不同碳源对生防木霉菌T23、T56生长和生防活性的影响

采用基础培养基添加不同碳源培养木霉菌T23、T56的方法,研究了不同碳源对木霉菌生长、产孢量和生防活性的影响,以期获得木霉菌理想的碳源。同时,评价生物炭作为碳源或基质的效果。结果表明,葡萄糖、淀粉、乳糖、生物炭单独为碳源时,木霉菌生长和产孢以葡萄糖为较优;与生物炭复配后,以生物炭和葡萄糖的复配为最佳,T23与T56的菌丝干重分别为1.24、1.41 g,孢子产量分别高达177.5×107个/m L、172.5×107个/m L,第3天的生长直径分别为75.30、79.20 mm。T23与T56对镰孢菌的抑制作用以生物炭和葡萄糖复配的培养基最好,抑制率分别为83.42%、72.00%。     

生防木霉菌Th2和T4对甜瓜根围土壤微生态的影响

利用平板计数、微生物量碳和末端标记限制性片段长度多态性分析(terminal restriction fragment length polymorphism,T-RFLP)相结合的方法研究了2株生防菌哈茨木霉Th2和T4对甜瓜根围土壤微生态的影响.结果表明,生防菌Th2和T4对根围土壤细菌有明显的促生作用,而对真菌和放线菌有明显的抑制作用.T-RFLP数据统计学分析表明,Th2和T4的引入可以增加土壤细菌的多样性.其中对开花期土壤细菌菌群影响最显著,Th2对TRF251和TRF513菌群有显著的促生作用,含量是对照的3.7倍和4.5倍,T4对TRF251和TRF513菌群含量也比对照高4倍和5.6倍,而2株生防菌对TRF60菌群均有显著的抑制作用,其含量分别为对照的51.6%和64.3%.在果熟期生防菌对土壤细菌菌群的影响减弱,表明土壤的缓冲功能发挥了作用.研究结果还表明,Th2和T4对土壤细菌菌群有着相似的促生和抑制作用,且不会破坏土壤微生态的稳定性.     

木薯炭疽病拮抗木霉菌筛选与室内防效研究

本文对木薯炭疽病的病原进行鉴定,筛选和鉴定拮抗该病原物的木霉菌,并测定了其室内防效.通过组织分离法和柯赫氏法则,明确了引起木薯炭疽病的病原菌为胶孢炭疽菌Colletotrichum gloeosporioides;采用平板对峙试验,从15株木霉菌中筛选获得3株对胶孢炭疽菌具有较强抑制作用的木霉菌LG004-52、LS0...     

土壤不同处理对木霉菌定殖及其生防效果的影响

重茬土经施肥改土、高温灭菌后,增施木霉菌的试验结果表明:木霉菌可以在甜瓜根表、根际定殖存活,同时不影响其他有益微生物的正常生长;土壤经高温灭菌后,土壤中的微生物呈真空状态,有利于木霉菌的快速定殖,促进甜瓜生长,降低甜瓜枯萎病的病情指数。甜瓜枯萎病发病率与土壤中镰刀菌孢子的存活量成正相关,与土壤中木霉菌、细菌数量的多少成负相关。     

木霉菌转化体对番茄灰霉病的防治效果

保护地的特殊条件使番茄灰霉病的发生越来越严重，同时由于生产上仍缺少抗病品种，且灰霉病菌已对多种化学杀菌剂产生了抗药性，严重制约了番茄的安全生产。因此筛选或构建具有高效生防活性的新型生物农药具有重要意义。利用木霉菌防治番茄灰霉病已有一些成功的例子，如Elad等、Eden等和Dik等利用339或Trichoderma harzianum成功防治了灰霉病。然而多数生防木霉菌的防效仍难超过化学农药，因此通过生物技术筛选出对番茄灰霉病具有高效生防活性的木霉菌具有重要意义。     

