利用木霉菌防治草坪镰刀枯萎病研究

为了筛选防治草坪镰刀枯萎病的木霉菌株,采用平板对峙法、盆栽试验、田间试验研究了木霉菌对草坪镰刀枯萎病的拮抗作用及其机制。结果表明,从草坪禾草根际分离获得木霉菌株150株,通过平板对峙法筛选获得20株对草坪镰刀枯萎病有明显拮抗作用的菌株。通过盆栽试验测定,获得8株能显著降低病害发生率和病情指数的菌株。在2008年和2009年2a的田间试验中,筛选到对草坪病害有良好防效的木霉菌株T12。体外拮抗试验显示,木霉菌株T12对引起草坪镰刀枯萎病的尖孢镰刀菌具有良好的拮抗活性。菌株T12的拮抗机制主要为生长竞争、重寄生及产生代谢物质等。木霉菌制剂被认为是化学农药的理想代替产品,它的应用为草坪病害的防治提供了另一条有效防控途径。     

镉对鸡免疫器官及ND免疫应答的影响及硒拮抗效应

在日粮中添加一定量浓度的镉及硒与之相拮抗,复制鸡慢性镉中毒及硒拮抗模型,探讨镉对鸡免疫器官及对免疫应答的影响.结果表明:(1)镉对免疫器官发育以及细胞免疫和体液免疫功能造成了广泛的抑制效应;(2)镉严重影响了ND抗体的产生,并在较低的水平上维持;(3)镉中毒组胸腺、脾脏、法氏囊免疫器官脏器指数均低于对照组,而硒镉拮抗组免疫器官脏器指数介于二者之间;(4)镉中毒组鸡的胸腺、脾脏、法氏囊免疫器官淋嘏组织广泛出现实质性的变化,包括淋巴细胞数量显著减少、变性、坏死、滤泡减少成元滤泡形成等,以及胸腺皮质萎缩变小.     

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。     

In vitro evaluation of fungal isolates for their ability to influence leaf rheology,production of pseudothecia,and ascospores of Venturia inaequalis

As part of an ongoing research project on biological control of apple scab, this study presents a novel approach for the in vitro selection of potential antagonists of the saprophytic phase of Venturia inaequalis. A collection of forty-two fungal isolates were tested for their in vitro ability to degrade apple leaf tissue, inhibit pseudothecia, and ascospore production. The inhibition of ascospore production cannot always be linked reliably with leaf degradation or the evaluation of pseudothecia production. Consequently, ascospore production was retained as the most useful screening parameter. Six isolates proved to significantly reduce the ascospore production of Venturia inaequalis. Two were as effective as Athelia bombacina, a previously reported antagonist of pseudothecia formation and inhibited over 98% of the ascospore production. These new organisms are now available for future field tests. Future selections from a large collection of fungal and bacterial saprophytes can now be based on a reliable and simple in vitro screening method.     

Endo-1,3-β-glucanase and cellulase from Trichoderma harzianum: purification and partial characterization, induction of and biological activity against plant pathogenic Pythium spp.

There were indications that endo-1,3--glucanase (1,3-(1,3;1,4)--D-Glucan 3(4)-glucanohydrolase (EC 3.2.1.6)) and cellulase (1,4-(1,3;1,4)--D-Glucan 4-glucanohydrolase (EC 3.2.1.4)) activity of Trichoderma harzianum Rifai isolate T3 were induced in sphagnum peat moss cultivations and dual culture experiments by the presence of Pythium ultimum. Further, P. ultimum stimulated the germination of Trichoderma conidia. Endo-1,3--glucanase and cellulase were purified from T. harzianum isolate T3, known to control Pythium damping-off of cucumber seedlings. The enzymes were purified from the culture filtrate of the fungus by gel filtration and isoelectric focusing. The purified endo-1,3--glucanase was a small protein with a molecular mass of 17 kilodaltons and a pI of 5.0. Two cellulases were purified to homogeneity and had molecular masses of 40 and 45 kilodaltons respectively, and pI's of 6.4 and 7.6 respectively. Germination of encysted zoospores and elongation of germ tubes of a plant pathogenic Pythium isolate were inhibited by low concentrations of the purified enzymes. A strong synergistic effect was observed on the inhibition of cyst germination by a combination of the endo-1,3--glucanase and the fungicide Fongarid. Finally, a time-course study of colonization of the rhizosphere of cucumber seedlings showed that the active fungal mycelial biomass of a GUS-transformant of T. harzianum isolate T3 increased over four weeks. Trichoderma appeared to colonize healthy roots only superficially, whereas the mucilage of the root hairs and of distal parts of wounded areas or broken parts of the roots, were extensively colonized.     

