Biocontrol of Late Blight Disease (Phytophthora capsici) of Pepper and the Plant Growth Promotion by Paenibacillus ehimensis KWN38

For field application of a bacterial strain used to control Phythophthora capsici, we will need a biologically and economically efficient carrier medium. The known antagonist Paenibacillus ehimensisKWN38 was grown in a grass medium where it showed high antifungal and lytic enzyme activities. To demonstrate the potential of P. ehimensisKWN38 for biocontrol of late blight disease in pepper, pot trials were conducted by treating the 1‐month‐old plants with water (W), a selected grass medium (G3), G plus P. ehimensisKWN38 inoculation (G3P) or synthetic fungicide (F). The shoot dry weight in G3P was higher than that in W and F treatments at 15 days after zoospore infection (DZI). The root dry weight in G3P was also higher than that in W. The root mortality of G3 and W increased over 58 and 80% at 15 DZI, and some plants in those treatments wilted due to the failure of root physiology. The plants in G3P and F survived well because of their better root health conditions. Soil cellulase activity of G3P was consistently higher than that of W and F at earlier observation times (0, 2 and 6 DZI). The root β‐1,3‐glucanase activity of G3P promptly increased to maximum shortly after zoospore infection and reached the maximum value of 51.12 unit g^?1 of fresh weight at 2 DZI. All these results indicate that inoculation of P. ehimensisKWN38 to the root zone of potted pepper plants increases plant growth, root and soil enzyme activities and alleviates the root death caused by infection with P. capsici zoospores.     

Bio-control activity of Purpureocillium lilacinum strains in managing root-knot disease of tomato caused by Meloidogyne incognita

The potential of 24 indigenous isolates of Purpureocillium lilacinum (Paecilomyces lilacinus) (Thom) Samson collected from different agro-climatic zones of India was investigated against the root-knot nematode, Meloidogyne incognita. The studies were conducted in vitro (larvicidal, ovicidal and egg-parasitising capacity) and under naturally infested field conditions with selected strains. Repeated field trials were conducted with talc-based preparations of fungal strains at 10 kg ha^?1, which were applied mixed in farm yard manure (FYM) at 1.5 t ha^?1. Results (in vitro) showed that all tested isolates were capable to parasitise eggs, inhibit egg hatching and cause juvenile mortality of M. incognita at various levels. Based on the performance under in vitro studies, eight isolates (NDPL-01, ANDPL-02, SHGPL-03, HYBPL-04, AHDPL-05, PTNPL-06, SNGPL-07 and VNSPL-08) were re-tested to confirm the results. HYBDPL-04 was found causing highest mortality (80%), inhibition of egg hatching (90%) as well as parasitisation of M. incognita eggs (75%). Under field trials also, the best protection of root-knot disease of tomato (Lycopersicon esculentum L.), in terms of reduction of galls (61%) and reproductive factor (P_f/P_i (RF) = 0.2) was achieved through application of HYBDPL-04 + FYM compared to control and other tested isolates. It also enhanced marketable yield of tomato up to 43%. It is concluded that the HYBDPL-04 strain of P. lilacinum is highly effective for management of root-knot disease of tomato under naturally infested field conditions. It is the isolate which produced the maximum number of metabolites which were extracted through high pressure liquid chromatography.     

Role of alternative pathway respiration in tomato roots attacked by Meloidogyne incognita

The effect of nematode infestation on the alternative pathway respiration of mitochondria isolated from resistant and susceptible tomato roots greatly depended on the oxidisable substrate tested. The percentage of alternative respiration in NADH, malate and succinate oxidation was markedly different between the resistant (Rossol) and the susceptible (Roma VF) cultivars before infestation. Only the percentage of malate alternative oxidation in mitochondria from the resistant roots was influenced by nematode invasion. Conversely, attacked roots showed consistent variations in the content of mitochondria per unit fresh weight and in the phosphorylation efficiency (ADP/O) of the organelles. Expression of the alternative pathway (ρ' value) was found to be unchanged in intact roots and isolated mitochondria six days after nematode inoculation.     

