Characterization of Botrytis cinerea and B. prunorum from Healthy Floral Structures and Decayed ‘Hayward’ Kiwifruit During Post-Harvest Storage

Plant Disease ◽  
2020 ◽  
Author(s):  
Danae Riquelme ◽  
Zdenka Aravena ◽  
Hector Valdes ◽  
Bernardo Antonio Latorre ◽  
Gonzalo A Díaz ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Central Valley ◽  
Gray Mold ◽  
Heat Shock Protein 60 ◽  
Post Harvest ◽  
Diseased Fruit ◽  
Apparently Healthy

Gray mold is the primary post-harvest disease of ‘Hayward’ kiwifruit (Actinidia deliciosa) in Chile, with a prevalence of 33.1% in 2016 and 7.1% in 2017. Gray mold develops during postharvest storage, which is characterized by a soft, light to brown watery decay that is caused by Botrytis cinerea and B. prunorum. However, there is no information related to the role of B. prunorum during the development and storage of kiwifruit in Chile. For this purpose, asymptomatic flowers and receptacles were collected throughout fruit development and harvest from five orchards over two seasons in the Central Valley of Chile. Additionally, diseased kiwifruits were selected after storage for 100 days at 0°C plus 2 days at 20° C. High (HCP) and low conidial production (LCP) colonies of Botrytis sp. were consistently obtained from apparently healthy petals, sepals, receptacles, styles, and diseased kiwifruit. Morphological and phylogenetic analysis using three partial gene sequences encoding glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) were able to identify and separate B. cinerea and B. prunorum species. Consistently, B. cinerea was predominantly isolated from all floral parts and fruit in apparently healthy tissue and diseased kiwifruit. During full bloom, the highest colonization by B. cinerea and B. prunorum was obtained from petals followed by sepals. In storage, both Botrytis species were isolated from the diseased fruit (n=644), of which 6.8% (n=44) were identified as B. prunorum. All Botrytis isolates grew from 0°C to 30°C in vitro and were pathogenic on kiwifruit leaves and fruit. Notably, B. cinerea isolates were always more virulent than B. prunorum isolates. This study confirms the presence of B. cinerea and B. prunorum colonizing apparently healthy flowers and floral parts in fruit and causing gray mold during kiwifruit storage in Chile. Therefore, B. prunorum plays a secondary role in the epidemiology of gray mold developing in kiwifruit during cold storage.

scholarly journals Characterization of Postharvest Fungicide-Resistant Botrytis cinerea Isolates From Commercially Stored Apple Fruit

2017 ◽  
Vol 107 (3) ◽  
pp. 362-368 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Eunhee Park ◽  
Jiujiang Yu ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
Apple Fruit ◽  
Sublethal Dose ◽  
Long Term Storage ◽  
Pose Control ◽  
First Time

Botrytis cinerea causes gray mold and is an economically important postharvest pathogen of fruit, vegetables, and ornamentals. Fludioxonil-sensitive B. cinerea isolates were collected in 2011 and 2013 from commercial storage in Pennsylvania. Eight isolates had values for effective concentrations for inhibiting 50% of mycelial growth of 0.0004 to 0.0038 μg/ml for fludioxonil and were dual resistant to pyrimethanil and thiabendazole. Resistance was generated in vitro, following exposure to a sublethal dose of fludioxonil, in seven of eight dual-resistant B. cinerea isolates. Three vigorously growing B. cinerea isolates with multiresistance to postharvest fungicides were further characterized and found to be osmosensitive and retained resistance in the absence of selection pressure. A representative multiresistant B. cinerea strain caused decay on apple fruit treated with postharvest fungicides, which confirmed the in vitro results. The R632I mutation in the Mrr1 gene, associated with fludioxonil resistance in B. cinerea, was not detected in multipostharvest fungicide-resistant B. cinerea isolates, suggesting that the fungus may be using additional mechanisms to mediate resistance. Results from this study show for the first time that B. cinerea with dual resistance to pyrimethanil and thiabendazole can also rapidly develop resistance to fludioxonil, which may pose control challenges in the packinghouse environment and during long-term storage.

scholarly journals Functional Analysis of the Cytochrome P450 Monooxygenase Gene bcbot1 of Botrytis cinerea Indicates That Botrydial Is a Strain-Specific Virulence Factor

2005 ◽  
Vol 18 (6) ◽  
pp. 602-612 ◽  
Author(s):  
Verena Siewers ◽  
Muriel Viaud ◽  
Daniel Jimenez-Teja ◽  
Isidro G. Collado ◽  
Christian Schulze Gronover ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Functional Characterization ◽  
Gray Mold ◽  
Infection Process ◽  
In Planta ◽  
P450 Monooxygenase ◽  
Α Subunit ◽  
Monooxygenase Gene

