植物根际促生菌对3种土传真菌病害病原的抑制作用

【目的】获取促生同时可防治3种土传真菌病害(Fusarium oxysporum、Sclerotinia sclerotiorum和Rhizoctonia solani)的生防菌,并明确其抑菌效果。【方法】利用前期研究获得的17株促生菌,采用平板对峙法测定其对病原真菌的拮抗作用及对菌丝生长的抑制作用。【结果】可有效拮抗立枯丝核菌的生防菌有6株,其中促生菌株FX2和LM4-3的抑制率达73.82%;拮抗尖孢镰刀菌的生防菌有7株,其中FX2的抑制率达到66.81%;拮抗油菜菌核病菌的生防菌有4株,其中菌株LHS11的抑制率高达85.71%。菌株LHS11和JM170通过次生代谢物抑制病原真菌。所有的生防菌对病原菌的菌丝生长均有一定的抑制作用。【结论】筛选得到对3种真菌病害病原具有较好生防作用的菌株LHS11和FX2。     

强化龙葵富集镉根际促生菌的分离、筛选与鉴定

【背景】植物-微生物联合修复土壤重金属污染日渐兴起,获取与超富集植物高效互作的微生物是实现联合修复技术进步的关键。龙葵在镉农田污染修复中广泛应用。【目的】筛选可促进龙葵生长及富集镉的耐镉根际促生菌。【方法】从龙葵根际土分离耐镉菌株,筛选具有良好促生特性的菌株,水培试验考察镉胁迫下菌株对植物生长及镉富集能力的影响,确定可促进龙葵生长及富集镉的微生物菌株,通过生理生化特性和16S rRNA基因序列分析进行菌株的初步鉴定。【结果】分离得到NT1、AXY1、AW2和AW1四株强化龙葵富集镉促生菌,经鉴定分别为Lysinibacillus sp.、Beijerinckia fluminensis、Achromobacter animicus和Herbaspirillum huttiense。上述菌株均可有效促进龙葵生长,增加其株高和干物质积累,提高地上部镉富集量。其中,NT1可使株高、地上部干重分别增加31.33%和62.65%,AW2可使地上部镉富集量增加37.29%。【结论】筛选所得菌株可为提高植物修复效率提供实践依据,为研制田间施用生态功能菌剂做铺垫,用于农田镉污染的微生物-龙葵联合原位修复。     

Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat

AIMS: Plant growth promoting rhizobacteria (PGPR) are commonly used as inoculants for improving the growth and yield of agricultural crops, however screening for the selection of effective PGPR strains is very critical. This study focuses on the screening of effective PGPR strains on the basis of their potential for in vitro auxin production and plant growth promoting activity under gnotobiotic conditions. METHODS AND RESULTS: A large number of bacteria were isolated from the rhizosphere soil of wheat plants grown at different sites. Thirty isolates showing prolific growth on agar medium were selected and evaluated for their potential to produce auxins in vitro. Colorimetric analysis showed variable amount of auxins (ranging from 1.1 to 12.1 mg l-1) produced by the rhizobacteria in vitro and amendment of the culture media with l-tryptophan (l-TRP), further stimulated auxin biosynthesis (ranging from 1.8 to 24.8 mg l-1). HPLC analysis confirmed the presence of indole acetic acid (IAA) and indole acetamide (IAM) as the major auxins in the culture filtrates of these rhizobacteria. A series of laboratory experiments conducted on two cv. of wheat under gnotobiotic (axenic) conditions demonstrated increases in root elongation (up to 17.3%), root dry weight (up to 13.5%), shoot elongation (up to 37.7%) and shoot dry weight (up to 36.3%) of inoculated wheat seedlings. Linear positive correlation (r = 0.99) between in vitro auxin production and increase in growth parameters of inoculated seeds was found. Based upon auxin biosynthesis and growth-promoting activity, four isolates were selected and designated as plant growth-promoting rhizobacteria (PGPR). Auxin biosynthesis in sterilized vs nonsterilized soil inoculated with selected PGPR was also monitored that revealed superiority of the selected PGPR over indigenous microflora. Peat-based seed inoculation with selected PGPR isolates exhibited stimulatory effects on grain yields of tested wheat cv. in pot (up to 14.7% increase over control) and field experiments (up to 27.5% increase over control); however, the response varied with cv. and PGPR strains. CONCLUSIONS: It was concluded that the strain, which produced the highest amount of auxins in nonsterilized soil, also caused maximum increase in growth and yield of both the wheat cv. SIGNIFICANCE AND IMPACT OF STUDY: This study suggested that potential for auxin biosynthesis by rhizobacteria could be used as a tool for the screening of effective PGPR strains.     