The potential of Beauveria bassiana and Metarhizium anisopliae in controlling the root-knot nematode Meloidogyne incognita in tomato and cucumber

Meloidogyne incognita is one of the most important plant parasitic nematodes. This study was conducted to determine the nematicidal potential of Beauveria bassiana and Metarhizium anisopliae against M. incognita. B. bassiana and M. anisopliae spore suspensions and bio -nematicide, Purpureocillium lilacinum were applied. B. bassiana and M. anisopliae revealed considerable nematicidal activity against M. incognita in tomato and cucumber. The gall index decreased gradually from 8.0 for control to 3.2, 2.0 and 2.2 for B. bassiana, M. anisopliae and P. lilacinum in tomato, respectively. The highest reduction (%) in gall formation (control index) was calculated as 75.2 % in M. anisopliae treated group for tomato. The gall index was 7.6 for control, but decreased to 3.6, 2.0 and 2.2 for B. bassiana, M. anisopliae and P. lilacinum in cucumber, respectively. The highest control index was 71.7 % for M. anisopliae in cucumber. The number of the second instar juveniles of M. incognita was recorded as 2240 for control. However, this value reduced to 508, 332 and 328 by B. bassiana, M. anisopliae and P. lilacinum in tomato, respectively. The highest control indexes for the second instar juveniles were 85.2 % and 85.3 % for M. anisopliae and P. lilacinum in tomato, respectively. Alike, the highest control indexes were 84.9 % and 85.7 % for the same applications in cucumber, respectively. B. bassiana and M. anisopliae displayed also positive effect on the number of leaves, dry and fresh root weights of tomato. The results showed that B. bassiana and M. anisopliae can be considered as an alternative.     

Use of some emulsified plant seed oils as a safe alternative for the management of Meloidogyne incognita infecting tomato

Abstract

The aim of the present study was to formulate six different plant seed oils namely canola, cotton, flax, olive, sesame and soybean as emulsifiable concentrates. The composition of the formulation comprises at least one organic solvent, one surfactant and one plant oil. Physico-chemical properties of the formulated oils (emulsion stability test, cold stability and heat stability tests) were measured. The successfully emulsified oils were evaluated for nematicidal activity against Meloidogyne incognita infecting tomato plants under greenhouse conditions. Emulsified canola oil proved to be the most effective oil as a protectant against M. incognita infection to tomatoes followed by soybean, cotton, flax and sesame oil. In addition, employing a high rate of the tested emulsified oils gave higher activity in suppressing nematodes both in the soil and in tomato roots than using a low rate. Moreover, all tested formulated oils at both rates of application had no adverse effect on the growth of tomato plants except sesame oil which significantly decreased the shoot length when compared to the control. The prepared plant oils might be used as potential sources for sustainable eco-friendly botanical nematicides to protect plants from nematode attack.     

Efficiency of Trichoderma harzianum as a biocontrol agent in combination with humate and chitosan against Meloidogyne incognita on tomato

Abstract

A pot trial was conducted to estimate the role of Trichoderma harzianum alone or in combination with two organic substances, potassium humate and chitosan in controlling Meloidogyne incognita on tomato. All treatments caused greater decreases in parameters of M. incognita in comparison to the control treatment (nematode only) and this led to noticeable enhancements in growth and yield of tomato. The lowest numbers of eggmasses, eggs/eggmass, galls/root, females/root, and second stage juveniles/250?g soil were recorded due to the combination of T. harzianum (10¹⁰ spore/ml) with chitosan and potassium humate after 120 days from the transplanting of tomato seedlings. Also, this treatment showed the best promotion for all tomato parameters (lengths and weights of shoots and roots, and productivity). So, mixing chitosan, potassium and T. harzianum is highly recommended to be used as an effective bio-nematicide against M. incognita on tomato plants.     

