scholarly journals In vitro Control of the Fungus Botrytis cinerea Pers. with Plant Extracts

Author(s):  
Raluca MICLEA ◽  
Carmen PUIA

The gray mould, caused by Botrytis cinerea Pers., is an economically important disease of plants throughout the world. It is common on vegetable crops, soft fruits, ornamental plants and grapevine, and occurs in greenhouses and in the field as well as in storage and transport. Plant extracts and their purified compounds have antibacterial, antiviral and fungicidal effects both in vitro and in vivo. Their properties were attributed mainly to alkaloids, several flavonoids and phenolic acids. This study was made in order to establish the materials and the methods most used for the in vitro control of the fungus Botrytis cinerea Pers. with plant extracts – types of plant extracts, the most appropriate culture media, methods of inoculation, methods of extraction and methods for testing the antifungal effect of the plant extracts against the phytopathogenic fungus. The most used medium was PDA (potato dextrose agar). Conform to the literature data this medium is the most suitable for the development of this fungus, the growth of the mycelium, the conidia growing and the formation of sclerotia. As plant extraction method, the hydrodistilation is the most ancient method of distillation and the most versatile. Reviewed methods used for testing the efficacy of natural products (plant extracts) were: the agar diffusion method, the poisoned food technique, the radial growth test, the conidial germination assay, the microathmosphere method, the technique of Thompson.

Plant Disease ◽  
1998 ◽  
Vol 82 (2) ◽  
pp. 165-170 ◽  
Author(s):  
J. F. Fourie ◽  
G. Holz

Sugars in exudates from Harry Pickstone plum and Sunlite nectarine fruit and from pollen of weeds commonly found in orchards were determined by gas-liquid chromatography, and their effect on the development of Botrytis cinerea was determined in vitro and in vivo. Fructose, glucose, and sorbitol were the only sugars detected in exudates of immature fruit. They occurred at low concentrations, but their concentration generally increased as fruit ripened. Sucrose was first detected during maturation. In nectarine, an increase in sugar concentration, especially sucrose, was pronounced during the period of rapid cell enlargement, which occurred approximately 2 weeks before harvest. Absorbance readings of culture media amended with sugar indicated that the hexose sugars (fructose and glucose) and sucrose did not markedly influence growth of B. cinerea at concentrations below 0.22 and 0.12 mM, respectively. The hexose sugars caused a steady increase in growth when supplied at concentrations in excess of 0.44 mM, and sucrose caused a steady increase in growth at 0.23 mM. The stimulatory effect of fruit exudates on growth of B. cinerea on glass slides coincided with the period of rapid sugar release from the fruit and the shift in susceptibility to decay. Only fructose (1.72 mM) and glucose (0.72 mM) were detected in nectarine pollen exudates. Pollen exudates from weeds stimulated fungal growth and significantly increased the aggressiveness of the pathogen on plum and nectarine fruit when added to conidia during the last 4 weeks prior to the picking-ripe stage. The study showed that changes in the composition of nectarine and plum fruit exudates may contribute to the late-season susceptibility of these fruit to B. cinerea infection.