植物根际促生菌的筛选及鉴定

【目的】植物根际促生菌(PGPR)和植物的互作关系往往不稳定,PGPR菌群有可能提高菌株对野外环境的适应性。为此,本文根据PGPR促生机制的多样性,从不同植物根际土壤进行了PGPR的筛选及鉴定。【方法】首先,按照固氮、解磷、解钾、拮抗6种常见病原真菌,同时能在植物根际定殖为基本初筛标准,然后在实验室条件下测定初筛菌株的多项促生能力(PGP),最后通过生理生化试验和16SrRNA基因序列分析对所筛菌株进行鉴定。【结果】从江苏扬州、盐城等地土壤样品筛选出14株PGPR,具有体外抑菌、产NH3、产IAA、产HCN、产嗜铁素、解磷、溶钾、固氮以及产抗生素等促生能力。分类鉴定结果显示:7株属于假单胞菌属(Pseudomonas)、3株属于类芽孢杆菌属(Paenibacillus)、2株为芽孢杆菌属(Bacillus)、1株为布克霍尔德氏菌属(Burkholderia)、1株为欧文氏菌属(Erwinia)。【结论】所筛细菌具有多种促生能力,且能在根际定殖,为进一步构建多功能PGPR广适菌群提供菌株资源。     

植物根际促生菌作用机制研究进展

植物在生长过程中可能会遭受许多生物和非生物因素胁迫,从而降低生物产量. 人们已知一些植物在不同因素的刺激诱导下,能系统化建立抵抗或忍受不利因素的机制,植物根际促生菌（PGPR）就是其中一类能定殖于根系并促进植物生长的细菌.本文对PGPR促生机制进行归纳和总结,系统阐述了诱导体系抗性和诱导体系产生忍耐力两大促生机制.PGPR的作用机制的多样性暗示着其可能在更多的农业生态系统中得到应用.     

The effect of plant growth-promoting rhizobacteria on the growth,physiology, and Cd uptake of Arundo donax L.

In this study, plant growth-promoting potential isolates from rhizosphere of 10 weed species grown in heavy metal-contaminated areas were identified and their effect on growth, antioxidant enzymes, and cadmium (Cd) uptake in Arundo donax L. was explored. Plant growth-promoting traits of isolates were also analyzed. These isolates were found to produce siderophores and enzymes such as 1-aminocyclopropane-1-carboxylate (ACC) deaminase, and aid in solubilization of mineral nutrients and modulate plant growth and development. Based on the presence of multiple plant growth-promoting traits, isolates were selected for molecular characterization and inoculation studies. Altogether, 58 isolates were obtained and 20% of them were able to tolerate Cd up to 400 ppm. The sequence analysis of the 16S rRNA genes indicates that the isolates belong to the phylum Firmicutes. Bacillus sp. along with mycorrhizae inoculation significantly improves the growth, the activity of antioxidants enzymes, and the Cd uptake in A. donax than Bacillus alone. Highly significant correlations were observed between Cd uptake, enzymatic activities, and plant growth characteristics at 1% level of significance. The synergistic interaction effect between these organisms helps to alleviate Cd effects on soil. Heavy metal-tolerant isolate along with arbuscular mycorrhizae (AM) could be used to improve the phytoremedial potential of plants.     

