木霉菌对植物寄生线虫防治作用机制

生物防治不仅作为植物寄生线虫病安全有效的防治措施，而且其具有降低化学农药施用量和减少环境污染的作用。木霉菌作为一种生防潜力巨大的线虫病防治真菌，已被广泛研究并应用于植物寄生线虫病的防治。本文主要从木霉防治植物寄生线虫病的直接和间接生防机制的视角，综述了木霉对线虫的重寄生作用、毒杀作用以及木霉对植物的诱导抗性、促生作用等多种协同防治植物寄生线虫的机制，展望了今后需要继续扩大对杀线木霉资源种类的调查和挖掘、木霉菌对植物寄生线虫的作用机理以及从分子水平上加深对木霉防治线虫病机制的研究，旨在为科学利用木霉菌防治植物线虫病提供参考。     

水稻纹枯病菌胞外蛋白酶特性研究

植物病原菌产生蛋白水解酶被认为是病原菌侵染寄主的一个重要策略。作者对水稻纹枯病菌(Rhizoctonia solani Kühn)产生的胞外蛋白酶的特性进行了研究。结果表明,水稻纹枯病菌产生一个分子量约为49.5ku的胞外蛋白酶。该酶对胰蛋白酶专一性底物Benz-Phe-Val-Arg-NA有较高的活性,并能从羧基端降解Arg和Lys。胰蛋白酶抑制剂对该酶活性有较强的抑制作用,表明该蛋白酶是一种类胰蛋白酶。在离体试验下,该蛋白酶可以降解水稻细胞壁蛋白并释放羟脯氨酸。SDS-PAGE电泳分析显示,至少有4个多肽被降解,分子量范围分别在14.4～20.1、20.1～31ku和31～43ku范围内。说明该胞外蛋白酶具有降解水稻细胞壁的能力,因此,该蛋白酶可能在水稻纹枯病菌的致病过程中起重要作用。     

玫瑰黄链霉菌对黄瓜根结线虫乙酰胆碱酯酶活性的影响

玫瑰黄链霉菌Men-myco-93-63是从马铃薯疮痂病自然衰退土壤中筛选出的对多种重要植物病原菌具有较强抑制作用的拮抗链霉菌。前期研究表明,该生防菌及其发酵液对蔬菜根结线虫具有较好的防治作用。为研究玫瑰黄链霉菌抑制根结线虫的作用机制,利用发酵液浸没法,检测了Men-myco-93-63代谢产物对根结线虫乙酰胆碱酯酶活性的影响。结果表明,Men-myco-93-63发酵液能够有效地抑制黄瓜根结线虫乙酰胆碱酯酶的活性,在36 h时抑制率最高,达到88.82%,之后抑制作用缓慢减弱,但72h时抑制作用仍可达47.32%。说明Men-myco-93-63发酵液能有效抑制根结线虫乙酰胆碱酯酶活性,且抑制作用持续时间长。     

木霉几丁质酶及其对植物病原真菌的拮抗作用

 木霉(Trichoderma spp.)作为重要的植病生防因子(biocontrol agents)一直受到普遍关注。木霉菌株产生的包括几丁质酶在内的细胞壁降解酶,在木霉重寄生中起重要作用。本文论述木霉几丁质酶的种类、诱导产生、理化特性及其对植物病原真菌的拮抗作用,并对木霉几丁质酶及其基因的生防应用进行讨论。     

高效产几丁质酶微生物的分离和筛选

正几丁质酶是具有生物催化活性、能水解几丁质的一类水解酶,许多微生物可通过胞外诱导产生该酶。几丁质广泛存在于植物病原菌、线虫卵和昆虫的细胞壁中,防治植物土传病害时,在土壤中加入几丁质可以刺激土壤微生物胞外分泌几丁质酶,从而抑制病虫害发生(郑秀丽等,2006;孙建波等,2010)。     

长枝木霉菌株TL16防治南方根结线虫的作用机理

为探明长枝木霉Trichoderma longibrachiatum菌株TL16防治南方根结线虫Meloidogyne incognita的作用机理,采用原生质体转化法获得绿色荧光蛋白(green fluorescent protein,GFP)标记菌株GFP-TL16,通过测定菌株TL16和GFP-TL16对南方根结线虫卵和2龄幼虫(2nd-stage juvenile,J2)的寄生与致死作用,其发酵液对卵孵化的抑制作用和对J2的致死作用,以及菌株GFP-TL16在黄瓜根系的定殖情况和菌株TL16对番茄根结形成的抑制作用来综合分析其作用机理。结果显示:菌株TL16菌丝对南方根结线虫卵无寄生作用,处理19 d后卵降解率为26.33%,致死作用较低;菌株TL16分生孢子悬浮液处理南方根结线虫J2后72 h的致死率为1.65%,且无寄生作用。菌株TL16发酵液处理南方根结线虫J2后48 h的校正死亡率为10.71%,处理卵15 d后对卵孵化的相对抑制率为77.11%。菌株GFP-TL16可定殖于黄瓜根系中,经菌株TL16处理后接种南方根结线虫J2,番茄根结减退率为55.88%。表明长枝木霉菌株TL16可通过抑制根结线虫卵孵化和诱导番茄产生抗病性来防治根结线虫病。     

