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1.
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. 相似文献
2.
ABSTRACT The effect of preliminary nutrient activation on the ability of conidia of the antagonist Trichoderma harzianum (atroviride) P1 to suppress Botrytis cinerea was investigated in laboratory, greenhouse, and field trials. Preliminary nutrient activation at 21 degrees C accelerated subsequent germination of the antagonist at temperatures from 9 to 21 degrees C; at >/=18 degrees C, the germination time of preactivated T. harzianum P1 conidia did not differ significantly from that of B. cinerea. When coinoculated with B. cinerea, concentrated inocula of preactivated but ungerminated T. harzianum P1 conidia reduced in vitro germination of the pathogen by >/=87% at 12 to 25 degrees C; initially quiescent conidia achieved this level of suppression only at 25 degrees C. Application of quiescent T. harzianum P1 conidia to detached strawberry flowers in moist chambers reduced infection by B. cinerea by >/=85% at 24 degrees C, but only by 35% at 12 degrees C. Preactivated conidia reduced infection by >/=60% at 12 degrees C. Both quiescent and preactivated conidia significantly reduced latent infection in greenhouse-grown strawberries at a mean temperature of 19 degrees C, whereas only preactivated conidia were effective in the field at a mean temperature of 14 degrees C on the day of treatment application. An antagonistic mechanism based on initiation of germination in sufficiently concentrated inocula suggests that at suboptimal temperatures the efficacy of Trichoderma antagonists might be improved by conidia activation prior to application. 相似文献
3.
ABSTRACT Grape berries are highly susceptible to powdery mildew 1 week after bloom but acquire ontogenic resistance 2 to 3 weeks later. We recently demonstrated that germinating conidia of the grape powdery mildew pathogen (Uncinula necator) cease development before penetration of the cuticle on older resistant berries. The mechanism that halts U. necator at that particular stage was not known. Several previous studies investigated potential host barriers or cell responses to powdery mildew in berries and leaves, but none included observation of the direct effect of these factors on pathogen development. We found that cuticle thickness increased with berry age, but that ingress by the pathogen halted before formation of a visible penetration pore. Cell wall thickness remained unchanged over the first 4 weeks after bloom, the time during which berries progressed from highly susceptible to nearly immune. Autofluorescent polyphenolic compounds accumulated at a higher frequency beneath appressoria on highly susceptible berries than on highly resistant berries; and oxidation of the above phenolics, indicated by cell discoloration, developed at a significantly higher frequency on susceptible berries. Beneath the first-formed appressoria of all germinated conidia, papillae occurred at a significantly higher frequency on 2- to 5-day-old berries than on 30- to 31-day-old fruit. The relatively few papillae observed on older berries were, in most cases (82.8 to 97.3%), found beneath appressoria of conidia that had failed to produce secondary hyphae. This contrasted with the more abundantly produced papillae on younger berries, where only 35.4 to 41.0% were located beneath appressoria of conidia that had failed to produce secondary hyphae. A pathogenesis-related gene (VvPR-1) was much more highly induced in susceptible berries than in resistant berries after inoculation with U. necator. In contrast, a germin-like protein (VvGLP3) was expressed within 16 h of inoculation in resistant, but not in susceptible berries. Our results suggest that several putative barriers to infection, i.e., cuticle and cell wall thickness, antimicrobial phenolics, and two previously described pathogenesis-related proteins, are not principal causes in halting pathogen ingress on ontogenically resistant berries, but rather that infection is halted by one or more of the following: (i) a preformed physical or biochemical barrier near the cuticle surface, or (ii) the rapid synthesis of an antifungal compound in older berries during the first few hours of the infection process. 相似文献
4.
J. Salinas D. C. M. Glandorf F. D. Picavet K. Verhoeff 《European journal of plant pathology / European Foundation for Plant Pathology》1989,95(1):51-64
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. 相似文献
5.
