scholarly journals Stability and Fitness of Anilinopyrimidine-Resistant Strains of Botrytis cinerea

2008 ◽  
Vol 98 (4) ◽  
pp. 443-450 ◽  
Author(s):  
G. A. Bardas ◽  
C. K. Myresiotis ◽  
G. S. Karaoglanidis
Keyword(s):  
Botrytis Cinerea ◽  
Spore Germination ◽  
Mycelial Growth ◽  
Spore Production ◽  
Vegetable Crops ◽  
Fitness Components ◽  
Development Of Resistance ◽  
Fitness Parameters

The fitness of anilinopyrimidine-resistant isolates of Botrytis cinerea compared with that of sensitive isolates, collected from vegetable crops in Greece during 2005, was investigated. Stability of resistance to anilinopyrimidine fungicides was determined after consecutive transfers of the fungal isolates on fungicide-free potato dextrose agar for 16 culture cycles or on fungicide-untreated cucumber seedlings for eight disease cycles. Results showed that after the consecutive transfers of the isolates either in vitro or in vivo sensitivity to cyprodinil was not changed significantly compared to the initial sensitivity in all the isolates tested, suggesting a stable genetically controlled trait. Fitness parameters measured were mycelial growth, spore production in vitro, osmotic sensitivity, virulence, spore production in vivo, percentage of spore germination, and competitive ability of the resistant isolates in four pairs with sensitive isolates both on artificial nutrient medium or on cucumber seedling plants. The measurements of the fitness components in individual isolates showed high variability within both sensitivity groups in all, except virulence, fitness components tested. As a group, resistant isolates showed significantly lower (P < 0.05) mycelial growth and virulence, while they were more osmotically sensitive than the sensitive isolates. In addition the resistant isolates showed higher (P < 0.05) spore production in vivo but there was no difference (P > 0.05) between the two sensitivity groups in spore production in vitro and in the percentage of spore germination. However, the correlation to test if there is any relationship between the values of each fitness component tested and the level of cyprodinil sensitivity of each isolate was for all, except the spore production in vivo, fitness components not significant (P > 0.05). This absence of significant correlation coefficient values suggests that the development of resistance to anilinopyrimidine fungicides did not affect the fitness of the resistant isolates. Competition of the resistant versus sensitive isolates was isolates-dependent, since in two of the isolate pairs the resistance frequency decreased significantly after five culture or disease cycles, while in the remaining two pairs resistance frequency increased significantly after five disease cycles or remained stable for one pair after five culture cycles on artificial nutrient media.

scholarly journals Competitive Ability and Fitness of Alternaria alternata Isolates Resistant to QoI Fungicides

Plant Disease ◽  
2011 ◽  
Vol 95 (2) ◽  
pp. 178-182 ◽  
Author(s):  
G. S. Karaoglanidis ◽  
Y. Luo ◽  
T. J. Michailides
Keyword(s):  
Alternaria Alternata ◽  
Mycelial Growth ◽  
Competitive Ability ◽  
Spore Production ◽  
Fitness Cost ◽  
Development Of Resistance ◽  
Fitness Parameters ◽  
Resistant Strains

Fungicides that act as quinone outside inhibitors (QoIs) constitute a fungicide group extensively used against Alternaria late blight of pistachio caused by Alternaria spp. However, developement of resistance to this fungicide class constitutes an important threat for the succesful control of the disease. This study was conducted to determine whether development of resistance to QoIs is associated with a fitness cost, by measuring several biological and epidemiological parameters and estimating the competitive ability in four QoI-resistant and four QoI-sensitive Alternaria alternata isolates. Fitness parameters measured were mycelial growth and spore production in vitro, disease latent period, aggressiveness, and spore production on detached pistachio leaves. The competitive ability of resistant isolates was assessed in coinoculation experiments with sensitive isolates on detached pistachio leaves, using a real-time polymerase chain reaction assay technique. Fitness parameters between grouped QoI-resistant and QoI-sensitive isolates were not significantly different (P = 0.13, 0.21, 0.31, and 0.27 for sporulation in vitro, mycelial growth, incubation period, and sporulation in vivo, respectively), while resistant isolates, as a group, showed a higher aggressiveness (P = 0.01) compared with the sensitive isolates. The data indicate that the resistant strains did not account for a fitness cost compared with the sensitive ones under the conditions of testing. The outcome of the competition experiments was isolate dependent. In two pairs, the resistance frequencies increased whereas, in the remaining two pairs of isolates, resistance frequency decreased, suggesting that the resistant isolates were competitive similarly to the sensitive isolates.

