致病疫霉拮抗菌筛选及复合发酵液的抑菌作用研究

为了利用微生物源活性物质控制马铃薯晚疫病,对显著抑制致病疫霉菌丝生长的拮抗微生物进行了筛选,并研究其单一发酵液和复合发酵液对致病疫霉的抑制作用。结果表明:从36种微生物中筛选出了16种对致病疫霉菌丝生长有显著抑制作用的微生物,其中B3282(细菌)、枯草芽孢杆菌和A32810(放线菌)这3种微生物的抑菌率均在70%以上;A4228、A32912、A5253和A32914等4种放线菌的抑菌率均在60%以上。对36种微生物单一发酵液的抑菌试验表明:仅白菜黑斑病菌、梨黑斑病菌和A32914的发酵液对致病疫霉表现出较强的抑制作用,抑菌率分别为80.51%、70.20%和48.91%;进一步将36种单一发酵液两两复合后发现,部分复合发酵液表现出了明显的增效作用,其中抑菌率最高及增效作用最强的是梨黑斑病菌和白菜黑斑病菌浓度为10 mg/ mL的复合发酵液,抑菌率达90.38%。     

不同茯苓菌株生物学特性和菌核主要成分比较

对6株茯苓(Poria cocos)菌株的菌丝生长速率、菌核产量、菌核中多糖和三萜含量以及菌株间亲和性进行比较.结果表明:供试菌株中,8号菌株的菌丝生长速率等生物学特性均显著优于其它茯苓菌株.该菌株在PDA培养基上的菌丝生长速率为2.10 cm/d;采用松树兜法栽培茯苓,该菌株的菌核产量达到每兜4.76 kg;菌核中还原糖含量为0.43％,总糖含量为7.37％,三萜含量为0.74％;拮抗试验表明6株茯苓菌株之间存在拮抗作用,说明茯苓菌株体细胞亲合基因间存在差异.     

Effect of benomyl on soil fungi associated with rye. 1. Effect on the incidence of sharp eyespot caused by Rhizoctonia cerealis

