Molecular, proteomic and morphological characterization of the ascomycete Guignardia bidwellii, agent of grape black rot: a polyphasic approach to fungal identification

Mycologia ◽  
2012 ◽  
Vol 104 (5) ◽  
pp. 1036-1045 ◽  
Author(s):  
B. Wicht ◽  
O. Petrini ◽  
M. Jermini ◽  
C. Gessler ◽  
G. A. L. Broggini
Plant Disease ◽  
2012 ◽  
Vol 96 (7) ◽  
pp. 1054-1059 ◽  
Author(s):  
Daniel Molitor ◽  
Cathleen Fruehauf ◽  
Ottmar Baus ◽  
Beate Berkelmann-Loehnertz

The duration of the incubation period of Guignardia bidwellii on leaves and clusters of Vitis vinifera strongly correlates to temperature. To describe this relationship mathematically, a new, cumulative degreeday- based model was developed. According to this model, first symptoms on leaves appear after reaching a threshold of 175 cumulative degree-days (calculated as the sum of average daily temperatures between 6 and 24°C starting on the day after the infection). On clusters, the duration of the incubation period is additionally affected by their respective developmental stages. For ‘Riesling’, the duration of the incubation period on clusters corresponds to the duration on leaves until reaching the phenological stage “berries beginning to touch“ but extends continuously with ongoing phenological development. Therefore, a correction factor recognizing cluster phenology was derived to calculate the cumulative degree-day thresholds for the occurrence of first symptoms on clusters after reaching “majority of berries touching”. Hence, this present model allows the estimation of fungal development and forecasts the appearance of new symptoms on leaves as well as on clusters, enabling growers to more precisely schedule curative as well as protective fungicide applications against grape black rot.


Plant Disease ◽  
2003 ◽  
Vol 87 (3) ◽  
pp. 273-281 ◽  
Author(s):  
Lisa Emele Hoffman ◽  
Wayne F. Wilcox

We studied several factors influencing the efficacy of the demethylation inhibitor (DMI) fungicide myclobutanil and the strobilurin fungicide azoxystrobin for control of grape black rot, caused by the pathogen Guignardia bidwellii (anamorph Phyllosticta ampelicida). The distribution of sensitivities to myclobutanil among G. bidwellii isolates from an “organic” vineyard (no previous exposure to synthetic fungicides, n = 50) and from a commercial vineyard with a history of DMI applications (n = 60) was determined in vitro. There was little difference between the two populations, and the range of sensitivities was narrow; for the composite population of 110 isolates, the value of the mean effective dose for 50% inhibition (ED50) was 0.04 mg/liter, and the most- and least-sensitive isolates were separated by a factor of 16. When applied from 2 to 6 days after inoculating grape seedlings with a suspension containing either 2 × 104 or 1 × 106 conidia per ml, myclobutanil (60 mg/liter) provided complete control of lesion development. When applied beyond 6 days after inoculation but prior to lesion appearance (9 to 11 days after inoculation, depending on temperature), it provided complete control of pycnidium production in those lesions that developed subsequently. In contrast, when applied 2 to 10 days after inoculation with 2 × 104 conidia per ml, azoxystrobin (128 mg/liter) provided only 78 to 63% control of lesion formation and erratic control of pycnidium formation, although conidium production was reduced by 85 to 68% across this range of treatments. Relatively little control was provided by azoxystrobin treatments following inoculation with 1 × 106 conidia per ml. On leaf disks treated with azoxystrobin at 20 mg/liter prior to inoculation, 8 to 43% of conidia from five G. bidwellii isolates germinated, and 4 to 19% formed appressoria. However, these processes were completely to near-completely inhibited when salicylhydroxamic acid (SHAM), which inhibits an alternative respiration pathway utilized to circumvent the activity of strobilurin fungicides, was added to the inoculum at 100 mg/liter. Thus, alternative respiration apparently allowed the conidia to germinate and form appressoria on azoxystrobin-treated leaves. When grape seedlings were sprayed with commercially formulated azoxystrobin at 200 mg/liter and inoculated the next day with G. bidwellii conidia, little or no disease was evident 4 weeks later. However, G. bidwellii pycnidia formed on up to 50% of the leaves from such plants when they were killed with paraquat 1 to 7 days after inoculation. These results suggest that latent infections became established on azoxystrobin-treated leaves and became active after the plants were killed with paraquat.