scholarly journals (E)-2-Hexenal Can Stimulate Botrytis cinerea Growth in vitro and on Strawberries in vivo during Storage

1998 ◽  
Vol 123 (5) ◽  
pp. 875-881 ◽  
Author(s):  
Elazar Fallik ◽  
Douglas D. Archbold ◽  
Thomas R. Hamilton-Kemp ◽  
Ann M. Clements ◽  
Randy W. Collins ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Spore Germination ◽  
Mode Of Action ◽  
Mycelial Growth ◽  
Titratable Acidity ◽  
Soluble Solids ◽  
In Vivo Studies ◽  
High Concentration

Some plant-derived natural volatile compounds exhibit antifungal properties and may offer an opportunity to control the causes of postharvest spoilage without affecting quality of, or leaving a residue on, fresh produce. The natural wound volatile (E)-2-hexenal has exhibited significant antifungal activity in earlier studies, but effects on spore germination and mycelial growth have not been separated, nor has the inhibitory mode of action been determined. To determine the efficacy of (E)-2-hexenal for control of Botrytis cinerea Pers. ex Fr. spore germination and mycelial growth, and to examine the mode of action, in vitro and in vivo studies were performed. Under in vitro bioassay conditions, spore germination was more sensitive to the compound than was mycelial growth. Vapor from 10.3 μmol of (E)-2-hexenal in a 120-mL petri dish completely inhibited spore germination. However, 85.6 μmol of (E)-2-hexenal was required to completely inhibit mycelial growth. Lower concentrations of the compound (5.4 and 10.3 μmol) significantly stimulated mycelial growth, especially when the volatile was added 2 days following inoculation. Mycelial growth did not occur as long as the vapor-phase concentration was 0.48 μmol·L-1 or greater. Light microscopy analysis indicated that a high concentration of volatile compound dehydrated fungal hyphae and disrupted their cell walls and membranes. Exposure of B. cinerea-inoculated and non-inoculated strawberry (Fragaria ×ananassa Duch.) fruit in 1.1-L low-density polyethylene film-wrapped containers to vapor of (E)-2-hexenal at 85.6 or 856 μmol (10 or 100 mL, respectively) per container for durations of 1, 4, or 7 days during 7 days of storage at 2 °C promoted the incidence of B. cinerea during subsequent shelf storage at 20 to 22 °C. Loss of fruit fresh mass and fruit firmness during storage at 22 °C was increased by (E)-2-hexenal treatment, but fruit total soluble solids, pH, titratable acidity, and color (L, C, and H values) were not affected. Thus, maintenance of a high vapor-phasel level of (E)-hexenal, perhaps >0.48 μmol·L-1, may be necessary to inhibit mycelial growth and avoid enhancing postharvest mold problems, while significantly higher levels may be necessary to completely eliminate the pathogen.

An Attempt at Biological Control of Blossom Blight of Rose Caused by Botrytis cinerea Using some Local Trichoderma spp. Strains

2020 ◽  
Vol 55 (1) ◽  
pp. 27-34
Author(s):  
G. Zadehdabagh ◽  
K. Karimi ◽  
M. Rezabaigi ◽  
F. Ajamgard
Keyword(s):  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Limiting Factor ◽  
Dual Culture ◽  
Trichoderma Spp ◽  
Khuzestan Province ◽  
Inhibitory Potential ◽  
Cut Rose

