果实褐腐病的调查与防治研究

 通过系统考察,标本采集,分离培养、鉴定,电镜扫描及酯酶同工酶电泳分析,明确我国丛梗孢属Monilia的种类及其寄主与分布,通过室内毒力测定,筛选出有效化学药剂并对桃褐腐病进行两年的综合防治,防效明显.     

环境条件对桃褐腐病菌生长和分生孢子萌发的影响

为了探明影响桃褐腐病发生和发展的因素,研究了温度、pH值、光照、营养条件等对桃褐腐病菌的生长和分生孢子萌发的影响。结果显示：桃褐腐病菌营养生长最适温度范围为20～25℃;最适pH值范围为6～7;光暗交替有利于该菌生长;供试的7种培养基中,PDA＋KNO3对生长最有利。分生孢子萌发的最适温度为25℃,最适pH值为8,最适营养液为1％蛋白胨液。     

鹿蹄草素对桃褐腐病菌的抑制作用及其抑菌机理

 【目的】探讨鹿蹄草素对桃褐腐病菌的抑制活性及作用机理。【方法】采用生长速率法和果实接菌的方法测定鹿蹄草素对桃褐腐病菌菌丝体生长的抑制作用;采用倍比稀释法测定鹿蹄草素最小抑菌浓度（MIC）和最小杀菌浓度（MFC）;观测接上鹿蹄草素和桃褐腐病菌对桃果实多酚氧化酶（PPO）、过氧化物酶（POD）和苯丙氨酸解氨酶（PAL）活性的诱导情况;并用扫描电镜和透射电镜观察鹿蹄草素对桃褐腐病菌超微结构的影响。【结果】鹿蹄草素对桃褐腐病菌有较强的抑制活性,且浓度越高,抑制作用越强,其MIC为0.032 mg?ml-1,MFC为0.156 mg?ml-1;桃果实接上鹿蹄草素＋病原菌后,PPO、POD和PAL活性开始升高,并在整个试验过程中一直保持较高的水平;扫描电镜下可见,菌丝扭曲变形,菌丝体之间相互粘连,细胞壁破裂,表面出现絮状凝集物。透射电镜下可见,细胞壁明显变薄,细胞内部组成紊乱,细胞结构遭到严重的破坏。【结论】鹿蹄草素对桃褐腐病菌具有较强的抗菌作用;可使菌丝体超微结构发生改变。     

Berberine as a natural compound inhibits the development of brown rot fungus Monilinia fructicola

Peach brown rot is a severe disease caused by ascomycete fungus Monilinia fructicola and is primarily controlled by chemical fungicides. However, the utilization of fungicides has caused many problems such as microbial resistance, environmental pollution, and fruit contamination. The growing concern for food safety requires alternative management methods that are safe to humans and benign to the environment. Here we report that the extract from Coptis chinensis (a Chinese herb named “Huang Lian”) demonstrates a strong inhibition to M. fructicola. The 50% effective concentration (EC₅₀) of C. chinensis extract to M. fructicola was only 0.91 mg/mL, while the EC₅₀s to other fungi such as Botrytis cinerea and Alternaria solani were 14.09 mg/mL and 27.35 mg/mL, respectively. These results indicate that the extract of C. chinensis has a specific inhibition to M. fructicola. Subsequent partitioning of the ingredients in C. chinensis extract revealed that berberine, the conventional anti-bacterial alkaloid, was the predominant ingredient that exerted robust inhibition against M. fructicola. The EC₅₀ and minimum inhibitory concentration (MIC) against M. fructicola were as low as 4.5 μg/mL and 46.9 μg/mL, respectively. Compared to berberine’s analogs in C. chinensis such as palmatine, berberine tannate and jatrorrhizine, berberine showed the strongest inhibition against M. fructicola. This finding provided insight into the structure–activity relationship between berberine and its analogs. Notably, not only can berberine prevent spore germination and hyphal growth, it also inhibits the activity of cutinase secreted by M. fructicola, implying the potential function of berberine in reducing the pathogenesis of M. fructicola. In addition, the strong in vivo inhibition of berberine against M. fructicola was observed with no visual cytotoxicity noted to peach fruits, even at berberine concentration of 400 μg/mL. Note that this was much higher than its MIC value (46.90 μg/mL). Overall berberine, as a natural compound, may be a promising candidate in control of brown rot.     

