Intercropping with aerobic rice suppressed Fusarium wilt in watermelon

Watermelon is susceptible to Fusarium wilt in successively mono-cropped soil. Pot experiments were carried out to investigate the effect of intercropping with aerobic rice on Fusarium wilt in watermelon. The tested soil was classified as a loam soil, previously planted with watermelon and collected from Hexian county, Anhui province, China. The results obtained are listed as follows: (1) 66.7% of watermelon plants were infected with wilt disease and 44.4% died on 40 days after transplanting in mono-cropped soil, but plants were much less susceptible to infection when intercropped with rice; (2) the density of Fusarium oxysporum f. sp. niveum decreased by 91% in soil from the intercropped watermelon rhizosphere when compared with that from the mono-crop 40 days after transplanting; (3) densities of bacteria and actinomycetes increased, but fungal density decreased in rhizosphere soil from the intercrops in comparison with the mono-crop control; (4) compared to the control, the germinated Fusarium spores were decreased by 41.0% in the treatment with addition of 1.5 ml rice root exudates. Adding 20 ml of root exudates decreased Fusarium spore production by 76.4%; and (5) the activities of defense enzymes in the leaves and roots of watermelons in the intercropped system were significantly lower than those in the mono-cropped system. It is suggested that intercropping with aerobic rice alleviated Fusarium wilt in watermelon, by restraining the spore production of Fusarium and by changing the microbial communities in rhizosphere soil through the production of rice root exudates.     

番茄枯萎病对植株维管束危害及抗氧化系统影响的研究

为了在早期诊断和确定番茄枯萎病的发生, 本文采用溶液培养方法研究了番茄(Lycopersicon esculentum Miller)幼苗剪根接种不同浓度枯萎病菌后染病植株维管束受害程度和抗氧化系统的响应。试验设4个病原菌梯度处理, B1(10⁴ cfu·mL^-1)、B2(10⁶ cfu·mL^-1)、B3(10⁷ cfu·mL^-1)、B4(10⁸ cfu·mL^-1), 以不接病原菌为对照; 分别在接种病原菌后4 d、8 d、12 d、16 d、20 d测定维管束褐变情况和抗氧化系统的变化。结果表明, 在水培条件下, 接种病原菌16 d植株维管束出现褐变, 其受害程度随病原菌接种浓度提高而增大; 维管束中病原菌只在B4处理中有检出。番茄叶片中丙二醛(MDA)含量随接种时间呈先降后升趋势, 12 d开始逐渐上升, 20 d达到最高, 各接菌处理均显著高于对照, 且B4处理显著高于其他处理; 过氧化物酶(POD)活性先缓慢下降, 12 d后回升, B4则急剧上升; 多酚氧化酶(PPO)活性逐渐上升, 接菌16 d时达到高峰; 随接菌浓度的提高, MDA含量、POD和PPO活性均有所增加, 尤以接菌浓度为10⁸ cfu·mL^-1时3种指标显著高于其他处理, 分别是未接菌植株的13.1倍、12.9倍和1.9倍; 而培养时间对番茄叶片中过氧化氢酶(CAT)活性的影响没有明显规律, 对照菌株CAT活性显著高于各接菌处理, 说明CAT活性对番茄枯萎病病原菌没有响应。本研究结果表明, 结合番茄茎的维管束褐变现象, 认为番茄叶片中MDA含量、POD和PPO活性可作为早期判断番茄是否感染枯萎病的重要指标。     

<Emphasis Type="Italic">Bacillus subtilis</Emphasis> SQR 9 can control <Emphasis Type="Italic">Fusarium</Emphasis> wilt in cucumber by colonizing plant roots