丛枝菌根真菌对不同番茄品种抗根结线虫病的影响

【背景】根结线虫病害严重制约我国设施蔬菜的生产。丛枝菌根真菌(Arbuscular Mycorrhizal Fungi,AMF)作为土壤中最重要的有益真菌之一,可以促进植物生长,提高植物抗病性,减轻土传真菌和线虫病害的发生。在蔬菜保护地栽培中,AMF对于植物线虫病防治作用的研究受到了广泛关注。【目的】针对番茄生产中危害最严重的南方根结线虫(Meliodogyne incognita)病害问题,研究AMF和番茄品种不同组合的抗线虫效应,以期为菌根真菌作为生物防治剂和生物菌肥应用于实际生产提供技术基础。【方法】在灭菌土壤中,人工接种根结线虫,比较不同菌种Rhizophagus intraradices(Ri)、Acaulosporamellea(Am)及菌种组合Rhizophagusintraradices+Acaulosporamellea(Ri+Am)在不同番茄品种(感病品种蒙特卡罗和抗线虫品种仙客1号)上对南方根结线虫侵染和繁殖的影响,研究AMF对根结线虫的拮抗效应。另外,采用南方根结线虫连作发病的土壤,在感病品种蒙特卡洛上接种AMF混合菌种Ri+Am,番茄苗移栽入连作土壤中,测定各生长指标和调查根结和卵块数量,评价接种AMF处理对根结线虫病的防治效果。【结果】在灭菌土壤中,普通番茄品种蒙特卡罗的菌根效应显著优于抗线虫番茄品种仙客1号,表现为前者单位根重的根结和卵块的数量均比对照显著降低,而后者仅降低了卵块数量;蒙特卡罗上接种Ri+Am混合菌种的效果优于接种单一菌种Am和Ri;而仙客1号上接种Ri的效果更好。接种线虫也显著影响了AMF的侵染,而且对抗性品种仙客1号的影响更为明显。但除了接种Am的处理,大多数处理收获时菌根侵染率仍维持较高的水平(70%以上)。在连作土壤中,感病品种蒙特卡罗接种混合菌种Ri+Am具有较好的抗/耐线虫效应,主要表现为促进植株生长,显著降低根结和卵块数量,但菌根侵染率较灭菌土壤低,约为40%。【结论】综合以上结果,表明菌根化苗能够在一定程度上减轻根结线虫病的危害。土壤灭菌条件下,在感病和抗线虫番茄品种上接种AMF能够减轻线虫的侵染和繁殖,而且在感病品种上的防治效果更加显著。在连作土壤中,在番茄感病品种上接种AMF也表现较好的抗线虫效果。     

Feasibility of growing Moringa oleifera as a mix-crop along with tomato for control of Meloidogyne incognita and Rotylenchulus reniformis in Egypt

Impact of nematicidal potential of drumstick, Moringa oleifera Lam. (Moringaceae), as a medicinal plant (1–4 plants per pot) is obtained when it is used as a mix-crop along with tomato cv. Super Strain B against Meloidogyne incognita and Rotylenchulus reniformis was evaluated under a greenhouse conditions (30?±?5?°C) at the National Research Centre, Egypt. The nematodes final population and their rate of build-up as well as the root gall index were significantly affected by the number of moringa plants when grown with tomato together. There was a negative correlation between the number of moringa seedlings and the final population of both nematodes. The lowest nematode final population and rate of build-up were determined at the highest number of moringa plants (four plants per pot). The highest number of root gall index (4.1) was found on roots of tomato grown alone, while, the lowest one (0.6) was found on roots of tomato grown with four plants of moringa. This type of control is considered easy, inexpensive and pollution-free.     

模拟氮沉降增加条件下土壤团聚体对酶活性的影响

氮沉降增加改变了森林土壤生态系统物质输入,影响土壤生物及酶活性,而土壤团聚体内相对稳定的微域生境可能减弱或延缓土壤生物和酶对氮沉降增加的响应强度。以广东省东莞大岭山森林公园荷木人工林为研究对象,用模拟N沉降方法,分析了2011年12月到2012年11月一年内氮沉降增加条件下表层混合土壤和土壤团聚体内脲酶、蔗糖酶和酸性磷酸酶活性的变化及影响因素,旨在理解氮沉降增加条件下土壤团聚体对酶活性的影响。结果表明:氮沉降增加对表层混合土壤中脲酶和蔗糖酶的抑制作用不显著,而酸性磷酸酶受氮沉降显著影响,表现为低氮(50 kg N hm-2a-1)促进,高氮(300 kg N hm-2a-1)抑制的规律。表层土壤团聚体内脲酶活性随氮沉降增加而降低,N300处理显著低于对照;蔗糖酶和酸性磷酸酶活性随氮沉降增加先降低后增加,N100处理最低,分别比其他处理降低了6.46%—25.53%和42.33%—68.25%。试验区内各粒径土壤团聚体内酶活性高于混合土壤,且随团聚体粒径增加酶活性均为先增加后降低。不同粒径土壤团聚体的3种酶活性均以2—5 mm最高,但脲酶、酸性磷酸酶在各团聚体粒径间差异不显著,蔗糖酶活性2—5 mm显著高于5—8 mm。土壤酶相对活性指数和相对活性综合指数结果显示,超过85%的团聚体粒径内的相对酶活性指数大于1,而土壤酶相对活性综合指数均大于1。以上结果表明,氮沉降增加条件下土壤团聚体对其团聚体内的土壤酶活性有隔离保护作用,但其隔离保护效果与酶的种类和土壤团聚体粒径有关。     