ABSTRACT Grape berries become resistant to powdery mildew early in development and are nearly immune to infection within 4 weeks after bloom. In this study, ontogenic resistance did not reduce attachment, germination, or appressorium formation of Uncinula necator on 3- to 4-week-old berries of Vitis vinifera 'Chardonnay' or 3-week-old berries of V. labruscana 'Concord'. Pathogen ingress halted at the cuticle before formation of a penetration pore. As berries aged, hyphal elongation and colony growth slowed until finally no secondary hyphae formed on fully resistant berries. More appressoria formed per unit of hyphal length as berries aged, indicating that failure to penetrate older berries led to increased attempts to penetrate resistant fruit. Additionally, hyphae within the colonies began to die as berries aged. Finally, the number of degree-hours between germination and sporulation of the colony (latent period) increased and sporophore density decreased with berry age at time of inoculation. Thus, ontogenic resistance both slows, and eventually halts disease development on grape berries, and limits the likelihood of spread by reducing absolute supply of conidia and delaying their formation. It furthermore has a consistent, stable, and predictable impact on grape powdery mildew and operates in a similar fashion and to a similar degree in both V. labruscana and V. vinifera, although at a slightly earlier phenological stage in V. labruscana. 相似文献
6.
ABSTRACT Resistance of mature berries of grapevine cultivars and selections to postharvest infection by Botrytis cinerea was assessed. Little or no resistance existed in most popular table grape Vitis vinifera cultivars, except in moderately resistant 'Emperor' and 'Autumn Black'. Highly resistant grapes were V. rotundifolia, V. labrusca, or other complex hybrids. Morphological, anatomical, and chemical characteristics of 42 genetically diverse cultivars and selections with various levels of resistance to B. cinerea were examined to determine which features were associated with resistance. We quantified the (i) density of berries within a cluster; (ii) number of pores and lenticels on the berry surface; (iii) thickness and number of cell layers in the epidermis and external hypodermis; (iv) amount of cuticle and wax; (v) berry skin protein content; (vi) total phenolic content of the skin before and after B. cinerea inoculation; and (vii) catechin and trans- and cis-resveratrol contents of the skin before and after inoculation. The number of pores was negatively correlated with resistance. Highly resistant cultivars had few or no pores in the berry surface. The number and thickness of epidermal and hypodermal cell layers and cuticle and wax contents were positively correlated with resistance. Other characteristics evaluated were not associated with resistance. trans-Resveratrol and cis-resveratrol were induced by B. cinerea inoculation only in sensitive and moderately resistant cultivars and selections. 相似文献
7.
ABSTRACT Brown rot, caused by Monilinia fructicola, is a destructive disease of stone fruit in California. Disease management requires information on inoculum dynamics and development of latent and visible fruit infections during the season to help make decisions on timing of fungicide treatments and choice of cultural practices. In this study, the daily spore concentration (ascospores and conidia) of M. fructicola in the air was monitored with spore traps in two prune orchards during the growing seasons in 2001 and 2002. The spore concentrations were low to moderate at early bloom, increased at full bloom, and decreased to the lowest level at the end of bloom. Improper timing of fruit thinning and irrigation in midseason increased spore concentration in the air and fruit infections late in the season. Artificial fruit inoculations were conducted periodically in 10 prune orchards in 2002 and 2004, and incidence of fruit rot at different inoculation dates was assessed. Fruit rot development rate increased linearly with inoculation date during the growing season. Natural blossom and fruit infections were monitored periodically in 10 prune orchards, and incidence of latent fruit infection was determined by using the overnight freezing-incubation technique. Incidence of fruit rot also was assessed 2 weeks before harvest in these orchards. The incidence of latent fruit infection at the pit hardening stage significantly correlated with that at the late stages and with the incidence of fruit rot at harvest. 相似文献
8.
The effects of different levels of inoculum of Aspergillus carbonarius and time of inoculation on berry infection and the development of aspergillus bunch rot on grapevines (cv. Sultana) were studied under field conditions. Inflorescences at full bloom were inoculated with aqueous spore suspensions of A. carbonarius containing 0 or 1 × 106 spores mL−1 in 2004/05 and 0, 1 × 102 or 1 × 105 spores mL−1 in 2005/06. In both years, the incidence of infection in inoculated berries was significantly higher than in uninoculated berries. Incidence of infection in berries from veraison until harvest was higher than at earlier stages of bunch development (berry set to berries that were still hard and green). Inoculation of bunches at veraison did not significantly increase A. carbonarius infection prior to harvest, at harvest, 6 days after harvest or when berries were over-ripe. Bunches inoculated at harvest did not significantly increase infection 6 days after harvest or when berries were over-ripe. Aspergillus carbonarius was isolated more frequently from the pedicel end (53·1%) than from the middle section (37·5%) and distal end (35·0%) of berries that were inoculated with 105 spores mL−1 . 相似文献
9.