绿色木霉菌T23对黄瓜枯萎病防治效果及其几种防御酶活性的影响

 本文通过生物测定的方法初步研究了绿色木霉菌T23分生孢子和厚垣孢子对黄瓜枯萎病防治效果及黄瓜幼苗几种防御酶活性的影响。结果表明:黄瓜幼苗经木霉菌处理后,病情指数由33.69分别降至13.12和10.28,经木霉菌处理的黄瓜体内与抗性反应相关的苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)及过氧化氢酶(CAT)活性有明显增加,其峰值分别为对照的2.75、2.49、2.42及15.84倍,说明生物防治过程中可能有木质素、植保素等抗性物质的参与。与分生孢子处理相比,木霉菌厚垣孢子处理的酶活高峰出现得晚,但酶活峰值高。     

棘孢木霉菌鉴定及其对花生白绢病生防机制的研究

为了利用生防菌有效防控花生白绢病Sclerotium rolfsii,本研究采用稀释培养法从花生根际土中分离木霉菌,采用平板对峙法和盆栽试验筛选对花生白绢病具有高效生防效果的菌株并测定其生防机制,通过形态学、分子生物学技术和环境因子对发育的影响确定生防菌株的分类地位及生物学特性。结果表明,从分离的132株木霉菌中筛选出1株对齐整小核菌S.rolfsii具有高效抑菌效果的菌株GH-20,抑菌率达89.93%,将其鉴定为棘孢木霉菌Trichodermaasperellum。对峙培养时,菌株GH-20对齐整小核菌的拮抗指数达Ⅰ级,当其菌落覆盖病原菌菌落后,其菌丝缠绕和侵入病原菌菌丝,并使病原菌菌丝体逐渐消解、减少,导致病原菌不能产生菌核;菌株GH-20产生的挥发性代谢产物（volatile gas,VOG）和非挥发性代谢产物（non-VOG）对病原菌的抑菌率分别为59.95%和79.49%。盆栽试验结果表明,同时接种菌株GH-20和齐整小核菌对花生白绢病生防效果最好,为79.64%。另外,该菌株具有在以果糖为碳源,以酵母膏为氮源和pH7的条件下生长发育较好,光照有利于生长而光暗交替有利于孢子...     

木霉菌REMI转化体对番茄灰霉病的防治及其机理的研究

研究不同木霉菌转化体对番茄灰霉病防治效果及机理,为木霉菌生物防治的合理利用奠定基础。利用限制性内切酶介导基因整合技术(restriction enzyme-mediated integration,REMI),通过插入线性化质粒DNA获得了生物防治番茄灰霉病(Botrytis cinerea)效果优于出发菌T21菌株(出发菌)的3个木霉菌转化体Ttrm31、Ttrm34和Ttrm55,对侵染花器和叶片的灰霉病防效分别比原生物防治木霉菌株提高了16.9%和8%。木霉菌转化体的产孢能力、分生孢子的萌发率、对碳氮源的利用能力及对高温的抵抗能力都有所提高;木霉菌转化体本身产生的几丁质酶和β-1,3-葡聚糖酶的活性均比出发菌高,因此通过REMI技术可以获得新的有益木霉菌转化体,在一定程度上提高了生物菌株防治番茄灰霉病的水平。说明REMI技术可以用于改良生防木霉菌株的功能,提高生物防治效果。     

微波诱变选育高效溶磷木霉菌株的研究

为了获得高效溶磷菌株,对木霉菌T6菌株进行微波诱变处理,测定并分析了微波输出功率和辐照时间等参数对诱变效果的影响。结果表明:最佳诱变条件是微波输出功率900 W,辐照时间70 s,且在此条件下得到8株具有较高溶磷能力的突变菌株T6-9、T6-33、T6-93、T6-126、T6-157、T6-188、T6-196和T6-203。经过多代转接培养最后得到一株遗传稳定的溶磷木霉菌株T6-157,该菌株在液体培养条件下溶磷量为204.46 mg·L~(-1),与出发菌株相比,溶磷量提高了107.97%,植酸酶活性提高了57.35%,且突变菌株对土传植物病原菌的拮抗能力也有所提高。     

