Growth Of Sclerotia Research Articles

ASAI KEMAMPUAN BAKTERI ENDOFIT DARI KACANG TANAH DALAM MENGHAMBAT PERTUMBUHAN Sclerotium sp. PADA KECAMBAH KACANG TANAH

Assay on ability of endophytic bacteria isolated from peanut to inhibit Sclerotium sp. growth in peanut seedlings. A study on assay of ability of endophytic bacteria to inhibit Sclerotium sp. in peanut seedling has been done. The bacteria were isolated from peanut healthy plants, while Sclerotium sp. was isolated from infected peanaut plant. Antagonistic assay was conducted by dual culture method. In vivo assay of inhibiting Sclerotium sp. was conducted by dipping peanut seed in bacterial solution, and planting the seed in soil:compost (3:1) growing media. Six endophytic bacterial isolates showed to inhibit the growth of Sclerotium sp. in vitro. LN1 seemed to inhibit more of Sclerotium sp., while LN5 showed to inhibit less. Two potential isolates LN1 of gram-negative and LN2 of gram-positive using for further study showed to decrease more of dumping off. It also seemed that the isolates increased the seedling height, number of leaves, and dry weight.

Antagonistic activity of cowshed Bacillus sp. bacteria against aflatoxigenic and sclerotic Aspergillus flavus

Mycotoxins produced by many food spoilage fungi cause serious damage to human as well as to livestock. Aflatoxins are one such group of mycotoxins produced by Aspergillus flavus in many agricultural and food products including rice. The fungus can virtually grow in any environment and also produces resistive structures such as sclerotia to overcome unfavourable environmental conditions. Hence, the need of the time is to control this toxigenic and sclerotic fungus through an eco-friendly approach. In the current study four biocontrol bacteria belonging to Bacillus species were isolated from Indian cow shed environment and these bacteria could efficiently control not only the mycelia growth of A. flavus but also the germination and growth of sclerotia. Both active bacterial culture broth and cell free culture filtrate could limit the growth of the fungus up to more than 90%. Older culture broth and filtrate lost their inhibition efficiency. Unlike many similar studies the current investigation emphasizes the importance of cowshed environment instead of cow dung and the control of sclerotia instead of fungal spores.

A landscape of transcriptome analysis of three sclerotia growth stages in Polyporus umbellatus

Polyporus umbellatus,a traditional Chinese precious medicine as long been used for eliminating dampness,diuresis and have effect on cancer,getting more and more popularly in China recently. And the developmental metabolic process of the medicinal fungus,P. umbellatus,has been gotten more attention. This study is for the first time to explore the three sclerotial growth stages in P. umbellatus,named " white Polyporus"( initial phase), " grey Polyporus"( developmental phase) and " black Polyporus"( mature phase),by utilizing the de novo transcriptome assembly analysis technology. Finally,we obtained 88. 12 Gb sequence containing85 235 unigenes( ≥200 bp) assembled and 100% were annotated. We identified genes differentially expressed among the three stages of the sclerotia and screened out MFSgst,ERG4/ERG24,WD40,Rho A,CYP450,PKS,GSase and CHS1,which may contribute to the production of medicinal secondary metabolites and the defense mechanism against the environmental stress and biological invasion. We did the qRT-PCR trial to verify our results,which is in line with expectations. Our results are purposed to unearth the molecular mechanism of the accumulation of active constituents in different stages of Polyporus sclerotia which can be applied in the production and protection of Polyporus effectively.