The northern of Khuzestan province in Iran is mainly considered as one of the major areas of miniature rose production. Blossom blight caused by Botrytis cinerea has recently become a serious limiting factor in rose production in pre and post-harvest. In current study, an attempt was made to evaluate the inhibitory potential of some local Trichoderma spp. strains against B. cinerea under in vitro and in vivo conditions. The in vitro results showed that all Trichoderma spp. strains were significantly able to reduce the mycelial growth of the pathogen in dual culture, volatile and non-volatile compounds tests compared with control, with superiority of T. atroviride Tsafi than others. Under in vivo condition, the selected strain of T. atroviride Tsafi had much better performance than T. harzianum IRAN 523C in reduction of disease severity compared with the untreated control. Overall, the findings of this study showed that the application of Trichoderma-based biocontrol agents such as T. atroviride Tsafi can be effective to protect cut rose flowers against blossom blight.

scholarly journals Effects of Origanum vulgare essential oil and its two main components, carvacrol and thymol, on the plant pathogen Botrytis cinerea

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9626
Author(s):  
Huiyu Hou ◽  
Xueying Zhang ◽  
Te Zhao ◽  
Lin Zhou
Keyword(s):  
Antifungal Activity ◽  
Essential Oil ◽  
Botrytis Cinerea ◽  
Spore Germination ◽  
Gray Mold ◽  
Origanum Vulgare ◽  
Mycelium Growth ◽  
Main Components

Background Botrytis cinerea causes serious gray mold disease in many plants. This pathogen has developed resistance to many fungicides. Thus, it has become necessary to look for new safe yet effective compounds against B. cinerea. Methods Essential oils (EOs) from 17 plant species were assayed against B. cinerea, of which Origanum vulgare essential oil (OVEO) showed strong antifungal activity, and accordingly its main components were detected by GC/MS. Further study was conducted on the effects of OVEO, carvacrol and thymol in vitro on mycelium growth and spore germination, mycelium morphology, leakages of cytoplasmic contents, mitochondrial injury and accumulation of reactive oxygen species (ROS) of B. cinerea. The control efficacies of OVEO, carvacrol and thymol on tomato gray mold were evaluated in vivo. Results Of all the 17 plant EOs tested, Cinnamomum cassia, Litsea cubeba var. formosana and O. vulgare EOs had the best inhibitory effect on B. cinerea, with 0.5 mg/mL completely inhibiting the mycelium growth of B. cinerea. Twenty-one different compounds of OVEO were identified by gas chromatography–mass spectrometry, and the main chemical components were carvacrol (89.98%), β-caryophyllene (3.34%), thymol (2.39%), α-humulene (1.38%) and 1-methyl-2-propan-2-ylbenzene isopropyl benzene (1.36%). In vitro experiment showed EC50 values of OVEO, carvacrol and thymol were 140.04, 9.09 and 21.32 μg/mL, respectively. Carvacrol and thymol completely inhibited the spore germination of B. cinerea at the concentration of 300 μg/mL while the inhibition rate of OVEO was 80.03%. EC50 of carvacrol and thymol have significantly (P < 0.05) reduced the fresh and dry weight of mycelia. The collapse and damage on B. cinerea mycelia treated with 40 μg/mL of carvacrol and thymol was examined by scanning electron microscope (SEM). Through extracellular conductivity test and fluorescence microscope observation, it was found that carvacrol and thymol led to increase the permeability of target cells, the destruction of mitochondrial membrane and ROS accumulation. In vivo conditions, 1000 μg/mL carvacrol had the best protective and therapeutic effects on tomato gray mold (77.98% and 28.04%, respectively), and the protective effect was significantly higher than that of 400 μg/mL pyrimethanil (43.15%). While the therapeutic and protective effects of 1,000 μg/mL OVEO and thymol were comparable to chemical control. Conclusions OVEO showed moderate antifungal activity, whereas its main components carvacrol and thymol have great application potential as natural fungicides or lead compounds for commercial fungicides in preventing and controlling plant diseases caused by B. cinerea.

scholarly journals Fitness and Competitive Ability of Botrytis cinerea Isolates with Resistance to Multiple Chemical Classes of Fungicides

