In Vitro Antifungal Activity of the Extract and Compound from Acorus tatarinowii Against Seven Plant Pathogenic Fungi

Acorus tatarinowii Schott is a traditional Chinese medicine plant and has multiple bioactivities in medicine and pesticide field. In this study, the antifungal compound 1,2-dimethoxy-4（2-propenyl） benzene was isolated from A. tatarinowii Schott by activity-directed isolation method, and the inhibitory activity of the extract and 1,2-dimethoxy-4（2-propenyl） benzene against seven plant pathogenic fungi was evaluated. The results showed that the extract and 1,2-dimethoxy-4（2- propenyl） benzene had high inhibitory activity against hyphal growth of Thielaviopsis paradoxa （de Seynes） V. Hohnel, Pestalotia mangiferae P. Henn., Fusarium oxysporum f. sp. niveum （E. F. Smith） Syn. et Hans., Alternaria alternate Tanaka, Colletotrichum musae （Berk et Curt） V. Arx, Sphaceloma fawcettii Jenk., and Mycosphaerella sentina （Fr.） Schroter. The EC50 values of extract were 1.6162, 1.6811, 1.1253, 3.5771, 1.7024, 2.2284, and 2.2221 g L^-1, respectively, and the EC50 values of 1,2-dimethoxy-4（2-propenyl） benzene were 0.1021, 0.0997, 0.0805, 0.1742, 0.1503, 0.1853, and 0.1924 g L^-1, respectively. 1,2-Dimethoxy-4（2-propenyl） benzene also inhibited spores germination of T. paradoxa （de Seynes） V. Hohnel and F. oxysporum f. sp. niveum （E. F. Smith） Syn. et Hans., with the inhibitory rates of 98.81 and 100% at a concentration of 0.4 g L^-1 after 8 h, respectively. 1,2-Dimethoxy-4（2-propenyl） benzene is a potential botanical antifungal agent for controling of plant fungal diseases.

Fungi Associated with Sand and Plants from Marine Coastlines: Potential Relevance for Human Health

The fungal community associated with beach sand and plants located along marine coasts are an under-studied area of research despite its potential relevance to human health. In this study, we isolated and identified the cultivable mycobiota associated with sand and plants collected along the coast of Gran Canaria (Spain) using culture-dependent and -independent methods. Clinically relevant species belonging to Cryptococcus spp. and related genera such as Naganishia and Papilotrema were isolated and identified from shoreline plants. Moreover, Candida tropicalis was isolated from beach sand, and Aspergillus fumigatus and Aspergillus terreus strains were associated with both types of samples (i.e., plants and beach sand). We conclude that beach sand and shoreline plants are potential reservoirs of fungi of high clinical interest. We recommend including beach sand and plants from the environment when assessing the quality of marine coastal systems. Our results open a framework for studying the natural marine environment and its role in the epidemiology of infectious diseases in order to more accurately manage public health.

Identification of Plant-Pathogenic Fungi Using Fourier Transform Infrared Spectroscopy Combined with Chemometric Analyses

Identification of plant-pathogenic fungi is time-consuming due to cultivation and microscopic examination and can be influenced by the interpretation of the micro-morphological characters observed.The present investigation aimed to create a simple but sophisticated method for the identification of plant-pathogenic fungi by Fourier transform infrared(FTIR)spectroscopy.In this study,FTIR-attenuated total reflectance(ATR)spectroscopy was used in combination with chemometric analysis for identification of important pathogenic fungi of horticultural plants.Mixtures of mycelia and spores from 27fungal strains belonging to nine different families were collected from liquid PD or solid PDA media cultures and subjected to FTIR-ATR spectroscopy measurements.The FTIR-ATR spectra ranging from 4 000to 400cm-1 were obtained.To classify the FTIRATR spectra,cluster analysis was compared with canonical vitiate analysis(CVA)in the spectral regions of3 050~2 800and 1 800~900cm-1.Results showed that the identification accuracies achieved 97.53%and99.18%for the cluster analysis and CVA analysis,respectively,demonstrating the high potential of this technique for fungal strain identification.