Biological control of Sclerotium fruit rot of snake fruit and stem rot of lettuce by Trichoderma sp. T76-12/2 and the mechanisms involved

Fruit rot of snake fruit and stem rot of lettuce caused by Sclerotium sp. SZ01 and LS01, respectively, are major diseases causing devastating crop losses. Tichoderma species has been known as an effective biological control agent (BCA) against several plant disease pathogens. This research aimed to select an effective Trichoderma species for controlling both diseases. Five Trichoderma spp. isolates were subjected to primary screening against Sclerotium sp. LS01 and SZ01 by dual culture test. Trichoderma sp. T76-12/2 showed significant inhibitory effects on mycelial growth of Sclerotium sp. LS01 and SZ01 with 81.48% and 78.33% inhibition. The Trichoderma sp. T76-12/2 caused discoloration of Sclerotium sp. mycelia at the contact point. Volatile antifungal bioassay revealed Trichoderma sp. T76-12/2 was superior in inhibiting the growth of Sclerotium sp. by up to 44.44% for LS01 and by 59.52% for SZ01. Gas chromatography-mass spectrometry (GC/MS) was conducted to resolve the volatile organic compounds emitted by Trichoderma sp. T76-12/2, and the results revealed presence of antifungal compounds including alcohol (phenylethyl alcohol) and sesquiterpene (epi-cubenol). Enzyme assay of cell-free culture filtrate showed that the activity of β-1,3-glucanase was higher than that in control. Application of Trichoderma sp. T76-12/2 spore suspension reduced disease severity index for Sclerotium sp. (LS01 and SZ01) in both lettuce and snake fruit in vivo. Based on these findings, Trichoderma sp. T76-12/2 is an excellent candidate to control fruit rot and stem rot diseases caused by Sclerotium sp., due to its capacity to compete, to produce volatile antifungal compounds and cell-wall degrading enzymes, as well as to reduce the disease severity.

Analysis of putative sclerotia maturation-related gene expression in Rhizoctonia solani AG1-IA

Rhizoctonia solani AG1-IA (R. solani AG1-IA) is a major soil-borne fungal pathogen of maize that causes significant yield losses in all maize-growing regions worldwide. The sclerotium produced by R. solani AG1-IA can overwinter in grass roots or diseased plants and infect crops the following year. The molecular mechanism underlying sclerotium formation in R. solani is poorly understood. In this study, we constructed the cDNA library of the R. solani AG1-IA pathogenic strain WF-9, from which 329 high-quality expressed sequence tags (ESTs) were obtained. Of the 250 clustered unigenes, 12 genes were selected for further expression analysis during the three stages of sclerotial growth (mycelium, initiation of sclerotium, and maturation of sclerotium). The results of expression analysis support the previously suggested roles of chitin synthase D and exo-beta-1,3-glucanase in facilitating sclerotial growth through preservation of water content and energy. In addition, cytochrome P450, NADPH oxidase, catalase (CAT), acyl-CoA oxidase 1 (ACOX1), mitogen-activated protein kinase (MAPK), mitogen-activated protein kinase HOG1 (HOG 1), and the G-protein ? subunit play important roles in balancing reactive oxygen species (ROS) levels during sclerotial development. The findings of this study can help understand the molecular mechanism of sclerotial development further.

The Kinome of Edible and Medicinal Fungus Wolfiporia cocos

Wolfiporia cocos is an edible and medicinal fungus that grows in association with pine trees, and its dried sclerotium, known as Fuling in China, has been used as a traditional medicine in East Asian countries for centuries. Nearly 10% of the traditional Chinese medicinal preparations contain W. cocos. Currently, the commercial production of Fuling is limited because of the lack of pine-based substrate and paucity of knowledge about the sclerotial development of the fungus. Since protein kinase (PKs) play significant roles in the regulation of growth, development, reproduction, and environmental responses in filamentous fungi, the kinome of W. cocos was analyzed by identifying the PKs genes, studying transcript profiles and assigning PKs to orthologous groups. Of the 10 putative PKs, 11 encode atypical PKs, and 13, 10, 2, 22, and 11 could encoded PKs from the AGC, CAMK, CK, CMGC, STE, and TLK Groups, respectively. The level of transcripts from PK genes associated with sclerotia formation in the mycelium and sclerotium stages were analyzed by qRT-PCR. Based on the functions of the orthologs in Sclerotinia sclerotiorum (a sclerotia-formation fungus) and Saccharomyces cerevisiae, the potential roles of these W. cocos PKs were assigned. To the best of our knowledge, our study is the first identification and functional discussion of the kinome in the edible and medicinal fungus W. cocos. Our study systematically suggests potential roles of W. cocos PKs and provide comprehensive and novel insights into W. cocos sclerotial development and other economically important traits. Additionally, based on our result, genetic engineering can be employed for over expression or interference of some significant PKs genes to promote sclerotial growth and the accumulation of active compounds.