U. Ma&#;olepsza† 《Plant pathology》2005,54(3):317-324
Pretreatment of detached tomato leaves with o -hydroxyethylorutin reduced the percentage leaf area affected, and delayed the appearance of necrosis, following inoculation with conidial suspensions in droplets of the grey mould fungus Botrytis cinerea . o -Hydroxyethylorutin delayed, but did not inhibit, in vitro germination of conidia, although overall percentage germination was reduced compared with water controls. Both the reactive oxygen species (ROS) – superoxide anions and hydrogen peroxide – increased twice as much in o -hydroxyethylorutin-treated leaf tissue 2 and 6 h postinoculation with B. cinerea conidia in tissues under inoculation drops, as well as in surrounding tissues, whereas in plants not pretreated with the compound ROS generation was noticed later, and only in tissues under inoculation drops. Compared with these compounds, changes in the levels of hydroxyl radicals, lipid peroxidation and the activity of the enzymes superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase and catalase were largely unchanged. In stimulating ROS in inoculated tomato tissue, B. cinerea appeared to be affected directly pre- and postinfection, but indirect effects increasing host resistance cannot be excluded. 相似文献
10.
A radioimmunosorbent assay was developed for Botrytis cinerea which can be used to detect as little as 100 ng of Botrytis mycelium (dry weight). Of 22 fungi tested in the assay, all significant reaction was restricted to members of the Sclerotiniaceae. No reaction was obtained with representative isolates of Aspergillus, Alternaria or Peiticillium. Other Botrytis species gave the most reaction relative to B. cinerea. and related genera such as Monilinia and Sclerotinia yielded small reactions. The assay can be used to detect fi. cinerea infection of grape berries when as little as 0.1 % of homogenate from infected fruit is mixed with homogenate from sound fruit to simulate a minor infection. The assay of free run juice from various lots of grapes was highly correlated with visual assessments of infection in the field. Pursuant to various theories concerning bloom time infection, the assay was used to analyse the B. cinerea content of immature berry tips and floral debris; however, no evidence of such colonization was obtained. 相似文献
11.
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. 相似文献
12.
Fungal development and plant responses were examined in detached leaves and mid-bulb scales of Allum cepa. Following inoculation with suspensions of 105 conidia/ml distilled water Botrytis squamosa consistently produced spreading lesions in leaves and bulb scales. B. allii produced spreading lesions at most sites in bulbs but was very inconsistent in its infection of leaves; lesions were often confined to inoculation sites. Limited lesions were usually produced by B. cinerea but R. fabae failed to produce symptoms at most sites. Extensive colonization by B. allii and B. tauamosa required rapid penetration and totally necrotrophic fungal growth. During development of a spreading lesion, plant cell walls became very swollen around intramural hyphae and wall swelling appeared to precede epidermal cell death. Resistance to colonization was due to poor germination, failure to produce distinct infection hyphae (associated with accumulation of deposits of granular reaction material [RM] in underlying live cells) or restriction of infection ryphae amongst small groups of dead cells (limited lesion formation). Only B. fabae germinated poorly, and germ-tubes produced often failed to attempt penetration but grew over the leaf or bulb scale surface. Reducing numbers of conidia increased the frequency of sites associated with RM accumulation; granular deposits being particularly common at sites inoculated with low numbers of B. allii conidia. Electron microscopy revealed that RM granules were osmiophilic aggregates formed between the plasma membrane and epidermal cell wall. In the absence of RM, growth of avirulent species was restricted within the swollen walls of dead epidermal cells. Results ae compared with those from studies on tulip and broad bean leaves. 相似文献
13.
梨叶叶龄与梨黑星病菌侵染发病的关系 总被引：6，自引：0，他引：6
田间试验结果表明，鸭梨叶片在生长发育期抗病性差，易感染黑星病；发育成熟后，抗病性逐渐增强。自然条件下，受叶片抗病性和孢子着落量的综合影响，鸭梨成熟叶片很少感染黑星病。鸭梨叶片从展叶到发育成熟，在梨树生长发育前期（盛花期后60d内）约需40d，后期（盛花期60d后）约需30d。正处在生长发育期的鸭梨叶片受侵染，潜育期为18－25d，发病后，病斑大，产孢期短；成熟叶片受侵染，潜育期多在40d以上，发病后，病斑小，产孢期长。梨树生长前期梨黑星病的流行主要由于大部分叶片正处在生长发育期抗病性较差；后期果实发病则主要源于成熟叶片内长期潜育和长期产孢的黑星病菌。 相似文献
14.
