突变哈茨木霉H-13对小白菜生长和品质的影响

用N＋离子束诱变哈茨木霉,得到促生长突变型哈茨木霉H-13,用不同浓度的哈茨木霉H-13发酵液浸种处理小白菜种子,对幼苗进行灌根处理发现,发酵液稀释100倍和200倍处理,提高了种子的活力指数、植株鲜重和叶绿素含量,增加了植株体内可溶性还原糖和可溶性蛋白的含量,降低了植株体内硝酸盐和亚硝酸盐的含量.     

芦荟内生菌内生哈茨木霉LH-7对植物病原菌的抗性

通过体外培养方法,研究了药用植物库拉索芦荟内生真菌内生哈茨木霉LH-7对9种常见植物病原菌的抑菌谱及其拮抗机理.结果表明： 内生哈茨木霉LH-7 对供试植物病原真菌具有显著的抑制作用(抑制率为62.4%～88.4%),且营养竞争和重寄生是其主要拮抗机制.该菌株的代谢产物中具有能够抑制病原菌生长及孢子萌发的活性物质,其作用可导致病原菌菌丝体生长的畸形、细胞壁破裂、产孢结构发育不健全、孢子形成及萌发率降低,从而有效抑制病原菌的生长和繁殖.
     

哈茨木霉的培养及其对烟草疫霉生长的抑制研究*

哈茨木霉是一类重要的植病生防因子。哈茨木霉TH-1分别在PDA培养基、麦芽糖培养基、查氏培养基和琼脂培养基上培养均能产孢,其中PDA培养基为最适培养基。PDA培养基上,菌丝生长适宜温度27.5℃~35℃,最适温度32.5℃,产孢最适温度27.5℃。菌丝生长适宜pH值为3~7,产孢适宜pH值为5~9,生长与产孢最适pH值为5。光照对菌丝生长影响不大但明显影响菌株的产孢数量,光照时间越长产孢量越大。对峙培养试验表明TH-1明显抑制疫霉菌的生长速率,其无菌滤液明显抑制烟草疫霉菌游动孢子的萌发,并抑制游动孢子芽管     

紫外光诱导苎麻疫霉抗甲霜灵突变及对苎麻疫霉生物学特性的影响

作者研究了紫外光对苎麻疫霉的抗甲霜灵诱变效应及对苎麻疫霉生物学特性的影响。结果表明,供试6个苎麻疫霉野生型菌株经菌丝块紫外光药剂诱变和菌丝块药剂驯化3周后,均获得抗甲霜灵突变体,且紫外光药剂诱变处理角变区出现频率明显高于药剂驯化处理,说明紫外光对苎麻疫霉抗甲霜灵突变有一定促进效应。紫外光显著地抑制苎麻疫霉游动孢子的萌发和菌丝的生长,在一定范围内,处理时间愈长,抑制率愈高。苎麻疫霉耐紫外光菌株(经亚致死剂量的紫外光照射处理后存活的游动孢子所形成的菌株)与野生型亲本相比,对温度和pH的敏感性大致相同,但菌落形态有一定变异,菌丝生长速率、卵孢子产生量均显著下降,表明紫外光对苎麻疫霉的菌丝生长和卵孢子产生量有明显的抑制作用。苎麻疫霉耐紫外光菌株EC50值比野生型亲本菌株EC50值提高了23.21%-56.70%,即耐紫外光菌株对甲霜灵敏感性比野生亲本菌株显著下降,这与紫外光诱变试验结果是相一致的。     

非洲哈茨木霉LMNS-M9的鉴定、生物学特性及其对藜麦的促生作用

【背景】藜麦作为风靡全球的功能性食品,其种子内生非洲哈茨木霉的研究尚未见报道。【目的】从藜麦种子中筛选出有抑菌活性的内生真菌,分析其抑菌和促生作用。【方法】通过形态学及ITS、tef1和rpb2基因序列等方法对筛选出的内生真菌进行鉴定;采用平板对峙和平板倒扣等方法测定内生真菌的抑菌活性;利用盆栽试验测定内生菌株对藜麦生长的影响。【结果】综合形态学特征和系统发育分析结果,确定菌株LMNS-M9为非洲哈茨木霉(Trichoderma afroharzianum)。30℃、pH 5.0和镁离子适宜菌株LMNS-M9菌丝生长和产孢;葡萄糖、乳糖、蛋白胨、磷酸二氢钾适宜菌株LMNS-M9菌丝生长;葡萄糖、硝酸铵、磷酸氢二钾适宜菌株LMNS-M9产孢。菌株LMNS-M9对Botrytis cinerea、Ascochyta caulina、Fusarium citri、Alternaria alternata和Trichothecium roseum的抑制率分别为12.53%、51.96%、52.38%、59.25%和62.04%,挥发物的抑制率分别为35.86%、61.54%、33.33%、41...     