Effectiveness of Ringworm Cassia and Turmeric Plant Extracts on Growth Inhibition against Some Important Plant Pathogenic Fungi

Crude plant extracts of ringworm cassia, Cassia alata L. and turmeric, Curcuma longa L. were prepared by either hot water or organic solvents such as ethanol and ether. Various concentrations of the crude extract were then subjected to an in vitro test for their effectiveness on mycelia growth inhibition against some important plant pathogenic fungi such as Alternaria alternata, Colletotrichum gloeosporioides, Fusarium oxysporum fsp. lycopersici, Sclerotium rolfsii, Phytophthora infestans and Pythium sp. in comparison to commercial fungicides such as copper oxychloride and mancozeb. Reduction of the fungal growth was significantly obtained with C. longa extracts and the best median effective inhibitory concentration (IC50) value of 6.07, 6.50 and 7.13 mg/ml was from the ethanol extract for S. rolfsii, C. gloeosporioides and F. oxysporum fsp. lycopersici respectively. While all extracts from C. alata were almost the least effective against these fungi. The efficacy of C. longa extracts therefore, provided an alternative regime for the control of the fungal diseases and a promising appreciable choice for a replacement of chemical fungicides.

Antimicrobial Activity of Wild Plant Seed Extracts against Human Bacterial and Plant Fungal Pathogens

Five wild plant species belonging to different families (Chenopodium album, Plantago major, Elytrigia elongata, Filipendula ulmaria and Nigella sativa) widely spread in Russian Federation and the former USSR were evaluated for their ability to inhibit growth of two important human food-borne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes strain EGD-e) and eight plant pathogens (Alternaria alternata, Alternaria tenuissima, Bipolaris sorokiniana, Stagonospora nodorum, Fusarium solani, Fusarium oxysporum, Fusarium culmorum and Phytophtora infestans). To isolate biologically active compounds from seeds, a step-wise procedure including extraction with hexane, ethyl acetate, ethanol, and 10% acetic acid followed by reversed-phase HPLC was developed. Using disc-diffusion assay, the highest activity against E. coli O157:H7 was observed with extracts from F. ulmaria (hexane and ethyl acetate extracts and the unbound RP-HPLC fraction) and P. major (ethyl acetate extract and the unbound RP-HPLC fraction);E. elongate (the unbound RP-HPLC fraction) was less active. The extracts from P. major and E. elongate (the unbound RP-HPLC fractions) were equally highly active against L. monocytogenes, while those of F. ulmaria (the unbound RP-HPLC fraction) and N. sativa (hexane and ethyl acetate extracts) were less active against this pathogen. The dynamics of L. monocytogenes EGD-е and E. coli O157:H7 growth in the presence of two most potent extracts (RP-HPLC-unbound fractions of P. major and E. elongate and the hexane extract of F. ulmaria) was studied.

Identification of candidate soil microbes responsible for small-scale heterogeneity in strawberry plant vigour

Studies were conducted to identify candidate soil microbes responsible for observed differences in strawberry vigour at a small spatial scale, which was not associated with visual disease symptoms. Samples were obtained from the soils close to the rhizosphere of ‘big＇ and ‘small＇ plants from small plots which exhibited large local heterogeneity in plant vigour. A metabarcoding approach was used to profile bacterial and fungal compositions, using two primer pairs for 16 S ribosomal RNA genes（16S r DNA） and one for the fungal internal transcribed spacer（ITS） region. Of the two 16 S r DNA primer sets, the 341F/805 R resulted in sequences of better quality. A total 28 operational taxonomic units（OTUs） had differential relative abundance between samples from ‘big＇ and ‘small＇ plants. However, plausible biological explanation was only possible for three fungal OTUs. Two were possible phytopathogens： Verticillium spp. and Alternaria alternata although the latter has never been considered as a main pathogen of strawberry in the UK. For samples from ‘small＇ plants, the abundance of these OTUs was much greater than from ‘big＇ plants. The opposite was true for a mycorrhizal OTU. These results suggest that soil microbes related to crop production can be identified using metabarcoding technique. Further research is needed to assess whether A. alternata and Verticillium spp. could affect strawberry growth in the field.

无子刺梨内生真菌拮抗活性菌株筛选及分子鉴定

为进一步开发利用无子刺梨枝条和叶片中具有生物防治潜力的内生真菌,以2种木腐菌和4种植物病原真菌为靶标,采用五点对峙法筛选对病原真菌具有拮抗活性的内生真菌菌株,并探讨活性菌株的分类地位。结果显示,菌株RSZF 36、RSZF 39和RSYF 34对2种木腐菌的抑制率均达80%以上,菌株RSZF 34和RSZF 36对4种植物病原真菌的抑制率均大于75%,菌株RSZF 36对6种病原菌的抑制率均高于75%,呈现强抑制能力。84株内生真菌中有12株对6种病原菌抑制率均在70%以上,属于活性菌株,其中9株来自枝条,3株来自叶片。对12株活性菌株进行分子生物学鉴定,将其归属为1门2纲3目3科4属,拟盘多毛孢属(Pestalotiopsis)为优势菌属,占活性菌株的41.67%。综上,无子刺梨内生真菌中广泛分布着抑菌活性菌株,且不同内生菌株对不同病原菌的拮抗能力表现出一定的选择性或专一性。本研究结果可为无子刺梨内生真菌资源的开发利用提供理论参考。