Interactions of air temperature,relative humidity and biological control agents on grey mold of bean

The interactions ofBotrytis cinerea and seven biological control agents (BCAs) were examined in controlled environments to determine the influence of selected relative humidities (RH) (90,95, and 100%) and air temperatures (20,24 and 28 C) on grey mold of bean. All main effects and interactions were significant (P0.05) among the 72 treatments. In the control, lesions of grey mold developed under all environmental conditions but were largest at 24 C×95 and 100% RH, and 28 C×95% RH. Interactions of environment, BCAs and grey mold were complex.Alternaria alternata, Drechslera sp.,Myrothecium verrucaria, Trichoderma viride, Gliocladium roseum and an unidentified pink yeast were all highly dependent on environment for biological control efficacy, and changes of 4 C or 5% RH were associated with variability in disease suppression that ranged from 15 to 100%. Efficacy ofEpicoccum purpurascens appeared independent of environment and this BCA suppressed disease by 100% in all of the environmental treatments. Suppression of grey mold by many of the BCAs was most effective under environmental conditions least conducive to disease. Therefore, evaluations of potential BCAs in environmental conditions that are marginal for disease can overestimate their efficacy in field environments. Assessments of biological control efficacy in various environments can be used to more accurately assess the potential of BCAs.     

哈茨木霉对几种病原菌的拮抗作用及液体产孢培养条件的研究

通过对峙培养法测定了哈茨木霉(Trichoderma harzianum Rifai Strain TXL051)对10种病原菌的拮杭作用,并对哈茨木霉在PDA液体培养基中产孢条件进行了研究。结果表明,哈茨木霉对10种病原菌均有抑制作用,是具有较好生防潜力的木霉菌。液体产孢最适条件pH 7,温度28℃,提高转数,增加光照均对产孢有促进作用。利用真菌18S rDNA通用引物扩增、测序获得了该哈茨木霉菌株的18S rDNA序列。     

多元抗生素与重金属混合物对蛋白核小球藻的时间依赖性协同与拮抗作用

为探讨水环境中典型污染物抗生素和重金属的联合毒性相互作用,以蛋白核小球藻(Chlorella pyrenoidosa,C.pyrenoidosa)为指示生物,采用96孔微板作为实验载体,以5种氨基糖苷类抗生素——硫酸阿米卡星(AMI)、硫酸庆大霉素(GEN)、硫酸卡那霉素(KAN)、硫酸巴龙霉素(PAR)、妥布霉素(TOB)和4种重金属——镉(Cd)、铜(Cu)、锰(Mn)和锌(Zn)为目标混合物,应用均匀设计射线法设计九元混合物体系,包括10条具有不同组分浓度配比的混合物射线。采用已建立的时间依赖微板毒性分析法系统测定了组分及其多元混合物体系的毒性,采用非线性最小二乘法拟合浓度-毒性数据,应用浓度加和参考模型分析在不同暴露时间的多元混合物体系的毒性相互作用。结果表明:9种污染物对C.pyrenoidosa的毒性具有明显的时间依赖性,但不同污染物的毒性随时间变化规律不同,毒性大小顺序也随时间不断发生变化,毒性大小顺序在96 h为:KANTOBZnMnCdAMIGENCuPAR;九元混合物射线的毒性也具有时间依赖性,但浓度配比不同射线的毒性随时间变化规律不同;10条混合物射线对C.pyrenoidosa的毒性具有协同和拮抗作用,且具有明显的时间依赖性,其中R5~R7三条混合物射线呈现明显的时间依赖性拮抗作用,且随暴露时间的延长而趋于明显,余下7条混合物射线呈现明显的时间依赖性协同作用,但随暴露时间的延长而逐渐减弱;对同一暴露生物,混合物毒性相互作用不仅与混合物的组分浓度配比有关,也与暴露时间有关。     

烟管菌M-1对柑橘炭疽病菌的抑制作用及发酵条件优化

本文优化了烟管菌Bjerkandera adusta M-1发酵液对柑橘炭疽病Colletotrichum gloeosprioides的抑制发酵条件,为柑橘炭疽病菌生物防治提供研究基础。通过采用平板对峙试验、单因素试验及响应曲面法,研究了烟管菌M-1菌株对柑橘炭疽病菌的拮抗作用。平板对峙试验表明,烟管菌M-1对柑橘炭疽病菌具有明显的抑制作用,抑菌率达65.7%,显著高于化学农药多菌灵和甲基托布津;发酵条件优化后,当C/N 7.5,初始pH 6.5,装瓶量36%,发酵时间22 d,转速180 r/min,温度30℃时,对柑橘炭疽病菌的抑菌率最高,达71.1%。本研究首次发现烟管菌M-1对柑橘炭疽病菌具有很好的拮抗作用,并确定了最优的发酵条件组合,这将为利用烟管菌防治柑橘炭疽病的实际应用奠定理论基础。