhizobacteria were isolated from potato roots and tubers obtained from fields with a history of high potato yields. Fluorescent pigment-producing rhizobacteria. identified as strains of Pseudomonas putida and P. fluorescens, were selected for their antibiosis against Erwinia carovotora ssp. carotovora and growth-promoting activity on potatoes. In greenhouse tests, treatments of potato seedpieces and stem cuttings increased shoot dry weight from 1.23- to 2.00-fold and root dry weight from 1.27- to 2.78-fold. Survival of PGPR in the rhizosphere was monitored using antibioticresistant strains. Populations of these strains decreased from 3.6 × 10⁹ cgu g^?1 dry root weight to 4.5 × 10⁵ cfu g^?1 dry root weight 4 weeks after treatment. In field trials, PGPR strains were applied to seedpieces of cultivars Kennebec, Pungo, Red Pontiac and Superior and planted in Cape Fear loam. Plymouth loamy sand or Delanco sandy loam. Significant yield increases of 1.17–1.37-fold over controls were observed in two of three field trials. Variability in plant growth-promoting activity was observed between greenhouse and field trials, and no given treatment combination of PGPR strain, potato cultivar and soil type was consistently better than another.     

Determination of bacterial DNA in soil

Two methods for the determination of DNA have been adapted for use on the bacterial fraction of an organic soil, obtained by fractionated centrifugation. The. soil contained about 1.1 × 10¹⁰ bacteria g^?1 dry weight when counted by fluorescence microscopy. One method was based on the reaction with 3,5-diaminobenzoic acid 2HCl, and the other on the specific reaction of the antibiotic mithramycin with double-stranded DNA. For the first method, about 8 × 10⁸ cells were required and for the second, about 4 × 10⁹. The analytical results with the two methods agreed well. As an average of several determinations, an amount of 8.4 fg (10^?15g) DNA per bacterial cell was found when the cells were counted by fluorescence microscopy. This is in the upper size range of bacterial genomes. showing that virtually all microscopically counted bacteria in this soil contain DNA. The total amount of DNA was about 90μg g^?1 soil dry weight.     

抗病性不同大豆品种根面及根际微生物区系的变化 Ⅰ.非连作大豆(正茬)根面及根际微生物区系的变化

采用平板计数法测定了3个抗病性不同的大豆品种在生育期内根面和根际微生物区系的变化情况,并应用荧光计数法直接测定了根际细菌和真菌的生物量。结果表明,土体的微生物种类最丰富、根际的次之、根面的较单一。播种后从三叶期到鼓粒初期,根面和根际的可培养细菌总数随生育期逐渐增加,鼓粒初期达最大值,而成熟期则有明显的下降;大豆根际细菌生物量也存在相同的变化规律。抗病性不同的大豆品种其根面、根际可培养细菌总数存在差异;抗病品种大豆的根瘤重明显高于感病品种。种植一季后感病品种根际积累的病原生物(镰孢霉Fusarium.sp.和大豆胞囊线虫Heterodera.glycines的胞囊数)明显高于抗病品种。说明大豆根系分泌物对微生物具有选择性的促进或抑制作用,不同大豆品种以及同一大豆品种在不同生育时期根系分泌物的组成和数量不同,从而使大豆根面及根际形成了特定的微生物区系组成。     

连续施用生物有机肥对烟草青枯病的防治效果

分离获得一株对烟草青枯病病原菌茄科劳尔氏菌(Ralstonia solanacearum,简称RS)具有较强拮抗能力的拮抗菌(SQR11)并制成生物有机肥,研究了连续施用该生物有机肥对烟草青枯病的防治效果。结合生理生化和16SrDNA技术鉴定,菌株SQR11被鉴定为解淀粉芽孢杆菌。施用该生物有机肥后第一季烟草青枯病的生物防治率达到47%以上,第二季为69%以上,第三季达到89%以上。第三批盆栽实验表明,当根际土中病原菌数量达到2×105cfu/g干土时,植株出现发病症状,随着病原菌数量的增加,发病症状加重。当根际土中拮抗菌活菌数量达到2×107cfu/g干土时,病原菌繁殖得到有效抑制,可有效阻止植株染病;若低于107cfu/g干土,则不能有效抑制病原菌增殖,植株表现发病症状。植株各组织内拮抗菌数量检测发现,未发病植株茎部拮抗细菌数量为4×104cfu/g(组织鲜重,下同)左右,而同处理中发病症状的植株茎部拮抗细菌数量仅为6×103cfu/g;相对应的病原菌数量分别为1.5×102cfu/g(健康植株)和3×103cfu/g(发病植株)。SQR11菌株制成的生物有机肥还具有较好的促生作用。总之,利用拮抗菌SQR11菌株制成的生物有机肥对烟草青枯病具有显著的生物防治作用,在根部进行大量定殖后可有效防止病原菌的侵入,能够获得显著的生防效果。     