2017 ◽  
Vol 107 (2) ◽  
pp. 173-183 ◽  
Author(s):  
G. Onesti ◽  
E. González-Domínguez ◽  
V. Rossi

Black rot, caused by the fungus Guignardia bidwellii, is a polycyclic disease affecting grape leaves and berries. In environmentally controlled experiments and in a 3-year field study, the effects of temperature and relative humidity (RH) were assessed on the following growth parameters of G. bidwellii: (i) formation of pycnidia and cirri in grape leaf lesions, (ii) production and germination of conidia, and (iii) length of the period between lesion appearance and pycnidia production. Pycnidia were produced between 5 and 35°C and at 90 to 100% RH but more pycnidia were produced between 20 and 30°C. No pycnidia were produced at RH < 90%. The first pycnidia were produced in approximately 2 days after lesion appearance at ≥20°C and in 8 days at 5°C; pycnidia continued to be produced on the same lesion for 5 to 16 days after lesion appearance, depending on the temperature. Models were developed to describe the effect of temperature and RH on pycnidia production, accounting for 95 and 97% of variability, respectively. Cirri were extruded only between 15 and 35°C and mainly at 100% RH. Field experiments confirmed that pycnidia are produced for several days on a leaf lesion and that the length of the period between lesion appearance and pycnidia production depends on temperature. Overall, the findings showed that production of conidia requires high humidity; under field conditions, some hours at high humidity, which usually occur at nighttime, rather than constant high humidity may be sufficient.


2002 ◽  
Vol 92 (10) ◽  
pp. 1068-1076 ◽  
Author(s):  
Lisa Emele Hoffman ◽  
Wayne F. Wilcox ◽  
David M. Gadoury ◽  
Robert C. Seem

The period of fruit susceptibility to Guignardia bidwellii (anamorph Phyllosticta ampelicida), the causal agent of grape black rot, was determined in the field. Intact fruit were inoculated weekly from bloom until 8 weeks later with a suspension containing 2 × 105 conidia per ml. Disease progress was monitored approximately every 2 days until 3 to 5 weeks after inoculation, depending on the year and variety. Fruit of Vitis × labruscana ‘Concord’ exhibited a period of maximum susceptibility from midbloom until 2 to 4 weeks later, although some berries became symptomatic when inoculated 4 to 5 weeks postbloom. Fruit of V. vinifera ‘Chardonnay’ and ‘Riesling’ exhibited a period of maximum susceptibility from midbloom until 3 to 5 weeks later, although some berries retained their susceptibility until 6 to 7 weeks postbloom. These susceptible periods were approximately 2 to 4 weeks shorter than previously assumed. Fruit age at the time of inoculation affected the length of the incubation period (time from inoculation until symptom appearance). When the incubation period was defined in terms of degree hours (base = 0°C) accumulated after inoculation, DH50 values (the number of degree hours required to reach 50% of final disease severity) increased by at least 50% as berries neared the end of their susceptible period. Newly symptomatic berries continued to appear for over 1 month after inoculation of older fruit. Thus, age-related or ontogenic, host resistance was manifested as both a decline in susceptibility and a significant increase in incubation period length. The control of black rot is likely to be improved by tailoring the intensity of fungicidal protection to the abbreviated period of fruit susceptibility defined in this study. Furthermore, the efficacy of management programs and the results of epidemiological studies are likely to be misinterpreted unless the variable effect of fruit age on incubation period length is recognized.