2018 ◽  
pp. 31-38

Actividad antibacteriana de extractos vegetales frente a cepas intrahospitalarias, Iquitos-Perú Antibacterial activity of plant extracts against nosocomial strains, Iquitos-Peru Ricardo E. Abadie, Ronald Medina O., Lastenia Ruiz, Alvaro Tresierra-Ayala Laboratorio de Microbiología. Centro de Investigación de Recursos Naturales de la Universidad Nacional de la Amazonía Peruana (CIRNA-UNAP). Psje. Los Paujiles S/N, San Lorenzo, distrito de San Juan Bautista, Iquitos-Perú DOI: https://doi.org/10.33017/RevECIPeru2014.0005/  Resumen La región amazónica es una de las áreas que posee la mayor biodiversidad del mundo, albergando varios miles de especies de plantas, muchas de las cuales son utilizadas por sus pobladores como plantas medicinales. Durante los últimos años, el empleo de estos recursos vegetales o de sus productos viene incrementándose de manera importante, lo cual podría deberse a una serie de factores, entre los que destacan el conocimiento de su composición química, y al hecho que en la actualidad se han realizado numerosos ensayos farmacológicos tanto in vivo como in vitro. La aparición de cepas resistentes a los antibióticos comerciales en los últimos tiempos, está creando la necesidad de buscar otras estrategias o alternativas para controlarlas, tal es el caso del uso de las plantas (medicina tradicional), debido a los principios activos que poseen. Se pretende con este trabajo, determinar probables alternativas para combatir infecciones bacterianas de aquellos agentes drogoresistentes, este problema reviste importancia crítica particular en los países en desarrollo, donde quizás no se dispone de antibióticos de segunda línea más costosos o, si los hay, su precio es inasequible. El estudio se realizó en la ciudad de Iquitos, Provincia de Maynas, Departamento de Loreto. Los ensayos microbiológicos se realizaron en el Laboratorio de Microbiología del Centro de Investigación de Recursos Naturales de la Amazonia (CIRNA) de la Universidad Nacional de la Amazonia  Peruana (UNAP). Se determinó la actividad antibacteriana de 6 extractos vegetales (Alchornea triplinervia, Annona muricata, Averrhoa carambola, Brunfelsia grandiflora, Caraipa grandifolia y Cedrela odorata) mediante la técnica de difusión en disco, y a aquellos que presentaron actividad se les determinó la Concentración Inhibitoria Mínima y la Concentración Bactericida Mínima mediante la técnica de macrodilución en caldo. Ninguno de los extractos tuvieron actividad frente a las cepas de E. coli; cuatro extractos tuvieron actividad frente a las cepas de P. aeruginosa, siendo los extractos de Cedrela odorata y Alchornea triplinervia los que tuvieron mayor actividad frente a esta bacteria, con CIM = 15.62 y 62.5 mg/ml, respectivamente; todos los extractos tuvieron actividad frente a las cepas de S. aureus, siendo los extracto de C. odorata, A. triplinervia y Caraipa grandiflora, los de mayor actividad con una CIM = 3.91 mg/ml para cada uno.  Se obtuvieron prometedores resultados de actividad antibacteriana de los extractos en estudio frente a cepas intrahospitalarias, mayormente contra S. aureus. Descriptores: Actividad antibacteriana, extractos vegetales, cepas intrahospitalarias Abstract The Amazon region is one of the areas with the largest biodiversity in the world, hosting several thousand species of plants, many of which are used by its people as medicinal plants. In recent years, the use of these plant resources or products has been increasing significantly, which could be due to a number of factors, among them the knowledge of their chemical composition, and the fact that at present there have been numerous pharmacological tests both in vivo and in vitro. The emergence of antibiotic-resistant strains in recent years, is creating a need for other strategies or ways to control them, as in the case of the use of plants (traditional medicine), because the active ingredients bearing. This work is intended to determine probable alternatives to combat bacterial infections of those agents antibiotic-resistant, this problem is particularly critical in developing countries, where perhaps there are no antibiotics or expensive second line, if any, price is unavailable. The study was conducted in Iquitos city, Province of Maynas, Department of Loreto. Microbiological tests were performed at Microbiology Laboratory of Research Center of Natural Resources of the Amazon (CIRNA) of the National University of the Peruvian Amazon (UNAP). The antibacterial activity of six plant extracts (Alchornea triplinervia, Annona muricata, Averrhoa carambola, Brunfelsia grandiflora, Caraipa grandifolia y Cedrela odorata) by the disk diffusion method was determined, and those that showed activity were determined Minimum Inhibitory Concentration and Minimum Bactericidal Concentration by macrodilution technique. None of the extracts were active against strains of E. coli; four extracts had activity against strains of P. aeruginosa, with Cedrela odorata and Alchornea triplinervia extracts which had greater activity against these bacteria, with MIC = 15.62 and 62.5 mg/ml, respectively; all extracts were active against strains of S. aureus, with the extract of C. odorata, A. triplinervia and Caraipa grandiflora, the most active with an MIC = 3.91 mg/ml for each. Was obtained Promising results of antibacterial activity of the extracts in study against nosocomial strains, mostly against S. aureus. Keywords: Antibacterial activity, plant extracts, nosocomial strains


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 ◽  
...  

