THE EFFECT OF ASSOCIATED SOIL MICROFLORA ON FUSARIUM UDUM BUTL., THE FUNGUS CAUSING WILT OF PIGEON-PEA (CAJANUS CAJAN (L.) MILLSP.)

Inoculation with Fusarium udum Butl. produced more wilt of pigeon-pea in sterilized than in unsterilized soils at the same pH. From unsterilized soils with low disease incidence, nine fungi, Bacillus subtilis and an Actinomyces were isolated. The number of isolations of a particular organism varied from month to month during the cropping season of pigeon-pea in Delhi. Interaction of Fusarium udum and other organisms isolated was studied. Aspergillus niger and A. terreus secreted inhibitory substances in potato-dextrose broth: Bacillus subtilis inhibited growth on solid medium and also produced a toxic substance in potato-dextrose broth. The nature of the medium employed and period of growth were important factors in the production of the inhibitory principle, which is thermostable. The low incidence of pigeon-pea wilt in unsterilized soils may result from the inhibitory activity of the associated microflora in the soil.     

INVESTIGATIONS OF THE INHIBITORY ACTION OF BACILLUS SUBTILIS ON FUSARIUM UDUM BUTL., THE FUNGUS CAUSING WILT OF PIGEON-PEA (CAJANUS CAJAN (L.) MILLSP.)

A suitable basic medium for the growth of Bacillus subtilis , particularly in relation to the production of an antibiotic inhibitory to Fusarium udum , is described. Organic nitrogen with carbohydrate and K₂HPO₄ is shown to be essential for the growth of Bacillus subtilis. Magnesium sulphate, manganese and iron are found to increase the production of the antibiotic. Effects of temperature, period of incubation, aeration, etc., on the production of the inhibitory substance are described. The active principle is adsorbed by soil and to a lesser degree by kaolin. The organic matter and the soluble salts in the soil do not appear to play an important role in the loss of activity of the filtrates during passage through a bed of soil.     

Biological activity of phenylpropionic acid isolated from a terrestrial Streptomycetes.

The strain ANU 6277 was isolated from laterite soil and identified as Streptomyces sp. closely related to Streptomyces albidoflavus cluster by 16S rRNA analysis. The cultural, morphological and physiological characters of the strain were recorded. The strain exhibited resistance to chloramphenicol, penicillin and streptomycin. It had the ability to produce enzymes such as amylase and chitinase. A bioactive compound was isolated from the strain at stationary phase of culture and identified as 3-phenylpropionic acid (3-PPA) by FT-IR, EI-MS, 1H NMR and 13C NMR spectral studies. It exhibited antimicrobial activity against different bacteria like Bacillus cereus, B. subtilis, Escherichia coli, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa, P. flourescens, Staphylococcus aureus and some fungi including Aspergillus flavus, A. niger, Candida albicans, Fusarium oxysporum, F. udum and Penicillium citrinum. The antifungal activity of 3-PPA of the strain was evaluated in in vivo and in vitro conditions against Fusarium udum causing wilt disease in pigeon pea. The compound 3-PPA is an effective antifungal agent when compared to tricyclozole (fungicide) to control wilt caused by F. udum, but it exhibited less antifungal activity than carbendazim.     

Antifungal potential of some higher plants against Fusarium udum causing wilt disease of Cajanus cajan

The fungitoxic effects of different plant extracts on Fusarium udum, which causes wilt disease of Cajanus cajan in vitro and in vivo, were examined. The complete arrest of the radial growth of the pathogen occurred at a 10% concentration of leaf extract from Adenocallyma alliaceum. A leaf extract of Citrus medica, a root extract of Asparagus adscendens, rhizome extracts of Curcuma longa and Zingiber officinale, and a bulb extract of Allium sativum inhibited up to 100% growth at higher concentrations. A. alliaceum controlled the disease up to 100% by amending its 4% powder in unsterilized soil and 2% in sterilized soil. The population of F. udum was found to be markedly reduced following treatments with plant powders.     

HPLC analysis of fusaric acid, 9,10-dehydrofusaric acid and their methyl esters,toxic metabolites from weed pathogenic Fusarium species

A simple and rapid HPLC method, using a high-density C18 column, has been developed for the quantitative analysis of fusaric and dehydrofusaric acids and their methyl esters in the methanol extract of lyophilised culture filtrates of species of Fusarium. The method has been used to determine the content of these metabolites in two strains of Fusarium oxysporum and in strains of F. nygamai and F. udum. Fusaric acid has been isolated and identified from a strain of F. udum for the first time.     

