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1.
Aureobasidium isolated from Vitis vinifera (cv Chardonnay) grapevine tissues were characterised using morphological and molecular techniques. Species level identification of 29 isolates was accomplished by partial amplification and sequencing of the ITS region (ITS1–5.8S–ITS2) using universal primers ITS1 and ITS4. A comparison of nucleotide sequences using BLAST followed by phylogenetic analysis revealed that all isolates examined were Aureobasidium pullulans. Strain level discrimination of a total of 100 epiphytic Aureobasidium isolates including three reference strains was successfully carried out using two inter simple sequence repeat (ISSR) primers, (AAC)5 and (GTG)5 and the Intron Splice Junction R1 (ISJ-R1) primer in which 24, 24 and 15 scorable bands were produced for each primer, respectively. The high level of genetic variation recorded among the isolates further highlighted the high levels of strain diversity among A. pullulans residing on grapevines. Thirty-two epiphytic Aureobasidium isolates were examined for their ability to inhibit the growth of Greeneria uvicola, responsible for bitter rot of grapes. Using an in-vitro dual-culture antagonism assay, all isolates inhibited the growth of G. uvicola (Isolates DAR 77272 and DAR 77273) with inhibition ranging from 15 to 85%. Three Aureobasidium isolates were then examined for their ability to inhibit G. uvicola when co-inoculated onto detached berries, leaves and grape bunches growing on potted vines in a glass house. All isolates reduced the severity of bitter rot infection. The results indicate that A. pullulans has the potential to suppress bitter rot of grapes.  相似文献   

2.
Infection of Malus x domestica cv. Royal Gala fruit by Colletotrichum acutatum causing bitter rot was studied in the temperate climate of New Zealand. Temperatures above 15 °C were required for lesions to develop on detached apple wound-inoculated or inoculated without wounding with C. acutatum spores, regardless of maturity. A wetness period of 72 h was required for infection of mature detached apple fruit without wounding. On wound-inoculated detached apple fruits, sporulation was related to temperature and followed a similar pattern. In the field, a mean temperature above 15 °C for 72 h after wound-inoculation was required for lesions to develop. Buds were a more important source of inoculum than twigs, and it was shown that C. acutatum could be isolated more frequently from outer bud scales than from inner scales. Asymptomatic infection of vegetative and reproductive buds was detected. C. acutatum was detected on asymptomic surface-sterilised petals and fruit, more commonly during summer than spring. Symptomless sterilised leaves generally yielded C. acutatum throughout the season, but isolations were more frequent in summer. Recovery of inoculum using a splash meter to detect vertical dispersal showed that in summer inoculum was primarily splashed up from the ground. In spring, inoculum was recovered in similar quantities from all heights up to a metre, suggesting that splash dispersal occurs from the canopy as well as from the ground. A disease cycle for C. acutatum infecting apples and causing bitter rot in New Zealand is suggested.  相似文献   

3.
Apple bitter rot caused by Colletotrichum acutatum sensu lato results in fruit decay before and after harvest. We investigated the epidemiology of the disease in terms of conidial formation and dispersal as well as the change in susceptibility of fruits in Iwate, Japan. Conidia of C. acutatum were detected in rainwater collected from inside the tree canopy from May to August with peaks in production in mid-May to early June and mid- to late July. The first peak corresponded to the most conidia being produced on fruit scars, but the second peak was due to conidiation on mummified fruitlets and peduncles collected in July. Inoculation experiments revealed that fruits were susceptible to the pathogen between 20 and 90 days after petal fall and that immature fruits infected as early as 20 days after petal fall frequently developed lesions on the lower fruit half as growth progressed. These results suggest that C. acutatum sporulates on infested fruit scars to infect immature fruits, resulting in bitter rot and that the fungus also colonizes mummified fruitlets and peduncles, contributing to survival of the pathogen on fruit scars. Thus, infested fruit scars represent the primary source of inoculum.  相似文献   

