菊芋叶片提取物抑菌活性与化学成分的研究

为了开发新型植物源杀菌剂和充分利用菊芋(Helianthus tuberosus L.)资源,本文尝试用石油醚、乙醚、乙酸乙酯和水等4种溶剂对菊芋叶片进行平行提取,采用生长速率法测定菊芋叶片提取物对水稻纹枯菌(Rhizoc-tonia solaniKühn)、小麦赤霉菌[Gibberella zeae(Schw.)Petch]、番茄早疫菌[Alternaria solani(Ellis et Martin)Jones et Grour]和番茄灰霉菌(Botrytis cinerea Pers.)生长量的抑制活性;并通过试管法和滤纸法对菊芋叶片内化学成分进行初步预试。抑菌实验结果表明:(1)菊芋叶片各溶剂提取物处理与对照处理相比差异显著;(2)菊芋叶片水提取物处理与各有机溶剂提取物处理差异也基本显著;(3)菊芋叶片各溶剂提取物同浓度处理对水稻纹枯菌、番茄早疫菌和番茄灰霉菌抑制效果较好;(4)菊芋叶片乙酸乙酯提取物抑菌效果最为显著,浓度为20mg/mL时对番茄早疫菌和番茄灰霉菌已达到完全抑制,对水稻纹枯菌抑制率也达到77.91%。初步化学预试结果说明,菊芋叶片中含有蛋白质、氨基酸、还原糖类、有机酸、酚类和鞣质、黄酮类、内酯类、强心甙以及油脂等化学成分。     

中国金孢属一新记录种

Fermentation broth of endophytic fungus Trichoderma taxi ZJUF0986 has high antifungal activity to the common 15 species of phytopathogenic fungi. Three bioactive metabolites I, II and III were obtained by extracted with petroleum ether, ethyl acetate and n-butanol and subsequent silica gel column chromatography and preparative HPLC. Among them, compound I, the main metabolite, has strong broad-spectrum antifungal activity, especially to Botrytis cinerea, Rhizoctonia solani and Sclerotinia sclerotiorum with IC50 1.06, 1.08 and 1.13 mg/L, respectively.     

紫杉木霉ZJUF0986抗真菌活性代谢产物分离提取及其活性研究

南方红豆杉Taxus chinensis var.maimi内生真菌紫杉木霉Trichoderma taxi菌株ZJUF0986发酵液对15种常见植物病原真菌具有很强的抑菌活性;用石油醚、乙酸乙酯和正丁醇对发酵液进行活性物质的提取分离,并通过硅胶柱层析和制备HPLC纯化,得到3个活性代谢产物Ⅰ、Ⅱ和Ⅲ,其中主要活性代谢产物Ⅰ具有广谱高效抑制植物病原真菌的特点,特别是对灰葡萄孢Botrytis cinema、立桔丝核菌Rhizoctonia solani、核盘菌Sclerotinia sclerotiorum具有很强的抑菌活性,其IC50为1.1mg/L.     

中国金孢属一新记录种

金孢属Chrysosporium是Corda于1833年以革质金孢C. corii Corda为模式种建立的一类有丝分裂产孢真菌.它的有性型主要隶属爪甲团囊菌目Onygenales中的爪甲团囊菌科Onygenaceae和裸囊菌科Arthrodermataceae(Kirk et al.2001;Oorschot 1980).此属真菌大多能分解角蛋白,极有应用价值.     

三种苔藓植物提取物对植物病原菌的抑菌性研究

以立枯丝核菌(Rhizoctoniasolani)、棉花枯萎病菌(Fusariumoxysporum)、苹果轮纹病菌(Physalosporapiricolanosa)、草莓灰霉病菌(Botrytiscinerea)四种植物病原菌为供试病原菌,对大镰刀藓Drepanocladusexannulatus、锐尖匍灯藓Plagiomniumacutum和疣小金发藓Pogonatumurnigerum三种藓类醇提液进行抑菌活性筛选,结果表明,大镰刀藓提取液对立枯丝核菌有较好的抑制作用,对立枯丝核菌的EC50为0.878mg/mL;而锐尖匍灯藓的提取液对立枯丝核菌的生长却有促进作用。     

Bacisubin, an antifungal protein with ribonuclease and hemagglutinating activities from Bacillus subtilis strain B-916

An antifungal protein, with a molecular mass of 41.9 kDa, and designated as bacisubin, was isolated from a culture of Bacillus subtilis strain B-916. The isolation procedure consisted of ion exchange chromatography on DEAE-Sepharose Fast Flow, and fast protein liquid chromatography on Phenyl Sepharose 6 Fast Flow and hydroxyapatite columns. The protein was adsorbed on all three chromatographic media. Bacisubin exhibited inhibitory activity on mycelial growth in Magnaporthe grisease, Sclerotinia sclerotiorum, Rhizoctonia solani, Alternaria oleracea, A. brassicae and Botrytis cinerea. The IC50 values of its antifungal activity toward the last four fungal species were 4.01 microM, 0.087 microM, 0.055 microM and 2.74 microM, respectively. Bacisubin demonstrated neither protease activity, nor protease inhibitory activity. However, it manifested ribonuclease and hemagglutinating activities.     

