Infection of flowers and fruits of red raspberry by Botrytis cinerea

The frequency and site of symptomless infection of flower buds, flowers, and fruits of red raspberry by Botrytis cinerea was studied using surface-disinfestation and culture techniques. Flower buds were rarely infected but open flowers were rapidly colonized and necrotic stamens and styles were an important source of infection for the developing fruit. The receptacle tissue within the drupelet cluster cavity was mostly sterile. Dichlofluanid sprays reduced symptomless infection of fruits and were more effective when applied early. Post-harvest rot tests indicated that incidence of grey mould was not related to symptomless infection unless fruits were surface-disinfested before subjecting to the rotting test.     

Gray mold of yacon and sunflower caused by Botrytis cinerea

Severe wilt and blight of entire plants of yacon (Smallanthus sonchifolius) grafts on sunflower (Helianthus annuus) were found in greenhouses in Kagawa Prefecture in the southwest region of Japan from April to May between 1997 and 1999. A mitosporic fungus, isolated repeatedly from the diseased plants and identified as Botrytis cinerea, was demonstrated to be virulent to both yacon and sunflower. The new disease on yacon was named gray mold of yacon. The disease on sunflower, gray mold of sunflower, is new to Japan.     

Expression of an oxalate oxidase gene in tomato and severity of disease caused by Botrytis cinerea and Sclerotinia sclerotiorum

A cDNA clone of wheat oxalate oxidase (OxO) under the control of the constitutive CAMV 35S promotor was expressed in tomato plants by Agrobacterium -mediated transformation. Twenty-six transgenic tomato lines were obtained and analysed. PCR experiments confirmed the incorporation of the OxO gene in all tested tomato lines. The transgenic tomato plants expressed a 124-kDa protein showing OxO activity, and were able to convert different oxalic acid (OA) concentrations in vitro . In a detached leaf assay, most of the transgenic lines showed reduced disease symptoms compared with controls, following inoculation with Botrytis cinerea . In addition, leaves of the line T15 showed a marked reduction in symptoms compared with the control following inoculation with Sclerotinia sclerotiorum .     

黄姜主要害虫--蛴螬的发生为害及防治技术

黄姜俗称“火头根” ,属盾叶薯蓣 ,是我国特有种。黄姜浑身是宝 ,我国利用黄姜根茎中皂甙元 (皂素) ,可合成 3大类 (皮质激素、性激素、孕激素 ) ,近2 0 0个产品系列 ,是生产甾体激素类药物最理想的物质 ,世界上 90 %的激素是利用其合成的 ,因此 ,被国家医药界誉为“药用黄金”。郧西县地处秦巴小区 ,是全国黄姜高产高含地区的中心 ,历史上曾形成庞大的野生黄姜资源带。1 9 87年开始进行大面积“野转家”栽培并获得成功 ,至今已发展到 1万hm2 以上 ,被农业部农业特产委员授予“全国黄姜之乡”。在栽培过程中 ,由于连年重茬及姜农们忽略了蛴…     

Fungal and plant gene expression during synchronized infection of tomato leaves by Botrytis cinerea

An inoculation procedure was developed to obtain efficient and synchronous infection on detached tomato leaves by Botrytis cinerea. In spray-inoculated leaves incubated at 20 °C, the infection process consisted of three phases: the formation of primary necrotic lesions (until 20 hpi), a quiescent phase (20-72 hpi), and the expansion of a proportion of the primary lesions (from 72 hpi onwards), resulting in full tissue maceration. At 4 °C, the infection progressed slowly but steadily without inducing necrotic responses in the host. The actin and -tubulin genes of B. cinerea were cloned, characterized and used as probes on blots containing RNAs from leaves at various stages of the infection. The genes displayed a similar expression pattern throughout the infection and the hybridization signal reflected the amount of fungal biomass. The actin mRNA accumulated to higher levels than the -tubulin mRNA. Tomato PR protein mRNAs (chitinase, -1,3-glucanase and PR-1) were induced during the infection, albeit with different kinetics and to different levels. At 20 °C, -1,3-glucanase and PR-1 mRNAs were induced more rapidly than chitinase mRNAs. At 4 °C, mRNAs encoding extracellular -1,3-glucanase and intracellular, as well as extracellular chitinase were hardly induced.     