4.
Botrytis bunch rot (BBR), caused by Botrytis cinerea, degrades wine grapes during ripening, even though infection can occur as early as flowering. Effective BBR management requires knowledge of whether some stages of fruit development are more important than others in relation to infection and BBR severity at harvest. Bunches of Vitis vinifera ‘Sauvignon blanc’ and/or ‘Pinot noir’ were inoculated in two vineyard trials and one glasshouse trial with nitrate non‐utilizing (nit) mutant strains at three phenological stages: early flowering, pre‐bunch closure (PBC) and veraison. Isolates recovered from symptomless berries at veraison and from bunches with symptoms at harvest were screened to measure the incidence of the nit strains used in the inoculations. It was found that latent infections, which resulted in BBR at harvest, could become established at all three phenological stages and no single stage was associated with greater latent incidence or harvest severity than any other stage. It was concluded that a proportion of BBR at harvest resulted from the expression of latent infections that had accumulated throughout the season. However, the time between infection and BBR symptom expression in near‐ripe grape berries was sufficiently short for polycyclic secondary infection to also contribute to epidemic development.  相似文献   

5.
云南葡萄产区葡萄炭疽病病原鉴定及致病力分析   总被引：11，自引：6，他引：5  
为了明确引起云南葡萄产区炭疽病的病原种类,利用形态鉴定和特异性引物分子检测相结合的方法对从云南省主要葡萄产区采集的60株炭疽病菌菌株进行了鉴定。葡萄炭疽病菌菌株的菌落形态和生长速率与对照菌株尖孢炭疽菌Colletotrichum acutatum差异不明显,但其分生孢子大小显著小于尖孢炭疽菌,附着胞深褐色,球形或不规则形。胶孢炭疽菌Colletotrichum gloeosporioides特异性引物CgInt/ITS4从供试葡萄炭疽病菌菌株基因组DNA中扩增出1条约500 bp的特异性条带,而尖孢炭疽菌特异性引物CaInt2/ITS4对葡萄炭疽病菌无扩增条带。研究表明,引起云南葡萄主产区炭疽病的病原为胶孢炭疽菌;供试胶孢炭疽菌对红提葡萄均有致病力,但菌株致病力差异较大,对番茄和草莓存在交叉侵染的能力,且对多菌灵的敏感性较尖孢炭疽菌高。  相似文献   

6.
Greeneria uvicola causes bitter rot on Vitis vinifera (bunch grapes) and Muscadinia rotundifolia (muscadine grapes) in warm moist temperate and subtropical regions. This study investigated the phylogenetic relationship of G. uvicola representatives from Australia (67 isolates), the USA (31 isolates), India (1 isolate) and Costa Rica (1 isolate) and compared their pathogenicity and fungicide sensitivity. Differences in cultural and conidial morphology were observed between the isolates from Australia and the USA. Phylogenetic relationships were determined based on three gene regions: the ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1–5?8S–ITS2), 28S large subunit (LSU) nuclear rDNA and β‐tubulin‐2. Greeneria uvicola isolates were clearly differentiated into four groups: isolates from Australia and India; USA isolates from V. vinifera; USA isolates from M. rotundifolia; and the isolate from Costa Rica. All isolates were pathogenic on V. vinifera (cv. Chardonnay) berries although those originating from M. rotundifolia were not as aggressive as isolates from V. vinifera, irrespective of geographical origin. Sensitivity to pyraclostrobin and salicylhydroxamic acid (SHAM) was studied. Despite differences in fungicide applications, hyphal growth inhibition was not significantly different for geographical location, cultivar, tissue, year of collection or different spray regimes. For the Australian and USA isolates, fungal growth inhibition was significantly greater for pyraclostrobin than for SHAM, and was significantly greater for the combined treatment than for each of the fungicides applied singly. The aetiological and epidemiological knowledge of bitter rot collected through this study will aid better prediction and management strategies of this pathogen.  相似文献   