哈茨木霉SKD-ZX-1的鉴定、发酵及其生防效果

哈茨木霉作为一类重要的生防菌,对蔬菜病害致病菌具有拮抗作用。为确定分离的哈茨木霉SKD-ZX-1的生防效果,利用气质联用技术对菌株的发酵液进行测定和分析,利用宏基因组测序方法分析该菌株对番茄根腐烂病土壤中的细菌菌群的影响,并且采用平板对峙培养法对该菌株与链格孢菌和茄链格孢菌进行了拮抗实验。结果显示,哈茨木霉SKD-ZX-1发酵液中有邻苯二甲酸二异丁酯、邻苯二甲酸二丁酯和邻苯二甲酸二(2-乙基己基)酯等抑菌成分,并且其发酵液增大了土壤中细菌菌群的种类,与对照组相比,实验组肠杆菌属提高了46%,而假单胞菌属和梭菌属降低了5.28%和36.1%。该哈茨木霉对两种病原菌的抑制率均达到100%,拮抗系数达Ⅰ级。研究表明哈茨木霉SKD-ZX-1对链格孢菌和茄链格孢菌具有显著的抑制作用,其发酵液具有一定的抑菌效果。     

梨树胶孢炭疽病生防菌筛选及其抑菌活性初步评价

采集安徽农业大学农业园果树实习基地梨园不同组织和根际土壤样本共83份,将分离纯化后得到的细菌、真菌和放线菌采用琼脂拮抗法,测定各微生物对梨胶孢炭疽病菌菌丝生长的抑制作用,并选用几种常用药剂分别对病原菌菌丝和分生孢子进行处理,观测其化学抑制效果。结果表明,菌株RT-30对梨胶孢炭疽病菌丝生长的抑制率达65.5%,明显高于80%波尔多液可湿性粉剂400倍液,将RT-30进行形态学观察及分子学序列分析,初步将RT-30菌株鉴定为甲基营养型芽孢杆菌Bacillus methylotrophicus。     

葡萄糖异构酶高产菌株的诱变选育

葡萄糖异构酶产生菌链霉菌B6经紫外线、亚硝基胍等诱变处理,经反复筛选获得T208变异株。测定了该菌产酶的适宜生长条件。该菌株比出发菌株酶活力提高2.75倍,可达550u/mol.该菌菌丝粗壮,易培养,有利于工业化生产。     

桔梗新病害——匍柄霉叶斑病的病原生物学特性及药剂敏感性分析

桔梗匍柄霉叶斑病是近年来发现的新病害,病原菌为桔梗匍柄霉Stemphylium platycodontis。为了更好地了解病害的发生规律,本研究对该病原菌的生物学特性及药剂敏感性进行了分析。结果表明：25℃为该病原菌的最适生长温度和产孢温度;菌丝生长最适pH值为4,而最适合产孢的pH值为8;24h黑暗处理,对病原菌的菌丝生长和产孢都最有利;PCA培养基对菌丝生长最有利,而桔梗汁液培养基对病原菌产孢最有利;可溶性淀粉对菌丝生长最有利,而蔗糖最利于病原菌产孢;乙酰铵对病原菌生长及产孢都最有利;26.8μg/mL氟硅唑对桔梗匍柄霉的抑制效果最好,毒力持效性最强。     

哈茨木霉的培养及其对烟草疫霉生长的抑制研究

哈茨木霉是一类重要的植病生防因子。哈茨木霉TH-1分别在PDA培养基、麦芽糖培养基、查氏培养基和琼脂培养基上培养均能产孢,其中PDA培养基为最适培养基。PDA培养基上,菌丝生长适宜温度27．5℃～35℃,最适温度32．5℃,产孢最适温度27．5℃。菌丝生长适宜pH值为3～7,产孢适宜pH值为5-9,生长与产孢最适pH值为5。光照对菌丝生长影响不大但明显影响菌株的产孢数量,光照时间越长产孢量越大。对峙培养试验表明TH-1明显抑制疫霉菌的生长速率,其无菌滤液明显抑制烟草疫霉菌游动孢子的萌发,并抑制游动孢子芽管的伸长,TH-1对游动孢子萌发的相对抑制率为12．7％,对芽管生长长度的相对抑制率为63．1％。水解酶平板活性测定显示,TH-1产生β-1,3葡聚糖酶与纤维素酶,从而使烟草疫霉菌细胞壁的消解,产生非挥发性抗生素抑制烟草疫霉菌孢子萌发,但对菌丝生长影响不大。     