Seed Bacterization with Bacillus subtilis AF 1 Increases Phenylalanine Ammonia-lyase and Reduces the Incidence of Fusarial Wilt in Pigeonpea

Cell free culture filtrate of a biocontrol PGPR strain of Bacillus subtilis showed a concentration-dependent growth and conidiation inhibition of Fusarium udum . the incitant of pigeonpea wilt. In vitro interaction of cell free filtrate of AF 1 induced bulb-like structures in the hyphae of F. udum . Seed treatment with B. subtilis significantly reduced the incidence of pigeonpea wilt, but not with the cell free culture filtrate of AF 1. Seeds bacterized with B. subtilis AF 1 showed an increase in phenylalanine ammonia-lyase (PAL) from day 1, while the peroxidase activity increased from day 6. The results are discussed in terms of induction of host plant resistance in biological control of plant pathogens.     

Influence of Fusarium wilt toxin(s) on carnation cells

The influence of culture filtrates of Fusarium oxysporum f.sp. dianthi which causes Fusarium wilt was investigated on growth and viability of carnation tissue cultures and leaf segments. Culture filtrates of avirulent race 1 of this fungus did not affect calli and leaf segments of cultivars both susceptible and resistant to Fusarium wilt. However, culture filtrates of virulent race 2 decreased viability and suppressed growth of callus of the susceptible cultivar. In contrast, callus of the resistant cultivar showed resistance to the culture filtrates. The results of these experiments may provide information on methods of selection of new wilt resistant carnation varieties.Abbreviations A270 absorbance at 270 nm - 2,4-d 2,4-dichlorophenoxyacetic acid - CF-MCD culture filtrate of 16064 grown in MCD medium - MCD medium modified Czapeck-Dox medium - MS medium basal medium of Murashige and Skoog - MW molecular weight - PD medium potato dextrose medium - TTC 2,3,5-triphenyl tetrazolium chloride     

Bio-activity of fungal culture filtrates against root-knot nematode egg hatch and juvenile motility and their effects on growth of mung bean (Vigna radiata L. Wilczek) infected with the root-knot nematode,Meloidogyne incognita

Culture filtrates of selected soil fungi, namely Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Fusarium oxysporum, Penicillium vermiculatum and Rhizopus nigricans exhibited variable response to egg hatching and mortality of the root-knot nematode, Meloidogyne incognita. Higher concentrations of the culture filtrates of all the fungi inhibited egg hatching and proved to be toxic to the juveniles of M. incognita. In addition, development of the gall and multiplication of M. incognita were also found adversely affected in varying degrees on all the plants of Vigna radiata treated with the filtrates. The culture filtrate of A. niger showed highest toxicity to the nematode than those of any other fungus tested. Soil drench application of the culture filtrates gave better seedling growth and least nematode multiplication in comparison to seed soaking treatment.     

Interactions of Meloidogyne javanica, Fusarium udum and Rhizobium on pigeon pea in the presence of nickel and cobalt as pollutants

The interactions between Meloidogyne javanica, Fusarium udum and Rhizobium on pigeon pea, in the presence of nickel (Ni) and cobalt (Co) as soil pollutants, were studied. In single inoculations, M. javanica caused a reduction in plant growth, particularly at higher inoculum levels (1000 and 10 000 JJ₂). Fusarium udum induced wilting symptoms and the higher concentrations of Ni and Co suppressed plant growth. Meloidogyne javanica and F. udum interacted to show more severe wilting symptoms than F. udum alone. Wilting was completely inhibited by Ni but reductions in plant growth and nodulation occurred. In the presence of Ni, M. javanica did not cause galling but nodulation was suppressed. Cobalt also suppressed wilting and nodulation but interacted with M. javanica to increase root galling and to reduce plant growth by a greater amount than the total of reductions caused by the individual treatments.     

Analysis of Pectic Enzyme Zymograms of Fusarium Species I. Fusarium lateritium and Related Species

Abstract Zymograms of the extracellular pectic enzymes (pectinesterase, polygalacturonase) of Fusarium lateritium and related species were prepared by electrophoresis from untreated culture filtrates. The zymogram technique gave evidence to conclude that F. lateritium, F. stilboides and F. xylarioides are distinct species. "pini" isolates are rather "subglutinans"-like fusaria than "lateritium"-like, while F. udum represents a genetical variant of F. oxysprorum .     

