Effectiveness of Coniothyrium minitans and Trichoderma atroviride in suppression of sclerotinia blossom blight of alfalfa

The effects of the mycoparasites Coniothyrium minitans and Trichoderma atroviride on the suppression of alfalfa blossom blight caused by Sclerotinia sclerotiorum were evaluated under indoor and field conditions. When T. atroviride (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) were applied to detached young alfalfa florets, T. atroviride effectively inhibited saprophytic growth of S. sclerotiorum, whereas C. minitans showed no inhibition under the same conditions. When T. atroviride (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) was applied to young alfalfa petals in vivo just after pollination, the percentage of pod formation was higher for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum, and the percentage of pod rot was lower for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum. However, when they were applied to senescent petals attached to developing pods of alfalfa at 9·2 × 10⁴ conidia/floret together with S. sclerotiorum at 4·5 × 10³ ascospores/floret at 14 days after pollination, C. minitans was more effective than T. atroviride in suppressing sclerotinia pod rot and seed rot of alfalfa. Field experiments showed that three applications of C. minitans (5·4 × 10⁶ conidia mL⁻¹) or T. atroviride (5·4 × 10⁶ conidia mL⁻¹) at a 7-day interval to blossoms of alfalfa effectively suppressed sclerotinia pod rot in two out of three annual trials. Coniothyrium minitans effectively suppressed sclerotinia seed rot in all three years, whereas T. atroviride was not effective against seed rot in any of the trial years. The efficacy of C. minitans was not significantly different (P > 0·05) from benomyl (250 µg ai mL⁻¹). This study suggests that C. minitans has potential as a biocontrol agent to control blossom blight of alfalfa caused by S. sclerotiorum.     

The process of antagonism of Sclerotium cepivorum in white rot affected onion roots by Trichoderma koningii

Trichoderma koningii (strain Tr5) grew in the epidermal mucilage of onion roots without entering healthy epidermal tissue. When placed on the epidermis of Sclerotium cepivorum -infected roots, T. koningii colonized epidermal passage cells, with little colonization of other epidermal tissues, then branched and spread throughout the root cortical tissues damaged by enzymes and toxins which diffused ahead of S. cepivorum hyphae, and impeded the path of the infection. When T. koningii colonized infected tissue, many S. cepivorum hyphae became detached at septa, cell walls dissolved and many hyphal apices burst. Contact between hyphae was not necessary for lysis to occur. T. koningii produced two endochitinases ( R _f 0·15 and 0·24) and two exo-acting chitinolytic enzymes ( R _f 0·46 and 0·62) during degradation of crabshell chitin and S. cepivorum cell walls. The R f 0·24 and 0·46 proteins were detected when T. koningii colonized S. cepivorum -infected roots and are likely to be a component of the antagonism process.     

Evaluation of Microbial Antagonists for Biological Control of Botrytis cinerea Stem Infection in Cucumber and Tomato

Of fifteen isolates of yeasts, filamentous fungi and bacteria and a commercial product, tested in a bioassay with stem segments, eleven isolates consistently reduced incidence of disease and sporulation of Botrytis cinerea Pers; Fr in tomato and seven isolates in cucumber. Several isolates reduced disease by more than 75% in all experiments. Six antagonists that performed well in the bioassays and that were fairly easy to produce in vitro, were selected for further testing in two glasshouse experiments with cucumbers. After application of spores of B. cinerea and the antagonists or the fungicide tolylfluanid to pruning wounds, disease incidence was reduced by 50–100% by all antagonists in both experiments and only in one experiment by tolylfluanid.For Trichoderma harzianum T39, Aureobasidium pullulans and Cryptococcus albidus, biological control efficacy in bioassays with cucumber stem segments was not strongly influenced by temperatures in the range between 18 and 30°C, but at 24°C the efficacy of the three antagonists strongly decreased at relative humidities of 90% and 80% (vapour pressure deficits 0.299 and 0.598kPa, respectively) compared to 100.     

The potential of nonpathogenic Fusarium oxysporum and other biological control organisms for suppressing fusarium wilt of banana

The aim of this study was to evaluate the ability of nonpathogenic F. oxysporum and Trichoderma isolates from suppressive soils in South Africa to suppress fusarium wilt of banana in the glasshouse. Several biological control agents and commercial biological control products were included in the study. The isolates were first screened in vitro on potato dextrose agar. In glasshouse evaluations, the fungal and bacterial isolates were established on banana roots before they were replanted in pathogen-infested soil, while the commercial biocontrol agents were applied as directed by the supplier. Banana plantlets were evaluated for disease development after 7 weeks. In vitro tests showed none of the nonpathogenic isolates suppressed Fusarium oxysporum f.sp. cubense ( Foc ), while slight suppression was observed with the two Trichoderma isolates. Results of the glasshouse evaluations revealed that two of the nonpathogenic F. oxysporum isolates, CAV 255 and CAV 241, reduced fusarium wilt incidence by 87·4 and 75·0%, respectively. The known biological control agent Fo47 did not suppress Foc significantly. Pseudomonas fluorescens strain WCS 417, known for its ability to suppress other fusarium wilt diseases (WCS 417), reduced disease incidence by 87·4%. These isolates should be further evaluated for potential application in the field, independently and in combination.     