7.
Colletotrichum gloeosporioides and C. acutatum are causal agents of grape ripe rot, but with available methods, sporulation of C. gloeosporioides on plate media has been unstable and inferior to that of C. acutatum. To facilitate studies on C. gloeosporioides, I developed an improved method to induce conidiation of this fungus. Isolates of C. gloeosporioides were pre-cultured in potato dextrose broth for 1 week, then pulverized in whole broth. The homogenate was then spread on diluted oatmeal agar (15–20% commercial oatmeal agar medium, 1.5% agar) plates. After the plates were cultured at 25°C under continuous light for another week, the C. gloeosporioides isolates sporulated stably on the plate medium.  相似文献   

8.
Anthracnose fruit rot, caused by Colletotrichum acutatum, is among the most important diseases of blueberries. Most cultivars are susceptible but ‘Elliott’ is resistant. Our objective was to identify possible antifungal compounds that play a role in the resistance response. Chemical fractions from freeze-dried, ripe fruit of ‘Elliott’ and a susceptible cultivar (Jersey) were extracted with methanol and ethyl acetate. Extracts were screened on solid media for suppression of microconidiation of C. acutatum. The methanolic extract was fractionated and the soluble methanolic fraction from ‘Elliott’ was the most biologically active. This fraction was dried, dissolved in water, and screened in vivo by pre-treating ripe ‘Jersey’ fruit with 0.5, 1, 2, and 4% solutions (w/v) and subsequently inoculating the fruit with C. acutatum. An 88% reduction in infection incidence was observed after 12 days with the 4% solution. Anthocyanins and other flavonoids were then quantified in fruit of the two cultivars using HPLC-MS. ‘Elliott’ fruit contained more anthocyanins (4.87 mg/g of freeze-dried tissue) than ‘Jersey’ (3.27 mg/g of freeze-dried tissue); however, the same compounds were found in both cultivars. ‘Elliott’ fruit also contained more non-anthocyanin flavonoids (0.18 mg/g of freeze-dried tissue) than ‘Jersey’ (0.12 mg/g of freeze-dried tissue), including two distinctive compounds in ‘Elliott’. The non-anthocyanin flavonoid fractions of both ‘Elliott’ and ‘Jersey’ significantly decreased the growth of C. acutatum in a liquid bioassay but the effect was more pronounced in the ‘Elliott’ fraction. The two distinctive compounds in ‘Elliott’ were further characterized by MS/MS and were identified as quercetin 3-O-rhamnoside and, putatively, syringetin rhamnoside. Additionally, we evaluated ‘Elliott’ and ‘Jersey’ methanolic extracts for their ability to inhibit lipid peroxidation, and the extract obtained from ‘Elliott’ was almost twice as active at inhibiting peroxidation. These results provide new insights into the role of antifungal compounds in the resistance response of ripe ‘Elliott’ blueberries to infection by C. acutatum.  相似文献   

9.
Fungi within the Colletotrichum acutatum species complex occur asymptomatically on plant parts of many different plant species. Leaves from apple orchards in southern Norway were sampled, frozen for five hours and incubated for six days to reveal presence of asymptomatic infections of C. acutatum. Number of leaves (incidence) and leaf area covered (severity) with conidial masses of C. acutatum were assessed biweekly on cv. Aroma from late May to late September during three growing seasons. The first finding of conidial masses occurred in the second half of July, and there was a higher incidence occurring in August and September. Sampling of leaves from fruit spurs and vegetative shoots of cvs. Aroma and Elstar showed that conidial masses of C. acutatum developed on leaves on both shoot types, and there was no difference in incidence between these two types. The fungus was detected on leaves from six of eight commercial orchards of cv. Aroma over three years, with a mean incidence of 5.5 %. After storage, bitter rot was found on apple fruit from all eight orchards. There was no correlation between incidence of conidial masses of C. acutatum on leaves and on fruit. In all orchards and seasons investigated, incidence and severity on leaves varied from 0 to 67 % and 0 to 85 %, respectively. The discovery of apple leaves containing conidial masses of C. acutatum clearly indicate for leaves as a potential source of inoculum for fruit infections.  相似文献   