儿茶素对棉枯萎病菌胞壁降解酶的抑制及在棉花抗病性中的作用

在含有儿茶素的培养液中棉枯萎病菌(Fusarium oxysporum f. sp. vasinfectum)的多聚半乳糖醛酸酶(PG)和果胶裂解酶(PL)活性明显降低。儿茶素可明显抑制初步纯化的PG和PL活性以及它们对棉苗组织的浸软作用。对棉苗组织中儿茶素含量的测定结果表明:抗病品种棉苗组织中儿茶素含量较高;氟乐灵处理可诱发棉苗产生对枯萎病的诱导抗性,同时也提高棉苗组织中的儿茶素的含量;枯萎病菌侵染后棉苗组织中儿茶素含量明显升高,以抗病品种棉苗和氟乐灵诱发处理棉苗组织中儿茶素含量的增加更为明显。棉苗组织提取液中的酚类物质可抑制PG和PL的活性,且证明这种抑制作用主要是由儿茶素引起。提取液对PG和PL活性的抑制作用与棉苗组织中儿茶素的含量呈直接的正相关关系。因此,作者认为棉苗组织中的儿茶素可能通过对病菌PG和PL等胞壁降解酶的抑制而与棉花对枯萎病的抗病性及氟乐灵诱发的诱导抗性有关。     

对黄瓜枯萎病具防效的海洋源芽孢杆菌Y3F的鉴定

【目的】对一株从连云港海域海藻样品中获得的拮抗细菌Y3F进行鉴定并且研究菌株Y3F对黄瓜土传枯萎病害的控制效果。【方法】对Y3F进行形态、生理生化特征和16S r RNA基因序列分析,采用平板对峙法测定菌株Y3F的活菌液和无菌滤液的抑菌活性,利用盆栽试验测定Y3F对黄瓜枯萎病害的防治效果。【结果】初步鉴定该菌属于蜡样芽孢杆菌(Bacillus cereus),用2216E培养基振荡培养24 h的Y3F的无菌滤液对黄瓜尖孢镰刀菌有较强的抑菌活性,表明菌株能分泌抑制病原菌生长的活性物质。盆栽试验表明,种植30 d后,浸种和灌根同时处理(JG)的防治效果达到50.46%,对黄瓜枯萎病害有明显的控制效果,显著提高黄瓜植株生物量,显著降低黄瓜根际土的真菌和尖孢镰刀菌数量,增加根际的细菌和放线菌数量。【结论】菌株Y3F能有效防治黄瓜枯萎病害,改善根际微生物结构,具有进一步开发应用的前景。     

Effect of cultival practices and soil treatments on incidence of wilt disease of pigeon pea

Summary The incidence of wilt disease of pigeon pea (Cajanus cajan (L.) Millsp.) caused byFusarium udum under soil treatments with various substances was studied under field conditions. The disease incidence was highly suppressed under mixed cropping withCrotalaria medicaginea. Phygon XL was found to be the most effective fungicide to reduce the incidence of the disease. The incidence of wilt disease also decreased in case of soil amended with the leaves ofC. medicaginea. Wilt incidence increased in the soil amended with the roots of pigeon pea.     

Toxigenic species of Penicillium, Fusarium, and Aspergillus from weevil-damaged pecans.

Of a total of 2392 fungi isolated from weevil-damaged pecans, 46.4% were Alternaria and Epicoccum, 23.9% were Penicillium, 12.4% were Pestalotia and Monochaeta, 6.5% were Cladosporium, 6.4% were Fusarium, and less than 2% each were Phoma, Aspergillus, Rhizopus, Trichothecium, and miscellaneous. Chloroform extracts of 34 of 105 representative Penicillium isolates, 3 of 28 Fusarium isolates, and 3 of 23 Aspergillus isolates were toxic to day-old cockerels during three bioassays. Eight of the toxic extracts from Penicillium spp. were tremorgenic. One tremorgenic isolate was identified as P. paxilli, four were identified as P. lanoso-coeruleum, and three as P. cyclopium. Nine of the non-tremorgenic isolates were identified as P. citrinum, five as P. aurantio-virens, three as P. oxalicum, and two as P. meleagrinum. Others were identified as one each of P. brevi-compactum (Series), P. nigricans (Series), P. roqueforti, P. rugulosum (Series), P. terrestre, and P. stoloniferium. One was unidentified. Toxigenic Aspergillus isolates were all A. flavus. Two of the toxic Fusarium isolates were F. moniliforme, and one was unidentified.     