产纳他霉素生防菌A01和A02与已知产生菌的RAPD比较

为了明确自主分离的天然抗真菌活性产物——纳他霉素产生菌A01和A02与已知纳他霉素产生菌的遗传相似性,采用RAPD技术对这2株菌和3个产纳他霉素的标准菌株进行了比较分析。利用筛选出的7个随机引物对5个菌株的PCR扩增共得到154条清晰稳定的DNA条带,其中同源性条带86条,多态性条带68条,分别占总条带数的55.8%和44.2%,平均每个引物扩增出9.7个多态性条带,得到了丰富的DNA指纹图谱。所得数据经NTSYS pc 2.10e软件聚类分析,表明5个菌株间的遗传距离为0.0991~0.4738,其中A02与其它菌株间的遗传距离均较远。结合前期的分类研究结果,确证了菌株A02为新的纳他霉素产生菌。     

Mycoparasitic Trichoderma isolates as a biocontrol agent against Valsa ceratosperma,the causal agent of apple valsa canker

European Journal of Plant Pathology - Apple valsa canker, caused by the fungus Valsa ceratosperma, is one of the most destructive diseases of this crop. Conventional fungicide treatments are not...     

Induced systemic resistance in Trichoderma harzianum T39 biocontrol of Botrytis cinerea

Biocontrol of Botrytis cinerea with Trichoderma spp. is generally believed to result from direct interaction of the biocontrol agent with the pathogen or from a Trichoderma-induced change in environmental conditions that affects B. cinerea development. In this work we provide arguments for the participation of induced plant defence in T. harzianum T39 control of B. cinerea. In tomato, lettuce, pepper, bean and tobacco, T. harzianum T39 application at sites spatially separated from the B. cinerea inoculation resulted in a 25–100%percnt; reduction of grey mould symptoms, caused by a delay or suppression of spreading lesion formation. Given the spatial separation of both micro-organisms, this effect was attributed to the induction of systemic resistance by T. harzianum T39. The observation that in bean the effect of T. harzianum T39 was similar to that of the rhizobacterium Pseudomonas aeruginosa KMPCH, a reference strain for the induction of systemic resistance, confirmed this hypothesis. Since B. cinerea control on tobacco leaves sprayed with T. harzianum T39 was similar to the control on leaves from T. harzianum T39 soil-treated plants, induction of plant defence might also participate in biocontrol on treated leaves.     

Honey bee dispersal of the biocontrol agent Trichoderma harzianum T39: effectiveness in suppressing Botrytis cinerea on strawberry under field conditions

Botrytis cinerea, which causes grey mould, is a major pathogen of many crops. On strawberry, isolates of Trichoderma spp. can effectively control B. cinerea, but frequent application is necessary. Bees can be used to disseminate biological control agents to the target crop. We tested the ability of honey bees to disseminate Trichoderma harzianum T39 to control B. cinerea in strawberry in the field during the winter in Israel over two consecutive seasons. We used the recently developed ‘Triwaks’ dispenser for loading the bees with the T. harzianum inoculum. During both years, grey mould developed in late January in untreated control plots; at low to medium disease levels it was partially controlled by fungicide treatment, and was best controlled in bee-visited plots. At high disease levels neither chemical nor biological control was effective. To assess the spatial distribution of inoculum by bees, we sampled flowers up to 200 m from the hives and found effective levels of T. harzianum even at 200 m. The approach used in this study provides an effective control of grey mould in strawberry in conditions of low to medium grey mould incidence.     

Trichoderma as a biostimulator and biocontrol agent against Fusarium in the production of cereal crops: Opportunities and possibilities

Despite technological advances in global agriculture in recent years, the problem of pathogenic fungi in the production of cereal crops continues to be an issue. Currently, the high variability of weather factors that are considered unusual for specific locations affect the growth and physiology of pathogens attacking cereal crops. One of the most common plant protection methods is the use of synthetic pesticides; however, there is growing controversy over this approach due to the build-up of pesticides in the environment and the presence of their residues in food. The purpose of this literature review is to explore the current state of knowledge regarding the potential of using Trichoderma species as a biostimulator and for the biological protection of cereal crops against pathogenic fungi. Trichoderma fungi—through mycoparasitism, antibiosis and competition for space and nutrients—help to inhibit the growth of pathogens and have a positive impact on the growth of plants, including their root system, which is considered a desirable effect during drought episodes. It has also been demonstrated that Trichoderma fungi can convert Fusarium toxins into new metabolites that can be of lower toxicity. However, the highly limited number of in vivo studies investigating the use of these fungi for biocontrol in cereal crops remains an obstacle to the commercialization of Trichoderma fungi. It appears that the determination of their effectiveness in the biocontrol of cereal crops under variable weather and climate conditions presents a considerable challenge.     