毒杀南方根结线虫的木霉种类鉴定及活性研究

木霉是国际上研究与应用非常普遍的生防真菌之一,具有重要的科研价值和广阔的应用前景。本研究采用形态学鉴定、ITS和β-tubulin序列分析法对4株具有杀线虫活性的木霉菌株进行种类鉴定,结果显示FT311和FT312为绿色木霉(Trichoderma viride),FT1937和FT85为哈茨木霉(T.harzianum)。利用贝氏皿浸没法测定,4株木霉发酵滤液均能显著降低南方根结线虫卵的孵化和2龄幼虫(J2)的活性,其中FT1937和FT85对卵孵化抑制率和J2致死率均达到85%~92%,而FT311能够达到80%~85%,但FT312毒性较低,仅达40%~60%;2株哈茨木霉的活性好于2株绿色木霉;同时观察到发酵液对线虫卵和幼虫体壁有消解作用、抑制和延迟卵孵化,并使幼虫体内空泡化。本研究为根结线虫病害的生防防治提供了更多的木霉资源信息。     

木霉在植物病害生物防治中的应用及作用机制

木霉是植物病害生物防治中应用和研究非常广泛的一类生防真菌。本文阐述了木霉在多种植物病害防治上的应用及防治效果，并概括了木霉在生防过程中对植物病原物的生防机制，包括竞争、重寄生、抗生作用等，以及木霉与植物互作中对植物促生和诱导植物抗性的机制。目前，世界上含木霉的商品化制剂已超过250种，在不同国家地区都取得了良好的防治效果，更多的优秀生防木霉菌株也在通过野生菌株筛选或遗传改良等方式开发。木霉生物防治及机制研究对推广生物防治和减少化学农药有重要意义。     

木霉对植物的促生及诱导抗性研究进展

木霉不仅能够直接抑制植物病原菌的生长,还可通过定殖于植物的根部引起植物新陈代谢的改变。本文综述了木霉在植物根部定殖后,通过产生植物生长调节剂、抑制或降解根际有害物质、增加养分利用率以促进植物生长以及通过产生激发子诱导植物形成胞壁沉积物、合成抗菌物质以诱导植物局部或系统抗性的研究进展,以期为进一步扩大木霉在农业生产中的应用提供参考。     

木霉生物学特性及拮抗作用研究进展

木霉（Trichoderma spp．）是一类广泛存在于土壤、根围、叶围、种子和球茎等环境中的拮抗性真菌,20世纪70年代以来关于木霉的拮抗作用及其机制有很多报道,木霉在植物病害生物防治中具有重要的地位。能用于防治多种真菌病害。研究木霉对病原真菌的拮抗作用,有利于木霉菌在植病生防领域的开发利用。木霉菌（Trichoderma spp．）属半知菌亚门，丝孢纲。     

Ultrastructure, Morphology, and Sporogenesis of Pasteuria penetrans

ABSTRACT Pasteuria penetrans is a bacterial parasite of root-knot nematodes that shows great potential as a biocontrol agent. Scanning and transmission electron microscopy were used to study the ultrastructure, morphology, and sporogenesis of four isolates of P. penetrans. The effects of different Meloidogyne spp. and tobacco cultivars on sporangium size and morphology of endospores attached to the cuticle of second-stage juveniles (J2) of root-knot nematodes also were investigated. The P. penetrans isolates and their origins were P-20 from M. arenaria race 1 in Levy County, FL; P-100 from Meloidogyne sp. in Pasco County, FL; B-4 from Pratylenchus scribneri in Seminole County, FL; and P-120 from Meloidogyne spp. in Alachua County, FL. Sporangia of the four isolates were identical morphologically and similar in their dimensions, ranging from 2.39 to 3.42 mum in diameter and from 1.38 to 2.38 mum in height. Different Meloidogyne spp. and tobacco cultivars had no effect on sporangium size. Endospores attached to J2 were visualized in three forms: endospores retaining the sporangium wall, endospores covered with a thin exosporium, and endospores without covering. Sporogenesis of P. penetrans was similar to that of other gram-positive bacteria and generally matched the seven-stage scheme reported for Bacillus thuringiensis.     

Biological Control of the Root-Knot Nematode Meloidogyne javanica by Trichoderma harzianum

ABSTRACT The fungal biocontrol agent, Trichoderma harzianum, was evaluated for its potential to control the root-knot nematode Meloidogyne javanica. In greenhouse experiments, root galling was reduced and top fresh weight increased in nematode-infected tomatoes following soil pretreatment with Trichoderma peat-bran preparations. The use of a proteinase Prb1-transformed line (P-2) that contains multiple copies of this gene improved biocontrol activity in the greenhouse experiments compared with the nontransformed wild-type strain (WT). All the Trichoderma strains showed the ability to colonize M. javanica-separated eggs and second-stage juveniles (J2) in sterile in vitro assays, whereas P-2 also penetrated the egg masses. This protease-transformed line presented the same nematicidal and overall proteolytic activity as the WT in in vitro tests in which concentrated soil extracts from Trichoderma-treated soils immobilized the infective J2. However, the J2 immobilization and proteolytic activities of both P-2 and the WT were higher than those obtained with strain T-203. Characterization of the activity of all Trichoderma strains soil extracts on J2 showed that it was heat resistant and restricted to the low-molecular-weight fraction (less than 3 kDa). It is suggested that improved proteolytic activity of the antagonist may be important for the biological control of the nematodes.     