独蒜兰提取物对植物病原真菌的抑菌活性

【目的】研究独蒜兰假鳞茎乙醇提取物对植物病原真菌的抑菌作用,为植物源杀菌剂的开发提供依据。【方法】采用菌丝生长速率法,研究独蒜兰假鳞茎乙醇提取物对15种植物病原菌的抑制活性;以西瓜尖孢镰刀菌作为供试菌,进一步研究该提取物对病原真菌的菌丝干重、细胞膜、过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)等的影响。【结果】独蒜兰假鳞茎提取物对辣椒疫霉病菌、西瓜尖孢镰刀菌、番茄灰霉病菌和非洲隐地疫霉菌的抑菌效果明显,其EC_(50)值分别为0.849、0.782、0.813和1.161 mg·mL^(-1);经独蒜兰提取物处理后的西瓜尖孢镰刀菌菌丝干重随着药剂浓度的增加而减少;细胞膜丙二醛含量和相对电导率增加;菌丝体细胞内CAT、POD和SOD 3种保护酶活性增加。【结论】独蒜兰假鳞茎提取物对植物病原真菌具有较好的抑菌活性,其抑菌作用可能与其干扰菌丝生长、使菌丝细胞膜正常功能受损等有关。

圆叶青藤内生真菌Phlebiopsis crassa次生代谢产物研究

本文对分离自圆叶青藤(Illigera orbiculata C.Y.Wu)的一株内生真菌Phlebiopsis crassa MYYJ-5-1的次生代谢产物进行了研究,从其土豆固体发酵物中分离得到9个化合物,分别鉴定为:benzophenone(1)、p-hydroxybenzoic acid(2)、p-hydroxybenzaldehyde(3)、indole-3-carboxaldehyde(4)、indole-3-carboxylic acid(5)、thymidine(6)、uracil(7)、thymine(8)、mono-methyl succinate(9)。同时采用二倍稀释法对化合物进行体外抗植物病原菌活性测定,结果显示:化合物1和3对黄瓜萎蔫球菌具有中等抑制活性,MIC值均为32μg·mL^(-1);化合物5对人参链格孢菌具有中等抑制活性,MIC值为32μg·mL^(-1);化合物6对黑曲霉菌具有显著抑制活性,活性等同于阳性对照制霉素,MIC值均为16μg·mL^(-1)。

植物病原真菌肌球蛋白及其抑制剂研究进展

肌球蛋白是一类在真核生物中广泛存在的功能性蛋白,在生物体细胞内的各项生命活动过程中扮演着重要的角色。此外,来源于植物病原真菌的肌球蛋白对真菌的生长发育和致病性也有着重要影响。基于这些特性,越来越多研究者把目光投入到了以肌球蛋白为靶标的农药创制研究当中。基于植物病原真菌来源的肌球蛋白进行抑制剂设计,从而开发具有新型作用机制的杀菌剂,将是防控植物病害、保障粮食安全的重要手段。本文重点介绍了植物病原真菌来源肌球蛋白及其抑制剂的研究进展,主要综述了植物病原真菌肌球蛋白的结构特征和生化功能,以及近年来已报道的肌球蛋白抑制剂类杀菌剂,可为植物病原真菌肌球蛋白靶标导向先导化合物的发现和新农药活性分子的合理设计提供参考。

利用植物病原真菌除草的研究进展

杂草对农业生产危害巨大,长期使用化学除草剂带来诸多问题,促使新型环保高效除草剂的开发需求迫切。植物病原真菌源除草剂是利用活体真菌或其代谢产物开发的生物除草剂,具有资源丰富、低毒性、环境污染小等优点,成为新型除草剂开发的重要资源。文章总结了具有除草活性的植物病原真菌及其代谢产物的种类,综述了利用植物病原真菌除草机理及应用现状,并对未来植物病原真菌源除草剂的发展进行了展望。