2016 ◽  
Vol 106 (9) ◽  
pp. 997-1005 ◽  
Author(s):  
S. N. Chen ◽  
C. X. Luo ◽  
M. J. Hu ◽  
G. Schnabel
Keyword(s):  
Botrytis Cinerea ◽  
Competitive Ability ◽  
Selective Pressure ◽  
Spore Production ◽  
Inoculum Potential ◽  
Eastern United States ◽  
Competitive Disadvantage ◽  
Multiple Chemical

Resistance to multiple chemical classes of fungicides in Botrytis cinerea isolates from eastern United States strawberry fields is common and strategies to control them are needed. In this study, we compared fitness and competitive ability of eight sensitive isolates (S), eight isolates resistant to five or six chemical classes of fungicides but not to phenylpyrroles (5CCR), and eight isolates resistant to six or seven chemical classes including phenylpyrroles (6CCR/MDR1h). The latter included the MDR1h phenotype due to overexpression of atrB based on Δ497V/L in mrr1. The 6CCR/MDR1h isolates grew more slowly at 4°C on potato dextrose agar, and both 5CCR and 6CCR/MDR1h isolates were hypersensitive to osmotic stress compared with S isolates. In contrast, no differences were found in oxidative sensitivity, aggressiveness, and spore production in vivo, and sclerotia production and viability in vitro. In competition experiments, the 5CCR and 6CCR/MDR1h isolates were both outcompeted by S isolates and 6CCR/MDR1h isolates were outcompeted by 5CCR isolates in the absence of fungicide pressure. Under selective pressure of a fludioxonil/pyraclostrobin rotation, the 6CCR/MDR1h isolates dominated after coinoculation with 5CCR and S isolates. The competitive disadvantage of 5CCR and especially 6CCR/MDR1h isolates suggest that, in the absence of fungicide selection pressure, S isolates may reduce inoculum potential of multifungicide-resistant isolates under field conditions.

scholarly journals Multiple Resistance of Botrytis cinerea from Vegetable Crops to Carbendazim, Diethofencarb, Procymidone, and Pyrimethanil in China

Plant Disease ◽  
2010 ◽  
Vol 94 (5) ◽  
pp. 551-556 ◽  
Author(s):  
Hai-Yan Sun ◽  
Han-Cheng Wang ◽  
Yu Chen ◽  
Hong-Xia Li ◽  
Chang-Jun Chen ◽  
...  
Keyword(s):  
Amino Acid ◽  
Botrytis Cinerea ◽  
Point Mutations ◽  
Amino Acid Position ◽  
Shandong Province ◽  
Jiangsu Province ◽  
Vegetable Crops ◽  
Kinase Gene

One hundred and eight isolates of Botrytis cinerea from greenhouse cucumber and tomato in two locations in Jiangsu Province (Nanjing and Huaiyin) and one location in Shandong Province were tested for their sensitivities to the four fungicides commonly used in China. Isolates with resistance to all four fungicides—carbendazim, diethofencarb, procymidone, and pyrimethanil (CarRDieRPrcRPyrR)—were found in all three regions in this study. High frequencies (52 and 53%) of resistance to all four fungicides were observed among the 62 isolates collected in Nanjing and the 36 isolates collected in Huaiyin in Jiangsu Province. The 10 isolates from Shandong Province were all resistant to the four fungicides. Pathogenicity and sporulation in vivo, and mycelial growth, sporulation, spore germination, and osmotic sensitivity to NaCl in vitro, were similar for the group of quadruple-resistant and wild-type isolates (P > 0.05). In the present study, the complete two-component histidine kinase gene (Bos1) was sequenced for 10 procymidone-resistant and 3 procymidone-sensitive B. cinerea isolates. Isolates representing four different procymidone-resistant phenotypes (CarRDieSPrcRPyrS, CarRDieRPrcRPyrS, CarRDieRPrcRPyrR, and CarRDieRPrcRPyrR) all had nucleic acid point mutations resulting in amino acid changes at position 369 (change from glutamine to proline) as well as at amino acid position 373 (asparagine to serine) in the Bos1 gene.

scholarly journals Baseline Sensitivity of Botrytis cinerea to Pyraclostrobin and Boscalid and Control of Anilinopyrimidine- and Benzimidazole-Resistant Strains by These Fungicides