Identification,characterization and optimization of phosphate solubilizing rhizobacteria (PSRB) from rice rhizosphere

Two billion people worldwide take rice (Oryza sativa L.) as a staple food. Phosphorus (P) and Nitrogen (N) are the major requirements of rice; although these are available in limited concentrations within rice growing regions. Among different types of Plant growth-promoting rhizobacteria (PGPR), Phosphate solubilizing rhizobacteria (PSRB) constitute an important class. These are known for plant growth promotion by enhancing P and N uptake. PSRB are nowadays used as biofertilizers to restore the soil health. Under the present investigation identification, characterization and optimization of phosphate solubilizing activity of these microbes at different pH, temperature and salt concentrations was carried out. Thirty-seven isolates were recovered from different regions of rice rhizosphere on Pikovskaya (PVK) agar among which 15 isolates were recovered from R.S. Pura, 12 isolates from Bishnah and 10 isolates were recovered from Akhnoor sector of Jammu, India. A prominent halo zone of clearance was developed around the colonies of 12 different isolates, indicating phosphate solubilization activity. Four distinct isolates were amplified, cloned and sequenced for taxonomic identification using 16S primers. The results indicated that PS 1, PS 2, PS 3, PS 4 were related to Pseudomonas aeruginosa, Bacillus subtilis strain 1, B. subtilis strain 2, B. subtilis strain 3, respectively. These strains when grown at a wide range of ecological factors showed maximum growth at pH between 6.8 and 8.8, temperature between 28 °C and 37 °C and salinity between 1% and 2%. Screening for phosphate solubilization activity revealed that the halo zone diameter formed by these isolates extended from 2.1 to 3.2 mm. The phosphate solubilizing efficiency (SE) ranged from 35.4 to 50.9 with highest value of 50.9 by PS4 and maximum P solubilization of 10.22 µg/ml was recorded by PS4 at 7th day. Phosphate solubilization activity of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus. MIC value for zinc sulphate heptahydrate in 12 isolates varied from 1 mg/ml to 6 mg/ml. Phosphate solubilization activity and MIC of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus.     

丛枝菌根真菌与根围促生细菌相互作用的效应与机制

丛枝菌根(arbuscular mycorrhiza,AM)真菌是植物活体营养专性共生菌,广泛存在于陆地各生态系统中.研究表明,AM真菌与根围促生细菌(plant growth promoting rhizobacteria,PGPR)之间的相互作用,尤其是它们之间的协同作用不仅影响植物养分吸收利用、病原物发生发展、土壤理化特性与生物修复等,而且对于可持续农、林、牧业生产、稳定生态系统都具有十分重要的意义.因此,近年来给予众多关注和研究.综述了AM真菌与PGPR之间的相互影响及其可能的作用机制,以及AM真菌与PGPR协同改善植物营养和生长、协同抑制病原菌、协同修复土壤方面的作用,旨在总结AM真菌与PGPR相互作用的效应与机制方面的最新研究进展,为今后研究发展提供依据.     

Effect of plant growth promoting rhizobacteria on bacterial canker of tomato

Use of plant growth promoting rhizobacteria in managing bacterial canker disease of tomato was studied in the present work. Tomato seeds were treated with PGPR strains viz., Bacillus pumilus INR7, Bacillus pumilus SE34, Bacillus pumilus T4, Bacillus subtilis GBO3, Bacillus amyloliquefaciens IN937a and Brevibacillus brevis IPC11 were subjected for seed germination and seedling vigor. Among the PGPR strains tested, only three strains (IN937a, GBO3 and IPC11) which showed enhancement in the seed quality parameters like seed germination and seedling vigor, were further subjected for estimation of one of the defence-related enzymes, Phenylalanine Ammonia Lyase (PAL) with total phenol contents. The same three strains were recorded for maximum disease protection under greenhouse conditions. The level of PAL and total phenol contents increased significantly upon the PGPR treatment. The rate of reduction in the bacterial canker disease incidence was directly proportional to the amount of increased level of PAL and total phenol content. The possible uses of these PGPR strains in effective management of bacterial canker of tomato were discussed in the present work.     