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.     

杀线虫植物内生细菌和根际放线菌对根结线虫的防效

为了获得防治植物寄生线虫的微生物菌种资源和防效稳定的生防菌剂,以松材线虫为靶标,采用直接触杀法对植物根际放线菌和内生细菌进行了筛选,通过盆栽试验测定了筛选到的菌株发酵液对根结线虫的防治效果,并将有初步防效的菌株制成固体菌剂,分别测定单菌剂和复合菌剂的防效。共筛选到16株放线菌和4株内生细菌具有杀线虫活性,其中2株放线菌和3株内生细菌的发酵液对根结线虫防效明显;放线菌和内生细菌混合发酵液均保持了对根结线虫的抑制作用,但是放线菌和细菌菌株的固体菌剂混合后对根结线虫的防效低于细菌单菌株的防效。植物内生细菌BCM2和CCM7单菌株的固体菌剂对番茄根结线虫的防效最高,分别达到了83.31%和83.07%。表明高杀线虫活性的植物内生细菌制备的固体菌剂具有应用潜力。     

生防菌株Snef85的鉴定及其发酵液对不同种类线虫的毒力

采用稀释分离法从人参土壤中分离获得一株生防真菌Snef85,采用培养性状、形态特征观察法及ITS序列分析法对菌株Snef85进行鉴定,并测定其发酵液对5种线虫的毒力作用。结果表明,该菌株为哈茨木霉Trichoderma harzianum,其发酵液对不同种类线虫的防治效果差异显著,对根结线虫Meloidogyne spp.、水稻干尖线虫Aphelenchoides besseyi和大豆胞囊线虫Heterodera glycines的毒力较强,对腐烂茎线虫Ditylenchus destructor的毒力次之,对小杆线虫Caenarhabditis spp.的毒力最弱。     

内蒙古赤峰市保护地蔬菜根结线虫种类鉴定

正根结线虫(Meloidogyne spp.)广泛分布于世界各地,是影响我国农业生产最大的一类植物病原线虫,寄主植物有3 000多种,可侵染几乎所有的蔬菜作物,其中,茄科(Solanaceae)、葫芦科(Cucurbitaceae)和十字花科(Brassicaceae)等蔬菜受害最重~([1])。常见的根结线虫种类有南方根结线虫     

我国蔬菜根结线虫发生、致害和绿色防控研究进展

根结线虫是威胁我国蔬菜生产的最重要病原物类群之一，寄主达3 000多种，每年造成的蔬菜损失价值高达5亿美元，目前缺乏安全高效的防治方法。近年来，随着保护地蔬菜种植面积的增加，根结线虫病对我国蔬菜生产的危害逐年加重，因此必须加强对蔬菜根结线虫病发生、致害和防治的深入研究。该文对近10来年我国根结线虫的发生分布和危害、主要种类、鉴定和快速诊断方法、成灾规律、与植物的互作及机理、绿色防控等方面的研究进展进行综述，并展望我国根结线虫的研究趋势。     

木霉对草坪褐斑病的拮抗效果及耐药性

通过木霉属5菌株与大连高尔夫球场上草坪褐斑病菌的对峙培养试验,研究结果表明:哈茨木霉Thar1菌株、哈茨木霉Thar2菌株、深绿木霉Tat菌株、钩状木霉Tha菌株及桔绿木霉Tci菌株对病原菌的抑制率为:66%、73%、71%、77%、55%,而相对抑菌效果分别为2.06、3.94、2.35、3.54、2.27,可以作为草坪褐斑病菌的生防菌加以利用。在这5株木霉中以哈茨木霉Thar2对草坪褐斑病菌的拮抗作用最强。观察结果表明,哈茨木霉Thar2对草坪褐斑病菌的拮抗机制主要表现为生长竞争、重寄生及产生抗菌物质。     

陕西省蔬菜根结线虫拮抗真菌的分离与初步鉴定

为了了解陕西省内的根结线虫生防资源,筛选更好的根结线虫生防菌,在陕西省9个县市采集了118份感染根结线虫的蔬菜植株根部样品,采用直接分离法从根结线虫卵、卵囊及雌成虫体上共分离到199株拮抗真菌.除43株不产孢及疑难种未鉴定外,其余分属于镰刀菌属、拟头孢霉属、芽枝霉属、拟青霉属、链格孢属、掘氏梅里霉属、单顶孢属、青霉属等22个属.其中镰刀菌属、拟头孢霉属及芽枝霉属真菌出现频率较高,约占总分离数的53%,且不同地区间拮抗真菌的种类与数量也存在较大差异.