Management of Meloidogyne incognita with Chemicals and Cultivars in Cotton in a Semi-Arid Environment

Management of Meloidogyne incognita (root-knot nematode) in cotton in the United States was substantially affected by the decision to stop production of aldicarb by its principle manufacturer in 2011. The remaining commercially available tools to manage M. incognita included soil fumigation, nematicide seed treatments, postemergence nematicide application, and cultivars partially resistant to M. incognita. Small plot field studies were conducted on a total of nine sites from 2011–2013 to examine the effects of each of these tools alone or in combinations, on early season galling, late-season nematode density in soil, yield, and value ($/ha = lint value minus chemical costs/ha). The use of a partially resistant cultivar resulted in fewer galls/root system at 35 d after planting in eight of nine tests, lower root-knot nematode density late in the growing season for all test sites, higher lint yield in eight of nine sites, and higher value/ha in six of nine sites. Galls per root were reduced by aldicarb in three of nine sites and by 1,3-dichloropropene (1,3-D) in two of eight sites, relative to the nontreated control (no insecticide or nematicide treatment). Soil fumigation reduced M. incognita density late in the season in three of nine sites. Value/ha was not affected by chemical treatment in four of nine sites, but there was a cultivar × chemical interaction in four of nine sites. When value/ha was affected by chemical treatment, the nontreated control had a similar value to the treatment with the highest value/ha in seven of eight cultivar-site combinations. The next “best” value/ha were associated with seed treatment insecticide (STI) + oxamyl and aldicarb (similar value to the highest value/ha in six of eight cultivar-site combinations). The lowest valued treatment was STI + 1,3-D. In a semi-arid region, where rainfall was low during the spring for all three years, cultivars with partial resistance to M. incognita was the most profitable method of managing root-knot nematode in cotton.     

金龟子绿僵菌对斜纹夜蛾幼虫的生防效果、抗氧化酶活性和肠道细菌群落的影响

【目的】测定金龟子绿僵菌(Metarhizium anisopliae)对斜纹夜蛾(Spodoptera litura) 2龄幼虫的毒力,研究金龟子绿僵菌侵染后寄主体内抗氧化酶活性和肠道内细菌群落的变化,探讨斜纹夜蛾对金龟子绿僵菌侵染的防御机制。【方法】采用浸渍法测定不同浓度金龟子绿僵菌对斜纹夜蛾2龄幼虫的毒力;应用IlluminaMiSeq高通量测序技术测定肠道细菌群落。【结果】不同浓度的孢悬液对斜纹夜蛾2龄幼虫均有一定的毒力,处理7 d时半致死浓度(LC₅₀)为3.944 107个孢子/mL;浓度为1.0×10⁹个孢子/mL时,半致死时间最短(LT₅₀)为4.6 d,校正后的死亡率为81.03%。处理后未致死的斜纹夜蛾幼虫体内抗氧化酶活性显著高于对照组。处理后致死的斜纹夜蛾幼虫肠道细菌群落多样性显著高于对照组;且处理后致死的斜纹夜蛾幼虫肠道细菌群落组成与对照组差异显著。【结论】金龟子绿僵菌对斜纹夜蛾幼虫的致死率和致死效率与金龟子绿僵菌的浓度呈正相关;斜纹夜蛾幼虫体内的抗氧化酶可能在抵抗金龟子绿僵菌侵染的过程中起重要作...     

Population Dynamics of Meloidogyne incognita on Corn Grown in Soil in Fested with Arthrobotrys conoides

Microplot and greenhouse experiments were conducted to evaluate the effects of soil incorporation of the nematophagous fungus Arthrobotrys conoides and green alfalfa mulch on the population dynamics of Meloidogyne incognita on corn. Reproduction of M. incognita and the incidence of root galling were reduced by the addition of A. conoides and/or green alfalfa in all tests. Numbers of juveniles were reduced by as much as 84%, and eggs were fewest in early to mid-season soil samples from microplots. Yields increased in treatments with A. conoides and/or green alfalfa in greenhouse tests and in the microplot tests in 1979. No interaction was found between the fungus and green alfalfa in the reduction of the nematode population.     