10.
Molecular sequencing (rpoB) and standard pathological and microbiological methods identified Pseudomonas syringae pv. syringae (Pss) as the causal agent of bacterial inflorescence rot of grapevines (Vitis vinifera) in three vineyards in Tumbarumba, NSW, Australia in 2006 and 2007. Pss strains from shrivelled berries and necrotic inflorescences of diseased grapevines were used to inoculate leaves and inflorescences of potted cv. Semillon grapevines. Pss caused disease symptoms similar to those experienced in the field, including angular leaf lesions, longitudinal lesions in shoot tissues and rotting of inflorescences from before flowering until shortly after fruit set. High humidity promoted symptom severity. The necrotic bunch stem and leaf lesions were susceptible to the development of Botrytis cinerea infections. Cryo‐scanning electron microscopy (cryoSEM) indicated that Pss entered leaves and inflorescence tissues via distorted, open, raised stomata surrounded by folds of tissue that appeared as ‘star‐shaped’ callose‐rich complexes when viewed by UV light microscopy. In necrotic tissues, cryoSEM revealed Pss within petiole parenchyma cells and air‐filled rachis xylem vessels. This is the first report of inflorescence and hence fruit loss caused by Pss in grapevines. The disease is described as ‘bacterial inflorescence rot’ and regarded as one that expands the previously reported pathology of grapevines caused by P. syringae. This study also indicated that infection by Pss might promote destructive B. cinerea infections when the fungus is already present but latent, although further experimentation is needed to prove such an interaction.  相似文献   

11.
Ripe rot is one of the most important bunch diseases of grapes in the highlands of southern Brazil and a matter of concern for winegrowers. Sixty-one strains isolated from berries of Vitis labrusca and V. vinifera varieties with symptoms were classified by phylogenetic analysis of the sequences of β-tubulin, glyceraldehyde-3-phosphate dehydrogenase, D1/D2 domain of 28S rDNA, and rDNA internal transcribed spacer. They were also characterized by morphology, and their pathogenicity was evaluated. The combined molecular data allowed identification of six Colletotrichum species: C. fructicola, C. kahawae and C. viniferum (gloeosporioides complex), C. limitticola, C. nymphaeae (acutatum complex), and C. karstii (boninense complex). This is the first report of C. karstii and C. limitticola associated with the ripe rot of grapes. Morphological characteristics varied within and among species, confirming their separation at the complex level. Pathogenicity tests on V. vinifera berries showed that the most prevalent species, C. viniferum (37.8%) and C. fructicola (36.1%), were more virulent than the less prevalent species C. limitticola and C. karstii. Our findings indicate that there is a high diversity of Colletotrichum species associated with ripe rot disease of grapes in Brazil. There were no clear differences in the distribution of Colletotrichum species between V. labrusca and V. vinifera varieties. The determination of fungal species responsible for grape ripe rot in Brazilian vineyards may contribute to further epidemiological studies and the development of more efficient prophylactic methods for ripe rot management.  相似文献   

12.
The study was carried out on phytoplasma susceptible grapevine variety ‘Chardonnay’ (Vitis vinifera L.). The changes in total and individual phenolics, with a focus on hydroxycinnamic acids, flavanols and flavonol contents, were studied in phytoplasma-symptomatic and non-symptomatic berries of Bois noir (BN) infected and uninfected vines. Evident responses to BN infection at veraison have been monitored in a decreased accumulation of caftaric and coutaric acids, p-coumaroyl hexose, procyanidin B1, procyanidin trimer as well as of quercetin-3-O-glucoside, quercetin-3-O-glucuronide and quercetin-3-O-xyloside. At berry softening BN infection statistically increased the content of total phenolics, hydroxycinnamic acids and flavanols, but decreased the flavonol contents, especially at phytoplasma-symptomatic berry skins. Later, at harvest, the BN infection caused an additional significant decrease of coutaric acid and p-coumaroyl pentose contents, moreover of procyanidin B1 and procyanidin dimmers (1, 2, and 3), trimer, kaempferol-3-O-glucoside and of most identified quercetins, except of quercetin-3-O-xyloside. At harvest, non-symptomatic berries from infected plants showed similar dynamics in the total phenolic content compared to berry skins from uninfected plants, but in total flavanols and flavonols content similarity to those symptomatic was observed. The latter decreases grape quality and its antioxidant potential. The Bois noir disease showed specific, local and growth-phase-induced responses regarding the content of phenolics in berry skins, where in particular the differences between phytoplasma-symptomatic and non-symptomatic grapes have to be underlined.  相似文献   