Exploration of Potential Actinomycetes from CIFOR Forest Origin as Antimicrobial, Antifungus, and Producing Extracellular Xylanase

<p>This study aimed to isolate and explore the actinomycetes of CIFOR forest origin as an antimicrobial and antifungal agent, to produce an extracellular xylanase, and to identify isolates based on 16S rRNA gene sequences. Actinomycetes were isolated using Humic-acid Vitamin-B agar (HV) media. Actinomycetes colonies that grow on the medium HV was subsequently purified by growing them on yeast malt agar (YMA) media), then an antagonistic test of selected bacteria against Bacillus sp., Escherichia coli, Fusarium oxysporum, and Sclerotium sp was performed. Xylanase activity test was detected by observing a clear zone, followed by identification. Total of 35 isolates of actinomycetes isolated based on their colony morphology characteristics and diverse types of spore chains showed Streptomyces spp. of isolates CFR-06, CFR-15, CFR-17, CFR-18, and CFR-19 were able to inhibit the growth of Bacillus sp.. The highest inhibition zone has a diameter of 10.1 mm (isolate CFR-17). Isolates CFR-01 and CFR-15 were able to inhibit the growth of E. coli with the highest inhibition zone diameter of 5.1 mm (isolate CFR-15). Isolates CFR-29 and CFR-12 were able to inhibit the growth of F. oxysporum while isolate CFR-35 were able to inhibit the growth of Sclerotium sp.. Xylanase activity test showed that isolates CFR-12, CFR-20, CFR-22, CFR-24, CFR-25, CFR-30, CFR-33, CFR-34 have an ability to produce extracellular xylanase enzyme. Actinomycetes isolate (Xyl_22) as a potential xylanase enzyme producer was closely related with Streptomyces drozdowicii by the maximum similarity of 99%.</p><p><strong>How to Cite</strong></p><p>Sipriyadi, S., Lestari, Y., Wahyudi, A., & Meryandini, A. (2016). Exploration Potential CIFOR Forest actinomycetes origin as Antimicrobial, Anti Fungus and Producing Enzymes Extracellular Xylanase. <em>Biosaintifika: Journal of Biology & Biology Education</em>, 8(1), 94-102.</p>

De Novo Analysis of Wolfiporia cocos Transcriptome to Reveal the Differentially Expressed Carbohydrate-Active Enzymes (CAZymes) Genes During the Early Stage of Sclerotial Growth.

The sclerotium of Wolfiporia cocos has been used as an edible mushroom and/or a traditional herbal medicine for centuries. W. cocos sclerotial formation is dependent on parasitism of the wood of Pinus species. Currently, the sclerotial development mechanisms of W. cocos remain largely unknown and the lack of pine resources limit the commercial production. The CAZymes (carbohydrate-active enzymes) play important roles in degradation of the plant cell wall to provide carbohydrates for fungal growth, development, and reproduction. In this study, the transcript profiles from W. cocos mycelium and 2-months-old sclerotium, the early stage of sclerotial growth, were specially analyzed using de novo sequencing technology. A total of 142,428,180 high-quality reads of mycelium and 70,594,319 high-quality reads of 2-months-old sclerotium were obtained. Additionally, differentially expressed genes from the W. cocos mycelium and 2-months-old sclerotium stages were analyzed, resulting in identification of 69 CAZymes genes which were significantly up-regulated during the early stage of sclerotial growth compared to that of in mycelium stage, and more than half of them belonged to glycosyl hydrolases (GHs) family, indicating the importance of W. cocos GHs family for degrading the pine woods. And qRT-PCR was further used to confirm the expression pattern of these up-regulated CAZymes genes. Our results will provide comprehensive CAZymes genes expression information during W. cocos sclerotial growth at the transcriptional level and will lay a foundation for functional genes studies in this fungus. In addition, our study will also facilitate the efficient use of limited pine resources, which is significant for promoting steady development of Chinese W. cocos industry.