Isolation and anti-oomycete activity of nyasol from Anemarrhena asphodeloides rhizomes

The methanol extract of Anemarrhena asphodeloides rhizomes exhibited strong antifungal activity against the plant pathogenic fungi Magnaphothe grisea, Rhizoctonia solani, and the plant pathogenic oomycete Phytophthora capsici. The antifungal substance isolated from the rhizomes of A. asphodeloides was identified to be nyasol, (Z)-1,3-bis(4-hydroxyphenyl)-1,4-pentadiene by NMR and mass spectral analysis. Nyasol effectively inhibited the mycelial growth of Colletotrichum orbiculare, P. capsici, Pythium ultimum, R. solani, and Cladosporium cucumerinum in a range of 1-50 mug/ml, but did not affect the growth of bacteria and yeast. In a greenhouse test, treatment with the antifungal compound nyasol was significantly effective in suppressing the Phytophthora blight on pepper plants.     

Effects of neolignans from the stem bark of Magnolia obovata on plant pathogenic fungi

Aims:  To characterize antifungal principles from the methanol extract of Magnolia obovata and to evaluate their antifungal activities against various plant pathogenic fungi.
Methods and Results:  Four neolignans were isolated from stem bark of M. obovata as antifungal principles and identified as magnolol, honokiol, 4-methoxyhonokiol and obovatol. In mycelial growth inhibition assay, both magnolol and honokiol displayed more potent antifungal activity than 4-methoxyhonokiol and obovatol. Both magnolol and honokiol showed similar in vivo antifungal spectrum against seven plant diseases tested; both compounds effectively suppressed the development of rice blast, tomato late blight, wheat leaf rust and red pepper anthracnose. 4-Methoxyhonokiol and obovatol were highly active to only rice blast and wheat leaf rust respectively.
Conclusions:  The extract of M. obovata and four neolignans had potent in vivo antifungal activities against plant pathogenic fungi.
Significance and Impact of the Study:  Neolignans from Magnolia spp. can be used and suggested as a novel antifungal lead compound for the development of new fungicide and directly as a natural fungicide for the control of plant diseases such as rice blast and wheat leaf rust.     

The production of antifungal volatiles by Bacillus subtilis

A strain of Bacillus subtilis which produces an antibiotic metabolite was also found to produce a volatile compound(s) which was antifungal to Rhizoctonia solani and Pythium ultimum.
Growth of the fungi was severely impaired in the presence of the volatiles and physiological abnormalities of the hyphae were observed, including hyphal distortion and vacuolation. A range of media were tested for volatile production and potato dextrose agar (PDA) was found to be the most active. Temperature had a considerable effect on antifungal volatile activity with the greatest inhibition occurring at 30°C. Addition of iron (III) chloride to Sabouraud's glucose agar (SGA) also enhanced the antifungal effect. The volatiles were found to be water soluble and remained active when trapped in SGA.     

Mycelial growth inhibition of plant pathogenic fungi by extracts of South African plant species

Eleven plant species were collected from the Blyde River Canyon area, Mpumalanga, South Africa. Methanolic crude extracts of the plant samples were concentrated to dryness, dissolved in sterile distilled water and bioassayed in the laboratory for their fungicidal potential against seven plant fungal pathogens, namely Botrytis cinerea, Fusarium oxysporum, Sclerotium rolfsii, Rhizoctonia solani, Verticillium dahliae, Botryosphaeria dothidea and Pythium ultimum, at equal concentrations. A modified agar dilution method was used to determine the inhibitory effect of the plant extracts on the mycelial radial growth of the plant pathogens. Antifungal activity was confirmed in all the plants, but not in all plant parts and also not against all the test organisms. The crude extract of Eucomis autumnalis performed best of all the plant extracts tested, as it showed significant antifungal activity against all seven of the plant pathogenic test organisms and compared favourably to the inhibition of the mycelial growth by a broad spectrum synthetic fungicide (carbendazim/ difenoconazole). The crude extract of Schrebera alata came out second best, in the sense that at least one plant part inhibited the mycelial radial growth of four of the seven test organisms by at least 50%.     

Histopathological studies of sclerotia of Rhizoctonia solani parasitized by the EGFP transformant of Trichoderma virens

Aims:  The aim of the study was to investigate the antagonistic interactions of Trichoderma species against Rhizoctonia solani sclerotia by enhanced green fluorescence protein (EGFP)-tagged transformant of Trichoderma virens TY009.
Methods and Results:  An EGFP was used as a report gene for transforming T. virens strain, and a stable EGFP transformant GF5 was obtained with the mycoparasitic activity against sclerotia of R. solani . Observation of parasitized sclerotia by fluorescence microscopy showed hyphae of transformant GF5 was able to invade into sclerotia and its colonization was mainly intercellar with uniformly distributed mycelium in sclerotia. The host cells were colonized, penetrated, and then the whole cells were replaced by transformant GF5 hyphae. Chlamydospores were seen after 10 days but mature ones after 20 days. Sclerotia became soft and decayed after 40 days but a few cells seemed not to be colonized completely.
Conclusions:  Trichoderma virens was able to parasitize sclerotia to make sclerotia soft and decayed, and its colonization was mainly intercellar in sclerotial tissues.
Significance and Impact of the Study:  This is first report of parasitism of sclerotia of R. solani by EGFP-tagged transformant, providing useful information for using T. virens as effective biocontrol agent.     