Effects of benomyl on incidence of pathogens affecting the culm base of rye were studied in field trials and growth chamber experiments. Spraying of the crop with the fungicide at a high dosage (2.4 kg.ha^–1) resulted in a tenfold increase of sharp eyespot caused byRhizoctonia cerealis and reduced foot rot symptoms caused by fusaria by 50%. In a field trial at a low dosage (0.24 kg.ha^–1) a slight increase of sharp eyespot was observed. In one year, probably because of wet conditions during the infection period, sharp eyespot did not occur in either benomyl-treated or untreated plots, but eyespot caused byPseudocercosporella herpotrichoides was abundant. Its occurrence was reduced from 74% affected culm bases in untreated plots to 8% and 1% in plots that received 0.24 and 2.4 kg.ha^–1 of the fungicide, respectively.In growth chambers seedlings were grown in two sandy soils inoculated withR. cerealis. The soil was kept dry at about 35% of the moisture holding capacity. In plots with benomyl (1 mg.kg^–1; moisture content 11% of fresh weight), fewer seedlings emerged than in plots without the fungicide. This result was highly significant (P<0.01) for one soil but not for the other. The number of seedlings that remained free of disease symptoms was higher (P<0.01) in untreated than in fungicide-treated plots of both soils.Isolates of pathogens obtained from diseased culms were tested for their sensitivity to benomyl. Growth of all of them includingR. cerealis was inhibited, although not always completely suppressed, at 10 g.ml^–1 on potato-dextrose agar. ED₅₀ values of most isolates ofR. cerealis were between 2.2 and 3.1 g.ml^–1. The fungus was slightly but consistently less sensitive thanF. culmorum. Mycelial growth ofF. nivale was appreciably more sensitive than that of the otherFusarium spp. from cereals.P. herpotrichoides andF. nivale were the most sensitive pathogens tested with ED₅₀ values of <1 g.ml^–1. Accordingly,F. nivale was absent on culms from treated plots. In a growth chamber experiment, seedlings were protected from infection by supplying the fungicide (1 mg.kg^–1) to previously inoculated soil.In a laboratory assay the effect of benomyl on microbial antagonism toR. cerealis was estimated for rhizosphere soil. Enhanced incidence of sharp eyespot in treated crops was associated with adverese effects of the fungicide on microbial antagonism. There is presumptive evidence thatR. cerealis is suppressed by bacteria after wet periods during the vegetation period of the crop and by fungi after dry periods. Only fungal antagonism, which may be less effective, is affected by benomyl. The response to benomyl of the microflora in different soils varied. Reasons for this inconsistency are suggested.Samenvatting In veldproeven en in een klimaatkamer werd de invloed van benomyl op het optreden van voetziekten in rogge onderzocht. In veldjes die bespoten waren met een hoge dosis van het fungicide (in totaal 2.4 kg.ha^–1) bleken tienmaal zoveel halmen met scherpe oogvlekken, veroorzaakt doorRhizoctonia cerealis, voor te komen dan in onbespoten veldjes. Daarentegen was voetrot veroorzaakt doorFusarium-soorten met 50% verminderd. In een volgende veldproef, waarbij een voor de praktijk geadviseerde dosis (0.24 kg.ha^–1) was toegepast, werd een lichte toename van scherpe oogvlekken waargenomen.In een ander jaar trad scherpe oogvlekkenziekte in het geheel niet op, ook niet in met benomyl behandelde veldjes. De vochtige omstandigheden tijdens de infectieperiode zijn daarvan waarschijnlijk de oorzaak. Daarentegen werd de oogvlekkenziekte, welke doorPseudocercosporella herpotrichoides werd veroorzaakt, veel aangetroffen. In de onbehandelde veldjes waren 74% van de halmen aangetast tegen 8 en 1% in de veldjes die met het fungicide waren behandeld in doseringen van 0.24 en 2.4 kg.ha^–1.De invloed van het fungicide op de aantasting van kiemplanten werd in klimaatkamerproeven onderzocht. Daartoe werden twee zandgronden metR. cerealis geënt. De grond werd droog gehouden (op 35% van het waterhoudend vermogen). In grond met fungicide (1 mg.kg^–1) was de opkomst minder dan in grond zonder fungicide. Dit was zeer significant (P<0.01) voor één van de beide zandgronden, maar niet voor de andere. Het aantal gezonde kiemplanten was in beide gevallen duidelijk hoger (P<0.01) voor de onbehandelde grond.De isolaten van ziekteverwekkers uit aangetaste halmen werden op hun gevoeligheid voor het fungicide getoetst. Op aardappel-glucoseagar werden alle isolaten in hun groei geremd bij een benomyl-concentratie van 10 g.ml^–1.R. cerealis was iets minder gevoelig danF. culmorum. Voor het overgrote deel van de isolaten vanR. cerealis lag de ED₅₀ waarde tussen 2,2 en 3,1 g.ml^–1. De myceliumgroei vanF. nivale werd meer geremd dan die van de andereFusarium-soorten.P. herpotrichoides enF. nivale waren met een ED₅₀ waarde van <1 g.m.^–1 de gevoeligste pathogenen die uit de halmvoeten werden geïsoleerd. Dat de populatie vanF. nivale in benomylhoudende grond wordt onderdrukt, blijkt uit (1) het feit dat de schimmel niet voorkwam op halmen uit behandelde veldjes en (2) de bescherming tegen infectie van kiemplanten als aan de besmette grond fungicide (1 mg.kg^–1) was toegevoegd.In laboratoriumproeven werd de invloed van benomyl op het microbiële antagonisme in rhizosfeergrond tegenR. cerealis bepaald. Een toename in het optreden van scherpe oogvlekkenziekte in behandelde gewassen bleek gepaard te gaan met een remming van het antagonisme tegen de ziekteverwekker. Er zijn sterke aanwijzingen datR. cerealis na vochtige perioden tijdens de vegetatieperiode door bacteriën wordt onderdrukt en na droge perioden door schimmels. Het antagonisme van de laatste groep lijkt minder effectief te zijn en alleen dit antagonisme wordt door benomyl verlaagd. Tenslotte wordt een mogelijke oorzaak aangegeven voor de ongelijke respons op het fungicide van het microbieel antagonisme in verschillende gronden.     

