Characterization of phosphate solubilizing fluorescent pseudomonads from the rhizosphere of Aloe vera (L.)

Soil phosphorous (P) deficiency is a major constraint to plant production which is overcome by adding inorganic-phosphate as chemical fertilizers. Fluorescent pseudomonads are the diverse group of bacteria able to mobilize sparingly soluble phosphate form. Total three hundred seven fluorescent Pseudomonas isolates were obtained from the Aloe barbadensis (Miller) rhizosphere. These Pseudomonas strains were further evaluated in vitro for their ability to solubilize phosphate and to produce indole acetic acid (IAA), hydrogen cyanide (HCN), siderophore and 1-aminocyclopropane 1- carboxylate (ACC) deaminase. Fifty three (36.8%) isolates produced IAA and 52 (36.1%) isolates produced siderophores whereas 36 (25.0%) and 31 (21.5%) isolates produced HCN and ACC deaminase, respectively. A positive correlation existed between siderophore and ACC deaminase producers. Cluster analysis showed rhizosphere as the major factor influencing the ecological distribution and physiological characterization of phosphate solubilizing bacteria (PSB). Based on partial 16S rRNA gene sequencing PSB were identified as Pseudomonas putida, Pseudomonas sp. and Pseudomonas plecoglossicida with highest phosphate solubilization ability. In conclusion, these phosphate solubilizing fluorescent pseudomonads would help in understanding their role in phosphorus solubilization and identification of potent phosphorus solubilizers from the rhizosphere of commercially grown A. barbadensis.     

盐胁迫油菜根际中含假单胞菌（Pseudomonas spp.1）的氨基环丙烷-1-羧酸 (ACC) 脱氨基酶特征研究

When exposed to biotic or abiotic stress conditions,plants produce ethylene from its immediate precursor 1-aminocyclopropane-1-carboxylate(ACC),leading to retarded root growth and senescence.Many plant growth-promoting rhizobacteria contain the enzyme ACC deaminase and this enzyme can cleave ACC to form α-ketobutyrate and ammonium,thereby lowering levels of ethylene.The aim of this study was to isolate and characterize ACC deaminase-producing bacteria from the rhizosphere of salt-stressed canola(Brassica napus L.).Out of 105 random bacterial isolates,15 were able to utilize ACC as the sole source of nitrogen.These 15 isolates were also positive for indole acetic acid(IAA) production.Phylogenetic analysis based on partial 16 S rDNA sequences showed that all isolates belonged to fluorescent Pseudomonas spp.In the canola rhizosphere investigated in this study,Pseudomonas fluorescens was the dominant ACC deaminase-producing species.Cluster analysis based on BOX-A1R-based repetitive extragenic palindromic-polymerase chain reaction(BOX-PCR) patterns suggested a high degree of genetic variability in ACC deaminase-producing P.fluorescens strains.The presence of indigenous ACC-degrading bacteria in the rhizosphere of canola grown in saline soils indicates that these bacteria may contribute to salinity tolerance.     

Rhizobacterium Bacillus amyloliquefaciens Strain SQRT3-Mediated Induced Systemic Resistance Controls Bacterial Wilt of Tomato

Tomato bacterial wilt caused by Ralstonia solanacearum seriously threats tomato growth in tropical and temperate regions around the world. This study reported an antagonistic bacterial strain, Bacillus amyloliquefaciens strain SQRT3, isolated from the rhizosphere soil of tomato plants, which strongly inhibited in vitro growth of pathogenic R. solanacearum. The suppression of tomato bacterial wilt by strain SQRT3 was demonstrated under greenhouse conditions. Additionally, induced systemic resistance (ISR) in tomato as one of the potential disease suppression mechanisms was investigated in the plants inoculated with the isolated bacterial strain SQRT3. The results showed that strain SQRT3 applied with R. solanacearum by drenching significantly reduced tomato bacterial wilt by 68.1% biocontrol efficiency (BE) and suppressed the R. solanacearum populations in the rhizosphere soil compared to the control only drenched with R. solanacearum. The BE of the isolated bacterial strain SQRT3 against tomato wilt increased to 84.1% by root-dipping. Tomato plants treated with both strain SQRT3 and R. solanacearum showed increases in activities of peroxidase and polyphenol oxidase compared with other treatments. The application of strain SQRT3 reduced membrane lipid peroxidation in tomato leaves. The expressions of marker genes for jasmonic acid-and salicylic acid-dependent signaling pathways were faster and stronger in tomato plants treated with both strain SQRT3 and R. solanacearum than in plants treated with either R. solanacearum or strain SQRT3 alone. Collectively, the findings indicated that strain SQRT3 can effectively control tomato wilt.     