13.
Brown lesions on red clover, resulting from infection by ascospores ofSclerotinia trifoliorum, were sometimes surrounded by a light grey ring. Clover rot developed rapidly from these rings under moist conditions. The beginning of rot in the field was often observed in the joints of the trifoliate leaves. Freezing plants at ?6° or ?8°C or freezing small parts of plants with carbon dioxide snow resulted in a quick spread of mycelium, with rotting, from the ascospore lesions. Weakening the plants by bruising did not promote this transition to active rotting. Very heavy infection by ascospores, however, may cause immediate rotting in the plants. The information obtained indicates that in the field clover rot originates mainly from ascospore lesions.  相似文献   

14.
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   

15.
Anthracnose, caused by Colletotrichum acutatum (species complex), has become a troublesome problem in strawberry production worldwide. This paper reports (i) an optimized sampling method combined with a real‐time PCR technique to detect the latent presence of C. acutatum in cold‐stored strawberry plants used as planting material in several European countries, and (ii) a study of the spread of C. acutatum following a point inoculation under field conditions. Screening of different parts of planting material suggested that C. acutatum is most likely to be present on runners and old petioles. In addition, in seven out of nine batches of planting material from different nurseries, latent infection by C. acutatum was detected in at least one of five replicate samples. Field experiments in 2009 and 2010 showed extensive latent within‐field spread of the pathogen on strawberry leaves, with a within‐row dispersal distance up to at least 1·75 m in 1 week. A straw ground cover between the rows did not decrease C. acutatum spread, probably because introduced (and/or subsequent) inoculum was confined to the plant bed (within the row) and was not present between the beds. Moreover, the number of C. acutatum spores on the symptomless leaves, as estimated using a real‐time PCR method, was significantly (P < 0·05) correlated with the incidence of fruit rot at harvest and post‐harvest (r = 0·56–0·66). These results illustrate the importance of detecting latent infections in planting material and strawberry leaves in the field.  相似文献   

16.
The efficacy of Trichoderma harzianum strain DGA01, sodium carbonate (SC), sodium bicarbonate (SBC) and sodium hypochlorite (SH) applied alone or in various combinations was evaluated in vitro against the most important postharvest pathogens of banana such as Lasiodiplodia theobromae, Thielaviopsis paradoxa, Colletotrichum musae, and Fusarium verticillioides. Trichoderma harzianum DGA01 was compatible with salts at a concentration of 1 % (w/v) as manifested by normal mycelial growth. The efficacy of the fungal antagonist in vitro was enhanced by 10.16–13.06 % in controlling mycelial growth of crown rot pathogens with the addition of salts. After a postharvest dip of fruit for 30 min in 106 conidia of DGA01/mL of 1 % SBC, the incidence of crown rot was reduced by 92 %, but DGA01 combined with SC or SH had no additive effect in controlling crown rot. Thus, DGA01 and SBC was the best combination for crown rot control, with an efficacy similar to synthetic fungicides, and maintained the overall quality of banana even at conditions favourable for the pathogens (22–25 °C, 90–95 % relative humidity).  相似文献   