石灰碳铵熏蒸与施用生物有机肥对连作黄瓜和西瓜枯萎病及生物量的影响

采用稀释涂布平板计数法,研究了石灰碳铵及碳铵熏蒸对黄瓜和西瓜连作土壤尖孢镰刀菌数量的影响,以及熏蒸后施用生物有机肥对黄瓜和西瓜枯萎病的防控效果及植株生长的影响.结果表明:与对照相比,石灰碳铵及碳铵熏蒸后,连作土壤中黄瓜尖孢镰刀菌的数量分别下降95.4%及71.4%,西瓜尖孢镰刀菌的数量分别下降87.2%及64.2%;多因素方差分析表明,熏蒸、施用有机肥及作物种类均对土壤中尖孢菌数量、枯萎病发病率、防控率及生物量有显著影响;与未熏蒸施用普通有机肥对照相比,石灰碳铵熏蒸后施用生物有机肥能显著减少后茬黄瓜或西瓜土壤中尖孢镰刀菌的数量并显著降低枯萎病发病率,防控率高达91.9%及92.5%,同时显著增加了植株的株高、茎粗、SPAD值及干质量.表明石灰碳铵熏蒸及施用生物有机肥能够降低土壤中尖孢镰刀菌数量,有效防控黄瓜和西瓜枯萎病的发生并促进其植株生长.     

Studies on antagonism betweenFusarium udum Butler and root region microflora of pigeon-pea

Antagonism betweenFusarium udum Butler causing wilt of pigeon-pea (Cajanus cajan (L.) Millsp.) and the saprophytic microflora of the root region of the host was studied with reference to colony interaction, hyphal interference, volatile and non-volatile metabolites and staling growth products. Studies were extended to screen potential antagonists against the wilt pathogen in soil. Aspergillus flavus, A. niger, A. terreus, Penicillium citrinum andMicromonospora globosa (an actinomycete) were antagonistic againstF. udum, whereas the pathogen parasitized and killedAspergillus luchuensis, Cunninghamella echinulata, Curvularia lunata, Mortierella subtilissima andSyncephalastrum racemosum. The pattern of growth of microorganisms on nutrient agar staled by rhizosphere soil inocula of healthy or wilted pigeon-pea plants was found to be different.F. udum colonized and grew on nutrient agar staled by the rhizosphere inoculum of the wilted plants upto 120h of incubation. However, it could not colonise and grow on the nutrient agar staled by rhizosphere microflora of healthy plants after 48h of incubation because of the presence of antagonists likeA. niger, A. flavus, A. terreus and a few species ofPenicillium in the soil inoculum. When pure cultures in soil ofF. udum was mixed with those of antagonists in different ratios,A. niger, A. flavus andM. globosa significantly suppressed the population ofF. udum, whereasA. terreus markedly reduced the population. When inoculated in soil, the antagonists exhibited a high fungistatic activity againstF. udum.     

Effect of Brassica carinata (L.) biofumigants (seed meal) on chickpea wilt (Fusarium oxysporum f.sp. ciceris), growth,yield and yield component in Ethiopia