Root-induced changes in the rhizosphere: Importance for the mineral nutrition of plants

Root-induced changes in the rhizosphere may affect mineral nutrition of plants in various ways. Examples for this are changes in rhizosphere pH in response to the source of nitrogen (NH₄-N versus NO₃-N), and iron and phosphorus deficiency. These pH changes can readily be demonstrated by infiltration of the soil with agar containing a pH indicator. The rhizosphere pH may be as much as 2 units higher or lower than the pH of the bulk soil. Also along the roots distinct differences in rhizosphere pH exist. In response to iron deficiency most plant species in their apical root zones increase the rate of H⁺ net excretion (acidification), the reducing capacity, the rate of Fe^III reduction and iron uptake. Also manganese reduction and uptake is increased several-fold, leading to high manganese concentrations in iron deficient plants. Low-molecular-weight root exudates may enhance mobilization of mineral nutrients in the rhizosphere. In response to iron deficiency, roots of grass species release non-proteinogenic amino acids (?phytosiderophores”?) which dissolve inorganic iron compounds by chelation of Fe^III and also mediate the plasma membrane transport of this chelated iron into the roots. A particular mechanism of mobilization of phosphorus in the rhizosphere exists in white lupin (Lupinus albus L.). In this species, phosphorus deficiency induces the formation of so-called proteoid roots. In these root zones sparingly soluble iron and aluminium phosphates are mobilized by the exudation of chelating substances (probably citrate), net excretion of H⁺ and increase in the reducing capacity. In mixed culture with white lupin, phosphorus uptake per unit root length of wheat (Triticum aestivum L.) plants from a soil low in available P is increased, indicating that wheat can take up phosphorus mobilized in the proteoid root zones of lupin. At the rhizoplane and in the root (root homogenates) of several plant species grown in different soils, of the total number of bacteria less than 1 % are N₂-fixing (diazotrophe) bacteria, mainly Enterobacter and Klebsiella. The proportion of the diazotroph bacteria is higher in the rhizosphere soil. This discrimination of diazotroph bacteria in the rhizosphere is increased with foliar application of combined nitrogen. Inoculation with the diazotroph bacteria Azospirillum increases root length and enhances formation of lateral roots and root hairs similarly as does application of auxin (IAA). Thus rhizosphere bacteria such as Azospirillum may affect mineral nutrition and plant growth indirectly rather than by supply of nitrogen.     

Biological nitrogen fixation associated with roots of field-grown barley (Hordeum vulgare L.)

Nitrogenase (C₂H₂) activity was measured in microbial media inoculated with barley root segments or corresponding rhizosphere soil. Three different media were used, Döbereiner's malate medium, a modified Ashby medium, and an acid nitrogen-free medium. Only Döbereiner's medium gave consistently positive results, and cultures inoculated with roots showed higher activity than cultures inoculated with corresponding rhizosphere soil. Similar experiments with roots and rhizosphere soil from wheat gave only negligible nitrogenase activity, whereas the tropical grass, Cynodon dactylon, gave higher activity than barley. Measurements on intact soil cores containing barley root systems showed an initial lag phase followed by a rather stable activity level over a period from 12 h to 48 h, and then the activity again decreased. The activity during the stable period corresponded to fixation of about 100 to 200 g N₂ ha^?1 24 h^?1. Measurements on isolated, washed barley roots showed only negligible nitrogenase activity.     