湿度调控对番茄灰霉病菌侵染的影响

 番茄灰霉病菌(Botrytes cinerea)的分生孢子萌发需要高湿的环境,一般要求相对湿度在80%以上或水滴中萌发最好。在适温下,随着湿度的增加,病菌致病性增强,孢子在相对湿度85%以上、菌丝在相对湿度80%以上的高湿环境能够发生侵染,并形成危害性的软腐斑。在一天中高湿的时间直接影响番茄灰霉病的发生,高湿(相对湿度超过85%)时间8h以上病菌才能够连续侵染。     

北京地区番茄灰霉病菌的多重抗药性检测

2009年12月-2010年5月,在北京12个郊区县采集番茄病标样150份,分离纯化得到109个灰葡萄孢（Botrytis cinerea）单孢菌株,用最低抑制浓度法（MIC）测定了其对苯并咪唑类（多菌灵）、二甲酰亚胺类（腐霉利）和氨基甲酸酯类（乙霉威）杀菌剂的抗药性。结果表明：番茄灰霉病菌对多菌灵、腐霉利和乙霉威产生抗性菌株的频率分别为96.3%、80.7% 和58.7%;所测菌株对3类杀菌剂的抗性类型有BenRDicSNPCS、BenSDicSNPCR、BenRDicRNPCS和BenRDicRNPCR 4种,所占比例分别是19.3%、3.7%、21.1%和56.0%,表明北京地区番茄灰霉病菌对苯并咪唑类、二甲酰亚胺类和氨基甲酸酯类三类杀菌剂的抗药性严重,在生产中需慎用,应选择一些替代的新型杀菌剂和生物农药。     

4种杀菌剂及其复配剂对番茄灰霉病菌的毒力

采用菌丝生长速率法测定了咯菌腈、氟啶胺、啶酰菌胺和苯醚甲环唑4种杀菌剂及其两元复配剂对番茄灰霉病菌的毒力。结果显示,咯菌腈、氟啶胺、啶酰菌胺和苯醚甲环唑对番茄灰霉病菌的有效抑制中浓度(EC50)分别为:0.018 0、0.018 1、1.896 8和2.087 4μg/mL。复配剂啶酰菌胺∶苯醚甲环唑1∶5、1∶3和1∶1,咯菌腈∶苯醚甲环唑1∶5增效作用最明显;复配剂咯菌腈∶苯醚甲环唑1∶3,咯菌腈∶氟啶胺1∶3,啶酰菌胺∶咯菌腈5∶1,啶酰菌胺∶苯醚甲环唑3∶1具有增效作用,SR值范围为1.5~4.05,其中以复配剂啶酰菌胺∶苯醚甲环唑1∶5增效作用最好,其SR值为4.05;其余不同配比的各组合复配剂具有相加作用,其SR值范围为0.5~1.46。表明咯菌腈、氟啶胺、啶酰菌胺和苯醚甲环唑4种不同作用机制的杀菌剂可以交替或复配使用,以阻止或延缓灰霉病菌抗药性的进一步加剧,为灰霉病的综合防控和抗药性治理提供理论依据。     

Assessing host specialization of Botrytis cinerea on lettuce and tomato by genotypic and phenotypic characterization

This study tested the hypothesis that Botyrtis cinerea shows host specialization on tomato and lettuce, using phenotypic and genotypic tools. Strains were isolated from tomato and lettuce grown together in the same greenhouse. Forty‐four lettuce strains and 42 tomato strains were investigated for their genetic diversity and their aggressiveness. Both gene diversity and allelic richness were significantly higher in lettuce strains than in tomato strains (P = 0·01). Cluster analysis revealed a clear division of the strains under study into two clusters. However, this structure did not separate the strains according to their host of origin. Tomato strains were significantly more aggressive than lettuce strains when inoculated on tomatoes (P = 0·001), but no significant differences in aggressiveness were observed when the strains were inoculated on lettuce (P = 0·17) or on apple (P = 0·87). The results suggest an absence of clear host specialization of B. cinerea on tomato and lettuce.     

Programming applications of dicarboximides to control bunch rot of grapes caused by Botrytis cinerea

When grapevine plants in a glasshouse were artificially inoculated with Botrytis cinerea and then treated with dicarboximides at different stages of bunch development, treatment at 80% cap-fall, with or without treatment before bunch closure, gave significantly better control than treatment at 3 weeks before harvest. In field trials, early applications of procymidone reduced natural infection by B. cinerea better than late applications. Spray application at pre-bloom followed by a second application during bloom is recommended. Procymidone, vmclozolin and iprodione at 1 g/l were significantly more effective in controlling natural infection than dichlozolinate, benomyl, captan, chlorothalonil and copper oxychloride.     