Comparative efficacy of arbuscular mycorrhizal fungi in combination with bioformulated Paecilomyces lilacinus against Meloidogyne incognita on tomato in two Ultisols of South-eastern Nigeria

Abstract

Greenhouse experiments were conducted to evaluate the efficacy of five arbuscular mycorrhizal fungi (AMF) in combination with bioformulated Paecilomyces lilacinus (PL Gold?) against Meloidogyne incognita on tomato in two Ultisols in Nigeria. Root galling and egg production were higher in Calabar than Nsukka Ultisols. P. lilacinus application did not inhibit tomato root colonisation by any of the AMF. Double application of PL Gold? in combination with Glomus etunicatum caused a significant change in the susceptibility of the tomato cultivar with a gall index (GI) =4.00 to (GI=2.00) in Nsukka Ultisol, while a similar result was obtained with Glomus mosseae, G. deserticola and G. etunicatum in Calabar Ultisols (GI=2.33). Application of both biocontrol agents significantly (P≤0.05) enhanced growth and yield of tomato plants. The highest fresh fruit yield was obtained in Calabar soil when Gi. gigantea was combined with double application of the bionematicide while the same result was obtained with G. deserticola in Nsukka soil.     

Mycorrhizosphere responsiveness to atmospheric ozone and inoculation with Phytophthora citricola in a phytotron experiment with spruce/beech mixed cultures

The aim was to analyze functional changes in the mycorrhizosphere (MR) of juvenile spruce and beech grown in a mixture under ambient and twice ambient ozone and inoculated with the root pathogen Phytophthora citricola. The phytotron experiment was performed over two vegetation periods, adding the pathogen at the end of the first growing season. Root biomass data suggest that the combined treatment affected spruce more than beech and that the negative influence of ozone on stress tolerance against the root pathogen P. citricola was greater for spruce than for beech. In contrast, beech was more affected when the pathogen was the sole stressor. The functional soil parameter chosen for studies of MR soil samples was activity of extracellular enzymes. After the first year of ozone exposure, MR soil samples of both species showed increased activity of almost all measured enzymes (acid phosphatase, chitinase, beta-glucosidase, cellobiohydrolase) in the O3 treatment. Species-specific differences were observed, with a stronger effect of P. citricola on beech MR and a stronger ozone effect on spruce MR. In the second year, the effects of the combined treatment (ozone and P. citricola) were a significant increase in the activity of most enzymes (except cellobiohydrolase) for both tree species. The results indicated that responsiveness of MR soils towards ozone and P. citricola was related to the severity of infection of the plants and the reduction of belowground biomass, suggesting a strong, direct influence of plant stress on MR soil enzyme activity. Additional research is needed using different species and combined stresses to determine the broader ecological relevance of shifts in rhizosphere enzymes.     

石羊河下游退耕地土壤微生物变化及土壤酶活性

采用时空替代法,对石羊河下游不同年限(1,2,3,4,5,8,15,24、31 a)退耕地土壤微生物(细菌、真菌和放线菌)数量、生物量(碳、氮和磷)及土壤酶(过氧化氢酶、蔗糖酶、脲酶和磷酸酶)活性变化及三者的相关性进行了测定和分析。结果表明,在退耕1—31 a的9个样地样方中土壤三大类微生物数量以细菌最高,放线菌次之,真菌最低。总体来看,三大土壤微生物数量的加权平均值最大值均在退耕后的前8 a。土壤微生物生物量碳在退耕初期随着退耕年限的增加而减小,退耕4 a后逐渐增大,退耕24 a期间达到了加权平均值的最大,最后趋于稳定;土壤微生物生物量氮在退耕初期随着退耕年限的增加而增加,退耕4 a加权平均值的最大值出现,随后逐渐减小的趋势,并且不同退耕年限土壤微生物生物量氮差异显著;土壤微生物生物量磷在退耕初期随着退耕年限的增加而增加,退耕8 a前后加权平均值达到最大值,随后逐渐减小,最终趋于稳定。土壤酶活性总趋势随着退耕地自然演替时间的增加呈波动式下降。不同土壤层次(0—10 cm,10—20 cm,20—30 cm及30—40 cm),土壤微生物数量、生物量及土壤酶活性随土层深度显著降低,并且表层土壤微生物生物量及土壤酶活性占有较大比例。土壤微生物及土壤酶活性的变化是一个极其缓慢的互动过程,存在着互相回馈的响应,特别是真菌与放线菌、微生物量氮及蔗糖酶,放线菌与过氧化氢酶、蔗糖酶,微生物量碳与磷酸酶,微生物量氮与脲酶,微生物量磷与蔗糖酶均存在极显著的相关性。总体来看在退耕年限4—5 a前,有利于土壤发育,退耕后期土壤肥力呈下降的趋势。     