Fusarium wilt is one of the major constraints on cucumber production worldwide. Several strategies have been used to control the causative pathogen, Fusarium oxysporum f. sp. cucumerinum J. H. Owen, including soil solarization, fungicide seed treatment and biological control. In this study, F. oxysporum f. sp. cucumerinum was successfully controlled by a newly isolated strain, Bacillus subtilis SQR 9, in vitro and in vivo. Greenhouse experiments were carried out to evaluate the effect of inoculation and solid fermentation of organic fertilizer with B. subtilis SQR 9, hereby defined as bio-organic fertilizer (BIO), on the control of Fusarium wilt. In comparison with the control, the wilt incidence was significantly reduced (49–61% reduction) by application of BIO. The rhizosphere population of F. oxysporum f. sp. cucumerinum, as detected both by selective plating and realtime PCR, was significantly lower in BIO-treated plants than the control. The localization of bacterial cells, pattern of colonization and survival of B. subtilis SQR 9 in the rhizsosphere of cucumber, was examined by fluorescent microscopy and explored following recovery of the green fluorescent protein (gfp)-labeled SQR 9 with the new gfp-marked shuttle vector pHAPII through selective plating. The preferential sites of the labeled strain were the differentiation and elongation zone, root hair and the lateral root junctions. The population of the strain was 10⁶ cfu/g root in rhizoplane. These results indicate that the strain was able to survive well in the rhizosphere of cucumber, suppressed growth of F. oxysporum in the rhizosphere of cucumber and protected the host from the pathogen.     

Funneliformis mosseae affects the root rot pathogen Fusarium oxysporum in soybeans

We analyzed the relationship between the dominant arbuscular mycorrhizal (AM) fungus Funneliformis mosseae and the dominant soybean root rot pathogen Fusarium oxysporum through the pot trials to help overcome obstacles to continuous cropping of soybean and to provide theoretical evidence that can be used to help prevent the reduced production induced by soybean root rot. Using qRT-PCR, we amplified the specific rDNA sequences of F. mosseae and F. oxysporum in soybean roots and rhizosphere soil and quantified the DNA contents of these fungi to determine the relationship between the dominant AM fungus F. mosseae and F. oxysporum. The DNA contents of F. oxysporum differed significantly depending on the presence of F. mosseae in both soybean roots and rhizosphere soil. Specifically, the DNA contents of F. oxysporum were reduced after inoculation with F. mosseae, suggesting that F. mosseae has a negative effect on the growth of F. oxysporum.     

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 hexachlorocyclohexane on rhizosphere fungal propagules and root colonization by arbuscular mycorrhizal fungi in Plantago lanceolata

Lindane ( γ‐hexachlorocyclohexane or γ‐HCH) is an organochlorine insecticide previously used extensively for the control of agricultural pests. We studied the effects of soil HCH contamination on vegetation and its associated arbuscular mycorrhizas (AM). The polluted and unpolluted plots had similar plant cover, with the same species richness and abundance. Plantago lanceolata plants were selected for mycorrhizal analysis because of their presence in both plots and known mycotrophy. The presence of HCH appeared to have no significant effect on the extent of colonization of Plantago roots by AM, suggesting a similar functionality of the fungal symbionts. However, infective AM propagules, the density of AM spores and viable AM hyphae in the rhizosphere were much less in the HCH‐polluted soil than in the unpolluted plot. Pre‐inoculation of four plant species with an isolate of Glomus deserticola obtained from the HCH‐contaminated soil resulted in increased growth and fungal colonization of roots compared with plants pre‐inoculated with the introduced fungus G. macrocarpum or colonized by the consortium of indigenous AM fungal species, when those plants were transplanted to an HCH‐contaminated soil. This suggests that the fungus increases the tolerance of plants to the toxic soil environment. We conclude that herbaceous and woody plants can grow in soil with little P contaminated with <100 mg HCH kg^?1 with the help of tolerant AM, despite the detrimental effect of HCH on AM fungal propagules in soil. The effects of AM fungi on plant growth and soil microbial community structure in HCH‐polluted sites could be important for remediation of the pollutant through the microbial activity in the rhizosphere.     

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.     

