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.     

Modeling Infection of Strawberry Flowers by Botrytis cinerea Using Field Data

ABSTRACT The incidence of strawberry flower infection by Botrytis cinerea was monitored in unsprayed field plots in three successive years together with meteorological data and numbers of conidia in the air. There were large differences in conidia numbers and weather conditions in the 3 years. Three sets of models were derived to relate inoculum and weather conditions to the incidence of flower infection; by inoculum only, by weather variables only, and by both inoculum and weather variables. All the models fitted the observed incidence satisfactorily. High inoculum led to more infection. Models using weather variables only gave more accurate predictions than models using inoculum only. Models using both weather variables and inoculum gave the best predictions, but the improvement over the models based on weather variables only was small. The relationship between incidence of flower infection and inoculum and weather variables was generally consistent between years. Of the weather variables examined, daytime vapor pressure deficit and nighttime temperature had the greatest effect in determining daily incidence of flower infection. Infection was favored by low day vapor pressure deficit and high night temperature. The accuracy and consistency of the weather-based models suggest they could be explored to assist in management of gray mold.     

Histopathology of the initial stages of the interaction between rose flowers and Botrytis cinerea

The early stages of the interaction between flowers of the cut rose cv. Sonia andBotrytis cinerea was investigated by scanning electron microscopy and light microscopy. Infection of petals by conidial germ tubes evoked a susceptible reaction. In contrast to general findings nutrient addition to the inoculum was not a prerequisite for this phenomenon. At the lower side of germ tube tips the cuticle was penetrated by infection pegs. Already at this early stage of the infection process, the infection sites were macroscopically visible as scattered white spots. After penetration, pegs enlarged to form infection hyphae, which invaded the periclinal wall of outer epidermal cells. At those sites, the petals formed outgrowths of variable appearance at their abaxial side. Thee outgrowths consisted of remanants of collapsed epidermal cells and of infection hyphae. Subsequent intra- and intercellular growth of hyphae led to a collapse of epidermal and mesophyll cells. The symptoms described generally developed within 24 h. After subsequent incubation the lesions became necrotic. Eventually, the necrosis would spread leading to the death of whole petals.     

Botrytis cinerea Infection of Grape Flowers: Light and Electron Microscopical Studies of Infection Sites

ABSTRACT Specific floral organs including the calyptra, stigma, and receptacle area of glasshouse-grown grapevines (Vitis vinifera cv. Cabernet Sauvignon) were inoculated with aqueous suspensions of Botrytis cinerea conidia, and the initial steps involved in colonization and infection of the host tissues were studied for several days postinoculation using light microscopy as well as scanning and transmission electron microscopy. Conidia germinated on all floral organs examined and became attached to the host surface within 48 h after inoculation. However, in all cases the vast majority of conidia accumulated in a channel-like gap between the ovary and the calyx that extended in a narrowing fashion into the flower interior where the ovary joined the receptacle. Very few conidial germ tubes were detected in the style following inoculation of the stigma, and no evidence for their growth toward the ovaries could be found. In contrast, hyphae were more abundant in the receptacle area, regardless of the site of inoculation. Tips of the calyx became necrotic and mycelium formed in the gap between ovary and calyx within 72 h following inoculation, providing a major point of colonization and infection. B. cinerea colonized dehiscent calyptras within 72 h of inoculation, providing a potential source of inoculum from calyptras that remained stuck in the cluster. The results suggest that the grape flower's receptacle area is the predominant site of infection for B. cinerea, although a minor portion of infections may also occur through the stigma and style.     

Infection of Commercial Hybrid Primula Seed by Botrytis cinerea and Latent Disease Spread Through the Plants

ABSTRACT Botrytis cinerea occurred commonly on cultivated Primula xpolyantha seed. The fungus was mostly on the outside of the seed but sometimes was present within the seed. The fungus frequently caused disease at maturity in plants grown from the seed, demonstrated by growing plants in a filtered airflow, isolated from other possible sources of infection. Young, commercially produced P. xpolyantha plants frequently had symptomless B. cinerea infections spread throughout the plants for up to 3 months, with symptoms appearing only at flowering. Single genetic individuals of B. cinerea, as determined by DNA fingerprinting, often were dispersed widely throughout an apparently healthy plant. Plants could, however, contain more than one isolate.     