Bacterial communities in the rhizosphere of Phragmites australis from an oil-polluted wetland

Although Phragmites australis is commonly planted in constructed wetlands, very little is known about its roots-associated bacterial communities, especially in wetlands used for the remediation of oil produced waters. Here, we describe the bacterial diversity, using molecular (illumina MiSeq sequencing) and cultivation techniques, in the rhizosphere soils of P. australis from an oil-polluted wetland in Oman. The obtained isolates were tested for their plant-growth promoting properties. Most sequences belonged to Proteobacteria, Bacteriodetes and Firmicutes. Sequences of potential hydrocarbon-degrading bacteria (e.g. Ochrobactrum, and Pseudomonas) were frequently encountered. All soils contained sequences of known sulfur-oxidizing (e.g. Thiobacillus, Thiofaba, Rhodobacter and Sulfurovum) and sulfate-reducing bacteria, although the latter group made up only 0.1% to 3% of total sequences. The obtained isolates from the rhizosphere soils were phylogenetically affiliated to Serratia, Acinetobacter, Xenorhabdus, Escherichia and Salmonella. All strains were able to solubilize phosphate and about half were capable of producing organic acids and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Around 42% of the strains had the ability to produce indole acetic acid and siderophores. We conclude that the rhizosphere soils of P. australis in oil polluted wetlands harbor diverse bacterial communities that could enhance the wetland performance through hydrocarbon degradation, nutrient cycling and supporting plant growth.     

大棚土壤接种丛枝菌根真菌增强黄瓜枯萎病抑制水平

A pot experiment was conducted to study the plant growth and fruit yields of cucumber (Cucumis sativus L.) on a greenhouse soil with or without inoculation of arbuscular mycorrhizal fungi (AMFs) and Fusarium oxysporum f. sp. cucumerinum under unsterilized conditions. Two AMF inocula were tested: only one AMF strain Glomus caledonium 90036 and an AMF consortium mainly consisting of Glomus spp. and Acaulospora spp. There were four treatments including no inoculation (control), inoculation with F. oxysporum but without mycorrhizae (FO), inoculation with F. oxysporum and G. caledonium (FO+M1), and inoculation with F. oxysporum and the AMF consortium (FO+M2). Cucumber plants were harvested at weeks 3 and 9 after transplanting. Compared with the control, the FO treatment without AMF inoculation had less biomass both at weeks 3 and 9 (P < 0.05) and had higher incidence of Fusarium wilt and produced no cucumber fruit at week 9. Both FO+M1 and FO+M2 treatments had higher mycorrhizal colonization than the treatments which received no AMF inoculation at week 3 (P < 0.05), but only the FO+M2 treatment elevated plant biomass, decreased the incidence of Fusarium wilt, and improved cucumber yields to the same level as the control at week 9. The results indicated that the AMF consortium could suppress Fusarium wilt of cucumber and, therefore, showed potential as a biological control agent in greenhouse agroecosystems.     

Evaluating crude extracts of Monsonia burkeana and Moringa oleifera against Fusarium wilt of tomato

Fusarium wilt is one of the major soil-borne diseases of tomato crop globally. The study aimed to evaluate the efficacy of medicinal plants in the control of Fusarium wilt in tomato. Methanolic extracts of Monsonia burkena and Moringa oleifera were assessed in vitro and under greenhouse conditions. The in vitro experiments evaluated the effect of both extracts on Fusarium oxysporum f. sp lycopersici growth and response to varying concentrations. In greenhouse experiment, tomato seedlings cv. HTX14 were inoculated with conidial suspension of F. oxysporum and transplanted into pasteurised growth media amended with plant extract. Seedlings were treated with aqueous extracts at varying concentrations with an interval of 7 days between applications. Control treatments were treated with sterile distilled water. Both plant extracts significantly reduced pathogen growth in vitro and reduced wilt severity under greenhouse conditions. The highest mycelial growth suppression was observed in Mon. burkeana treatments. Under greenhouse conditions, both plant extracts significantly (P?≤?0.05) reduced Fusarium wilt severity and had a positive effect on plant growth parameters. A significant increase in soil-pH was also recorded in extract treated soil resulting in reduction in disease severity. The results further provide new scientific information on how their effect on soil pH can be beneficial in the control of Fusarium wilt.     