Fusarial wilt suppression and crop improvement through two rhizobacterial strains in chick pea growing in soils infested with Fusarium oxysporum f.sp. ciceris

 Bacterization of chick pea seeds with a siderophore-producing fluorescent Pseudomonas strain RBT13 and an antibiotic-producing Bacillus subtilis strain AF1, isolated from tomato rhizoplane and pigeon pea rhizosphere repectively, increased the shoot height, root length, fresh weight, dry weight and yield in soils infected with Fusarium oxysporum f.sp. ciceris. Seed bacterization also resulted in a significant reduction in chick pea wilt caused by the same pathogen. Addition of iron to the soil completely eliminated disease suppression by RBT13 but not by AF1. Dual drug-resistant mutant strains derived from the rhizobacteria were used to monitor and confirm root colonization. The results indicate the potential for development of both strains for the biological control of chick pea wilt. Received: 29 April 1998     

Application of bio-organic fertilizer can control Fusarium wilt of cucumber plants by regulating microbial community of rhizosphere soil

Fusarium wilt, caused by Fusarium oxysporum f. sp. cucumerinum J. H. Owen, results in considerable yield losses for cucumber plants. A bio-organic fertilizer (BIO), which was a combination of manure composts with antagonistic microorganisms, and an organic fertilizer (OF) were evaluated for their efficiencies in controlling Fusarium wilt. Application of the BIO suppressed the disease incidence by 83% and reduced yield losses threefold compared with the application of OF. Analysis of microbial communities in rhizosphere soils by high-throughput pyrosequencing showed that more complex community structures were present in BIO than in OF treated soils. The dominant taxonomic phyla found in both samples were Proteobacteria, Firmicutes, Actinobacteria and Acidobacteria among bacteria and Ascomycota among fungi. Abundance of beneficial bacteria or fungi, such as Trichoderma, Hypoxylon, Tritirachium, Paenibacillus, Bacillus, Haliangium and Streptomyces, increased compared to the OF treatment, whereas the soil-borne pathogen, Fusarium, was markedly decreased. Overall, the results of this study demonstrate that the application of the BIO was a useful and effective approach to suppress Fusarium wilt and that the high-throughput 454 pyrosequencing was a suitable method for the characterization of microbial communities of rhizosphere soil of cucumber.     

Seed priming with Pseudomonas putida isolated from rhizosphere triggers innate resistance against Fusarium wilt in tomato through pathogenesis-related protein activation and phenylpropanoid pathway

Pathogenesis-related (PR) proteins are one of the major and preliminary proteins accumulated as a defense against biotic stress. This defense response can be induced by using beneficial rhizobacteria, which has been studied in various host-pathogen interactions. In the present study, eleven Pseudomonas isolates were assessed for their potential to ferment sorbitol, reduce nitrate, and produce mycolytic enzymes, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, phenazine antibiotics, and N-acyl homoserine lactones (AHLs). All isolates were tested against the host-specific pathogen Fusarium oxysporum MTCC1755 in tomato under greenhouse conditions, and shortlisted isolates were tested for their rhizosphere competence. In-vitro test results showed that the isolates were able to produce mycolytic enzymes, including protease, lipase, chitinase, cellulase, and amylase, and the antibiotic phenazine and were negative for pyoluteorin. All the isolates except two were positive for ACC deaminase production. Greenhouse results showed that the isolates M80, M96, and T109 significantly reduced symptoms of Fusarium wilt. Extended greenhouse tests under autoclaved and unautoclaved soil conditions showed that M80, M96, and T109 were excellent rhizosphere competitors and were identified as Pseudomonas putida. In brief, the defense-specific biochemical variations in the host could describe the improved defense against Fusarium wilt occurring in the primed plants. These three Pseudomonas strains could be used as potential biocontrol agents, along with their rhizosphere competence.     

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.     