内蒙古白刺属植物遗传多样性的RAPD分析

利用随机扩增DNA多态性 (RAPD)技术对4种不同地理种群的白刺属(Nitraria L.)植物进行了遗传多样性分析.15个引物扩增出118个位点,其中多态性位点95个(80.5%),各引物的Nei's指数和Shannon's 多样性指数差别较大,利用UPGMA方法构建分子系统树.结果表明:白刺属植物的遗传分化程度和基因流因种而异,除球果白刺以外,其余3种白刺的种群间有很大的基因流,其遗传一致度在0.8561～0.9694之间,而种间遗传分化程度较低,故4种白刺有着相近的祖先,这与形态学和细胞学的分析结果一致,说明RAPD分析方法可从分子生物学角度为白刺属植物的系统学分类提供更为有利和可靠的证据.     

我国植物青枯菌菌株的遗传多样性和组群划分

 采用15条随机引物对我国11个省(市、区)6种不同寄主植物的43个青枯菌代表性菌株和4个国外青枯菌菌株,进行了PCR扩增.引物OPB11、OPA15、OPE1和OPZ10对上述所有菌株扩增获得了相似的产物电泳图谱,分别具1~5条谱带不等;引物OPB7、OPA10和OPF1对马铃薯菌株获得了相同的产物图谱,但对其它寄主菌株的产物间有明显差别;引物OPA14、OPC,6、OPG14、OPF5、OPK14、OPK20和OPK17对于不同菌株的扩增产物多态性很强.供试菌株被聚类为2个组群,即组群A和组群B.组群A中又可分为7个亚组(A1、A2、A3、A4、A5、A6、A7),其中A1含有2个类型(A1-1、A1-2);组群B中也可分为2个亚组(B1、B2),其中B1含有3个类型(B1-1、B1-2、B1-3),B2也含有3个类型(B2-1、B2-2、B2-3).RAPD组群A中包含了27个来自我国不同地区的马铃薯菌株,主要是3号小种、生化变种2;组群B中含有20个来自不同地区、不同寄主的菌株,分属于其它不同的小种和生化变种.研究结果表明,我国青枯菌菌株RAPD组群的划分与菌株的地理来源关系不大,而与寄主来源有明显相关性.此外,通过对我国青枯菌菌株组群进行同源性PCR分析表明,来源自马铃薯的3号小种菌株属于美洲分支"Americanmm",而来自其它寄主的青枯菌1号、5号小种菌株属于亚洲分支"Asiaticum",与本研究RAPD组群A和组群B的划分是一致的.     

广东茄科青枯菌致病力分化及其DNA多态性分析

 分别采自广州、增城、东莞、花都、三水、清远、电白、高要8个菜区的番茄、茄子和辣椒上的31个青枯病菌株,经人工接种于10个鉴别寄主植物上,结果表明,它们的致病力存在明显的差异。聚类分析这31个菌株,可以聚为3个组:第I组菌株主要来自种植番茄和茄子历史较长的广州、东莞、增城老菜区,其致病力较强;第Ⅱ组菌株主要分离自茄子和辣椒上,其致病力中等;第Ⅲ组菌株主要来自近年来新发展的三水市各番茄产区,它们的致病力较弱。从200个随机引物中筛选出17个引物用于上述31个菌株DNA的RAPD分析,共扩增出523条带,其中468条为多态性带,占89.5%。聚类分析这31个菌株,又可聚为4个簇群:第I簇群主要分离自已推广种植多年的丰顺、金丰等抗病番茄品种上;第Ⅱ簇群分离自抗病的番茄新品种新星、年丰和石碣紫红茄上;第IV簇群主要分离自辣椒和茄子;而第Ⅲ簇群来源包括番茄、茄子和辣椒这3种寄主植物。上述试验结果说明,广东茄科青枯菌的致病力存在明显的分化现象,其DNA存在较高的遗传多样性。