串珠霉利用稻草产纤维素酶的研究

纤维素酶是一类可以将纤维素水解成葡萄糖的酶,其利用对人类可持续发展具有非常重要的意义。本文对串珠霉利用稻草产纤维素酶的工艺条件进行了一系列优化实验,最终得出最佳产酶条件为:稻草粉6%,麸皮3%,豆饼粉3%,玉米粉2%,KH2PO40.5%,CaCl22.03%,pH 4.5,接种量3mL,28℃,发酵7天。     

大樱桃褐腐病菌生物学特性研究

褐腐病在辽宁大樱桃产区严重发生,对大樱桃褐腐病菌(Monilia fructigena)生物学特性进行了研究。试验从温度、光照、pH以及营养条件几方面对病菌菌丝生长、孢子产生及萌发的环境条件进行测定。结果表明,大樱桃褐腐病菌菌丝在PSA培养基、25℃、pH 6.0及黑暗条件下生长最好;产孢最佳条件为PDA、木糖、NH4H2PO4、25℃、pH 6.0;分生孢子在可溶性淀粉、NH4H2PO4、25℃、pH 7.0的条件下萌发率最高。病菌菌丝致死温度为56℃、10 min。     

Influence of storage approaches on instability of propiconazole resistance in Monilinia fructicola

BACKGROUND: The long‐term preservation of interesting phenotypes in plant pathogenic fungi allows for follow‐up studies in the future. Twelve storage approaches were investigated to determine their effects on instability of propiconazole resistance for three demethylation inhibitor (DMI) fungicide‐resistant and two DMI‐sensitive isolates of Monilinia fructicola. They included mycelium in PDA slants under mineral oil, in PDA plugs under 10% glycerol, on dried filter paper and conidia on silica gel, each stored for 36 weeks at 4, ? 20, and ? 80 °C. RESULTS: None of the storage approaches prevented the rapid decline of EC₅₀ values for propiconazole in the three resistant isolates, and no significant differences were found among storage approaches (P = 0.787) or between storage approaches and consecutive transfers (P = 0.053). Most of the decline in resistance occurred during the first 4 weeks of storage. The DMI resistance‐associated genetic element Mona, located in the immediate upstream region of the MfCYP51 gene, was still present in the three resistant isolates after 36 weeks of storage and weekly transfers. Furthermore, the Mona element and a portion of the MfCYP51 gene, which encodes the target enzyme for DMIs, did not reveal signs of DNA methylation. Resistance to propiconazole was partially regained in resistant isolates after two growth cycles on fresh peach fruit. CONCLUSIONS: Obtained data indicate that the decline of DMI resistance in M. fructicola cannot be prevented using commonly employed storage methods at various temperatures. The number of consecutive transfers and the storage duration prior to fungicide sensitivity tests in M. fructicola should be indicated in scientific papers. Copyright © 2011 Society of Chemical Industry     

瑞香狼毒根的超临界萃取物抑菌活性和成分分析

瑞香狼毒(Stellera chamaejasme)为瑞香科(Thymelaeaceae)植物,广泛分布于我国西北、华北等地(刘欣等,2004)。据《神农百草经》记载,瑞香狼毒与大戟科(Euphorbiaceae)植物狼毒大戟(Euphorbia fischeriana)、月腺大戟(E.ebracteolata)同作狼毒入药,其性味苦平,全草有毒,有逐水祛痰、破积杀虫之功效(张慧文等,2006)。瑞香狼毒也是一种     

桃褐腐病菌对多菌灵抗性的AS-PCR检测技术

由于杀菌剂的长期大量使用, 植物病原菌对杀菌剂抗性问题日趋突出, 严重影响病害防治的效果?褐腐病是一种在全世界范围内广泛发生和流行的重要果树病害?本文以桃褐腐病菌Monilinia fructicola为例, 基于已知的多菌灵抗性机理, 即靶标β微管蛋白基因TUB2的点突变E198A, 建立了一种桃褐腐病菌对多菌灵抗性的等位基因专化性PCR检测技术?结果表明, 碱基错配?内参引物?退火温度?dNTPs?Taq DNA聚合酶?专化性引物浓度及配比等因素均对检测效率有影响?经过对以上因素的优化, 并对优化后的检测技术进行专化性及灵敏度评价, 证实该检测技术能特异性地鉴别桃褐腐病菌M. fructicola对多菌灵抗性基因型E198A, 且检测灵敏度可达到4.342 pg/μL病菌DNA, 有望在实践中推广使用?     

Molecular characterization of the two-component histidine kinase gene from Monilinia fructicola

Brown rot of stone fruit caused by Monilinia fructicola (G. Wint) Honey is one of the most common fungal diseases in California. In this study, two laboratory-induced iprodione-resistant (LIR) mutants of M. fructicola were characterized by osmotic sensitivity, virulence on prune and sequence of the two-component histidine kinase gene (Mfos-1). The LIR mutants showed more sensitivity to osmotic stress and lower virulence on prune than their wild-type parent. Analysis of deduced amino acid of Mfos-1 showed that this protein exhibited all the characteristic features of the two-component histidine kinase genes, including osmotic sensing domain, six 90-amino-acid repeat motifs (coiled coil region) and kinase core and response regulator domains. Comparison of DNA sequences of the Mfos-1 from LIR mutants and the wild-type sensitive (S) isolate showed that LIR mutants had single point mutations in the coiled coil region of Mfos-1.