拮抗木霉耐多菌灵菌株的筛选

该文进行了耐多菌灵生防木霉菌株的筛选和生防效果的测定研究。为获得对灰葡萄孢霉等病原菌有显著拮抗能力的耐多菌灵木霉菌株,通过含药平板诱导和紫外线诱变,获得5株耐药性菌株,在多菌灵2.0g/L浓度下生长较好,Ec50值提高了40多倍;抗性菌株在含药培养基上连续继代转移培养10次,抗性程度未见下降;经过平板对峙和活体拮抗性测定,确定T42-2为最佳拮抗性耐多菌灵菌株,其对黄瓜灰霉病的防效达92.13%,并且对多菌灵、速克灵和三唑铜存在交互抗性。     

茶树内生木霉种的鉴定及其在植物体内的定殖

采用分离自健康茶树叶片组织的一株长枝木霉Trichoderma longibrachiatum菌株CSN-18,研究茶树内生木霉的人工接种、再分离及其在茶树地上部组织中的内生性。该菌在PDA培养基上生长速度快,产孢量大。用CSN-18回接茶苗,接种后一个月可以从茶苗的茎和叶组织中再分离获得该真菌。接种后的茶苗没有观察到明显的病害表现;与对照相比,接种后6个月组培苗生长良好,叶色更绿;接种后1个月,实生苗生长正常。通过石蜡切片和苯胺蓝染色,在已接种的茶树组培苗的叶片内部组织中可观察到木霉的存在,从而证明了木霉能够在茶树地上部组织内定殖,是茶树的一种内生真菌。     

几种虫生真菌菌株的培养性状及其对褐飞虱的毒力

从褐飞虱Nilaparvata lugens (St(a)l)罹病虫体上新分离出的一种黄绿绿僵菌菌株Metarhizium flavoviride (Mf82),与实验室保存的黄绿绿僵菌、金龟于绿僵菌和白僵菌3种菌种5个菌株作为对比,研究了在SDAY和PDA培养基上的培养性状,并测定了它们对褐飞虱成虫的毒力.结果表明:...     

两种生防木霉菌的种群动态监测及回收毒力测定

目的:对施入人参地的两株木霉菌的种群动态进行长期监测及回收毒力测定,为应用木霉菌安全、长期、有效防治根部病害提供理论依据。方法:采用木霉菌选择性培养基筛选法和平板对峙培养法,对木霉菌进行回收测定及毒力评价。结果:Th3080与Tv04-2在人参地土壤环境中定殖后可长期大量存活。150d检测时,木霉菌种群数量与初始投入量的比率平均为16.2%,最高达44.5%。两种木霉菌对3种人参病原菌的抑制率较原始菌株有所增强或无显著差异。结论:木霉菌引入土壤微生态后可以建立种群。木霉菌不仅可以长期大量存活,而且能很好的保持毒力。     

红托竹荪菌种快速分离培养基优化

通过对红托竹荪快速分离培养基优化,提高红托竹荪菌种分离与评价效率。采用响应面分析法,以菌种生长速度为响应值拟合二次多元回归方程,确定培养基配方;测定优化培养基与PDA对照培养基菌丝生长速度和菌丝直径,以菌丝形态、锁状联合和菌落形态等指标评价优化培养基;测定优化培养基与PDA培养基培养菌丝在木屑培养基中菌丝生长速度,验证应用效果。通过试验,筛选出快速分离培养基配方为葡萄糖20.71 g/L、全麦粉8.36 g/L、玉米粉8.07 g/L、琼脂粉18.00 g/L、木屑水1.06 L。快速分离培养基与PDA培养基对比,培养的菌落直径平均增加66.25%,快速分离培养基菌丝日平均生长速度增加33.33%,木屑培养基菌丝日平均生长速度增加44.22%。由于优化培养基中含有淀粉、纤维素等有效成分,其刺激了菌种分泌淀粉酶、纤维素酶等,维持了胞外酶系的完整性。还可根据菌丝培养基过程形成的透明圈大小判定菌种胞外酶产生能力,达到快速评价菌种质量,保障菌种质量的目的。     

Influence of Culture Conditions on Growth and Survival of Conidia of Trichoderma spp. Coated on Seeds