表观遗传调控在植物病原真菌发育和致病过程中的作用与分子机制

经过长期进化,植物病原真菌发展出多种复杂而精妙的侵染宿主策略。在植物与病原菌互作过程中,病原菌致病基因的精准表达与调控起至关重要的作用。表观遗传调控是指通过化学修饰改变染色体上的DNA和蛋白质,从而调控基因表达的过程,包括DNA甲基化、组蛋白甲基化、组蛋白乙酰化以及染色质重塑等。越来越多的证据表明,表观遗传调控在植物病原真菌基因的转录重编程中发挥重要作用,进而影响其发育、环境应激反应、次生代谢物的生物合成和致病性等方面。本文综述了近年来表观遗传调控因子方面的研究进展,包括组蛋白修饰、染色质重塑等在植物病原真菌发育和致病性中的作用,并对当前的研究进行了展望,以期为未来的病害防治研究奠定坚实的理论基础。

一株红树林来源放线菌的抗菌活性物质

红树林沉积物微生物资源丰富,是新型天然产物以及药物先导化合物的重要来源之一.为获得具有新颖结构的微生物次级代谢产物,以活性为导向对红树林来源菌株进行筛选,对表现出抗细菌、生物膜和真菌活性的GP3T5(Streptomyces sp.)菌株的次级代谢产物进行研究.通过萃取、C18反相硅胶柱层析、Sephadex LH20凝胶柱层析和制备高效液相色谱,从菌株发酵液中分离出3种化合物,分别记为化合物1、2和3.利用高分辨电喷雾质谱、核磁共振波谱和紫外光谱等技术,鉴定化合物1、2和3依次为pyrisulfoxin A、collismycin A和echinomycin A.对化合物1、2和3进行活性评价,结果表明,3种化合物均有不同程度的抗菌和抗生物膜活性.其中,化合物3的抗菌活性最好,对食源性病原菌有很好的抗菌活性,对单增李斯特菌和无乳链球菌的最小抑菌质量浓度均为0.06μg/mL.化合物1、2和3对鲍曼不动杆菌的最小生物膜抑制质量浓度分别为32、16和1μg/mL.化合物2和3具有良好的抗植物病原真菌活性,化合物2的半数抑制质量浓度IC50≤10μg/mL,化合物3的IC50≤1μg/mL.以上红树林来源菌株GP3T5的研究结果可为深入挖掘红树林土壤微生物生物活性物质提供参考.

SSR和ISSR分子标记技术应用研究

近年来,SSR和ISSR技术在生命科学领域中得到广泛应用。从亲缘关系和遗传多样性研究、DNA指纹库建立及分子标记辅助育种3个方面介绍这两种技术在植物研究中的应用情况,从植物抗病基因的筛选、生理小种的鉴定及遗传多样性研究、病原菌致病性3个方面介绍这两种技术在植物病原真菌研究中的应用情况,并对这两种技术未来的发展方向进行展望。

IN VITRO ACTIVITY OF EXTRACTS OF FIVE MEDICINAL PLANT SPECIES ON PLANT PATHOGENIC FUNGI

The antifungal effectiveness of extracts of five medicinal plant species was determined.The inhibitory activity of extracts of Eucalyptus tereticornis,Xanthium sibiricum,Artemisia argyi,Tupistra chinensis and Pyrola calliantha were evaluated against the mycelial growth of the plant pathogenic fungi Aspergillus niger,Botrytis cinerea,Penicillium digitatum,P.expansum,P.italicum and Rhizopus stolonifer.All plant extracts were prepared at 60°C using solvents(either water,50%ethanol(v/v),95%ethanol(v/v),ethyl acetate or petroleum ether).Fungicidal effects of all plants tested were confirmed.Different extracts from the same plant species gave different degrees of inhibition.All aqueous extracts had weak or no activity on all fungi tested.Ethyl acetate and 95%ethanol extracts from T.chinensis rhizomes gave greater inhibition and a broader spectrum inhibition than the other extracts.T.chinensis may have potential as a new natural fungicide and may be used for the preservation of agricultural and forestry products such as fruits and vegetables.