田菁根际促生菌的筛选及其促生耐盐效果

【目的】为探究盐生植物田菁及其根际功能微生物改良盐碱地的效果,本研究从黄河三角洲盐碱区田菁根际土壤中分离促生菌,并明确其耐盐促生效果。【方法】采用选择培养方法从田菁根际土壤中分离固氮菌、解磷菌以及解钾菌,并进行16S rRNA分子生物学鉴定。之后对菌株的耐盐及促生特性进行测定,筛选性状优良菌株进行玉米促生作用研究。【结果】共分离得到105株根际促生菌,其中N102兼具多种促生特性且耐盐性达15%。田菁种子发芽试验表明,N102可显著提高田菁发芽率(47%,P<0.05)、芽长(48.5%,P<0.05)和根长(60%,P<0.05);玉米盆栽试验结果表明,N102对盐胁迫下玉米的株高、根长、叶绿素含量、地上部干重以及根干重具有显著的促进作用。经系统发育分析,N102与Enterobacter soli ATCC BAA-2102 (NR117547)序列相似度为99.30%,鉴定属于Enterobacter属。【结论】菌株N102具有多种植物促生耐盐特性,具有开发成有效促进盐碱地作物生长的微生物肥的良好前景。     

Induction of resistance against downy mildew on sunflower by rhizobacteria

Abstract

Induction of resistance to downy mildew caused by Plasmopara halstedii in sunflower was studied after treatment with PGPR (plant growth promoting rhizobacteria) strain INR7 (Bacillus spp). Treatment of sunflower seeds with 1×10⁸cfu/ml of PGPR strain INR7 resulted in decreased disease severity and offered 51 and 54% protection under green house and field conditions, respectively. The induction of resistance to P. halstedii by PGPR strain INR7 was accompanied by the accumulation of various host defense-related enzymes in susceptible sunflower seedlings. Enhanced activation of catalase (CAT), phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO) and chitinase (CHI) was evident at 6, 9, 12, 12 and 12h post inoculation, respectively, in sunflower seedlings raised from seeds treated with PGPR strain INR7. This enhanced and early activation of defense-related responses in the susceptible cultivar after treatment with PGPR strain INR7 was comparable to that in the resistant cultivar. The results indicate that PGPR strain INR7 induced resistance against P. halstedii in sunflower is mediated through enhanced expression of defense mechanism.     

A novel bacteriocin, thuricin 17, produced by plant growth promoting rhizobacteria strain Bacillus thuringiensis NEB17: isolation and classification

AIMS: The aim of this study was to identify and characterize a compound produced by the plant growth promoting bacterium, Bacillus thuringiensis non-Bradyrhizobium Endophytic Bacterium 17. METHODS AND RESULTS: The bacterial peptide was analysed and purified via HPLC. Using the disk diffusion assay this peptide inhibited the growth of 16/19 B. thuringiensis strains, 4/4 Bacillus cereus strains, among others, as well as a Gram-negative strain Escherichia coli MM294 (pBS42). Both bactericidal and bacteristatic effects were observed on B. cereus ATCC 14579 and bactericidal effects were observed on B. thuringiensis ssp. thuringiensis Bt1267. The molecular weight of the peptide was estimated via SDS-PAGE and confirmed with Matrix Assisted Laser Desorption Ionization Quadrapole Time of Flight mass spectrometry; its weight is 3162 Da. The peptide is biologically active after exposure to 100 degrees C for 15 min, and within the pH range 1.00-9.25. Its activity disappeared when treated with proteinase K and protease, but not with alpha-amylase or catalase. CONCLUSIONS: We conclude that this is the first report of a bacteriocin produced by a plant growth promoting rhizobacteria (B. thuringiensis) species and have named the bacteriocin thuricin 17. SIGNIFICANCE AND IMPACT OF THE STUDY: Our work has characterized a bacteriocin produced by a plant growth promoting bacterium. This strain is previously reported to increase soya bean nodulation.     