Five Trichoderma strains were grown on rice, on vermiculite plus potato-dextrose broth (PDB), on potato-dextrose agar (PDA) or in liquid cultures supplemented with glycerol, KCl or polyethylene glycol (PEG) at -1 MPa or - 2 MPa. Conidia were coated on seeds through a methyl cellulose coating or through an industrial film-coating process. The conidial yield decreased with glycerol, KCl or PEG compared with PDB alone. The percentage viability was from 23 to 44% after methyl cellulose coating, regardless of the culture conditions for conidial production. In general, the industrial coating resulted in lower numbers of living conidia. The viability during storage was enhanced when vermiculite, rice or PDA were used as substrates for fungal growth. Nevertheless, temperature of storage was found to be more critical to spore survival than the substrate used for spore production; conidial viability on seeds did not exceed 4 months at 15 C. Solid and liquid cultures produced conidia able to control R. solani and P. ultimum when applied to seeds through industrial film coating. The level of disease suppression varied with the number of viable conidia/seed and with the culture medium used for conidial production. The three main conditions for further industrial application-high yields, longevity and biocontrol effectiveness-might be optimized by selecting the appropriate medium (liquid or solid), water potential and solutes used.     

Regeneration and molecular characterization of an intergeneric hybrid between Graphium putredinis and Trichoderma harzianum by protoplasmic fusion

The fungal strains Graphium putredinis and Trichoderma harzianum were selected as parents for fusant development. Protoplasts were isolated using the combination of lysing enzymes Novozym 234 and cellulase with 0.6 M KCl as osmotic stabilizer. The optimum conditions for release of viable protoplasts from the fungal mycelium viz. age of the mycelium, lytic enzymes, osmotic stabilizers, pH, incubation period and regeneration medium were determined. Intergeneric protoplast fusion was carried out using 50% polyethylene glycol with calcium chloride (CaCl₂) and glycine buffer and the conditions for effective protoplast fusion, viz. fusogen, osmotic stabilizer, pH, incubation period and regeneration medium were optimized. At optimum conditions, the regeneration frequency of the fused protoplasts on potato dextrose agar (PDA) medium and fusion frequency were calculated. The regeneration frequency on non-selective (PDA) and selective media (PDA amended with starch) was determined for the parental and fusant strains in which, fusant showed a higher rate of regeneration. Fusant formation was confirmed by morphological markers (colony morphology and spore size and shape) and genetical markers like, mycelial protein pattern, restriction digestion pattern and random amplified polymorphic DNA (RAPD) analysis. The efficiency of these parental strains and their intergeneric fusant in the production of hydrolytic enzymes – amylases (treatment plant for sago factory effluent), cellulases (bioethanol), xylanases (bleaching agents for waste paper pulp) and proteases (additives in commercial detergents) – have probable applications in various industrial processes.     

脱落酸产生菌的遗传育种

从南京蔬菜研究所植物致病株中分离到3株脱落酸（ABA）产量较高菌株,经初步鉴定均为灰葡萄孢（BolrytiscinereaPers．）。分别命名为A23,A101,A160通过紫外诱变及菌株生长状况比较,选出一株菌A101作为出发菌,经紫外,硫酸二乙酯多次诱变,在ANA合成代谢关键酶抑制剂平板上定向选育出一株ANA高产菌UUD－1,该菌固体发酵可产生ABA374μg／ml,是出发菌株产量的14倍,且其产量连续培养5代稳定。     

Effect of spore concentration on germination and autotropism inTrichoderma hamatum

Summary The effect of spore concentration on spore germination and germtube growth ofTrichoderma hamatum on water agar and on potato dextrose agar (PDA) was studied. Increasing inoculum size up to 10⁹ spores/plate on PDA and up to 10⁷ spores/plate on water agar shortened the incubation period required for germtubes emergence and increased germination rate. However, on water agar germination was inhibited at 10⁸ and was completely arrested at 10⁹ spores/plate. Inhibition in germination of 10⁷ spores/plate was observed on water agar when the plates were preincubated with 10⁹ spores/plate for 5 h or more. Addition of glucose and ammonium nitrate to the water agar medium allowed only 25% of the spores to germinate at 10⁹ as compared to 78% at 10⁷ spores/plate after 8 h of incubation. Addition of polysaccharides to the C+N supplemented medium, significantly increased germination up to 84% as compared to 100% on PDA, after 8 h of incubation. Germlings ofTrichoderma hamatum phialospores exhibited positive autotropism and anastamosis on both media. The phenomenon was positively related to inoculum size, being most pronounced at 10⁷ spores/plate.     

耐热木霉菌株筛选及其对热作区香蕉促生效应的研究

[目的]为筛选安全、高效的耐热木霉功能菌株,有效促进热作区香蕉的生产,本研究从热作区土壤分离筛选适温范围较宽的木霉菌株,研制木霉生物有机肥,并研究其对香蕉枯萎病发病率、果实产量及品质的影响.[方法]通过稀释涂布法筛选出木霉菌株,根据菌株在不同温度下的生长情况、拮抗尖孢菌能力及酶活强弱进行菌株复筛;将复筛所得菌株试制成生...