ABSTRACT Monilinia vaccinii-corymbosi infects open blueberry flowers via the gynoecial pathway, leading to mummification of the developing fruit. To determine the effect of flower age on infection, stigmata were inoculated with conidia of M. vaccinii-corymbosi between 0 and 5 days after anthesis, fungal growth rates through the stylar canal were measured in detached flowers in the laboratory, and fruit disease incidence was determined in plants grown in the greenhouse. Hyphal growth rates were greatest in flowers inoculated on the day of anthesis, declined linearly with increasing flower age at inoculation (r = 0.921; P < 0.0001; n = 12), and were unaffected by the presence or absence of pollen applied at the time of inoculation. In greenhouse-grown plants, the percentage of infected fruit decreased exponentially with increasing flower age at inoculation (R = 0.878; P = 0.0057; n = 10), with disease incidence ranging from 76.4% for flowers inoculated on the day of anthesis to 15.5% for those inoculated 4 days later. Fruit disease incidence in the greenhouse was linearly correlated with hyphal growth rates in detached flowers (r = 0.985; P < 0.0001; n = 9), justifying the use of detached flowers when investigating gynoecial infection by M. vaccinii-corymbosi. In separate experiments, the effects of timing and sequence of pollination and inoculation on hyphal growth rates through the stylar canal and on disease incidence were investigated. Application of pollen to detached flowers 1 or 2 days before inoculation reduced hyphal growth rates by between 14.0 and 42.9% compared with flowers that received pollen and conidia simultaneously. Similarly, reductions in fruit disease incidence by between 9.5 and 18.3% were observed on greenhouse-grown plants for pollination-to-inoculation intervals ranging from 1 to 4 days. These results document that newly opened flowers are most susceptible to infection by M. vaccinii-corymbosi and that fruit disease incidence is reduced if pollination occurs at least 1 day before inoculation. Strategies that lead to early pollination of newly opened flowers may be useful for managing mummy berry disease in the field. 相似文献
15.
Infestation of kiwifruit flowers by Thrips obscuratus , and inoculation with Botrytis cinerea , increased the amount of flower infection, external contamination and internal infection of fruit at harvest. Combination of these treatments was synergistic. The mechanisms of this role of T. obscuratus in the epidemiology of B. cinerea and development of stem-end rot after cool storage are discussed in relation to commercial production, harvesting and handling procedures. Significant reductions in inoculum available for fruit infection may be achievable by control of populations of T. obscuratus in orchards during flowering. 相似文献
16.
J. Salinas K. Verhoeff 《European journal of plant pathology / European Foundation for Plant Pathology》1995,101(4):377-386
The formation of lesions on ray florets of gerbera flowers caused by single conidia ofBotrytis cinerea was studied in two cultivars infected by two isolates of the pathogen. No differences in reaction after inoculation with conidia of either isolate were seen on either cultivar. The conidia produced usually one germ tube not longer than 10 m, but conidia with five germ tubes were also seen. Direct penetration of germ tubes through the upper cuticle of ray florets was observed. No appressoria or other specialised structures were observed before penetration, and degradation of the cuticle did not occur. Germination of conidia and subsequent flower infection was dependent on the availability of free water, but not on the addition of external nutrients.Between 18 to 25°C, fungal development usually stopped after cuticle penetration, two to four cells around the site of penetration becoming necrotic. This number did not increase when inoculated flowers were subsequently placed at 4°C, conditions conductive for the formation of spreading lesions. When flowers were incubated constantly at 4°C, lesions became visible 3 days after inoculation as a group of 10 to 14 cells. Initially from a vesicle-like structure, mycelium spread subcuticularly or in the lumen of epidermal cells resulting in the death of 40 to 50 cells at 18 days after inoculation. Ungerminated conidia and conidial germlings which has not yet penetrated the cuticle did not cause any visible symptoms in underlying epidermal cells. 相似文献
17.