Antagonistic effects of bacteriocin and lactobacillales on stem rot causing Sclerotium rolfsii in betel

Economic potentiality of betel vine ( Piper betle L.) is always under threat of fungal diseases caused by various soil borne phytopathogenic fungi. Betel vine production accounts for extensive economic loss due to collar rot caused by Sclerotium rolfsii . With a view to search an eco-friendly biocontrol system for the management of S. rolfsii , we have studied the antagonistic effect of a bacteriocinogenic Lactic Acid Bacteria (LAB) strain, Lactococcus lactis JC10 and produced bacteriocin on this pathogen. Significant decrease in mycelia growth of Sclerotium in dual culture plate experiment was observed. There is also noteworthy amount of decrease in growth rate of Sclerotium mycelium in presence of viable L. lactis JC10 in medium. Crude bacteriocin (25% v/v) in a PDA plate can totally seize the mycelia growth. It was also found that crude bacteriocin treatment of Sclerotium spore can delay germination considerably.

Inhibitory Effects of Powdered Caraway and Peppermint Extracts on Pea Root Rot under Greenhouse Conditions

Inhibitory Effects of Powdered Caraway and Peppermint Extracts on Pea Root Rot under Greenhouse ConditionsThe effects of caraway and peppermint extracts was evaluated at concentrations of 2, 4, 6, 8 and 10%, respectively on the radial mycelial growth ofSclerotium rolfsii.High significant inhibitory effect on radial fungal growth was observed with different concentrations of each of plant extracts. Concentration of 6% and more of the two extracts in combination were able to cause complete growth inhibition of the tested fungus. In greenhouse, the efficacy of plant extracts in addition to the fungicide Rizolex-T as seed dressing on pea root rot incidence was evaluated in pot experiment using soil artificially infested with the disease agent. All treatments showed a significant reduction in disease incidence compared with the control treatment. Rizolex-T followed by combination of caraway and peppermint extracts as a mixture showed superior reduction effect on root rot disease incidence at pre-, and post-emergence growth stages than individual treatment with each of extracts. The usage of caraway and peppermint extracts for seed dressing before sowing might be applied as control measure for controlling root rot diseases.

English

Investigation was carried out on Sclerotium rolfsii isolated from four plant parts on PDA media for differences in the mycelia and sclerotia growth, in the tropical humid lowlands of Southeastern Nigeria. The result reveals that there were variation in the number and size (in diameter) of the sclerotia produced by the isolates. The mycelia growth rate was same in all the isolates covering the 9.0 cm plates at the 7 and 8 day after inoculation. These sclerotia constitute the major source of inoculum for the organism in nature. Key words: Sclerotium rolfsii, sclerotia, mycelia, inoculum, infection loci.

In vitro growth of four isolates of Sclerotium rolfsii Sacc in the humid tropics

Investigation was carried out on Sclerotium rolfsii isolated from four plant parts on PDA media for differences in the mycelia and sclerotia growth, in the tropical humid lowlands of Southeastern Nigeria. The result reveals that there were variation in the number and size (in diameter) of the sclerotia produced by the isolates. The mycelia growth rate was same in all the isolates covering the 9.0 cm plates at the 7 and 8 day after inoculation. These sclerotia constitute the major source of inoculum for the organism in nature.