生物质炭介导生防微生物抑制辣椒疫霉的作用

生物质炭可有效防控土传病害,筛选并鉴定出生物质炭介导下的生防微生物,可为研究生物质炭防病机理和强化生物质炭防病效果提供理论依据。本研究首先进行秸秆生物质炭防控辣椒疫病盆栽试验,利用定量PCR和平板计数明确生物质炭在防控辣椒疫病时可富集的已知生防微生物,再通过选择性培养基初筛和定殖复筛筛选出生物质炭可富集的潜在生防微生物菌株,最后研究各菌株在土壤中对辣椒疫霉的抑制作用。结果表明,秸秆生物质炭使根际辣椒疫霉数量显著降低95.1%、辣椒疫病发生率显著降低91.1%,并使具有生防功能的木霉菌、青霉菌、曲霉菌、芽孢杆菌、假单胞菌和鞘氨醇单胞菌数量显著增加2.22倍、4.09倍、3.89倍、2.45倍、1.45倍和1.30倍。通过平板初筛得到可能被生物质炭富集的22株潜在生防菌株。定殖复筛剔除部分假性生物质炭介导菌株,获得可明确被生物质炭富集的2株木霉菌、3株青霉菌、2株曲霉菌、3株芽孢杆菌、3株假单胞菌、3株链霉菌和2株鞘氨醇单胞菌。木霉菌（TR1和TR3）、青霉菌（PE1）、曲霉菌（AS1和AS2）、芽孢杆菌（BA1、BA2和BA3）、假单胞菌（PS1和PS3）、链霉菌（ST1、ST4和ST5）13个菌株可显著削减土壤辣椒疫霉数量。其中,所有木霉菌和曲霉菌菌株（TR1、TR3、AS1和AS2）及芽孢杆菌（BA1和BA2）、假单胞菌（PS1和PS3）和链霉菌（ST1）9个菌株与生物质炭具有显著的协同抑制辣椒疫霉效果。因此,防控辣椒疫病时,木霉菌、曲霉菌、芽孢杆菌、假单胞菌和链霉菌是生物质炭介导下的主要防病微生物。     

间作小麦和接种AM真菌协同提高蚕豆抗枯萎病能力和根际微生物碳代谢活性

【目的】接种丛枝菌根 (arbuscular mycorrhizal,AM) 真菌和间作均是防治蚕豆枯萎病的有效方法,从土壤微生物学角度研究两者协同减轻蚕豆枯萎病的机理,对控制蚕豆枯萎病传播具有重要意义。【方法】利用盆栽试验方法,进行了间作和接种AM真菌摩西管柄囊霉 (Funneliformis mosseae,Fm) 和扭形球囊霉 (Glomus tortuosum,Gt) 试验。设蚕豆单作对照 (MF)、蚕豆小麦间作 (IF)、蚕豆单作接种Fm (MFFm)、蚕豆小麦间作接种Fm (IFFm)、蚕豆单作接种Gt (MFGt)、蚕豆小麦间作接种Gt (IFGt) 6个处理。于蚕豆开花期 (生长70天) 取土壤样品,测定蚕豆幼苗生长、枯萎病发生、根际镰刀菌数量和微生物碳代谢活性。【结果】间作显著增加蚕豆幼苗干重93.0%、降低蚕豆枯萎病病情指数71.4%,接菌显著增加蚕豆幼苗干重55.3%、降低病情指数76.6%,其中接种Fm真菌对蚕豆幼苗干重的影响更大,对病情指数的抑制效果更好。间作接菌显著增加蚕豆幼苗干重100%、降低病情指数89.8%。Biolog微平板测试结果显示,间作提高根际微生物碳代谢活性32.3%;接菌提高微生物活性85.4%;间作接菌提高微生物活性122%。主成分分析结果表明,间作和接菌均明显改变了根际微生物的群落结构,并主要改变了对碳水化合物类、氨基酸和羧酸类碳源的利用。相关性分析结果显示,枯萎病发病率和病情指数与根际镰刀菌数量呈极显著正相关关系,与AWCD值、Shannon多样性指数和丰富度指数均呈极显著负相关。【结论】蚕豆与小麦间作和接菌对抑制蚕豆枯萎病和促进蚕豆生长均具有积极效应,间作显著提高了AM真菌的定殖率,二者协同提高了根际微生物活性,改变了微生物群落结构,并抑制了病原菌增殖,进而控制蚕豆枯萎病发生。     

