木霉拮抗黄瓜枯萎病菌菌株的筛选

通过对峙培养、拮抗作用、耐性的测定,筛选出对黄瓜枯萎菌(Fusarium oxysporumf. sp. cucum erinum)拮抗作用强的木霉菌株(Trichoderma spp.)有13株,其中T55的拮抗作用最强。该菌株对峙培养对黄瓜枯萎菌抑菌率为80.16%,拮抗作用测定其圆盘滤膜法抑菌率大于50%,其它方法的抑菌率低于30%。该菌在5、10、35℃,pH4、9、10的条件下生长良好。     

烟草尖孢镰刀菌拮抗真菌的筛选鉴定及促生作用研究

为获得烟草镰刀菌根腐病拮抗真菌菌株,试验以河南省豫中烟区烟草根际土壤为试材,以主要致病菌尖孢镰刀菌Fusarium oxysporum为主要靶标,采用稀释涂布平板法分离获得土壤真菌371株,平板对峙培养法测定获得1株具有明显抑制效果的拮抗菌株,利用形态学和分子生物学的方法将其鉴定为棘孢木霉Trichoderma asperellum Tr-0111。抑制效果测定表明,该菌株对尖孢镰刀菌的抑制率最高可达93.13%,对根串珠霉菌、拟茎点霉、链格孢菌等其他8种烟草病原菌均具有较强的抑制效果。促生作用测定结果表明,菌株发酵液使烟草种子萌发率提高5.34%,烟苗根长增长率为62.39%,对盆栽烟草的最大叶面积和地上部鲜重也具有显著的促生效果。为进一步相关机理的研究和生防菌剂的研制奠定了基础。     

生物刺激素协同对草莓炭疽病防治及其生长发育的影响

草莓炭疽病是草莓定植时期最易造成死苗的真菌病害。本文研究了生物刺激素（木霉菌、海藻酸、聚谷氨酸）协同灌根对草莓炭疽病的防治以及促生作用。结果表明,与对照相比,木霉菌、海藻酸、聚谷氨酸单剂灌根和木霉菌+海藻酸、木霉菌+聚谷氨酸的协同灌根处理均能显著降低炭疽病造成的死苗率,其中木霉菌+海藻酸的防效最高,可达78.26%。相比对照,所有处理均能显著提高净光合速率、1,5-二磷酸核酮糖羧化酶活性和加氧酶活性等光合指标,以及土壤脲酶活性。协同灌根处理显著提高草莓株高、根茎粗、叶柄长、叶面积等生长指标达15.62%～47.00%。其中木霉菌+海藻酸处理的净光合速率、土壤脲酶活性最高。综合而言,木霉菌与海藻酸协同灌根在促进生长、促进光合、提高土壤供氮水平以及防控草莓炭疽病方面均表现出显著的增效作用,可作为生物防治的一种有效手段。     

Silicon,biological seed treatment and cutting reduce the intensity of leaf spot diseases affecting Lolium multiflorum

Italian ryegrass is a major pasture crop cultivated mainly for feeding cattle in southern Brazil. The choice of crop is based on nutritional quality and the shortage of pastures that occur during the winter in the southernmost part of Brazil. In this region, the production of biomass and seeds are threatened by diseases, in particular fungal diseases that affect the foliage. In this study we identified and monitored the natural occurrence of a complex of leaf spotting diseases and evaluated the effects of combined management practices to reduce the intensity of foliar diseases: seed treatment with Trichoderma atroviride, soil amendment with silicon and cutting frequency (none, one or two cuts). Three diseases were present in a two-season experiment: grey leaf spot (Pyricularia oryzae), brown leaf spot (Bipolaris sorokiniana) and parrot's eye leaf spot (Cercospora sp.). Grey leaf spot had the highest severity, regardless of treatment, during the first season, and was the only disease present in the following season. Grey leaf spot dominated, and plants receiving only one cut were more prone to P. oryzae infection. The treatments with silicon or T. atroviride reduced the relative disease by 40% to 80%, in relation to untreated and uncut regimes. This study showed the main leaf spot diseases associated with L. multiflorum. Moreover, it demonstrates for the first time in the region the effectiveness of reducing leaf spot via the application of silicon in the soil, inoculation of seeds with T. atroviride and managing the cutting regime.     

Evaluation of Trichoderma spp. as a Biocontrol Agent Against Soilborne Fungi and Plant-parasitic Nematodes in Israel

The optimal conditions required to market Trichoderma as a biocontrol agent against soilborne fungi and nematodes are discussed. These include a proper formulation, an efficient delivery system, and alternative methods for Trichoderma's application.The implementation of Trichoderma in integrated pest management (IPM) can be achieved using a soil treatment which combines reduced amounts of biocides/fungicides and the Trichoderma preparation. Biocontrol activity can be increased by combining two (or more) types of biocontrol agents. Moreover, the construction of a genetically modified Trichoderma can lead to the improvement of certain traits which are absent or not highly expressed in the native microorganism isolated from its natural habitat.Different Trichoderma harzianum and T. lignorum isolates were tested for their nematicidal activity against the root-knot nematode Meloidogyne javanica. In short-term experiments, improved growth of nematode-infected plants and decreases in the root-galling index and the number of eggs per gram of root were achieved when nematode-infested soils were pre- exposed to the T. harzianum preparations. A long-term experiment resulted in improved growth and higher yield of nematode-infected plants, but no significant change in the galling index, either by pre-exposure of the fungus to the soil or by enrichment in the root-ball.As biocontrol is an integral part of the IPM philosophy, judicious use of Trichoderma against soilborne pathogens, when demonstrated to be consistently effective, practical and economic, can serve as a model for the introduction and implementation of other biocontrol means into IPM.     