Characterization of cultivable methanotrophs from paddy soils and rice roots

Abstract

Characterization of methanotrophs isolated from paddy soils and rice (Oryza sativa) roots was investigated in the present study. The number of methanotrophs in root homogenates of the rice cultivar Mutsuhomare was 4.9 × 10⁷ most-probable-number (MPN) g^?1 dry roots, in Yumeakari it was 2.0 × 10⁸ MPN g^?1 dry roots and in Kirara it was 4.6 × 10⁷ MPN g^?1 dry roots. Although bacterial cells were observed infrequently on the surface and in the interiors of roots before incubation, a large number of colonies, measuring 0.5–5 mm in diameter, were observed on the sterilized roots after incubation on nitrate mineral agar plates under methane in air. In particular, a large number of colonies were observed at the emergence sites of lateral roots and root hairs. Strains MD5-1 and M1 were isolated from the roots of Mutsuhomare and strain R62 was isolated from the root homogenate of Yumeakari. All isolates were catalase-positive and oxidase-positive, Gram-negative, straight-rod-shaped and curved-rod-shaped bacteria, and formed exospores. The isolates were able to fix nitrogen and grew in the absence of copper. In addition, all were found to be positive for naphthalene-oxidizing activities (corresponding to soluble methane mono-oxygenase activities). Strains MD5-1, M1 and R62 were closely related to Methylosinus sporium. Methanotrophic strains W3-6, SD3-5 and 2-19, isolated previously from paddy field soils, were classified into Methylosinus (W3-6) and Methylocystis (SD3-5 and 2-19) type II methanotrophs. Isolates from the rice roots (MD5-1, R62 and M1) grew logarithmically when casamino acid was used as the nitrogen source; however, the growth of these strains was reduced on the nitrate medium. These strains preferred amino acids over inorganic nitrogen as a nitrogen source for growth.     

Suppression of wheat take-all and ophiobolus patch by fluorescent pseudomonads from a fusarium-suppressive soil

Fluorescent pseudomonads isolated from a soil suppressing Fusarium wilt significantly reduced take-all (Gaeumannomyces graminis var. tritici) in wheat and Ophiobolus patch (G. graminis var. avenae) in Agrostis turfgrass. The bacteria were mixed into a conducive soil at a concentration of 10⁷ colony-forming units (cfu)g^?1 soil at sowing. There were significantly fewer (P ? 0.05) diseased wheat roots in the treatments with the bacteria and pathogen than in those with the pathogen alone. Dry weights of the tops of wheat and Agrostis turfgrass were significantly greater (P ? 0.01) in treatments inoculated with the bacteria in the presence of the pathogens compared to controls with the pathogens alone. Dry weights of the tops of plants from treatments inoculated with the bacteria alone were not significantly different to those of healthy wheat non-inoculated with the bacteria, showing that the fluorescent pseudomonads did not stimulate plant growth. At the end of the experiments, the bacterial isolates (genetically-marked with rifampicin resistance) were recovered from wheat roots and rhizosphere soil at concentrations of 10⁵–10⁷cfu g^?1 fresh weight of roots or oven-dried rhizosphere soil.Many of the fluorescent pseudomonads and some non-fluorescent pseudomonads showed in vitro antibiosis on quarter-strength potato dextrose agar (QPDA) against the pathogens. However, there was no correlation between in vitro antibiosis on agar plates and suppression of disease in pot experiments. Further, while some isolates of G. graminis var. tritici and var. avenae were inhibited by certain bacterial isolates, other isolates of the same fungus were not similarly inhibited by the same isolates of bacteria. Most of the fluorescent pseudomonads that produced inhibition zones (>5mm) against G. graminis var. tritici on QPDA did not do so on King's medium B, where fluorescent siderophores were formed. In vitro antibiosis is, therefore, a poor criterion for selecting effective bacterial antagonists of the wheat take-all fungus. All of the fluorescent pseudomonads tested produced siderophores in low-Fe media while a non-fluorescent pseudomonad and the fungal pathogens did not produce siderophores of comparable activity. The addition of 500 μg FeEDTA g^?1 with a lower stability constant did not. The evidence suggests that iron competition at the rhizoplane or in the rhizosphere is one mechanism of suppression.