Effects of Arbuscular Mycorrhizal Colonization on Microbial Community in Rhizosphere Soil and Fusarium Wilt Disease in Tomato

Fusarium wilt is caused by soil-borne pathogen Fusarium oxysporum. Tomato (Lycopersicon esculentum Mill.) is susceptible to Fusarium oxysporum f. sp. lycopersici race 1 and was infected with wilt disease. A pot experiment was conducted to investigate effects of inoculating arbuscular mycorrhizal (AM) fungus (Glomus etunicatium) on the microbial community in the rhizosphere soil and Fusarium wilt in tomato (cv. Oogatafukuju). The results indicated that AM fungal inoculation suppressed the Fusarium number in the rhizosphere soil of tomato and decreased the Fusarium wilt disease index. Compared to the control, AM fungal inoculation increased the actinomycete number but increased bacterial number. Bacterial and fungal numbers were high but actinomycetes number was low when tomato basal stems became discolored brown. Fusarium inoculation significantly suppressed development of AM colonization and decreased polyphenol oxidase (PPO) activity in leaves and roots of tomato. Inoculation with AM fungi and Fusarium maintained high PPO activity in leaves and roots. The AM colonization increased root growth of tomato, whereas Fusarium inoculation had no significant effect on tomato growth. These findings suggest that because AM fungal inoculation changes microbial communities and enhances PPO activity, it should suppress occurrence of Fusarium wilt in tomato.     

Antagonistic effect of Pseudomonas spp. on pathogenic fungi and enhancement of growth of green gram (Vigna radiata)

species were isolated from the rhizosphere of green gram [Vigna radiata (L.) Wilczek] and some of the rhizobacterial isolates were found to have a wide range of antifungal activity inhibiting growth of the phytopathogenic fungi Aspergillus sp., Curvularia sp., Fusarium oxysporum and Rhizoctonia solani in culture. These isolates also showed slight inhibition of the growth of a Bradyrhizobium strain (Vigna) in a spot test which was mainly a result of nutrient competition as culture supernatants of the Pseudomonas isolates did not inhibit the growth of bradyrhizobia but inhibited the growth of fungi. The rhizobacterial isolates produced siderophores in Fe-deficient succinate medium. However, the inhibition of fungal growth by different Pseudomonas isolates in Luria Bertani and King's medium B which were not limiting in Fe³⁺ ions suggested that, besides siderophores, other antifungal compounds (antibiotics) produced by these rhizobacteria were involved in antagonism. On coinoculation of green gram with Pseudomonas strains MRS13 and MRS16 and Bradyrhizobium sp. (Vigna) strain S24, there was a significant increase in nodule weight, plant dry weight and total plant N as compared to inoculation with Bradyrhizobium strain S24 alone, suggesting that the nodule-promoting effects of Pseudomonas sp. lead to an increase in symbiotic N fixation and plant growth. Received: 27 October 1997     

不同盐分条件下植物促生菌对燕麦和黑麦草幼苗的促生效应

从盐生植物根际土中分离得到4株含1-氨基环丙烷-1-羧酸(ACC)脱氨酶的植物促生菌(PGPR),通过无菌育种袋栽培试验,考查其在不同盐分条件下对燕麦和黑麦草幼苗的促生效应。结果表明,4株菌对5 g/kg或10 g/kg NaCl盐分胁迫下的燕麦和黑麦草幼苗均表现出显著地缓解促生效应,其中假单胞菌属S1最显著,10 g/kg NaCl比无NaCl时促生作用更大。4株PGPR的ACC脱氨酶活性与植物生长参数(根长和下胚轴长)之间具有极显著的正相关性(Pearson相关系数>0.81)。     

Biocontrol efficiency of Fusarium wilt diseases by a root-colonizing fungus Penicillium sp.