Increased rhizosphere populations of Trichoderma asperellum strain T34 caused by secretion pattern of root exudates in tomato plants inoculated with Botrytis cinerea

Root exudates secreted from plants can modify rhizosphere microbiota by enhancing or inhibiting the growth of biological control agents (BCAs) and/or pathogens. Similarly, microorganisms can modify the secretion of plant root exudates. The aim of this study was to analyse the effect of a Botrytis cinerea leaf infection on the secretion of tomato root exudates and on the populations of the BCA Trichoderma asperellum strain T34 (T34). This study found that the secretion pattern of root exudates in tomato plants was influenced by B. cinerea infection in plant leaves. An increase in the levels of gluconic acid was observed, while levels of sucrose and inositol decreased. A decrease in the severity of B. cinerea by the induction of systemic resistance triggered by T34 was also observed. Tomato plants infected with B. cinerea maintained the populations of T34 in the roots, while populations of T34 decreased in plants not inoculated with the pathogen. Samples exposed to media containing gluconic acid (as the only carbon source or at the same concentration found in roots exudates) saw an increase in the in vitro growth of T34 compared to media without gluconic acid. In conclusion, a change in the secretion pattern of root exudates caused by B. cinerea, together with the enhanced growth of T34 in the presence of gluconic acid, indicates the existence of leaf to root communication. The result of this is enhanced populations of T34, and in turn induced disease resistance and a consequential reduction in disease severity.     

北京地区番茄灰霉病菌对嘧霉胺的抗药性检测

为了检测北京地区番茄灰霉病菌对嘧霉胺的抗药性,指导生产上科学用药,从北京12个区、县采集番茄灰霉病样150份,分离纯化获得109个番茄灰霉病菌(Botrytis cinerea)的单孢菌株,采用菌丝生长速率法测定了番茄灰霉病菌对嘧霉胺(pyrimethanil)的抗药性。所测菌株中嘧霉胺抗性菌株出现的比例高达82.57%,且以高抗菌株为主,占78.89%,抗性水平最高的菌株EC50是最敏感菌株的5 096倍;不同区县番茄灰霉病菌对嘧霉胺的抗药性存在差异,门头沟区、密云县、延庆县、怀柔区和平谷区、大兴、通州和顺义8个区县的高抗菌株所占比例达66.67%～100%,海淀、朝阳和房山3个区未检测到高抗菌株。上述结果说明北京地区番茄灰霉病菌对嘧霉胺的抗药性非常普遍,且以高抗菌株为主,生产上应替换新型杀菌剂防治番茄灰霉病。     

Preventive and post‐infection control of Botrytis cinerea in tomato plants by hexanoic acid

The antifungal activity of hexanoic acid on the phytopathogen Botrytis cinerea was studied. This chemical inhibited both spore germination and mycelial growth in vitro in a concentration‐ and pH‐dependent manner, and stopped spore germination at a very early stage, preventing germ‐tube development. The minimum fungicidal concentration (MFC) for in vitro spore germination was 16 mm . Hexanoic acid also inhibited in vitro mycelial growth of germinated spores at an MFC of 12 mm . Studies performed to characterize the mechanisms underlying the antimicrobial effect of hexanoic acid showed that it alters fungal membrane permeability. In addition, hexanoic acid treatment increased the levels of spermine, spermidine, putrescine and cadaverine in B. cinerea mycelia. Spray application of hexanoic acid at fungicidal concentrations on 4‐week‐old tomato plants prior to fungal inoculation reduced necrosis diameter by approximately 60%. Application of the same hexanoic acid concentrations on previously infected plants reduced further necrosis expansion by around 30%. The results suggest that this chemical acts as a preventive and curative fungicide. Interestingly, treatments with hexanoic acid at concentrations below the MFC in hydroponic solution prior to fungal inoculation significantly reduced necrosis area. These results suggest an inducer effect of plant responses for hexanoic acid treatments at these concentrations. Hexanoic acid is a good candidate for safe antifungal treatments for the control of B. cinerea, which is responsible for many economic losses on fruits, vegetables and flowers.     

In vitro and in vivo production of cell wall degrading enzymes by Botrytis cinerea from tomato

The production and activity of pectolytic and cellulolytic enzymes byBotrytis cinerea in tomato plants, as well as by conidia of this fungus in some nutrient media was studied. In inoculated tomato plants, the activity of these enzymes varied. PME, endo-PG and exo-PG were produced in petioles and fruits, while cellulase was only found in those parts which were softened by the invading mycelium. PGTE could only be detected in the softened parts of the petiole stumps. In suitble substrates, PME, endo-PG and exo-PG are produced rapidly with the onset of germination of the conidia. At some temperatures, endo-PG activity was found before germination could be observed. Some endo- and exo-PG was detected in washing water of conidia. The significance of the production of these enzymes by germinating conidia in relation to the infection process on tomato plants is discussed.Samenvatting In geïnoculeerde delen van een tomateplant blijkt de activiteit van deze enzymen afhankelijk te zijn van de aard van het aangetaste weefsel en de mate van aantasting (Tabel 1). Conidiën blijken in bepaalde media zeer snel pectolytische enzymen te vormen (Tabel 2, Fig. 1). Endo-PG activiteit kon bij 26° en 30°C in een natriumpolypectaatmedium worden gemeten voordat kieming van de conidiën kon worden aangetoond, zij het in zeer geringe hoeveelheid, lijkt aanwezigheid van deze twee enzymen in niet gekiemde conidiën aannemelijk. PME blijkt in een pectinebevattend medium te kunnen worden aangetoond op het moment, waarop kiembuizen zichtbaar worden. Cellulase kon daarentegen in een medium met kiemende sporen niet worden gevonden.     