17.
Phytopthora root rot in pepper (C. annuum) is caused by Phytophthora capsici L., which exhibits a high level of pathogenic diversity. Resistance to this disease is conditioned by a number of quantitative trait loci. Pyramiding resistance alleles is desirable and could be simplified by the use of molecular markers tightly linked to the resistance genes. The purpose of this study was development of molecular markers linked to Phytophthora root rot resistance. An F8 recombinant inbred line (RIL) population derived from a cross between YCM334 and a susceptible cultivar ‘Tean’ was used in combination with bulk segregant analysis utilizing RAPD and conversion of AFLP markers linked to Phytophtora root rot resistance into sequence-characterized amplified region (SCAR) markers. In conversion: one marker was successfully converted into a co-dominant SCAR marker SA133_4 linked to the trait. In bulked segregant analysis (BSA): three RAPD primers (UBC484, 504, and 553) produced polymorphisms between DNA pools among 400 primers screened. Genetic linkage analysis showed that the SCAR and RAPD markers were located on chromosome 5 of pepper. Quantitative trait locus (QTL) analysis showed that the SA133_4 and UBC553 were linked to Phytophtora root rot resistance. These markers were correctly identified as resistant or susceptible in nine promising commercial pepper varieties. These markers will be beneficial for marker-assisted selection in pepper breeding.  相似文献   

18.
为探明东方百合茎腐病的发病机制及抗病机理,以湖南株洲地区的东方百合为材料,分别采用升汞消毒、刺伤浸苗接种、超声波甲醇浸提、混液平板等方法对其病原菌进行了分离、抗病性鉴定、皂苷含量及活性测定。结果显示：尖孢镰刀菌Fusarium oxysporum为湖南株洲地区东方百合茎腐病病原菌;30份自主培育的东方百合材料通过聚类分析可划分为3类,包含抗病材料3份、中抗材料16份、感病材料11份,其中y、ZN-12和ZN-27的抗病指数都在65.00以上;抗病材料的总皂苷含量明显高于感病材料,且皂苷含量与抗性水平之间极显著相关;总皂苷含量为1.194 mg/mL的提取液对茎腐病病原菌菌丝生长的抑制率为29.39%。表明y、ZN-12和ZN-27具有较好的开发潜力;总皂苷含量与东方百合抗茎腐病水平密切相关,可作为大规模筛选抗病材料的一个重要指标。  相似文献   

19.
Oranges and lemons infected with Phytophthora citrophthora and P. syringae were placed among healthy fruits in boxes kept at 5, 10 and 14°C. The spread of the rot by contact infection was assessed after 3,6,12 and 18 days and again after 7 days' shelf-life at 17°C. P. citrophthora infected adjacent fruits after 12 days at 10°C and 3 days at 14°C. No infection occurred at 5°C. P. syringue infected the fruits at all the temperatures tested. On fruits kept for 6 days at 5°C, the rot developed after shelf-life. The presence of Phytophthora-infected fruits in orange and lemon packing boxes enhanced the development of penicillium rots. Post-harvest dips in metalaxyl at 0.05 and 0.1% concentrations and fosetyl-Al at 0.1,0.2 and 0.3% concentrations prevented the spread of P. citrophthora in packing boxes kept for 30,40 and 60 days at 11°C, but had no effect on penicillium rots. The mixture of the above fungicides with thiabendazole at 0.1%] or imazalil at 0.05%, concentrations gave best control against phytophthora and penicillium rots.  相似文献   

20.
Eucalyptus diversicolor (karri) is the second most important commercial timber tree in Western Australia. Sawlogs from regrowth trees often have a discoloration in the heartwood that is more abundant than in sawlogs from mature trees. Other symptoms in regrowth logs include white rot, white pocket rot and brown rot. Fungal isolations and pathogenicity tests were conducted to determine whether this discoloration was incipient rot, and if so, what caused it and which rot(s) would eventually develop. A combined sample of 329 discs from recently felled trees and freshly cut scantling had discoloration in 48%, white rot in 14%, white pocket rot in 12% and brown rot in 4% of pieces of wood. Hymenochaete semistupposa was isolated from 22% of discoloured wood and 39% of white pocket rot samples. Stereum hirsutum was isolated from 4% of discoloured wood and 13% of white rot samples. Koch's postulates in regrowth karri trees showed that H. semistupposa caused extensive discoloration, with white pocket rot developing in the heartwood within 4 years, while S. hirsutum caused extensive discoloration, with white rot developing in both the heartwood and sapwood within 2 years. It was concluded that the discoloration was incipient rot, which would eventually develop into either white rot or white pocket rot.  相似文献   

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