Soil-inhabiting fungal pathogen Fusarium oxysporum often causes severe yield losses in many crops. We investigated the effect of a plant growth-promoting fungus, Penicillium sp. EU0013 on Fusarium wilt disease. In dual culture experiments, EU0013 inhibited the growth of Fusarium wilt pathogens by producing an inhibition zone. In experiments using sterile potting medium under controlled conditions, EU0013 significantly reduced the severity of Fusarium wilt on tomato (Solanum lycopersicum L.) and cabbage (Brassica oleracea L. var. capitata). In non-sterile soil, benomyl-resistant mutants of EU0013 were selected by exposing the conidial solution of EU0013 to ultraviolet light. The selected mutant EU0013_90S isolate did not show any distinct differences from EU0013 in colony characteristics, growth rate or antifungal activity against Fusarium wilt pathogens in dual culture. The effect of EU0013_90S on tomato wilt was studied under greenhouse conditions using non-sterile soil. Two-weeks old tomato seedlings were dipped in four different concentrations of EU0013_90S conidial suspension (1?×?10³, 1?×?10⁴, 1?×?10⁵, and 1?×?10⁶ conidia mL^–1). Seedlings were then planted in soil inoculated with either F. oxysporum f. sp. lycopersici race 1 CU1 or race 2 JCM 12575 (1?×?10⁶ bud-cells g^–1). We found the greatest disease suppression occurred when seedlings were dipped in the highest concentration of EU0013_90S conidia. This same inoculum concentration of EU0013_90S also resulted in the highest disease reduction in soil infested with JCM 12575. Higher root colonization with EU0013_90S showed a significant reduction in Fusarium wilt disease, suggesting that colonization by Penicillium sp. EU0013_90S is important for efficient biocontrol of these diseases.     

Nodulation competition among Bradyrhizobium japonicum strains as influenced by rhizosphere bacteria and iron availability

Summary Bacteria isolated from the root zones of field-grown soybean plants [Glycine max (L.) Merr.] were examined in a series of glasshouse experiments for an ability to affect nodulation competition among three strains of Bradyrhizobium japonicum (USDA 31, USDA 110, and USDA 123). Inocula applied at planting contained competing strains of B. japonicum with or without one of eleven isolates of rhizosphere bacteria. Tap-root nodules were harvested 28 days after planting, and nodule occupancies were determined for the bradyrhizobia strains originally applied. Under conditions of low iron availability, five isolates (four Pseudomonas spp. plus one Serratia sp.) caused significant changes in nodule occupancy relative to the corresponding control which was not inoculated with rhizosphere bacteria. During subsequent glasshouse experiments designed to verify and further characterize these effects, three fluorescent Pseudomonas spp. consistently altered nodulation competition among certain combinations of bradyrhizobia strains when the rooting medium did not contain added iron. This alteration typically reflected enhanced nodulation by USDA 110. Two of these isolates produced similar, although less pronounced, effects when ferric hydroxide was added to the rooting medium. The results suggest that certain rhizosphere bacteria, particularly fluorescent Pseudomonas spp., can affect nodulation competition among strains of R. japonicum. An additional implication is that iron availability may be an important factor modifying interactions involving the soybean plant, B. japonicum, and associated microorganisms in the host rhizosphere.Paper No. 10648 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7601, USA     

拮抗菌强化的生物有机肥对西瓜枯萎病的防治作用

由西瓜专化型尖孢镰刀菌(Fusarium oxysporum f.sp.niveum)引起的西瓜枯萎病是导致西瓜生产毁灭性损失的土传病害,当前对该病尚无有效的防治措施.为了探索该病的生物防治效果,本研究从土壤中分离筛选西瓜枯萎病的拮抗菌,制成生物有机肥,通过温室盆栽试验检验防病效果,并对与拮抗相关的拮抗菌葡聚糖酶进行分子生物学检测.从不同土壤中分离纯化到对西瓜枯萎病菌有潜在拮抗作用的细菌172株,通过平板对峙法筛选出抑菌率在60％以上的拮抗细菌13株,从中挑选出2株抑菌率最高的菌株Cy5和CR38,分别用其与已腐熟的有机肥制成生物有机肥BIO5和BIO38.盆栽试验结果表明,BIO5在防病和促进西瓜生长方面表现优于BIO38.与对照相比,BIO5和BIO38对西瓜枯萎病的相对防治率分别为75％和25％.BIO5处理植株的株高、地上部鲜重、地上部干重分别比对照增加64.8％、63.0％和50％.施用生物有机肥还能显著改变根际土壤的微生物组成.BIO5处理根际土壤的细菌和枯草芽孢杆菌数量分别比对照增加48.5％和61.1％,真菌和尖孢镰刀菌的数量比对照分别下降52.1％和70.2％.分子生物学分析表明,菌株Cy5属于Paenibacillus jamilae菌株,并含有类似于P.polymyxa的β-1,3-1,4-葡聚糖酶.本研究结果说明,拮抗菌强化的生物有机肥对西瓜枯萎病有防治潜力.     