Pot experiments were carried out in the green house at Amhara Regional Agriculture Research Institute (ARARI) Bahirdar, Ethiopia to evaluate the potential of Brassica carinata cultivars namely; Holleta-l, S-67 and Yellow Dodola in 2007 and 2008. The treatment effects of B. carinata (L.) cultivars Holleta–1, S-67 and Yellow Dodola seed meals on chickpea fusarium wilt (Fusarium oxysporum f.sp. ciceris) were studied. Six rates of seed (0, 5, 10, 15, 20 and 25 g/kg of infested soil) were used. Infested soil without B.carinata cultivars amendments as a control and susceptible check variety JG-62 also without amendments were used in all the experiments. For each seed meal experiment, the treatments were arranged in factorial randomised complete block design in three replications. Data on seedling emergence, wilt incidence, fresh weight, dry weight, pod per plant, seed per pod, hundred seed weight and yield per hectare were collected. The amendments of infested soil with B.carinata cultivars seed meal reduced the incidence of chickpea fusarium wilt and increased yield per hectare. The interaction of the seed meal Holleta-1, S-67 and Yellow Dodola at 10–25 g/kg infested soil were effective in reducing wilt incidences on chickpea. However, the interaction of Yellow Dodola with 20 and 25 g seed meal per kg infested soil were the best combination in reducing significantly wilt incidence. The three cultivars incorporated at different level of doses significantly affected the influence of Fusarium wilt on the fresh weight, dry weight, pod per plant, seed per pod, hundred seed weight and yield per hectare. The highest yield kg/ha was recorded in combination of Yellow Dodola seed meal at 20 and 25 g followed by S-67 and Holleta-1 at 25 g /kg infested soil, respectively. The interaction of Holleta-1 at 5–25 infested soil significantly reduced disease incidence up to 16.7–43.3% and increased yield per hectare with mean by (30%) over the control. Seed meal amendment S-67 significantly reduce disease incidence 26.7–46.7% and increased yield kg/ha with mean by (36.7%) from the unamended control. Yellow dodola reduces disease incidence with 26.7–63.3% and increased yield kg/ha with mean by (45%) from the unamended control. The result indicates the potential of using Brassica crop seed meal amendment as useful component of integrated chickpea wilt management.     

Toxigenic Fungi in Food

Forty-five fungal isolates from moldy supermarket foods were tested for toxicity to brine shrimp, and twenty-two of these isolates were subsequently tested for toxicity to chicken embryos. Highly toxigenic fungi were Cladosporium sphaerospermum from a bakery product, Fusarium oxysporum from carrots, F. solani from cabbage, Aspergillus niger and Penicillium corylophilum from bread, P. cyclopium and P. herguei from corn meal, P. lanosum from onions,P. steckii from chocolate syrup, Penicillium sp. from jelly, and Rhizopus nigricans isolates from sweet potato, applesauce, and strawberries. Approximately one-third of the fungal cultures were moderately to highly toxigenic to brine shrimp and chicken embryos, while several additional cultures were slightly toxigenic.     

Influence of microbial co-inhabitants on aflatoxin synthesis of Aspergillus flavus on maize kernels

The co-inhabiting mycoflora with Aspergillus flavus observed on individual maize kernels was evaluated for its influence on aflatoxin synthesis. All 13 types of associations of different fungal species inhibited aflatoxin B₁ and G₁ production at different levels (34·3–100%). Inhibition of radial growth of A. flavus by Fusarium moniliforme (59·8%), Trichoderma viride (72·5%) and Rhizopus nigricans (42%) could be directly correlated to the per cent inhibition of aflatoxin production. High levels of inhibition of aflatoxin elaboration were noted in competition of A. flavus with other toxigenic moulds.     

Non-pathogenic Fusarium solani represses the biosynthesis of nematicidal compounds in vitro and reduces the biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato

AIMS: The aim of the present investigation was to determine the influence of various Fusarium solani strains on the production of nematicidal agent(s) in vitro and biocontrol of Meloidogyne javanica in tomato by Pseudomonas fluorescens strains CHA0 and CHA0/pME3424. METHODS AND RESULTS: Culture filtrates (CF) of P. fluorescens strain CHA0 and its diacetylphloroglucinol-overproducing derivative CHA0/pME3424 caused substantial mortality of M. javanica juveniles in vitro. Bacterial growth medium amended with the growth medium of F. solani repressed the nematicidal activity of the bacteria. Methanol extract of F. solani CF resulting from Czapek's Dox liquid (CDL) medium without zinc amendment repressed the nematicidal activity of the bacteria while the CF obtained from CDL medium amended with zinc did not. Conidial suspension of F. solani strain Fs5 (repressor strain for the biosynthesis of nematicidal compounds in P. fluorescens) reduced biocontrol potential of the bacterial inoculants against M. javanica in tomato while strain Fs3 (non-repressor) did not. CONCLUSIONS: Fusarium solani strains with increased nematicidal activity repress the biosynthesis of nematicidal compounds by P. fluorescens strains in vitro and greatly alter its biocontrol efficacy against root-knot nematode under natural conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium solani strains are distributed worldwide and found in almost all the agricultural fields which suggest that some mycotoxin-producing strains will also be found in almost any soil sample taken. Besides the suppressive effect of these metabolite-producing strains on the production of nematicidal compound(s) critical in biocontrol, F. solani strains may also affect the performance of mycotoxin-sensitive biocontrol bacteria effective against plant-parasitic nematodes.