357细菌及其拮抗物质对植物病原真菌的抑制

Bacillus cereus357细菌及其分泌的拮抗物质对水稻纹枯病菌和草莓灰霉病菌有较好的拮抗作用。它们可抑制或杀死这两种病原菌的菌丝,可使菌丝枯萎和产生畸形。357细菌分泌的拮抗物质还可抑制草莓灰霉病菌孢子的萌发和孢子的形成。该拮抗物质对热、酸、碱等有较高的稳定性。     

Isolation,characterisation of major secondary metabolites of the Himalayan Trichoderma koningii and their antifungal activity

Ethyl acetate extracts of two strains of Trichoderma koningii were evaluated for antifungal activity against soilborne pathogenic fungi, Rhizoctonia solani, Sclerotium rolfsii, Macrophomina phaseolina and Fusarium oxysporum by poisoned food technique. Secondary metabolites, namely δ-decanolactone, 6-pentyl-α-pyranone and 6-(4-oxopentyl)-2H-pyran-2-one were isolated from T. koningii (T-8) extract while palmitic acid, 6-pentyl-α-pyranone and stigmasterol were isolated from T. koningii (T-11) extract. Secondary metabolites were identified by ¹H NMR, ¹³C NMR and mass spectroscopic methods. These metabolites were evaluated for antifungal activity against the test pathogens 6-Pentyl-α-pyranone and 6-(4-oxopentyl)-2H-pyran-2-one exhibited excellent antifungal activity against S. rolfsii. The antifungal activity though slightly less was comparable to that of hexaconazole, a commercial fungicide.     

藓类提取物对植物病原真菌的抑菌性研究

以立枯丝核菌(Rhizoctonia)、尖孢镰孢菌(Fusarium oxysporum)、苹果轮纹病菌(Macrophoma kawatsukai)、梨黑星病菌(Fusicladium pirina)、草莓灰霉菌(Botrytis cinerea)等七种植物病原真菌为供试菌,通过生长速率法测定了24种藓类的乙醇提取物对病原真菌的抑制作用.结果表明,在提取物浓度为1.2mg干样/mL,大镰刀藓提取物对立枯丝核菌的抑制作用最强,抑菌率为67.8%.藓类提取物不仅对植物病原真菌有抑菌活性,而且还有多种藓类提取物对病原真菌菌丝有促进生长活性,如锐尖匍灯藓对立枯丝核菌菌丝生长的促进率为95.1%.选出大镰刀藓、扁灰藓、紫萼藓、山地水灰藓、疣小金发藓、橙色净口藓、锐尖匍灯藓和羊角藓对立枯丝核菌和灰霉菌的抑菌性或菌丝生长促进性进行了进一步研究,藓类提取物对病原菌的抑菌性或菌丝生长促进性在一定范围内随浓度的增加而增强,并且抑菌性随时间的延长呈下降趋势.     

First isolation of a novel thermostable antifungal peptide secreted by Aspergillus clavatus

A novel antifungal peptide produced by an indigenous fungal strain (VR) of Aspergillus clavatus was purified. The antifungal peptide was enriched in the supernatant after heat treatment at 70 degrees C. The thermostable character was exploited in the first purification step, as purified peptide was obtained after ultrafiltration and reverse phase-HPLC on C18 column application. The purified peptide named "AcAFP" for A. clavatus antifungal peptide, has molecular mass of 5773Da determined by MALDI-ToF spectrometry. The N-terminal sequence showed a notable identity to the limited family of antifungal peptides produced by ascomycetes fungi. The AcAFP activity remains intact even after heat treatment at 100 degrees C for 1h confirming its thermostability. It exhibits a strong inhibitory activity against mycelial growth of several serious human and plant pathogenic fungi: Fusariuym oxysporum, Fusarium solani, Aspergillus niger, Botrytis cinerea, Alternaria solani, whereas AcAFP did not affect yeast and bacterial growth.     

两种测定化合物对真菌菌丝生长抑制活性方法的比较

采用带毒平板法和液体培养法测定两种植物抗菌化合物血根碱和白屈菜红碱对植物病原真菌水稻纹枯病菌（Rhizoctonia solani）、番茄枯萎病菌（Fusarium oxysporium f．sp．lycopersici）和棉花枯萎病菌（F.oxysporium f.sp．vasinfectum）的抑制活性。论文对两种测定方法进行了比较和讨论,结果发现：和带毒平板法相比,采用液体培养法获得的化合物半抑制浓度要低。