Influence of inocula and grains on sclerotia biomass and carotenoid yield of Penicillium sp. PT95 during solid-state fermentation.

Various inocula and grains were evaluated for carotenoid production by solid-state fermentation using Penicillium sp. PT95. Millet medium was more effective in both sclerotia growth and carotenoid production than other grain media. An inoculum in the form of sclerotia yielded higher sclerotia biomass compared to either a spore inoculum or a mycelial pellet inoculum. Adding wheat bran to grain medium favored the formation of sclerotia. However, neither the inoculum type nor addition of wheat bran resulted in a significant change in the carotenoid content of sclerotia. Among grain media supplemented with wheat bran (wheat bran:grain =1:4 w/w, dry basis), a medium consisting of rice and wheat bran gave the highest sclerotia biomass (15.10 g/100 g grain), a medium consisting of buckwheat and wheat bran gave the highest content of carotenoid in sclerotia (0.826 mg/g dry sclerotia), and a medium consisting of millet and wheat bran gave the highest carotenoid yield (11.457 mg/100 g grain).

Sclerotia growth and carotenoid production of Penicillium sp PT95 during solid state fermentation of corn meal

Using corn meal as fermentation substrate, the effect of some factors, fermentation time and supplementation of saccharide and nitrogen sources as well as vegetable oil, on the sclerotia growth and carotenoid production of Penicillium sp PT95 during solid state fermentation were studied. When PT95 strain was grown on the amended medium by supplementing of 3g NaNO3, 10g maltose and 2.5g soybean oil per liter of salt solution to basal medium for 20 days, the dry sclerotia weight and carotenoid yield reached 9.70 g and 5260 μg / 100 g of substrate, respectively. Without supplementation only 5.36g dry sclerotia and 2149μg carotenoid / 100 g of substrate was attained. © Rapid Science Ltd. 1998

The effect of aeration and agitation on Claviceps purpurea dimorphism and alkaloid synthesis during submerged fermentation

Varying the air flow rate (vvm) in a fermentor under constant drive speed, Claviceps purpurea dimorphism as well as alkaloid biosynthesis were greatly influenced. At a high flow rate (2.5 vvm) sclerotial growth was favoured in seed and in production media, while at a low air flow rate (1.0 vvm) sphacelial growth dominated. When using high flow rates the oxygen uptake rate was small, but at low flow rates it increased markedly. In both cases the alkaloid production was lower than at the intermediate value of 1.5 vvm of air flow rate, which proved to be optimal. This could be explained by the difference in the air/water interface and two-phase oxygen uptake. At a high air/water interface direct oxygen uptake from the gaseous phase prevails, while at a low air/water interface uptake is due to the oxygen liquid-phase only. Thus for optimal fungal development and alkaloid production a compromise between uptake from the liquid and the gaseous phase has to be established by a defined ratio between aeration and agitation.

Polysaccharide cytochemistry in maturing Aspergillus flavus sclerotia

Polysaccharide content and the dynamics of its accumulation was studied during sclerotial growth and differentiation in Aspergillus flavus. Using cytochemical methods and enzymic digestion with α-amylase on ultrathin sections, it was confirmed that glycogen represented the main polysaccharide storage material. During sclerotial development glycogen progressively accumulated in the cytoplasm in the form of α- and β-particles, while in the vacuolar system it appeared only as β-particles. In mature sclerotia most of the glycogen was found in the cytoplasmic rather than the vacuolar form and it is therefore suggested that the cytoplasmic form of glycogen is the main storage polysaccharide accumulated for survival and germination.

Biochemical characterization of the inoculum of Claviceps purpurea for submerged production of ergot alkaloids

Biochemical and morphological changes during the growth and development of Claviceps purpurea seed cultures under submerged conditions were established. The fall in RNA content at about the middle of the exponential phase of growth has been found to be associated with the transition from sphacelial to sclerotial growth and may be considered the major indicator of changes in the metabolism of the fungus. It may also serve as a biochemical indicator for the optimal activity of Claviceps purpurea seed cultures.