Mineral nutrition, soil factors and growth rates of Gmelina arborea plantations in the humid lowlands of northern Costa Rica

The purpose of this study was to determine the factors which are responsible for clearly visible growth irregularities in Gmelina arborea stands by relating growth rates to soil and site properties. All Gmelina plantations under study, established on degraded pasture soils in the Atlantic lowlands of Costa Rica, showed the same growth pattern: very poor growth and chlorotic foliage of trees in mid-slope positions, and fast growth and healthy leaves of trees on hilltops and hillbases. The variation in site and soil properties and tree growth rates was analysed, investigating 24 plots in eight different plantations. Leaf samples were taken from 120 trees (five trees per site). Foliar analysis revealed that the tree growth is highly dependent on the supply of N, P, K and S, indicating that poorly growing trees suffer from a multiple nutrient disorder. To identify the most restricting soil factor, simple correlations between growth rates and soil chemical and physical properties were applied to the entire data set. The best correlation was obtained with exchangeable soil K (r=0.78, P<0.001). Subdivision of the data set into plots on brown soils (eight) and plots on red soils (16) and subsequent correlation analysis resulted in much stronger relationships. Growth depressions in mid-slope positions had other causes on brown soils than on red soils. On the acid brown soils the combination of the variables Al saturation and bulk density could explain 82% of the variation of tree growth. Al saturation of up to 80% in brown soils inhibits nutrient uptake, particularly of N and P. The red soils dominate on Ca---Mg-enriched, alluvial terraces and were among the first soils to be cultivated in the region. During the agricultural use and at the establishment of the tree plantations, they received dolomitic lime to reduce Al toxicity. Here, very low K/Mg ratios (less than 0.03) may induce K deficiency. Therefore, the best multiple regression model for tree growth rates on red soils is obtained with K/Mg ratio and thickness of the humic A-layer (r²=0.75, P<0.001). Inclusion of the variable bulk density resulted in a clear improvement of the model, explaining 91% of the growth variability.     

杏鲍菇单孢诱变杂交及其子代的鉴定

将杏鲍菇单核菌丝进行紫外诱变,挑选优良单核菌株杂交配对,获得47个长势较好的子代菌株。通过拮抗试验与脂酶同工酶分析预测子代菌株杂种优势,筛选出有异于亲本的7个杏鲍菇新菌株。     

多功能木霉的筛选及鉴定

测定了60株分离自土壤的木霉菌株的解磷解钾、拮抗和对有机磷农药毒死蜱的耐受、降解等功能。结果表明,5株木霉具有很好的解磷能力,其中木霉T-404的解磷效果最好,为109.22 mg·L-1。6株木霉具有明显的解钾透明圈,但是液体培养后测定发现木霉的解钾能力不强。木霉具有不同程度的拮抗作用,其中6株木霉的拮抗能力很好。大部分木霉可以耐受高浓度的毒死蜱,但是不能降解该农药。对5株具多功能潜力的优势木霉菌株进行形态学和分子生物学鉴定表明:T-403、T-404、T-440为Trichoderma koningiopsis,T-400、T-450为Trichoderma koningii。     

短小芽孢杆菌D82对小麦根腐病原菌拮抗的研究

平板培养结果表明，短小芽孢杆菌Ｄ８２对小麦根腐病有强抗生作用，共同培养至第４天，抑菌圈直径为４－５．５ｃｍ；发酵液稀释平板培养结果表明，稀释浓度１００倍以下的抑菌效果为１００％。１０００倍、２０００倍、４０００倍稀释液的抑菌效果分别为６４％、５４％和３４％。     

木霉菌和腐霉菌对棉铃疫菌的作用研究

用玻璃纸对峙法观察到4种木霉(S-327,BT-12,BT-95,T-594)和寡雄腐霉(P.O.)对棉铃疫霉的营养生长均有一定抑制作用,但没有看到明显的抑菌圈。生物测定试验表明,拮抗菌不仅对棉铃疫霉有直接作用,而且还可以保护棉铃不受疫菌侵染。在5种拮抗菌中,P.O.和BT-12可能是防治棉铃疫病比较有潜力的生防菌。     

拮抗细菌B20-006对玉米根系内生细菌种群的影响

 Treatment of corn seeds with Bacillus subtilis strain B20-006, a potential biocontrol agent, had no effect on type of endophytic bacteria on corn. However, it stimulated endophytic bacterial reproduction in the plants. B20-006 did not affect the original population of endophytic bacteria in corn roots in early stage. In late stage, population of endophytic bacteria in corn roots treated with the bacteria was significantly higher than CK. Colonization of the bacteria in corn roots would benefit the micro-ecological resistance of cultivars against the diseases. The results suggested that strain B20006 could enhance the percentage of antagonistic endophytic bacteria against Rhizoctonia solani. The percentage in the resistant cultivar, Lianyu 11 was higher than in the susceptible one, Lianyu 13.