3株木霉(Trichoderma spp.)对华山松疱锈病菌锈孢子的破坏作用

从不同来源的茶藨生柱锈锈孢子堆和疱锈病枝干上分离获得3株木霉,室内生测结果表明,3株木霉的菌丝和培养滤液对锈孢子壁有较强的破坏作用;菌株TR2和TR3在锈孢子堆上生长速度快,锈孢子死亡率随孢子壁破坏率的增加而升高,锈孢子壁对孢壁降解酶有强烈诱导作用;TR1对锈孢子壁的破坏作用弱于TR2和TR3,但在PD培养液中有较强的产毒能力,锈孢子壁能抑制TR1菌株产毒。     

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.     

木霉菌和番茄叶表主要微生物间关系

对番茄叶表分离到的几种真菌和木霉菌BTW41(T.atroviride)之间相互关系作了初步研究,研究发现木霉菌具有较强的营养竞争能力,几种叶面真菌和木霉菌混合处理叶片试验中,黑曲霉菌混合木霉菌的处理会影响木霉菌在番茄叶表的定殖,使木霉菌在番茄叶表的定殖量减少,而其它几种真菌未使木霉菌定殖量减少。     

康宁木霉固态发酵改善茶渣营养价值

本试验旨在研究康宁木霉固态发酵茶渣,提高茶渣营养价值,并筛选出最佳的发酵条件。用单因素试验优化发酵茶渣的基质比例（茶渣∶玉米粉=6∶4、7∶3、8∶2、9∶1）、料液比（3∶7、4∶6、5∶5、6∶4、7∶3）、接种量（2%、4%、6%、8%、10%）、发酵温度（25、28、31、34、37 ℃）和发酵时间（0、2、4、6、8、10、14、22 d）。以基质比例、发酵温度、接种量和发酵时间为影响因素,进行L9（3⁴）正交试验确定茶渣最优发酵条件。测定各组发酵产物粗蛋白质、粗脂肪、还原糖、黄酮、皂苷和咖啡因含量,计算各组合的综合评分。确定最优条件后进行对比试验,比较了发酵前后茶渣的营养成分、活性物质和游离氨基酸含量。结果表明：1）单因素试验中,以下条件的综合评分最高：基质比例为7∶3,料液比为5∶5,接种量为8%,发酵温度为31 ℃,发酵时间为6 d。2）正交试验表明,康宁木霉发酵茶渣（料液比为5∶5）的最佳条件是：基质比例为7.0∶2.5,发酵温度为31 ℃,接种量为7%,发酵时间为6或7 d。3）与未发酵茶渣相比,最优条件下发酵茶渣的粗蛋白质、还原糖、黄酮、咖啡因、多种游离氨基酸含量和必需氨基酸/总氨基酸、风味氨基酸/总氨基酸均显著提高（P<0.05）,皂苷含量显著降低（P<0.001）,粗脂肪含量与未发酵茶渣无显著差异（P>0.05）。发酵6和7 d的茶渣营养成分和活性物质含量无显著差异（P>0.05）。由此可见,康宁木霉发酵茶渣能够改善茶渣的营养价值。     

挤压对麦糟纤维结构和固态发酵过程中pH值影响的研究

低成本纤维素酶的生产是酶法转化纤维质生物量为酒精的关键。尽管已有很多有关纤维素酶的研究报告，但底物的预处理对固态发酵过程中pH值和产酶的影响很少见诸于报道。作者研究了在固态发酵过程中，里氏木霉在未处理和经挤压处理的麦糟培养基上pH值的变化。经单螺杆和双螺杆挤压的麦糟，其纤维的结晶度变化较小。单螺杆挤压撕开了纤维结构，而双螺杆破坏并摧毁了纤维细胞的细胞壁，同时麦糟的颗粒也变得很小。两种经挤压处理的麦糟均有利于菌体的生长，并提高了产酶。以单螺杆挤压处理的麦糟为培养基时，最高FPA酶活力为182．8IU／g纤维素。     

两种产纤维素酶菌株分离的初步研究

本研究对采自贵州林区中的响叶杨(Populusadenopoda Maxim)腐朽木和白蚁(Odontotermes formosanus(Shiraki))进行分离培养,分离自杨树腐朽木的产纤维素酶菌株为木霉属康宁木霉(Trichodermakoningii Oud);分离自白蚁肠的产纤维素酶菌株为芽孢杆菌属(Bacillus sp.)。对所分离的两个菌株进行滤纸降解观察与测定。所分离的菌株(Trichoderma koningii Oud.)滤纸培养30天后,有菌丝生长,培养基浑浊,滤纸呈絮状,培养7天后该菌对滤纸降解强度为7.13%(校正值)。刚果红法初步证明该菌能产纤维素酶。从白蚁肠分离的菌株(Bacillus sp.)滤纸培养15天后,滤纸呈絮状,培养基变浑浊,有细菌菌体沉积;培养7天后,对滤纸的降解强度为8.18%(校正值)。刚果红法初步证明该菌能产生纤维素酶。用五种培养基质培养芽孢杆菌属(Bacillus sp.)菌株,并测定其生长量,其中以滤纸为主要基质的培养液中,最高可检测到0.3×107cfu/ml菌株量。