Colonization of red raspberry flowers and fruit by Botrytis cinerea under commercial production conditions in northwestern Washington,USA

Colonization of red raspberry flowers and fruit by Botrytis cinerea was determined during 2017–2018 growing seasons under commercial fungicide application programmes used for grey mould management in northwestern Washington, USA. Colonization of flowers and fruit was assessed qualitatively (incidence, %) and quantitatively (abundance, number of colonies) by recovering B. cinerea from surface-disinfested samples. Both incidence and abundance of flower colonization were significantly lower than fruit colonization in both untreated and fungicide-treated plots. Incidence of flower colonization did not differ significantly between untreated and fungicide-treated plots (43% vs. 45%, respectively). In contrast, significantly greater colonization incidence was detected at green fruit stage in untreated compared to fungicide-treated plots (96% vs. 77%, respectively). Ripe fruit had the greatest colonization incidence among the three stages sampled and colonization was not significantly different between untreated and fungicide-treated plots (100% vs. 92%, respectively). Similarly, colonization abundance of flowers did not differ significantly between untreated and fungicide-treated plots (1.0 colonies per colonized flower in both treatments), but colonization abundance of green and ripe fruit was decreased 2.3- and 2.1-fold, respectively, in fungicide-treated plots. DNA fingerprinting analysis of the pathogen revealed that different multilocus genotypes colonized flowers and fruit within the same inflorescence and that genotypic diversity increased through time, suggesting independent infection events. Overall, our results demonstrate that under current environmental conditions, raspberry flowers may not be the exclusive or major route of infection for grey mould of red raspberry in northwestern Washington. Implications of current findings for management and further research are discussed.     

Spotting of tomato fruits caused by Botrytis cinerea

Botrytis spot or ghost spot on tomato fruits occurs after penetration of germ tubes ofB. cinerea into epidermal cells. A few days after the penetration a halo appears around the infected necrotic cells. These symptoms can be reproduced by inoculating young fruits with a few dry conidia. When many conidia alight on the epidermis of the fruit, scab-like symptoms develop, while under conditions of high humidity, blisters can be formed on the fruit surface before the fungus spreads through the fruit parenchyma. Under conditions of low humidity, necrotic areas are formed.In the necrotic cells, developed after inoculation with many or with a few conidia, no mycelium could be found by the histological methods so far used. However,B. cinera can be reisolated, implying that theBotrytis spot is a latent infection by the fungus. No renewed growth takes place when the fruit is fully ripe.Samenvatting Botrytis-stip op tomatevruchten ontstaat na binnendringen van kiembuizen vanB. cinerea in epidermiscellen van een jonge vrucht. Enkele dagen na de infectie treden celdelingen op in het onderliggende parenchymweefsel. De geïnfecteerde cel en enkele aangrenzende cellen sterven af, terwijl rondom dit necrotische plekje een zilverwit gekleurde ring ontstaat. Worden veel conidiën bijeen op een vruchtwand gebracht, dan krijgt de epidermis binnen 24 uur een schurftig uiterlijk. Blijft de luchtvochtigheid na de inoculatie hoog, dan kunnen daarentegen kleinere en grotere blaasjes in de vruchtwand ontstaan. Deze barsten na enkele dagen open, terwojl mycelium zich door de vrucht verbreidt. Daalt de luchtvochtigheid circa 16 uur na de inoculatie, dan blijft de aantasting beperkt tot de epidermis. Hoewel in het necrotische weefsel geen mycelium vanB. cinerea kon worden aangetoond, blijkt herisolatie van de schimmel gemakkelijk te zijn. Ook uitBotrytis-stippen van uit de praktijk afkomstige vruchten kanB. cinerea geïsoleerd worden. Dit betekent, dat het hier om een latente infectie gaat, waarbij evenwel geen hernieuwde groei plaats vindt als de vrucht geheel rijp is.     

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.     

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.     

Biosuppression of Botrytis cinerea in grapes

There is increasing interest in the use of biological control agents (BCAs) and plant resistance stimulants to suppress botrytis bunch rot in grapes, caused by Botrytis cinerea . Numerous different filamentous fungi, bacteria and yeasts have been selected as potential BCAs for control of grey mould based upon demonstrated antagonism towards B. cinerea. Biological suppression of the pathogen arises via competition for nutrients and space, the production of inhibitory metabolites and/or parasitism. Preformed and inducible grapevine defence mechanisms also contribute to disease suppression by preventing or delaying pathogenic infection. Furthermore, various biotic and abiotic agents can stimulate grapevine defence mechanisms and so elevate resistance to B. cinerea infection. Biosuppression of B. cinerea in vineyards, using BCAs and resistance stimulants, has been inconsistent when compared with that observed in controlled glasshouse or laboratory conditions. This may be attributable, in part, to the innate variability of the field environment. Research to improve field efficacy has focused on formulation improvement, the use of BCA mixtures and combinational approaches involving BCAs and plant resistance stimulants with complementary modes of action.     