Plant Disease ◽  
2008 ◽  
Vol 92 (10) ◽  
pp. 1427-1431 ◽  
Author(s):  
C. K. Myresiotis ◽  
G. A. Bardas ◽  
G. S. Karaoglanidis
Keyword(s):  
Electron Transport ◽  
Botrytis Cinerea ◽  
Spore Germination ◽  
Mycelial Growth ◽  
Cross Resistance ◽  
Vegetable Crops ◽  
Baseline Sensitivity ◽  
Transport Chain ◽  
Resistant Strains ◽  
And Control

Fifty-five isolates of Botrytis cinerea collected from vegetable crops were used to determine the pathogen's baseline sensitivity to two new fungicides: boscalid, which inhibits the enzyme succinate dehydrogenase in the electron transport chain, and pyraclostrobin, which blocks electron transport between cytochrome b and cytochrome c1. Measurement of sensitivity to boscalid was based on both inhibition of mycelial growth and spore germination, while measurement of sensitivity to pyraclostrobin was based only on inhibition of spore germination. For both fungicides, the sensitivity distribution was a unimodal curve, with a mean EC50 value (effective concentration that reduces mycelial growth or spore germination by 50%) of 0.033 μg ml-1 for pyraclostrobin and 2.09 and 2.14 μg ml-1 for boscalid based on the inhibition of mycelial growth and spore germination, respectively. No cross-sensitivity relationship was observed between the two fungicides (r = 0.09). In addition, no cross-resistance relationship was observed between these two fungicides with other botryticides: cyprodinil, pyrimethanil, fenhexamid, fludioxonil, and iprodione. Moreover, the control efficacy of the two fungicides was tested against two anilinopyrimidine-resistant and two benzimidazole-resistant isolates, and two of wild-type sensitivity. Both pyraclostrobin and boscalid provided satisfactory control of all six isolates that was independent of the isolate sensitivity to benzimidazoles and anilinopyrimidines. In contrast, carbendazim failed to control sufficiently the benzimidazole-resistant isolates, while cyprodinil failed to provide satisfactory control of the anilinopyrimidine-resistant isolates.

scholarly journals Parasitic Fitness of Fungicide-Resistant and -Sensitive Isolates of Alternaria solani

Plant Disease ◽  
2018 ◽  
Vol 102 (3) ◽  
pp. 666-673 ◽  
Author(s):  
Mitchell J. Bauske ◽  
Neil C. Gudmestad
Keyword(s):  
Spore Germination ◽  
Point Mutations ◽  
Alternaria Solani ◽  
The United States ◽  
Wild Type ◽  
Parasitic Fitness ◽  
Fitness Parameters ◽  
Multiple Chemical

Resistance to chemistries of the succinate dehydrogenase inhibiting (SDHI) and quinone outside inhibiting (QoI) fungicides has developed rapidly in populations of Alternaria solani, the cause of early blight of potato. Reduced sensitivity to the anilinopyrimidine (AP) fungicide pyrimethanil has also been identified recently, determining that resistance to three chemical classes of fungicides is present within the A. solani population. Although no mutations have been characterized to confer resistance to APs, in A. solani five point mutations on three AsSdh genes have been determined to convey resistance to SDHIs, and the substitution of phenylalanine with leucine at position 129 (F129L) in the cytb gene confers resistance to QoIs. The objective of this study was to investigate the parasitic fitness of A. solani isolates with resistance to one or more of these chemical classes. A total of 120 A. solani isolates collected from various geographical locations around the United States were chosen for in vitro assessment, and 60 of these isolates were further evaluated in vivo. Fitness parameters measured were (i) spore germination in vitro, (ii) mycelial expansion in vitro, and (iii) aggressiveness in vivo. No significant differences in spore germination or mycelial expansion (P = 0.44 and 0.51, respectively) were observed among wild-type and fungicide-resistant isolates in vitro. Only A. solani isolates possessing the D123E mutation were shown to be significantly more aggressive in vivo (P < 0.0001) compared with wild-type isolates. These results indicate that fungicide-resistant A. solani isolates have no significant fitness penalties compared with sensitive isolates under the parameters evaluated regardless of the presence or absence of reduced sensitivity to multiple chemical classes. Results of these studies suggest that A. solani isolates with multiple fungicide resistances may compete successfully with wild-type isolates under field conditions.