Effect of arbuscular mycorrhizal fungi on rhizosphere organic acid content and microbial activity of trifoliate orange under different low P conditions

ABSTRACT

Arbuscular mycorrhizal (AM) fungi can improve plant phosphorus (P) uptake; however, information about how AM fungi affect rhizosphere organic acid and microbial activity to alleviate citrus low P stress is limited. Here, a pot experiment was conducted to evaluate the effect of AM fungi (Rhizophagus intraradices, Ri) inoculation on rhizosphere organic acid content, microbial biomass (MB) and enzyme activity of trifoliate orange (Poncirus trifoliata L. Raf.) seedlings grown under three low P conditions. The results showed that mycorrhizal seedlings all recorded higher P concentrations, plant biomass and better root morphology with more lateral and fine roots, but lower root mass ratios, irrespective of P conditions. Mycorrhizal P absorption contribution did not differ significantly among three P conditions. Mycorrhizal seedling rhizosphere soil exhibited lower organic acid content, soil organic P content and ratio of MB-carbon (C)/MB-P, but higher MB and enzyme activity. Additionally, the main organic acids showed a negative relationship with mycorrhizal colonization rate and hyphal length; however, phosphatase and phytase activity had a significantly positive relationship with MB. Therefore, the results suggest that AM fungi inoculation may help citrus to efficiently utilize organic P source by improving microbial activity under low available P conditions.     

Control of Fusarium Wilt of Cucumber Seedlings by Inoculation with an Arbuscular Mycorrhical Fungus

A glasshouse pot experiment was conducted to investigate the impact of inoculation of cucumber at the germination stage with Glomus etunicatum BEG168 on plant yield and incidence of Fusarium oxysporum f.sp. cucumerinum inoculated 28 days after the start of the experiment. Inoculation with the AM fungus decreased both disease incidence and disease index. Mycorrhizal inoculation also increased P concentrations in the cucumber seedlings. The mycorrhizal seedlings had higher concentrations of proline and polyphenol oxidase activity but lower malondialdehyde than non-mycorrhizal seedlings, indicating that AM inoculation may have protected membrane permeability and reduced the extent of the damage caused by F. oxysporum. The results indicate that the mycorrhizal fungus may influence plant secondary metabolites and increase resistance to wilt disease in cucumber seedlings and may therefore have some potential as a biological control agent.     

Pineapple–banana rotation reduced the amount of Fusarium oxysporum more than maize–banana rotation mainly through modulating fungal communities

Continuous cropping with banana results in an enrichment of Fusarium oxysporum f. sp. cubense race 4 (FOC) in soil, causing the soil-borne disease Fusarium wilt. Crop rotation has been an effective method of controlling various soil-borne diseases. However, no information is currently available concerning variations in soil microbial community structure in banana crop rotations. Thus, the influence of two-year crop rotation systems of pineapple–banana and maize–banana on the population density of FOC and soil microbial community structure was investigated to identify which rotation system is more effective in FOC suppression and differences in microbial community composition among different rotations. Bacterial and fungal communities were interrogated by pyrosequencing of the 16 S RNA gene and the internal transcribed spacer (ITS) region. The pineapple–banana rotation was more effective than maize–banana in reducing FOC abundances and suppressing Fusarium wilt disease incidence. Allelopathic effects of pineapple root exudates on FOC were not observed. Greater fungal community variations than bacterial were identified between the two rotation systems, suggesting that fungal communities may play a more important role in regulating FOC abundances. Furthermore, in the pineapple–banana rotation, Acidobacteria, Planctomycetes, Chloroflexi phyla, Gp1, Gp2 and Burkholderia bacterial genera increased while the fungal phyla Basidiomycota, (esp. Gymnopilus) increased and Sordariomycetes decreased. Such changes may be important microbial factors in the decrease in FOC.     