Effects of temperature,relative humidity and age of conidia on the incidence of spotting on gerbera flowers caused by Botrytis cinerea

In recent years, spotting of ray florets of gerbera flowers has become an important problem. This type of small necrotic lesions may occur before, but especially shortly after harvesting the flowers.Botrytis cinerea was easily isolated from such lesions. Inoculation withB. cinerea only gave typical necrotic lesions, when dry conidia were dusted on the flowers with a short period of high rh after inoculation. At 18–25 °C a high rh for at least 5 hours was necessary. Rotting of ray florets and receptacles byB. cinerea occurred when inoculated flowers were kept wet for a few days. Spots consist of one to several necrotic, usually epidermal cells. A single conidium could give rise to a necrotic lesion after germination. Germination of conidia and lesion formation occurred between 4 and 25 °C; at 30 °C, germination and lesion formation did not occur. Between 18 and 25 °C, many lesions became visible within 1 day after inoculation; at 4 °C it took 2 to 3 days before lesions could be seen. If kept dry, conidia ofB. cinerea remained ungerminated on ray florets of gerbera flowers and could be removed from the ray florets. Within 1 day at high rh, germination occurred and lesions were produced. Conidia ofB. cinerea, stored dry, were able to survive much longer than the lifetime of a gerbera flower. Even after storage at room temperature for up to 14 months, some conidia were able to germinate in vitro and on ray florets and induce the formation of lesions. Addition of gerbera pollen diffusate stimulated germination and lesion formation.     

Electrophoretic karyotype analysis of Botrytis cinerea

The karyotypes of five strains ofBotrytis cinerea were analysed by pulsed field gel electrophoresis (PFGE). None of the five chromosome patterns were identical and all five strains contained one or several minichromosomes of variable length. Considering that some of the chromosomal bands represent unresolved doublets, the total genome size of the five strains was estimated to vary between at least 33.9 and 39.7 megabasepairs (Mbp). Hybridization of blots of pulsed field gels with rDNA and ß-tubulin probes revealed that these hybridize with the same chromosomal band. Chromosome-specific probes were isolated hybridizing to chromosomal bands 9 and 11 of strain SAS56. The latter two probes hybridized to only two of the five strains tested. The results are discussed in relation to the genome plasticity and variability of filamentous fungi.     

Botrytis cinerea Infection in Grape Flowers: Defense Reaction, Latency, and Disease Expression

ABSTRACT Inflorescences of field-grown grapevines (Vitis vinifera L. cv. Gamay) were inoculated with a Botrytis cinerea conidia suspension or dried conidia at different stages during bloom in moist weather. Approximately 10% of the conidia germinated within 72 h, resulting in two to three times more latent infections than uninoculated controls in pea-size (7 mm in diameter) berries. In surface-sterilized pea-size berries, latent B. cinerea was present predominantly in the receptacle area. After veraison, latent B. cinerea also was found in the style and, in mature berries, latent colonies were distributed throughout the pulp. Inoculation at full bloom led to the highest disease severity (66%) at harvest, compared with 38% in controls. Stilbene stress metabolites in the flowers were measured by high-performance liquid chromatography. Resveratrol accumulated mainly after pre-bloom and full-bloom inoculation, but did not prevent infection. Piceid levels did not change following inoculation, while epsilon-viniferin was found in necrotic tissues only, and pterostilbene and alphaviniferin were not detected at all. B. cinerea conidia suspensions also were applied to various locations on flowers of pot-grown cvs. Pinot Noir and Chardonnay. Inoculation of the receptacle area, but not that of the stigma and ovary, resulted in latent infections. Stilbene synthesis was similar to the field results, with resveratrol accumulating mainly in the calyptra and receptacle area. Constitutive soluble phenolic compounds (mainly derivatives of quercetin and hydroxy-cinnamic acid) were present at high concentrations in the calyptra but at low levels in the receptacle area. These experiments confirmed bloom as a critical time for B. cinerea infection in grapes and suggest that the most likely site of infection is the receptacle area or cap scar exposed at anthesis. Stilbenes may have a limited role in inhibition of flower infection and latency in susceptible grape cultivars, and epsilon-viniferin may be a by-product rather than a deterrent of infection.     