Latent infection by Botrytis cinerea was frequently detected in young Primula × polyantha (horticultural hybrid polyanthus) plants. Genetically marked isolates were used to demonstrate that conidial inocula applied to young plants generally did not result in disease appearing on the leaves until plants flowered, regardless of when plants were inoculated. The severity of disease was proportional to the length of each day spent at 100% relative humidity during the latent period, regardless of time or form of inoculum. Plants with a prior B. cinerea infection were more susceptible to further infection and also showed symptoms of B. cinerea infection more quickly. Bacteria found to be associated with B. cinerea increased the likelihood of B. cinerea symptoms being seen on the plant, but did not directly assist infection. 相似文献
18.
The ability of Botrytis cinerea to infect plum and nectarine flowers was studied in moist chambers and in the orchard. Within 36 h of inoculation, the pathogen penetrated and colonized the stamens, styles and petals on shoots placed in moist chambers, causing blossom blight. Similar lesion development was observed following inoculation with dry and wet conidia, Hyphae were usually distorted in stylar tissue, but grew normally in petals and filaments. Growth of the fungus through filaments into the sepals or floral tubes, or through the transmitting tissue of the style into the ovary, was never observed. Symptoms of blossom blight were not observed on inoculated shoots in the orchard. The floral tube, bearing the sepals and stamens, dehisced within 14 days of fruit set and infected floral parts did not remain attached to young developing fruit. No relation was found between post-harvest decay and flower infection. Losses following post-harvest decay might have been caused by direct penetration of ripening fruit and not by flower infection. The importance of infected floral parts as a source of secondary inoculum on ripening fruit is discussed. 相似文献
19.
ABSTRACT Trichoderma biocontrol isolates are most effective as highly concentrated inocula. Their antagonism to other fungi may be a result of pregermination respiration. In a nutrient-rich medium, almost all Trichoderma atroviride P1 (P1) conidia initiated germination processes and increased respiration, even in dense suspensions. When 1 x 10(7) P1 conidia/ml were coinoculated with 1 x 10(5) Botrytis cinerea conidia/ml, dissolved oxygen fell to <1% within 2 h and the pathogen failed to germinate. More dilute P1 suspensions consumed oxygen slowly enough to allow coinoculated B. cinerea to germinate. On nutrient-poor media, fewer P1 conidia initiated germination. Oxygen consumption by the inoculum and inhibition of B. cinerea were enhanced when P1 conidia were nutrient activated before inoculation. Pregermination respiration also affected competitive capacity of the antagonist on solid substrates, where respiratory CO(2) stimulated germination rate and initial colony growth. These parameters were directly correlated with inoculum concentration (R(2) >/= 0.97, P < 0.01). After initiating germination, Trichoderma conidia became more sensitive to desiccation and were killed by drying after only 2 h of incubation on a nutrient-rich substrate at 23 degrees C. These results indicate that nutrient-induced changes preceding germination in Trichoderma conidia can either enhance or decrease their biological control potential, depending on environmental conditions in the microhabitat. 相似文献
20.
ABSTRACT Experiments were conducted in three prune orchards in California. In each orchard, inoculations with Monilinia fructicola, the causal agent of brown rot of stone fruits, were performed on branches of trees at bloom and fruit developmental stages. Five inoculum concentrations were used in each inoculation. Six and four wetness durations were created for each inoculum concentration at bloom and fruit developmental stages, respectively. Fruit were harvested 3 weeks before commercial harvest. The overnight freezing incubation technique was used to promote sporulation and to determine incidence of latent infection (ILI) of fruit brown rot. No differences in ILI among locations were found. A seasonal pattern of bloom and fruit susceptibility to latent infection was determined. Susceptibility to latent infection at bloom stage was at a moderate level and increased to reach the highest level at pit hardening stage. Subsequently, fruit susceptibility to latent infection decreased, reaching the lowest level in early June at embryo growth stage. Thereafter, the susceptibility increased again with fruit development and maturity until harvest. Linear relationships between ILI and inoculum concentration were obtained for most combinations of growth stage and wetness duration. Incidence of latent infection increased linearly with increased wetness duration at bloom stage and increased exponentially with increased wetness duration at early and late fruit developmental stages. The optimum temperatures for latent infection at pit hardening stage ranged from 14 to 18 degrees C, but the effect of temperature on latent infection was reduced at resistant stages. The temperature range favorable to latent infection varied for different wetness durations. 相似文献