Reduction in dose of chemical fertilizers and growth enhancement of sesame (Sesamum indicum L.) with application of rhizospheric competent Pseudomonas aeruginosa LES4

Pseudomonas aeruginosa LES4, an isolate of tomato rhizosphere was found to be positive for several plant growth-promoting attributes like production of indole acetic acid, HCN and siderophore, solubilization of inorganic phosphate along with urease, chitinase and β-1-3-glucanase activity. In addition, it showed strong antagonistic effect against Macrophomina phaseolina and Fusarium oxysporum. P. aeruginosa LES4 caused halo cell formation and other morphological deformities in mycelia of M. phaseolina and F. oxysporum. Root colonization was studied with Tn5 induced streptomycin resistant transconjugants of spontaneous tetracycline-resistant LES4 (designated LES4^tetra+strep+) after different durations. The strain was significantly rhizospheric competent, as 17.4% increase in its population was recorded in sesame rhizosphere. Seed bacterization with LES4 resulted in significant increase in vegetative growth parameters and yield of sesame over non-bacterized seeds. However, application of LES4 with half dose of fertilizers resulted in growth equivalent to full dose treatment, without compromising with the growth and yield of sesame. Moreover, the oil yield increased by 33.3%, while protein yield increased by 47.5% with treatment of half dose of fertilizer along with LES 4 bacterized seeds, as compared to full dose of fertilizers.     

Diversity and antagonistic potential of Pseudomonas spp. associated to the rhizosphere of maize grown in a subtropical organic farm

Pseudomonas spp. are one of the most important bacteria inhabiting the rhizosphere of diverse crop plants and have been frequently reported as biological control agents (BCAs). In this work, the diversity and antagonistic potential of Pseudomonas spp. in the rhizosphere of maize cultivars Nitroflint and Nitrodent grown at an organic farm in Brazil was studied by means of culture-dependent and -independent methods, respectively. Sampling of rhizosphere soil took place at three different stages of plant development: 20, 40 and 106 days after sowing. A PCR-DGGE strategy was used to generate specific Pseudomonas spp. fingerprints of 16S rRNA genes amplified from total community rhizosphere DNA. Shifts in the relative abundance of dominant populations (i.e. PCR-DGGE ribotypes) along plant development were detected. A few PCR-DGGE ribotypes were shown to display cultivar-dependent relative abundance. No significant differences in diversity measures of DGGE fingerprints were observed for different maize cultivars and sampling times. The characterisation and assessment of the antagonistic potential of a group of 142 fluorescent Pseudomonas isolated from the rhizosphere of both maize cultivars were carried out. Isolates were phenotypically and genotypically characterised and screened for in vitro antagonism towards three phytopathogenic fungi and the phytopathogenic bacterium Ralstonia solanacearum. Anti-fungal activity was displayed by 13 fluorescent isolates while 40 isolates were antagonistic towards R. solanacearum. High genotypic and phenotypic diversity was estimated for antagonistic fluorescent Pseudomonas spp. PCR-DGGE ribotypes displayed by antagonists matched dominant ribotypes of Pseudomonas DGGE fingerprints, suggesting that antagonists may belong to major Pseudomonas populations in the maize rhizosphere. Antagonists differing in their genotypic and phenotypic characteristics shared the same DGGE electrophoretic mobility, indicating that an enormous genotypic and functional diversity might be hidden behind one single DGGE band. Cloning and sequencing was performed for a DGGE double-band which had no corresponding PCR-DGGE ribotypes among the antagonists. Sequences derived from this band were affiliated to Pseudomonas stutzeri and P. alcaligenes 16S rRNA gene sequences. As used in this study, the combination of culture-dependent and -independent methods has proven to be a powerful tool to relate functional and structural diversity of Pseudomonas spp. in the rhizosphere.     