Use of response surface methodology to optimize protease synthesis by a novel strain of Bacillus sp. isolated from Portuguese sheep wool

Aims: To investigate the influence of yeast extract, peptone, temperature and pH upon protease productivity by Bacillus sp. HTS102 – a novel wild strain isolated from wool of a Portuguese sheep breed (Merino). Methods and Results: A 2⁴ full factorial, central composite design together with response surface methodology was used to carry out the experiments and analyse the results, respectively. Among the individual parameters tested, temperature and peptone concentration produced significant effects upon protease productivity. A high correlation coefficient (R² = 0·994, P < 0·01) indicated that the empiric second‐order polynomial model postulated was adequate to predict said productivity, with the optimum loci characterized by: temperature of 43°C, peptone content of 1·4 g l^?1, pH of 5·1 and yeast extract concentration of 10·0 g l^?1. Conclusions: Protease synthesis depends chiefly on temperature and peptone level. The maximum protease activity was more than twice that obtained with the basal medium, so the experimental design and analysis undertaken were effective towards process optimization. Significance and Impact of the Study: Rational choice of processing conditions for maximum protease productivity will be relevant if an economically feasible fermentation process based on Bacillus sp. HTS102 is intended.     

Activity profiles of bacterioplankton in a eutrophic river

1. The significance of microbial diversity in processing dissolved organic matter (DOM) is largely unknown. We investigated the range of functional diversity in the bacterioplankton from a eutrophic river by profiling extracellular enzyme activities (EEA) and substrate-induced respiration (SIR) patterns.
2. The EEA profiles consisted of assays for 21 hydrolases, measured using fluorogenic substrates arrayed on 96-well microplates. Commercially available BiOLOG^® GN and ECO plates, which contained 120 different substrates, were used for the SIR profiles.
3. The EEA data were more dynamic than the SIR. Five enzymes, leucine aminopeptidase, alkaline phosphatase, alanine aminopeptidase, arginine aminopeptidase and β-glucosidase, showed consistently high activity; ten others were ubiquitous at lower activity levels; the remainder were detected intermittently. The SIR data showed less temporal variability. With one exception (citrate), the 20 substrates that generated the largest responses were all saccharides or their derivatives.
4. The EEA and SIR data did not generally correlate. Both methods were effective for ordinating bacterioplankton although, unlike the SIR, the EEA ordination followed a clear temporal trajectory.
5. Because the SIR profiles are based on a culture response, whilst the EEA profiles measure activity of the extant community, the latter appear to be more directly linked to the mechanics of DOM processing.     

Phenylacetic acid-producing Rhizoctonia solani represses the biosynthesis of nematicidal compounds in vitro and influences biocontrol of Meloidogyne incognita in tomato by Pseudomonas fluorescens strain CHA0 and its GM derivatives