Changes in respiration patterns of Claviceps purpurea during parasitic development

During parasitic development of Claviceps purpurea (Fr.) Tul. on rye the initial sporulating sphacelial mycelium had an endogenous respiration consistent with oxidation of carbohydrate. Exogenously supplied sugars were fermented. Differentiation to the plectenchymatic sclerotial growth form was accompanied by a large decrease in endogenous respiration and in the ability to utilize exogenously supplied sugars. However, the greatest fermentative capacity of all tissues was found in actively growing sclerotial mycelia. With increasing age sclerotial tissues resorted to oxidation of lipid. Sphacelial and sclerotial tissues grown in axenic culture did not exhibit these distinctive capacities for carbohydrate utilization. It is concluded that the marked changes during parasitism can be ascribed primarily to the morphological and organisational differences between the two types of tissue.

Differentiation of Claviceps purpurea in Axenic Culture

The growth form of a strain of Claviceps purpurea in axenic culture has been controlled by the amino nitrogen source. Within the pairs asparagine/aspartic acid or glutamine/glutamic acid the amide promoted sphacelial growth of the colony whereas the acid supported differentiation of plectenchymatic sclerotial tissue and synthesis of ergot alkaloids. Sclerotial colonies showed purple pigmentation. The mycelium had a greater lipid content, rich in ricinoleic acid, and sporulation was much less than in sphacelial colonies. Changes in the relative distribution of amino acids between the free and peptidyl components of the cells was most marked with respect to lysine; growth on asparagine resulted in more than half remaining free, whereas less than 10% remained free on aspartic acid. Lysine supplied exogenously as a nitrogen source did not promote sclerotial differentiation. Frequent transfers to fresh medium of colonies grown on dialysis membrane accentuated the extent of sphacelial or sclerotial growth; four transfers during an 18-day growth period yielded mycelia with an alkaloid content (0-4%, w/w) similar to that of parasitic ergot sclerotia. Asparagine and glutamine were taken up more rapidly from liquid media than their corresponding acids, but each acid exerted a dominant effect over the amide, in a mixture providing equivalent nitrogen, resulting in differentiation from sphacelial to sclerotial growth analogous to that occurring during parasitism. The apparently greater (w/w) proportion of total amino acids in sphacelial mycelia than in sclerotial mycelia mainly reflected the lower lipid content of these tissues, but this factor was insufficient to account for the persistence of a significant proportion of the total lysine amongst the free amino acids. The promotion of sclerotial growth by aspartic and glutamic acids was not confined solely to the experimental strain of the fungus. Although some isolates failed to differentiate into plectenchymatic mycelia on these nitrogen sources, the extent of their sphacelial growth, as indicated by the degree of sporulation, was always much reduced with respect to that promoted by asparagine.

Effect of ethanol on production of sclerotia by Sclerotium rolfsii

Mycelial growth of Sclerotium rolfsii Sacc. was prolonged, initiation and maturation of sclerotia were delayed, and fewer sclerotia were formed, when ethanol was added to the growth medium at concentrations from 0·5 to 3% (v/v). Similar changes were induced by exposing colonies to ethanol vapour at various stages before the initiation of sclerotia. On liquid glucose-containing medium, total dry weight was little affected by ethanol levels from 0·5 to 3% (v/v). Lower levels of ethanol (0·5–1·5 %, v/v) caused stimulation of total growth, but the relative amount of sclerotial growth decreased with increasing ethanol concentration. The fungus grew sparsely with ethanol as the sole source of carbon. The growth of three other isolates of S. rolfsii and one of Sclerotinia sclerotiorum (Lib.) de Bary was similarly affected by ethanol.