Plant growth promoting rhizobacteria improve growth and essential oil yield in Origanum majorana L.

Effects of root colonization by plant growth promoting rhizobacteria (PGPR) on biomass, and qualitative and quantitative composition of essential oils, were determined in the aromatic crop Origanum majorana L. (sweet marjoram). PGPR strains evaluated were Pseudomonas fluorescens, Bacillus subtilis, Sinorhizobium meliloti, and Bradyrhizobium sp. Only P. fluorescens and Bradyrhizobium sp. showed significant increases in shoot length, shoot weight, number of leaf, number of node, and root dry weight, in comparison to control plants or plants treated with other PGPR. Essential oil yield was also significantly increased relative to non-inoculated plants, without alteration of oil composition. P. fluorescens has clear commercial potential for economic cultivation of O. majorana.     

Perspective of plant growth promoting rhizobacteria (PGPR) containing ACC deaminase in stress agriculture

Ethylene is a gaseous plant growth hormone produced endogenously by almost all plants. It is also produced in soil through a variety of biotic and abiotic mechanisms, and plays a key role in inducing multifarious physiological changes in plants at molecular level. Apart from being a plant growth regulator, ethylene has also been established as a stress hormone. Under stress conditions like those generated by salinity, drought, waterlogging, heavy metals and pathogenicity, the endogenous production of ethylene is accelerated substantially which adversely affects the root growth and consequently the growth of the plant as a whole. Certain plant growth promoting rhizobacteria (PGPR) contain a vital enzyme, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which regulates ethylene production by metabolizing ACC (an immediate precursor of ethylene biosynthesis in higher plants) into α-ketobutyrate and ammonia. Inoculation with PGPR containing ACC deaminase activity could be helpful in sustaining plant growth and development under stress conditions by reducing stress-induced ethylene production. Lately, efforts have been made to introduce ACC deaminase genes into plants to regulate ethylene level in the plants for optimum growth, particularly under stressed conditions. In this review, the primary focus is on giving account of all aspects of PGPR containing ACC deaminase regarding alleviation of impact of both biotic and abiotic stresses onto plants and of recent trends in terms of introduction of ACC deaminase genes into plant and microbial species.     

Identification of rhizobacteria from maize and determination of their plant-growth promoting potential

During the growing season of 1986, the rhizobacteria (including organisms from the ectorhizosphere, the rhizoplane and endorhizosphere) of 20 different maize hybrids sampled from different locations in the Province of Quebec were inventoried by use of seven different selective media. Isolates were characterized by morphological and biochemical tests and identified using the API20E and API20B diagnostic strips.Pseudomonas spp. were the prominent bacteria found in the rhizoplane and in the ectorhizosphere.Bacillus spp. andSerratia spp. were also detected, but in smaller numbers. In the endorhizosphere,Bacillus spp. andPseudomonas spp. were detected in order of importance. Screening for plant growth-promoting rhizobacteria was carried out in three soils with different physical and chemical characteristics. The results depended on the soil used, but two isolates (Serratia liquefaciens andPseudomonas sp.) consistently caused a promotion of plant growth.Contribution no. 350 of the Research Station, Agriculture Canada, Sainte-Foy, Quebec.     

The effect of plant growth-promoting rhizobacteria on the phytoextraction of Cd and Zn by Brassica napus L.