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.     

Biological Control of Botrytis cinerea by Volatiles of 'Isabella' Grapes

ABSTRACT The effect of volatiles from cv. Isabella (Vitis labrusca) on the growth of Botrytis cinerea was tested in vitro and in situ, in the latter case on 'Roditis' grapes (V. vinifera), at various temperatures. The goal of the research was to determine whether the volatiles emitted by Isabella grapes could be effective biocontrol agents of Botrytis cinerea. The closed Mariotte system was used as a bioassay method to analyze quantitatively the biological action of these volatiles on fungal growth and disease development. The in vitro experiments revealed the inhibitory action of the Isabella volatiles on the sporulation and sclerotia formation of the fungus, as well as the stimulating action of the Roditis volatiles on the sporulation of the fungus. The in situ study confirmed the antifungal action of the Isabella volatiles as they reduced the inoculum and pathogenicity of B. cinerea. The antibiotic action was more pronounced at 21 degrees C. The study indicates that Isabella volatiles act as biocontrol agents of B. cinerea.     

灰霉菌的抗药性与适合度测定

室内监测了北京、沈阳、保定等地区番茄灰霉菌 (Botrytis cinerea)对不同杀菌剂的抗药性 ,发现抗多菌灵菌株占监测总数的 74 .6 7% ,抗乙霉威菌株占 2 5.82 % ,还监测到 3株抗多菌灵、乙霉威菌株——即双抗菌株 ;利用菌落直径法和番茄叶片法测定了 3种抗药类型的番茄灰霉菌在离体和活体条件下的适合度 ,结果表明 ,不同抗药类型菌株在生长速率、致病力、繁殖力等方面差异不明显。     

Hypovirulence and Double-Stranded RNA in Botrytis cinerea

ABSTRACT Twenty-one strains of Botrytis cinerea isolated from 13 species of plants grown in China were compared for pathogenicity on Brassica napus, mycelial growth on potato dextrose agar, and presence of double-stranded (ds)RNA. The results showed that the strain CanBc-1 was severely debilitated in pathogenicity and mycelial growth, compared with the 20 virulent strains. A dsRNA of approximately 3.0 kb in length was detected in CanBc-1 and 4 hypovirulent single-conidium (SC) isolates of CanBc-1, but was not detected in the 20 virulent strains of B. cinerea and 4 virulent SC isolates of CanBc-1. Results of the horizontal transmission experiment showed that the hypovirulent trait of CanBc-1 was transmissible and the 3.0-kb dsRNA was involved in the transmission of hypovirulence. Analysis of a 920-bp cDNA sequence generated from the 3.0-kb dsRNA of CanBc-1 indicated that the dsRNA element was a mycovirus, designated as B. cinerea debilitation-related virus (BcDRV). Further analyses showed that BcDRV is closely related to Ophiostoma mitovirus 3b infecting O. novo-ulmi, the causal agent of Dutch elm disease. Mitochondria and cytoplasm in hyphal cells of CanBc-1 became degenerated, compared with the virulent isolate CanBc-1c-66 of B cinerea. This is the first report on the occurrence of Mitovirus-associated hypovirulence in B. cinerea.     

Influence of environmental conditions on dispersal of Botrytis cinerea conidia and on post-harvest infection of gerbera flowers grown under glass

Dispersal of Botrytis cinerea in a gerbera crop grown in two glasshouses 30 km apart was studied over a period of 18 months, in 1988 and 1989. Conidia were caught in spore traps consisting of agar in petri dishes exposed at different heights in the crop in each glasshouse. No seasonal patterns could be identified in the spore catches, assessed as colonies on the agar traps after incubation. The number of lesions caused by conidial infection of gerbera flowers following incubation, however, showed a distinct pattern. In spring and early summer few lesions were recorded whereas at other times of the year many lesions appeared. In linear regression analysis, variation in numbers of colonies (spore catches) could not be explained by environmental factors recorded during the experiments. Linear regression accounted for 77% and 81% of the variation in the number of lesions on flowers in the two glasshouses, in terms of relative humidity (postively correlated), global radiation outside the glasshouse (negatively correlated) and age of the crop (positively correlated). Despite differences in the systems by which the gerbera crop was produced and in the spore catches, the numbers of lesions on gerbera flowers in the two glasshouses were significantly correlated though not significantly different from each other.