scholarly journals Antifungal Activity of Alpha-Mangostin against Colletotrichum gloeosporioides In Vitro and In Vivo

Molecules ◽  
2020 ◽  
Vol 25 (22) ◽  
pp. 5335
Author(s):  
Huochun Ye ◽  
Qin Wang ◽  
Fadi Zhu ◽  
Gang Feng ◽  
Chao Yan ◽  
...  
Keyword(s):  
Spore Germination ◽  
Inhibitory Activity ◽  
Mycelial Growth ◽  
Colletotrichum Gloeosporioides ◽  
Crop Protection ◽  
Ultrastructural Observation ◽  
Garcinia Mangostana ◽  
Mitochondrial Energy Metabolism

We investigated alpha-mangostin (α-mangostin, α-MG), a xanthone natural product extracted from the pericarp of mangosteen (Garcinia mangostana), for its antifungal activities and possible mechanism against Colletotrichum gloeosporioides, which causes mango anthracnose. The results demonstrated that α-MG had a relatively high in vitro inhibitory activity against C. gloeosporioides among 20 plant pathogenic fungi. The median effective concentration (EC50) values of α-MG against mycelial growth were nearly 10 times higher than those of spore germination inhibition for both strains of C. gloeosporioides, the carbendazim-sensitive (CBD-s) and carbendazim-resistant (CBD-r). The results suggested that α-MG exhibited a better inhibitory effect on spore germination than on the mycelial growth of C. gloeosporioides. Further investigation indicated that the protective effect could be superior to the therapeutic effect for mango leaves for scab development. The morphological observations of mycelium showed that α-MG caused the accumulation of dense bodies. Ultrastructural observation further revealed that α-MG caused a decrease in the quantity and shape of the swelling of mitochondria in the mycelium cells of C. gloeosporioides. In addition, bioassays disclosed that the inhibitory activity of α-MG on spore germination was reduced by adding exogenous adenosine triphosphate (ATP). These results suggested that the mode of action of α-MG could be involved in the destruction of mitochondrial energy metabolism. The current study supports α-MG as a natural antifungal agent in crop protection.

scholarly journals E-2-Hexenal Can Both Stimulate and Inhibit Botrytis Growth in Vitro and on Strawberry Fruit in Vivo

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 505E-506
Author(s):  
Elazar Fallik ◽  
Douglas D. Archbold ◽  
Thomas R. Hamilton-Kemp
Keyword(s):  
Volatile Compounds ◽  
Vapor Phase ◽  
Spore Germination ◽  
Mycelial Growth ◽  
Storage Period ◽  
Strawberry Fruit ◽  
High Concentration ◽  
Fungal Cell

Some plant-derived natural volatile compounds exhibit antifungal properties and may offer a tremendous opportunity to control the causes of postharvest spoilage without affecting fresh produce quality or leaving a residue on the produce. E-2-hexenal has shown significant potential for use as a fumigant for controlling Botrytis cinerea in prior studies. In in vitro studies on the mode of action of E-2-hexenal, mycelial growth and percent spore germination were inversely proportional to concentrations of the compound. Spore germination was found to be more susceptible to the compound then mycelial growth. Much higher concentrations of E-2-hexenal were required to inhibit mycelial growth than spore germination. Lower concentrations of the compound significantly stimulated mycelial growth, especially when the volatile was added 2 days following inoculation. Light microscopy analysis revealed that a high concentration of the volatile damaged fungal cell wall and membranes. Treatment with a high vapor phase level of E-2-hexenal during postharvest storage of strawberry fruit at 2°C prevented botrytis development in a subsequent storage period at 15°C. However, treatment with a low vapor phase level enhanced botrytis development. The implications of these results with respect to the practical use of E-2-hexenal and other natural volatile compounds will be discussed.

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