产纤维素酶原生质体形成和再生条件选择

研究了4种营养缺陷型产纤维素酶的原生质体形成和再生条件。结果表明,它们原生质体形成条件均以0．2mol.L^-1（pH5.8)磷酸缓冲液为好。绿色木霉（Trichoderma viride)UV2-11和另一未定木霉（T．sp.)UV2-2以0．2mol.L^-1KCl的渗透稳定剂为好,菌丝菌龄分别为20,18－20h;康宁木霉（T.Toningii)UV2-15以0．6mol.L^-1NaCl的渗透稳定剂为好,菌丝菌龄均为16－20h,酶解时间均为2－3h。原生质体再生培养基以加入0．5％酵母膏和0．5％泛酸钙钙的改良Czapek培养基效果好,用0．7mol.L^-1KCl和NaCl作为渗透稳定剂。菌株原生质体的生率较高。     

短密木霉菌株GAS1-1的分离鉴定、拮抗作用及其生物学特性

为了解短密木霉Trichoderma brevicompactum对植物病害的生防作用及其生物学特性,利用稀释平板分离法从甘肃省景泰县马铃薯连作田植株根际土壤中分离获得1株木霉菌株GAS1-1,经形态观察、rDNA-ITS和EF-1α序列分析明确其分类地位;用生物学方法研究该菌的营养生长和产孢条件;采用对峙培养法测定该菌株对5种植物病原真菌的抑制作用。结果表明,基于形态学特征和基因序列分析结果,菌株GAS1-1被鉴定为短密木霉Trichoderma brevicompactum,为甘肃省木霉新记录种。该菌株对玉米镰孢茎腐病菌Fusarium graminearum、玉米穗腐病菌F. verticillioides、棉花枯萎病菌F. oxysporum、玉米腐霉茎腐病菌Pythium inflatum和灰葡萄孢霉Botrytis cinerea均具有较好的拮抗效果,尤其对玉米腐霉茎腐病菌的抑制作用最好,抑菌率达100.00%。该菌株营养生长和产孢的最适碳源和氮源分别为葡萄糖和酵母膏;其在15～35℃均可生长,菌丝最适生长温度为30℃,最佳产孢温度为25℃;在pH 5～12的培养基上菌丝均可生长,菌丝最适生长和产孢的pH均为5;24 h黑暗条件下菌丝营养生长最快,12 h光暗交替条件有利于产孢;孢子致死温度为69℃,10 min。表明短密木霉菌株GAS1-1具有较好的生防应用潜力。     

4种木霉对核盘菌的抑制作用

通过对峙培养以及对木霉菌(Trichoderma spp.)次生代谢物对病原菌的抑菌率进行了测定,结果表明,供试哈茨木霉(T.harzianum)、绿色木霉(T.viride)、康氏木霉(T.koningii),以及一株未知种名的木霉菌株TY(Trichoderma sp.)均对核盘菌(Sclerotinia sclerotiorum (Lib) de Bary)有明显的抑制作用.在对峙培养中,木霉菌菌落与核盘菌的菌落相向生长一定时间后,挑取核盘菌菌落内部的菌丝进行镜检,发现有木霉菌孢子存在,且核盘菌发生菌丝变形,如原生质浓缩,菌丝断裂,或菌丝消解等现象.并发现不同种木霉所产生的次生代谢物的活性不同,对核盘菌的抑制作用也存在显著差异.     

木霉的营养生长及发酵条件

 木霉是一类分布广泛的土壤习居菌,而且对一些病原真菌有很大的生防潜力。大部分木霉对营养的需求并不十分严格,可以分解利用纤维素、半纤维素、几丁质等多种多聚物作为其生长的碳源物质,也可以利用多种简单的和复杂的氮源化合物生长。为了应用木霉防治真菌病害,研究者摸索了各种培养条件,以求在低成本的前提下获得高的培养产物。其中包括应用糖蜜、麦麸、植物秸杆、虫草头孢废液和玉米粉等进行的固体培养、液体培养和固液两相培养,为大规模工业化生产奠定了一定的基础。     

Eight New Peptaibols from Sponge-Associated Trichoderma atroviride

Eight new and four known peptaibols were isolated from a strain of the fungus, Trichoderma atroviride (NF16), which was cultured from an Axinellid sponge collected from the East Mediterranean coast of Israel. The structures of the pure compounds were determined using HRMS, MS/MS and one- and two-dimensional NMR measurements. The isolated compounds belong to the trichorzianines, a family of 19-residue linear hydrophobic peptides containing a high proportion of α-aminoisobutyric acid (Aib), an acetylated N-terminus and a C-terminal amino alcohol. These new peptaibols exhibited antimicrobial activity against environmental bacteria isolated from the Mediterranean coast of Israel.