Preparation of dry flowable formulations of Clonostachys rosea by spray drying and application for Sclerotinia sclerotiorum control

A dry flowable formulation of Clonostachys rosea with fungicidal activity against Sclerotinia sclerotiorum was prepared by spray drying. The formulation was optimized by a four-factor, three-level orthogonal experiment to screen inert ingredients and spray-drying conditions. The optimal dry flowable formulation of C. rosea included 30% C. rosea （ratio of conidia powder and its fermentation broth is 1：3）, 3% Morwet EFW, 4% K12, 10% Morwet D425, 9% sodium salt of polynaphthalene sulphonic acid （NNO）, 5% croscarmellose sodium, 5% （NH4）2SO4, 0.5% carboxymethyl cellulose sodium （CMC-Na）, 1% oxalic acid and palygorskite （carrier） up to 100%. The formulation exhibited good physical characteristics, such as high dispersibility, viability and a long shelf life. Plate antagonism tests and pot trials indicated that the dry flowable formulation was very effective against S. sclerotiorum, with control efficiency of up to 88.30%. This dry flowable formulation of C. rosea is a new potential commercial fungicide for spray drying to control S. sclerotiorum.

薄叶卷柏内生真菌Clonostachys rogersoniana次生代谢产物研究

目的研究薄叶卷柏内生真菌Clonostachys rogersoniana的次生代谢产物。方法利用硅胶柱色谱、Sephadex LH-20凝胶柱色谱和半制备型高效液相色谱等现代色谱技术分离纯化Clonostachys rogersoniana的大米发酵产物,运用HR-ESI-MS、NMR等技术鉴定化合物结构。采用微量肉汤稀释法评价所有单体化合物对4种临床常见的致病菌(金黄色葡萄球菌Staphylococcus aureus、粪肠球菌Enterobacter faecalis、铜绿假单胞菌Pseudomonas aeruginosa和大肠埃希菌Escherichia coli)的抗菌活性。结果从Clonostachys rogersoniana大米发酵产物的乙酸乙酯萃取部位中分离得到11个化合物,分别鉴定为indole-3-acetic acid(1)、N-acetyltryptamine(2)、(1S,3S)-1-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid(3)、(1R,3S)-1-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid(4)、4-methoxyphenylacetic acid(5)、4-hydroxybenzoic acid(6)、hydroquinone(7)、4-hydroxyphenylacetic acid(8)、clonostachydiol(9)、2,3-dihydroxypropyl pentadecanoate(10)和daucosterol(11)。体外抗菌实验结果显示化合物3和4对Staphylococcus aureus具有中等抑制活性,MIC值分别为16和8μg·mL^(-1)。结论化合物1~11均为首次从该属真菌中分离得到。

Enhancement of defense responses by <i>Clonostachys rosea</i>against Botrytis <i>cinerea</i>in tomatoes

Clonostachys rosea (C. rosea) is a biocontrol agent that is used to combat and prevent phytopathogenic fungi attacks because of its ability to involve many factors and diverse modes of action. The reactions of C. rosea on the control of gray mold disease in tomato leaves were investigated in this study. To investigate the reactions of C. rosea in inducing resistance to tomato plants, three treatments, including Botrytis cinerea treatment (treatment B), C. rosea treatment (treatment C), C. rosea and B. cinerea treatment (treatment C + B) and water (control), to be applied on tomato leaves were set up. Disease severity was subsequently evaluated and compared with the control. The treatment of tomato leaves with C. rosea (15 μg/ml) significantly reduced the disease index after inoculation and severity of gray mold caused by Botrytis cinerea. The results indicated that the C. rosea treatment stimulated the activity of the defense related enzymes: Peroxidases (POX), lipoxygenases (LOX) and glutathione S-transferases (GST), and the treatment C + B reduced the incidence and severity of the gray mold. Furthermore, C. rosea treatment increased the activity of pathogenesis related proteins PR1. Therefore, our results suggest that C. rosea could enhance the resistance of tomato plants to gray mold through the activation of defense genes and via the enhancement of defense-related enzymatic activities.

拮抗微生物防治仙客来灰霉菌Botrytis cinerea的评估(英文)

以仙客来叶圆片方法对灰霉菌Botrytis cinerea的生防效果进行了评估,结果发现Clonostachys rosea和Trichoderma koningii抑制B.cinerea的效果达94%～100%,其效果优于所测试的从仙客来上分离的真菌和细菌的多数菌株。用整株仙客来测试生防菌效果表明,C.rosea在叶片、花瓣、叶柄和花梗上抑制B.cinerea效果分别达96%、58%、53%、77%;而Trichoderma koningii的效果分别达100%、42%、100%、90%。以析因试验设计对B.cinerea和C.rosea或T.koningii施用浓度在生防效果上的交互效应进行了测定,结果表明:当C.rosea的接种浓度是病原菌浓度的10倍或10倍以上时,其在叶片、花瓣、叶柄和花梗上抑制B.cinerea的效果分别达73%～100%、52%～86%、100%、54%～68%;当T.koningii的接种浓度为106个/mL或以上时,其抑制B.cinerea在仙客来(接种浓度为103～106个/mL时)各部分发生的效果分别为:叶片100%、叶柄62%～100%、花瓣38%～96%、花梗56%～100%。C.rosea在叶龄为2、3、5、6周的叶片上的生防效果更好,而在叶龄为4周的叶片上效果差些,在叶龄为1周的叶片上无效果。T.koningii在叶龄为1～6周的叶片上均可有效地抑制B.cinerea的发生,抑制效果达89%～100%。C.rosea和T.koningii均是在温室内防治仙客来灰霉菌B.cinerea的具潜力的有效生防菌,但T.koningii的防治效果显著地优于C.rosea的效果。

Bionectria pseudochroleuca,a new host record on Prunus sp.in northern Thailand

This study presents the first report of Bionectria pseudochroleuca(Bionectriaceae)on Prunus sp.(Rosaceae)from northern Thailand,based on both morphological characteristics and multilocus phylogenetic analyses of internal transcribe spacer(ITS)and Beta-tubulin(TUB2).

一种由粉红粘帚霉引起的青稞根腐病

[背景]根腐病在青稞生产中的危害日趋严重,阻碍了青稞根腐病的有效防控及青海省青稞产业的发展。然而人们对青稞根腐病的研究甚少且病原菌不详。[目的]明确青稞根腐病发生的危害、病原及致病性,为青稞根腐病的防控提供理论依据。[方法]采用常规的组织分离法分离青稞根腐病病原,通过形态鉴定与分子鉴定结合的方法对病原进行鉴定,并采用烧杯水琼脂法测定其致病性。[结果]共分离得到4株青稞根腐病病原菌,鉴定为Clonostachys rosea,有较强的致病性且致病性差异显著,经柯赫氏法则验证为青稞根腐病病原菌,并且是一种新的青稞根腐病病原,该类根腐病也是一种新的根腐类病害,在国内外属首次发现。[结论]Clonostachys rosea可引起青稞根腐病且致病性强。