温室及田间条件下解淀粉芽孢杆菌菌株W19对香蕉生长、产量的促进和对香蕉镰刀菌萎蔫病的抑制研究

Plant growth-promoting rhizobacteria (PGPR) are considered to be the most promising agents for cash crop production via increasing crop yields and decreasing disease occurrence. The Bacillus amyloliquefaciens strain W19 can produce secondary metabolites (iturin and bacillomycin D) effectively against Fusarium oxysporum f. sp. cubense (FOC). In this study, the ability of a bio-organic fertilizer (BIO) containing W19 strain to promote plant growth and suppress the Fusarium wilt of banana was evaluated in both pot and field experiments. The results showed that application of BIO significantly promoted the growth and fruit yield of banana while suppressing the banana Fusarium wilt disease. To further determine the beneficial mechanisms of the strain, the colonization of green fluorescent protein-tagged strain W19 on banana roots was observed using confocal laser scanning microscopy and scanning electron microscopy. The effect of banana root exudates on the formation of biofilm of strain W19 indicated that the banana root exudates may enhance colonization. In addition, the strain W19 was able to produce indole-3-acetic acid (IAA), a plant growth-promoting hormone. The results of these experiments revealed that the application of strain W19-enriched BIO improved the banana root colonization of strain W19 and growth of banana and suppressed the Fusarium wilt. The PGPR strain W19 can be a useful biocontrol agent for the production of banana under field conditions.     

Effect of chloropicrin fumigation on microbial communities evaluated by community-level physiological profile and the resistance against fusarium wilt

Different microbial communities characterized by the Biolog pattern were developed in the rhizosphere of radish grown on a rockwool hydroponic system treated with chloropicrinfumigated and non-fumigated soil suspensions although no differences were observed in their viable counts. Different microbial communities also were developed in the rhizosphere and non-rhizosphere. After the development of microbial communities in the rhizosphere, bud cells of Fusarium oxysporum that causes vascular wilt of radish plants were inoculated, and disease symptoms were examined. Treatment with the non-fumigated soil suspension was much more effective than that with the fumigated one in controlling the disease, indicating that the Biolog method might be applicable to characterize microbial communities that control the disease caused by F. oxysporum.     

Response of alfalfa (Medicago sativa L.) to single and mixed inoculation with phosphate-solubilizing bacteria and Sinorhizobium meliloti

The objectives of this work were to phenotypically and genetically characterize alfalfa rhizosphere bacteria and to evaluate the effect of single or mixed inoculation upon nodulation and biological nitrogen fixation. Thirty-two strains showed tricalcium phosphate solubilization ability, and two of them caused bigger or equal solubilization halos than the control strain P. putida SP22. The comparison of the 16S ribosomal DNA sequences indicated that these strains are phylogenetically related to Bacillus spp. and Pseudomonas spp. A beneficial effect of both isolates on alfalfa growth was observed in coinoculation assays. Pseudomonas sp. FM7d caused a significant increase in root and shoot dry weight, length, and surface area of roots, number, and symbiotic properties of alfalfa plants. The plants coinoculated with Sinorhizobium meliloti B399 and the Bacillus sp. M7c showed significant increases in the measured parameters. Our results indicating that strains Pseudomonas sp. FM7d and Bacillus sp. M7c can be considered for the formulation of new inoculants.     

Trichoderma harzianum T-E5 significantly affects cucumber root exudates and fungal community in the cucumber rhizosphere

The Fusarium wilt in cucumbers, caused by the pathogenic fungus Fusarium oxysporum f. sp. cucumerinum, is a serious and destructive disease worldwide. An effort was made to explore the role of Trichoderma harzianum T-E5 in reducing the incidence of Fusarium wilt. Three treatments (Control, T1, and T2) were established in the greenhouse experiment. The effects of T-E5 on the composition of root exudates and fungal community in the cucumber rhizosphere were measured. Compared with the control, the application of a bio-organic fertilizer (BIO) enriched with T-E5 was found to decrease the incidence of Fusarium wilt notably and promote the growth of cucumber plants. Based on real-time PCR, the population of F. oxysporum in the control without T-E5 increased from 10³ to 10⁴ ITS copies g⁻¹ soil, whereas the population decreased from 10³ to 10² ITS copies g⁻¹ soil in the T1 and T2 treatment groups when T-E5 was included. Significant difference in fungal community was also found among the treatment groups. HPLC analysis showed that the detected levels of phenolic compounds in control were significantly higher than the levels in the samples subjected to T1 and T2 treatments. The root exudates from the control group significantly increased the numbers of germinating spores of the pathogen compared with those from the samples treated with T1 and T2. In conclusion, the modification of root exudates and the fungal community by the application of BIO might account for the effective suppression of Fusarium wilt disease in cucumbers.