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.


1997 ◽  
Vol 43 (9) ◽  
pp. 835-840 ◽  
Author(s):  
Yael Meller-Harel ◽  
Anat Argaman ◽  
Yair Aharonowitz ◽  
David Gutnick ◽  
Dafna Ben-Bashat ◽  
...  

Polygalacturonase activity from the phytopathogenic fungus Botrytis cinerea was inhibited in vitro by extracellular polyphosphate from Streptomyces sp. A50, as well as other polyphosphates of biological and chemical origin. The extent of inhibition increased with polyphosphate chain length between 20 and 100 Pi residues. Although the activity of polygalacturonase from B. cinerea appeared not to depend on the presence of cations, inhibition was partially blocked by divalent cations such as Mg2+or Ca2+. Production of polyphosphate in Streptomyces sp. A50 was followed by chemical measurements, as well as by in vivo 31P-NMR analysis. During the first 2 days of growth, polyphosphate accumulated within the cells, after which it appeared in the broth as an extracellular product. A maximum concentration of extracellular polyphosphate (1 mM Pi equivalent) was reached, corresponding to about 25% of the input Pi. NMR analysis suggested that the intracellular form of polyphosphate exists as a mobile soluble pool. In contrast, the extracellular form of polyphosphate appears to be complexed with cations.Key words: polygalacturonase, polyphosphate, Botrytis cinerea, Streptomyces, 31P-NMR.


2020 ◽  
Vol 73 (2) ◽  
Author(s):  
Hala Abdel Wahab ◽  
Ahmed Malek ◽  
Mohamed Ghobara

<em>Botrytis cinerea</em> and <em>Sclerotinia sclerotiorum</em> are necrotrophic fungi and are closely related pathogenic fungi that infect hundreds of plant species worldwide. In this study, the natural botryticidal/scleroticidal efficacy of some plant extracts, bioagents, and organic compounds known to possess antifungal activity was evaluated. Pathogenicity tests of the fungal molds showed virulence divergence, depending on the isolate and host plant. All <em>B. cinerea</em> isolates, except the BF isolate that infected only broad bean leaves, demonstrated ability to infect detached lettuce and broad bean leaves. Moreover, all <em>Sclerotinia sclerotiorum</em> isolates, except for SSP, demonstrated ability to infect the two plant species, whereas the SSB isolate did not cause any infection in broad bean leaves. The efficacies of <em>Moringa oleifera</em> (Mor), <em>Cinnamomum zeylanicum</em> (Cin), amino acid derivatives (Aad), <em>Trichoderma harzianum</em> (TH), <em>Cactus</em> spp. (Agr), and <em>Bacillus subtilis</em> (BS) were tested either in vitro or in vivo against the highly virulent isolates of the two pathogenic fungi. The efficacy of most potential biofungicides was consistent in vitro as well as in vivo, and the inhibitory efficacy of TH, BS, Cin, Mor, and Aad treatments was significantly high against <em>Botrytis cinerea</em> and <em>Sclerotinia sclerotiorum</em> in vitro and ranged from 62% to 100%, depending on the isolate. In addition, BS, Aad, TH, and Mor treatments had significant inhibitory effects ranging from 53% to 100% against most of the isolates on lettuce leaves. The Agr and Cin treatments exhibited low or no inhibitory effects against many isolates in vivo, and they reduced the mold infection caused by only BCC and SSB isolates. Most of the tested potential biofungicide treatments tended to reduce mold infections, and some of them, such as Cin, exhibited a higher inhibitory effect in vitro than the others. Real-time PCR was conducted for some symptomatic/asymptomatic samples, and the results showed either consistent molecular/symptomatic patterns or latency of <em>B. cinerea</em>. The results confirmed the suitability of the studied natural compounds as effective biofungicides, and they could be the best choice to safely control the most destructive fungal molds.