施用生物有机肥抑制香蕉镰刀菌萎蔫病的研究

Fusarium wilt is one of the most serious diseases of banana plants caused by soil-borne pathogen Fusarium oxysporum f.sp. cubense(FOC). In this study a pot experiment was conducted to evaluate the effects of different bio-organic fertilizers(BIOs) on Fusarium wilt of banana, including the investigations of disease incidence, chitinase and β-1,3-glucanase activities of banana plants, and FOC populations as well as soil rhizosphere microbial community. Five fertilization treatments were considered, including chemical fertilizer containing the same N, P and K concentrations as the BIO(control), and matured compost mixed with antagonists Paenibacillus polymyxa SQR-21 and Trichoderma harzianum T37(BIO1), Bacillus amyloliquefaciens N6(BIO2), Bacillus subtilis N11(BIO3), and the combination of N6 and N11(BIO4). The results indicated that the application of BIOs significantly decreased the incidence rate of Fusarium wilt by up to 80% compared with the control. BIOs also significantly promoted plant growth, and increased chitinase andβ-1,3-glucanase activities by 55%–65% and 17.3%–120.1%, respectively, in the banana roots. The population of FOC in the rhizosphere soil was decreased significantly to about 104 colony forming units g-1with treatment of BIOs. Serial dilution plating and denaturing gradient gel electrophoresis analysis revealed that the application of BIOs increased the densities of bacteria and actinomycetes but decreased the number of fungi in the rhizosphere soil. In general, the application of BIOs revealed a great potential for the control of Fusarium wilt disease of banana plants.     

Interactions between the arbuscular mycorrhizal fungus Glomus intraradices and the plant growth promoting rhizobacteria Paenibacillus polymyxa and P. macerans in the mycorrhizosphere of Cucumis sativus

Interactions between the arbuscular mycorrhizal (AM) fungus Glomus intraradices and bacteria from the genus Paenibacillus (P. macerans and P. polymyxa) were examined in a greenhouse pot experiment with Cucumis sativus with and without organic matter amendment (wheat bran). P. polymyxa markedly suppressed AM fungus root colonization irrespective of wheat bran amendment, whereas P. macerans only suppressed AM fungus root colonization in combination with wheat bran amendment. Dual inoculation with P. macerans and G. intraradices in combination with wheat bran amendment also caused severe plant growth suppression. Inoculation with G. intraradices was associated with increased levels of dehydrogenase activity and available P in the growth substrate suggesting that mycorrhiza formation accelerated the decomposition of organic matter resulting in mobilization of phosphorus. Inoculation with both Paenibacillus species increased all measured microbial fatty acid biomarkers in the cucumber rhizosphere, except for the AM fungus biomarker 16:1ω5, which was reduced, though not significantly. Similarly, inoculation with G. intraradices increased all measured microbial fatty acid biomarkers in the cucumber rhizosphere, except for the Gram-positive bacteria biomarker 15:0 anteiso, which was overall decreased by G. intraradices inoculation. In combination with wheat bran amendment G. intraradices inoculation caused a 39% reduction in the amount of 15:0 anteiso in the treatment with P. polymyxa, suggesting that G. intraradices suppressed P. polymyxa in this treatment. In conclusion, plant growth promoting species of Paenibacillus may have suppressive effects of AM fungi and plant growth, especially in combination with organic matter amendment. The use of an inert plant growth media in the present study allowed us to study rhizosphere microbial interactions in a relative simple substrate with limited interference from other soil biota. However, the results obtained in the present work mainly show potential interactions and should not be directly extrapolated to a soil situation.     

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.     

Interaction between Alternaria alternata or Fusarium equiseti and Glomus mosseae and its effects on plant growth

The effect of inoculation with the saprophytic fungi Alternaria alternata or Fusarium equiseti on maize (Zea mays) and lettuce (Lactuca sativa) with or without arbuscular mycorrhizal (AM) colonization by Glomus mosseae was studied in a greenhouse trial. Plant dry weights of non-AM-inoculated maize and lettuce were unaffected by the presence of A. alternata and F. equiseti. In contrast, A. alternata and F. equiseti decreased plant dry weights and mycorrhization when inoculated to the rhizosphere before G. mosseae. The saprophytic fungi inoculated 2 weeks after G. mosseae did not affect the percentage of root length colonized by the AM endophyte, but did affect its metabolic activity assessed as succinate dehydrogenase activity. Although F. equiseti inoculated at the same time as G. mosseae did not affect mycorrhization of maize roots, its effect on AM colonization of lettuce roots was similar to that with A. alternata. In the rhizosphere of both plants, the population of saprophytic fungi decreased significantly, but was not affected by the presence of G. mosseae. Our results suggest that there may have been a direct effect of the saprophytic fungi on the mycorrhizal fungi in the extramatrical phase of the latter, and when the AM fungus was established in the root the AM fungus was less affected by the saprophytic fungi. Received: 16 January 1996     