武夷菌素对番茄灰霉菌的抑制作用及对番茄抗病性相关酶活性的影响

采用凹陷载玻片法测定了武夷菌素对番茄灰霉菌分生孢子萌发的抑制作用,在离体番茄叶片上测定了被武夷菌素处理后的番茄灰霉菌菌丝和分生孢子致病性的变化以及武夷菌素对番茄幼苗体内抗病性相关酶活性的影响。结果表明:武夷菌素对番茄灰霉菌的分生孢子有较强的抑制作用,其EC₅₀为14.1μg／mL。浓度为100μg／mL的武夷菌素可完全抑制孢子的萌发。武夷菌素能使灰霉菌菌丝和分生孢子的致病性明显下降,同时还能诱导番茄体内抗病性相关酶(SOD、POD、PPO、PAL)活性的增强,提高番茄幼苗的抗病性。     

辽宁省不同地区番茄灰霉病菌对咯菌腈的敏感性

为明确辽宁省不同地区间番茄灰霉病菌对咯菌腈的敏感性是否存在差异,以及咯菌腈与其他杀菌剂之间是否存在交互抗性,采用菌丝生长速率法测定番茄灰霉病菌对咯菌腈、腐霉利、多菌灵的敏感性。结果表明,从辽宁省6个不同地区分离的番茄灰霉病菌菌株对咯菌腈的平均EC50差异不显著,番茄灰霉病菌菌株对咯菌腈的敏感性频率呈正态分布,咯菌腈对不同地区共117株番茄灰霉病菌的EC50平均值为(0.007 3±0.004 5)μg/mL,且咯菌腈与腐霉利和多菌灵之间均不存在交互抗药性。     

Epidemiology and control of Botrytis cinerea in tomato crops grown under cover in Greece

Out-of-season tomatoes grown in unheated glasshouses in Greece annually suffer considerable losses from grey mould induced by Botrytis cinerea. Characteristic soft rots of young fruit by the calyx end, flower abortions and stem lesions or cankers are the most commonly occurring symptoms of the disease. Severe ghost spots on fruit only appear when fungicides inhibiting spore germination are not applied. Conidia produced on plant debris of the previous year's crop serve as primary inoculum. Disease starts in late autumn, increases rapidly during the winter period and ceases abruptly at the end of April. Infections caused by isolates resistant to dicarboximides or to both dicarboximides and benzimidazoles are uncommon at the beginning of each new crop cycle. These increase substantially as the season progresses and decline after cessation of fungicide sprays. By contrast, infections caused by benzimidazole-resistant strains are stable throughout the growing season. Efficient disease control implies use of proper glasshouse covering material, adequate indoor air circulation, partial night heating, strict indoor and outdoor hygiene and regular fungicide sprays. When disease pressure is low, application only of conventional compounds like dichlofluanid and chlorothalonil is recommended. Under conditions favourable to Botrytis infections, mixtures of conventional compounds with reduced strength specific botryticides should preferably be applied. Such regimes efficiently minimize losses from fungicide resistance development and ghost spot.     

Virulence factors of Botrytis cinerea

Botrytis cinerea is responsible for gray mold disease in more than 200 host plant species. The infection of host plants is mediated by numerous extracellular enzymes, proteins and metabolites. Each of these compounds may play a role in different stages of the infection process. Cell wall-degrading enzymes may facilitate the penetration into the host surface, while toxins, oxalic acid and reactive oxygen species may contribute to killing of the host cells. Cell wall-degrading enzymes contribute to the conversion of host tissue into fungal biomass. On the other hand, B. cinerea infection induces biosynthesis of phytoalexins. Therefore, the ability to overcome a wide spectrum of phytoalexins contributes to the pathogenicity of the fungus with a broad host range. The cloning of the corresponding genes has facilitated studies on gene expression and targeted mutagenesis. This review gives an overview of the research performed on virulence factors that play the roles in pathogenesis.