2004 ◽  
Vol 94 (6) ◽  
pp. 641-650 ◽  
Author(s):  
Lisa Emele Hoffman ◽  
Wayne F. Wilcox ◽  
David M. Gadoury ◽  
Robert C. Seem ◽  
Duane G. Riegel

The epidemiology and control of black rot (Guignardia bidwellii) was studied from 1995 to 1999 in vineyards in Dresden and Naples, NY, where disease pressure was moderate and extreme, respectively. The efficacy of serial applications of myclobutanil, provided at 2-week intervals and varied with respect to their number and time of initiation, was examined within the context of host phenology, inoculum availability, and sanitation. At Dresden, sprays applied over 4 weeks through the immediate prebloom stage provided only 13 to 91% control of diseased clusters, despite the release of 95% of the season's ascosporic inoculum during the period of fungicidal protection. However, applications immediately prior to bloom plus 2 and 4 weeks later, which afforded protection while fruit are highly susceptible to infection, provided virtually complete control. At Naples, where mummified berries were retained in the canopy after mechanical pruning, this same regime provided only approximately 80% disease control, but applying a fourth spray 2 weeks prebloom generally improved control. Hand-pruning mummies to the ground in selected plots significantly (P ≤ 0.05) improved control in some spray regimes. Although this sanitation practice did not affect inoculum dynamics through bloom, very few spores were recovered thereafter from mummies collected from the ground, whereas abundant ascospores and conidia were recovered from mummies in the trellis for an additional 6 to 8 weeks.


Author(s):  
B. L. Soloff ◽  
T. A. Rado

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.


Planta Medica ◽  
2010 ◽  
Vol 76 (05) ◽  
Author(s):  
APPR Amarasinghe ◽  
RP Karunagoda ◽  
DSA Wijesundara

2020 ◽  
Vol 13 (11) ◽  
pp. 1
Author(s):  
A. R. B. Zanco ◽  
A. Ferreira ◽  
G. C. M. Berber ◽  
E. N. Gonzaga ◽  
D. C. C. Sabino

The different integrated production systems can directly interfere with its bacterial community. The present study aimed to assess density, bacterial diversity and the influence of dry and rainy season in different integrated and an exclusive production system. The fallow and a native forest area was assessed to. Samples were collected in 2012 March and September. The isolation were carried out into Petri dishes containing DYGS medium. The number of colony forming units (CFU) was counted after 48 hours and. The bacterial density ranged between 106 and 107 CFU g-1 soil. The crop system affected the dynamics of the bacterial community only in the rainy season. The rainy season showed greater density of total bacteria when compared to the dry period regardless of the cropping system. The dendrograms with 80 % similarity showed thirteen and fourteen groups in the rainy and dry seasons. Isolates with the capacity to solubilize phosphate in vitro were obtained from all areas in the two seasons, but this feature has been prevalent in bacteria isolated during the rainy season


2020 ◽  
Vol 20 (4) ◽  
pp. 448-454
Author(s):  
Rahmita Burhamzah ◽  
Gemini Alam ◽  
Herlina Rante

Background: Endophytic fungi live in plants’ tissue and can produce the same bioactive compounds as its host plant produces. Syzygiumpolyanthum leaves have known to be one of the antibacterial compound producers. Aim and Objective: This study aimed to characterize morphologically, microscopically, and molecularly the antibacterial-producing endophytic fungi of Syzygiumpolyanthum leaves. Methods: The isolation of endophytic fungi was done by fragment planting method on PDA medium. The antibacterial screening was performed using the antagonistic test as the first screening followed by the disc diffusion test method. The morphological characterization was based on isolate’s mycelia color, growth pattern, margin, and surface texture of the colony, while the microscopic characterization was based on its hyphae characteristics. The molecular characterization of the isolate was done by nitrogen base sequence analysis method on nucleotide constituent of ITS rDNA genes of the isolate. Results: The results found that isolate DF1 has antibacterial activity against E.coli, S.aureus, P.acne, and P.aeruginosa, with the greatest inhibition at 10% concentration of broth fermentation extract on S.aureus with a diameter of inhibition of 13.77 mm. Conclusion: Based on macroscopic, microscopic, and molecular characterization, DF1 isolate is similar to Ceriporialacerate.


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