Interactions of arbuscular-mycorrhizal fungi and Bacillus strains and their effects on plant growth,microbial rhizosphere activity (thymidine and leucine incorporation) and fungal biomass (ergosterol and chitin)

The effects of two Bacillus strains (Bacillus pumillus and B. licheniformis) on Medicago sativa plants were determined in single or dual inoculation with three arbuscular-mycorrhizal (AM) fungi and compared to P-fertilization. Shoot and root plant biomass, values of thymidine and leucine incorporation as well as ergosterol and chitin in rhizosphere soil were evaluated to estimate metabolic activity and fungal biomass, respectively, according to inoculation treatments. For most of the plant parameters determined, the effectiveness of AM fungal species was influenced by the bacterial strain associated. Dual inoculation of Bacillus spp. and AM fungi did not always significantly increase shoot biomass compared to single AM-colonized plants. The most efficient treatment in terms of dry matter production was the dual Glomus deserticola plus B. pumillus inoculation, which produced similar shoot biomass and longer roots than P-fertilization and a 715% (shoot) and 190% (root length) increase over uninoculated control. The mycorrhizas were more important for N use-efficiency than for P use-efficiency, which suggests a direct mycorrhizal effect on N nutrition not mediated by P uptake. Both chemical and biological treatments affected thymidine and leucine incorporation in the rhizosphere soil differently. Thymidine was greater in inoculated than in control rhizospheres and B. licheniformis was more effective than B. pumillus in increasing thymidine. Non-inoculated rhizospheres showed the lowest thymidine and leucine values, which shows that indigenous rhizosphere bacteria increased with introduced inocula. The highest thymidine and leucine values found in P-fertilized soils indicate that AM plants are better adapted to compete with saprophytic soil bacteria for nutrients than P-amended plants. Chitin was only increased by coinoculation of B. licheniformis and G. intraradices. B. pumillus increased ergosterol (indicative of active saprophyte fungal populations) in the rhizosphere of AM plants and particularly when colonized by G. mosseae. The different AM fungi have different effects on bacterial and/or fungal saprophytic populations and for each AM fungus, this effect was specifically stimulated or reduced by the same bacterium. This is an indication of ecological compatibilities between microorganisms. Particular Glomus–bacterium interactions (in terms of effect on plant growth responses or rhizosphere population) do not seem to be related to the percentage of AM colonization. The effect on plant growth and stimulation of rhizosphere populations, as a consequence of selected microbial groups, may be decisive for the plant establishment under limiting soil conditions.     

Reducing chemical fertilizer use mitigates obstacles in intensive monocropping of cucumber: a probable role of Pseudomonads in the process

A five-year continuous monocropping of cucumber in a greenhouse receiving 50% NPK and 100% NPK was conducted to explore if reducing chemical fertilizer use can mitigate obstacles in intensive monocropping, and the microbial mechanism of the process. Significant decreases in living seedling rates and total yields with time were exhibited in the 100% NPK treatments, showing an obvious obstacles event, which can be mitigated by reducing fertilizer use. It was found that both total yields and living seedling rates were negatively correlated with the proportion of Fusarium oxysporum f. sp. cucumerinum (a known cucumber wilt pathogen) in F. oxysporum, suggesting the two fungal groups out of balance in the rhizosphere, was the key factor for the decline in cucumber growth in the monocropping system. Furthermore, the abundance of pseudomonas group, not hydrogen cyanide- or phenazine-producing pseudomonas, can act as the defender against possible fungal pathogens. The opposite relationships between HCN-producing pseudomonas and two plant indicators (cucumber yield and living seedling rate) suggested that HCN-producing pseudomonas exerted positive effects only when cucumber plants’ density was above a certain threshold. The Redundancy Analysis (RDA) showed that changes in soil microbial population size, especially in pseudomonas groups, contributed to the mitigation process.