2008 ◽  
Vol 98 (4) ◽  
pp. 443-450 ◽  
Author(s):  
G. A. Bardas ◽  
C. K. Myresiotis ◽  
G. S. Karaoglanidis

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.


Biology ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 492
Author(s):  
Hernando José Bolivar-Anillo ◽  
Victoria E. González-Rodríguez ◽  
Jesús M. Cantoral ◽  
Darío García-Sánchez ◽  
Isidro G. Collado ◽  
...  

Plant diseases are one of the main factors responsible for food loss in the world, and 20–40% of such loss is caused by pathogenic infections. Botrytis cinerea is the most widely studied necrotrophic phytopathogenic fungus. It is responsible for incalculable economic losses due to the large number of host plants affected. Today, B. cinerea is controlled mainly by synthetic fungicides whose frequent application increases risk of resistance, thus making them unsustainable in terms of the environment and human health. In the search for new alternatives for the biocontrol of this pathogen, the use of endophytic microorganisms and their metabolites has gained momentum in recent years. In this work, we isolated endophytic bacteria from Zea mays cultivated in Colombia. Several strains of Bacillus subtilis, isolated and characterized in this work, exhibited growth inhibition against B. cinerea of more than 40% in in vitro cultures. These strains were characterized by studying several of their biochemical properties, such as production of lipopeptides, potassium solubilization, proteolytic and amylolytic capacity, production of siderophores, biofilm assays, and so on. We also analyzed: (i) its capacity to promote maize growth (Zea mays) in vivo, and (ii) its capacity to biocontrol B. cinerea during in vivo infection in plants (Phaseolus vulgaris).


2020 ◽  
Vol 55 (1) ◽  
pp. 27-34
Author(s):  
G. Zadehdabagh ◽  
K. Karimi ◽  
M. Rezabaigi ◽  
F. Ajamgard

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.


Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1503
Author(s):  
Carla Guijarro-Real ◽  
Mariola Plazas ◽  
Adrián Rodríguez-Burruezo ◽  
Jaime Prohens ◽  
Ana Fita

Antiviral treatments inhibiting Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication may represent a strategy complementary to vaccination to fight the ongoing Coronavirus disease 19 (COVID-19) pandemic. Molecules or extracts inhibiting the SARS-CoV-2 chymotripsin-like protease (3CLPro) could contribute to reducing or suppressing SARS-CoV-2 replication. Using a targeted approach, we identified 17 plant products that are included in current and traditional cuisines as promising inhibitors of SARS-CoV-2 3CLPro activity. Methanolic extracts were evaluated in vitro for inhibition of SARS-CoV-2 3CLPro activity using a quenched fluorescence resonance energy transfer (FRET) assay. Extracts from turmeric (Curcuma longa) rhizomes, mustard (Brassica nigra) seeds, and wall rocket (Diplotaxis erucoides subsp. erucoides) at 500 µg mL−1 displayed significant inhibition of the 3CLPro activity, resulting in residual protease activities of 0.0%, 9.4%, and 14.9%, respectively. Using different extract concentrations, an IC50 value of 15.74 µg mL−1 was calculated for turmeric extract. Commercial curcumin inhibited the 3CLPro activity, but did not fully account for the inhibitory effect of turmeric rhizomes extracts, suggesting that other components of the turmeric extract must also play a main role in inhibiting the 3CLPro activity. Sinigrin, a major glucosinolate present in mustard seeds and wall rocket, did not have relevant 3CLPro inhibitory activity; however, its hydrolysis product allyl isothiocyanate had an IC50 value of 41.43 µg mL−1. The current study identifies plant extracts and molecules that can be of interest in the search for treatments against COVID-19, acting as a basis for future chemical, in vivo, and clinical trials.


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