一种快速简便的植物病原真菌基因组DNA提取方法

尿素提取法是最新报道的一种植物拟南芥基因组DNA的提取方法,但是还没有应用于植物病原真菌DNA的提取。本研究采用改进的4种不同的方法(尿素提取法、CTAB提取法、氯化苄提取法以及SDS-CTAB提取法),分别对5种分类地位不同的植物病原真菌(草莓疫霉病菌、西瓜蔓枯病菌、草莓炭疽病菌、西瓜枯萎病菌和水稻纹枯病菌)的基因组DNA进行提取和比较。结果表明,尿素提取法和SDS-CTAB法均能提取到所有5种真菌的基因组DNA,而尿素提取法提取DNA的效率更高、质量更好,操作步骤更为快速简单。

植物病原真菌对甾醇生物合成抑制剂类(SBIs)杀菌剂的抗药性研究进展

甾醇生物合成抑制剂类(SBIs)杀菌剂通过抑制植物病原真菌甾醇生物合成途径中不同环节的酶,干扰或阻断病原菌麦角甾醇生物合成而发挥抗真菌作用。综述了植物病原真菌对SBIs杀菌剂的抗药性发生现状、遗传机制、生理生化机制、分子机制及治理策略等方面的最新研究进展。室内及田间有关SBIs杀菌剂抗药性的研究结果表明,植物病原菌对该类杀菌剂的抗药性可能是由1种或多种机制共同作用的结果。ABC和MFS运输蛋白基因及CYP51蛋白基因是植物病原真菌对SBIs杀菌剂产生抗药性的主要分子机制。其中ABC运输蛋白基因能够通过翻转酶将药剂从膜内层转移至外层而排出细胞体外;MFS运输蛋白基因的超表达和本底表达则是导致病原菌产生抗药性的关键因素;而CYP51蛋白基因与药剂作用时易在病原菌体内发生基因点突变或基因超表达,造成编码蛋白与药剂亲和力下降,导致病原菌产生抗药性。随着分子生物学的迅速发展,可从基因水平上寻找出与抗药性直接相关的基因、蛋白及调控途径等信息,同时与其他学科结合,合理设计新的、多作用位点的高效甾醇生物合成抑制剂,从而延长该类杀菌剂的使用寿命。

几种植物提取物对4种植物病原真菌的抑制作用

以链格孢菌、芭蕉炭疽菌、尖孢镰刀菌、灰葡萄孢菌等4种植物病原真菌为供试菌,对采自福建省福州地区16种植物乙醇提取物的抑菌活性进行测试,结果表明,0.0lg·mL-1下,有8种植物对至少1种病原真菌孢子的萌发抑制率达60%以上,大飞扬、杨梅、桃金娘3种植物的乙醇提取物对孢子萌发与菌丝生长都有较好的抑制效果.对这3种植物用极性从小至大的溶剂依次提取,提取物抑菌活性表明,活性物质多只分布在其中1至3种极性溶剂层中,不同极性提取物的抑菌能力与抑菌范围存在较大差异.大飞扬的丙酮提取物对芭蕉炭疽菌孢子可能具有毒害作用,乙酸乙酯提取物使芭蕉炭疽菌和链格孢菌的孢子芽管生长不正常.

11种植物精油对6种植物病原真菌的抑菌活性研究

为筛选有效的植物杀菌成分,采用菌丝生长法,测定了香茅油、薰衣草油、菊花油、月桂油、柠檬油、广藿香油、肉桂油、天竺葵油、迷迭香油、茶树油、薄荷油对6种植物病原真菌的抑菌活性。发现在2 g/L的浓度下,上述11种精油对6种供试病菌均有明显的抑制作用,其中香茅油、肉桂油、天竺葵油、月桂油、茶树油和薄荷油对6种病原真菌的抑制率均为100%。剂量效应试验表明,肉桂油对灰葡萄孢(Botrytis cinerea)和禾谷镰孢菌(Fusariumgraminearum)的EC50值分别为29.05μg/mL和42.96μg/mL,而天竺葵油对灰葡萄孢(Botrytis cinerea)和禾谷镰孢菌(Fusarium graminearum)的EC50值分别为34.02μg/mL和68.48μg/mL。

海洋细菌B-9987菌株产生的抑菌物质及对几种植物病原真菌的作用

从海洋生境中分离得到 317株菌株 ,筛选获得 1株芽孢杆菌B 9987菌株 ,其胞外分泌的代谢产物对植物病原真菌有显著的抑制作用 ,其中对茄交链孢菌、大丽轮枝菌、黄枝孢菌、立枯丝核菌主要是造成真菌孢子或菌丝末端膨大成球状 ,继而胞壁崩解 ,原生质外泄 ;对尖孢镰刀菌和莴苣霜霉菌的抑制作用 ,则表现为孢子萌发率降低和芽管长度明显变短。该抑菌物质经乙醚萃取 ,硅胶G薄层层析纯化 ,官能团显色反应初步推断为酚类化合物。