AIMS: The aim of the present investigation was to determine the influence of Rhizoctonia solani and its pathogenicity factor on the production of nematicidal agent(s) by Pseudomonas fluorescens strain CHA0 and its GM derivatives in vitro and nematode biocontrol potential by bacterial inoculants in tomato. METHODS AND RESULTS: One (Rs7) of the nine R. solani isolates from infected tomato roots inhibited seedling emergence and caused root rot in tomato. Thin layer chromatography revealed that culture filtrates of two isolates (Rs3 and Rs7) produced brown spots at Rf-values closely similar to synthetic phenylacetic acid (PAA), a phytotoxic factor. Filtrates from isolate Rs7, amended with the growth medium of P. fluorescens, markedly repressed nematicidal activity and PhlA'-'LacZ reporter gene expression of the bacteria in vitro. On the contrary, isolate Rs4 enhanced nematicidal potential of a 2,4-diacetylphloroglucinol overproducing mutant, CHA0/pME3424, of P. fluorescens strain CHA0 in vitro. Therefore, R. solani isolates Rs4 and Rs7 were tested more rigorously for their potential to influence biocontrol effectiveness of the bacterial agents. Methanol extract of the culture filtrates of PAA-producing isolate Rs7 resulting from medium amended with phenylalanine enhanced fungal repression of the production of nematicidal agents by bacteria, while amendments with zinc or molybdenum eliminated such fungal repression, thereby restoring bacterial potential to cause nematode mortality in vitro. A pot experiment was carried out, 3-week-old tomato seedlings were infested with R. solani isolates Rs4 or Rs7 and/or inoculated with Meloidogyne incognita, the root-knot nematode. The infested soil was treated with aqueous cell suspensions (10(8) CFU) of P. fluorescens strain CHA0 or its GM derivatives or left untreated (as a control). Observations taken 45 days after nematode inoculation revealed that, irrespective of the bacterial treatments, galling intensity per gram of fresh tomato roots was markedly higher in soil amended with isolate Rs4 than in Rs7-amended soils. Soil amendments with R. solani and the bacterial antagonists resulted in substantial reductions of the number of galls per gram of root. These results are contradictory to those obtained under in vitro conditions where culture filtrates of PAA-positive Rs7 repressed the production of nematicidal compounds. Plants grown in Rs7-amended soils, with or without bacterial inoculants, had lesser shoot and root weights than plants grown in nonamended or Rs4-amended soils. Moreover, amendments with Rs7 substantially retarded root growth and produced necrotic lesions that reduced the number of entry sites for invasion and subsequent infection by nematodes. Populations of P. fluorescens in the tomato rhizosphere were markedly higher in Rs7-amended soils. CONCLUSIONS: PAA-producing virulent R. solani drastically affects the potential of P. fluorescens to cause death of M. incognita juveniles in vitro and influences bacterial effectiveness to suppress nematodes in tomato roots. SIGNIFICANCE AND IMPACT OF THE STUDY: As most agricultural soils are infested with root-infecting fungi, including R. solani, it is likely that some PAA-producing isolates of the fungus may also be isolated from such soils. The inhibitory effect of PAA-producing R. solani on the biosynthesis of nematicidal agent(s) critical in biocontrol may reduce or even eliminate the effectiveness of fluorescent pseudomonads against root-knot nematodes, both in nursery beds and in field conditions. Introduction of bacterial inoculants, for the control of any plant pathogen, should be avoided in soils infested with PAA-producing R. solani. Alternatively, the agents could be applied together with an appropriate quantity of fungicide or chemicals such as zinc to create an environment more favourable for bacterial biocontrol action.     

Response of rhizosphere microbial diversity and soil physico-chemical properties in a rotation of cucumber with Volvariella volvacea

Crop rotation can significantly increase yield by improving soil conditions through reducing soil pathogens and increasing the microbial diversity near the root system of the plant. Cucumber is widely consumed across the world, and monoculture of cucumber under greenhouse conditions is often threatened by Fusarium wilt, which is caused by fungi belonging to the genus Fusarium. In Jiangsu province of China, rotating cucumber crops with Volvariella volvacea is used as a way to control Fusarium wilt. Here, we tested how rotating cucumber with V. volvacea influences soil physico-chemical and biological properties. We found that rotating with V. volvacea improved bacterial diversity near the cucumber root system. We also found an increase in nitrogen-fixing bacteria that have been used for biological control of plant pathogens, and a decrease in Fusarium species. In the treated soil, we found increased activity of the soil enzymes catalase, dehydrogenase, polyphenol oxidase and alkaline phosphatase by 2–22%, 14–29%, 16–58% and 9–64%, respectively. Lastly, the total and available nitrogen, phosphorus and potassium content significantly increased one to three weeks after V. volvacea residual substrate was mixed into the soil, up to 39%, 24%, 17%, 28%, 18% and 70, but declined slowly afterwards. These results suggested that rotating cucumber with V. volvacea could aid in resetting the structure of the microbial community of the cucumber root system. Increase in beneficial microbes and improved soil conditions are likely to contribute to improved yield of this commercially important crop.