The test strains Bacteroidetes bacterium (Ba), Pseudomonas fluorescens (Pf) and Variovorax sp. (Va) were selected in advance for their in vitro capability for growth promotion of rapeseed in the presence of increased concentrations of Cd, Cu, Pb and Zn in the medium. In the pot experiment, the strains were used for single Ba, Pf, Va or combined Ba + Pf, Ba + Va, Pf + Va, and Ba + Pf + Va inoculation of B. napus growing in contaminated soil from alluvial deposits. The positive effect of bacterial strains on plant growth was observed in vitro, but was not confirmed in situ in the contaminated soil, where the tested strains inhibited biomass production, rather than stimulating it. However, single inoculation with Ba significantly increased the chlorophyll content and K⁺ concentration in the leaves. The inoculation of rapeseed with Ba and Va strains was indicated to be the most promising combination for phytoextraction of Cd and Zn from contaminated soil. Combined inoculation with Pf+Va and Pf + Ba+Va significantly decreased the concentration of heavy metals in the roots of rapeseed. We conclude that suitable combinations of PGPR can control the metal uptake of B. napus, selectively increasing either metal extraction or metal stabilization in the rhizosphere and offering promising applications in soil remediation.     

Effect of plant growth promoting rhizobacteria containing ACC-deaminase on maize (Zea mays L.) growth under axenic conditions and on nodulation in mung bean (Vigna radiata L.)

AIMS: This study was conducted to test the hypothesis that the bacterial strains possessing 1-aminocyclopropane-1-carboxylic acid (ACC)-deaminase activity may also promote growth of inoculated plants and could increase nodulation in legumes upon co-inoculation with rhizobia. METHODS AND RESULTS: Several rhizobacteria were isolated from maize rhizosphere through enrichment on ACC as a sole N source. Purified isolates were screened for growth promotion in maize under axenic conditions and for in vitro ACC-deaminase activity. A significant positive correlation was observed between in vitro ACC-deaminase activity of bacterial cells and root elongation. None of the isolates produced auxins. Bradyrhizobium japonicum produced less amount of auxins but did not carry ACC-deaminase activity. Results of pot experiment revealed that co-inoculation with Bradyrhizobium and plant growth promoting rhizobacteria (PGPR) isolates enhanced the nodulation in mung bean compared with inoculation with Bradyrhizobium alone. CONCLUSIONS: It is highly expected that inoculation with rhizobacteria containing ACC-deaminase hydrolysed endogenous ACC into ammonia and alpha-ketobutyrate instead of ethylene. Consequently, root and shoot growth as well as nodulation were promoted. SIGNIFICANCE AND IMPACT OF THE STUDY: The ACC-deaminase trait could be employed as an efficient tool to screen effective PGPR, which could be successfully used as biofertilizers to increase the growth of inoculated plants as well as nodulation in legumes.     

Factors associated with detrimental effects of rhizobacteria on plant growth

Some of the factors interfering with the specific response of young common bean plants to two rhizosphere fluorescent pseudomonads were studied. These two bacterial strains produced symptoms in foliar plant parts and reduced yield in beans and several other plant species when inoculated on roots. Sensitivity in the plants subjected to bacterial application was highest at early growth stages (up to ten days old plants) giving typical symptoms for each strain in first expanding leaves and typically stunted plant growth. Symptoms to some extent also appeared on plants treated at the age of 2–3 weeks and the fresh shoot weight was affected also in such plants. Deleterious effects of the tested bacteria were found to be related to the level of inoculum (cfu/ml) used and presence or absence of certain nutrients in the bacterial suspensions used for inoculation. The two tested strains differed in nutritional requirements for affecting plant growth. One strain needed sucrose only. The other tested strain required peptone or yeast extract in addition to sucrose to induce typical symptoms and significant yield reductions. Supply of peptone as the only nutrient source to the bacterial suspensions eliminated the deleterious effects of both bacterial strains as tested on beans under nonsterile conditions. The two strains are assumed to affect test plants by different modes of action.