The micrographic phytopathogen Botrytis cinerea causes gray mold diseases in a large number of dicotyledonous crop plants and ornamentals. Colonization of host tissue is accompanied by rapid killing of plant cells ahead of the growing hyphen, probably caused by secretion of nonspecific phytotoxins, e.g., the sesquiterpene botrydial. Although all pathogenic strains tested so far had been shown to secrete botrydial and although the toxin causes comparable necrotic lesions as infection by the fungus, the role of botrydial in the infection process has not been elucidated so far. Here, we describe the functional characterization of bcbot1, encoding a P450 monooxygenase and provide evidence that it is involved in the botrydial pathway, i.e., it represents the first botrydial biosynthetic gene identified. We show that bcbot1 is expressed in planta and that expression in vitro and in planta is controlled by an α-subunit of a heterotrimeric GTP-binding protein, BCG1. Deletion of bcbot1 in three standard strains of B. cinerea shows that the effect on virulence (on several host plants) is strain-dependent; only deletion in one of the strains (T4) led to reduced virulence.

scholarly journals First Report of Post-Harvest Gray Mold on Hyacinth bean (Lablab purpureus) Caused by Botrytis cinerea in Korea

Plant Disease ◽  
2021 ◽  
Author(s):  
Md Aktaruzzaman ◽  
Tania Afroz ◽  
Sung Kee Hong ◽  
Byung Sup Kim ◽  
Hyo-Won Choi
Keyword(s):  
Botrytis Cinerea ◽  
Petri Dish ◽  
Gray Mold ◽  
Distilled Water ◽  
Conidial Suspension ◽  
Heat Shock Protein 60 ◽  
Lablab Purpureus ◽  
First Report ◽  
Post Harvest ◽  
Hyacinth Bean

Hyacinth bean (Lablab purpureus L.) is a highly proteineous legume under the family Fabaceae. It is native to Africa, cultivated throughout the world, and recently introduced vegetable in Korea. In April 2020, approximately 10 to 15% of the total harvested pods showed gray mold rot symptoms after 3–5 days of storage at 4 °C in Jeonju, Jeonbuk province, Korea. The symptoms observed were irregular, water-soaked spots become brown or gray with white hyphae were appeared on the infected pods. Diseased tissue was excised, and surface sterilized by immersing in 1% sodium hypochlorite (NaOCl) for 1 min, rinsed three times with sterilized distilled water, placed on potato dextrose agar (PDA) plates, and incubated at 20 ± 2°C for 7 days. A total of five morphologically similar fungal isolates (HBGM001 to HBGM005) were obtained from diseased samples; isolate HBGM002 and HBGM005 were selected for identification. The fungus produced initially white colonies, after 7 days it changes to gray to dark colonies with dark mycelium that sporulated abundantly on PDA at 20ºC. The conidia (n = 50) were single-celled, ellipsoid or ovoid in shape, and 6.11 to 13.9 × 4.8 to 9.4 μm in size for HBGM001 isolate and 5.81 to 14.1× 4.5 to 9.6 μm in size for HBGM005. Conidiophores (n = 15) arose solitary or in groups, straight or flexuous, septate, with an inflated basal cell brown to light brown, and measured 103 to 420× 7 to 25 μm for HBGM001 isolate and 101 to 415 × 5 to 23 μm for HBGM005 isolate. After two weeks, the fungus formed several black sclerotia (n = 20) ranging from 0.5 to 4.2 × 0.5 to 3.4 mm for HBGM001 isolate and 0.4 to 4.4 × 0.3 to 3.3 mm for HBGM005 isolate near the edge of the Petri dish. Morphological characters were consistent with those of Botrytis cinerea Pers.: Fr. (Ellis 1971). As for molecular identification, the internal transcribed spacer (ITS) and three nuclear protein-coding genes (glyceraldehydes-3-phosphate dehydrogenase gene [G3PDH], heat-shock protein 60 gene [HSP60], and DNA-dependent RNA polymerase subunit gene [RPB2]) were amplified using primer pairs ITS1/ITS4 (White et al. 1990), G3PDH-F/G3PDH-R, HSP60-F/HSP60-R, and RPB2-F/RPB2-R (Staats et al. 2005), respectively. The ITS, G3PDH, HSP60, and RPB2 sequences of HBGM002 and HBGM005 isolates (GenBank accession number MT439648 and MT968495 for ITS; MT439649 and MT968496 for G3PDH; MT439650 and MT968497 for HSP60; MT439651 and MT968498 for RPB2 respectively) were 99% to 100% identical to those of B. cinerea (KY364366, KF015583, KJ018758, and KJ018756, respectively). To determine pathogenicity, five disinfected pods were pinpricked (3 sites per pod) with sterile needles and 50 µl of conidial suspension (1 × 105 conidia/ml) was inoculated by pipetting into the wounds. An analogous five pods, serving as controls, were inoculated with sterile distilled water. All the pods were placed in a growth chamber and maintained a temperature of 20±2ºC and a relative humidity >80%. After 5 days, gray mold symptoms developed on the inoculated pods, whereas no symptoms appeared on control pods. The pathogen was re-isolated from the inoculated pods, fulfilling Koch’s postulates. B. cinerea has been reported causing gray mold in Hyacinth bean in China, Taiwan and India (Farr and Rossman 2021). To our knowledge, this is the first report of B. cinerea causing post-harvest gray mold on hyacinth bean in Korea. The disease could represent a threat for hyacinth bean post-harvest and storage and management strategies should be investigated and applied.

scholarly journals Potencial de extratos etanólicos de propólis e extratos aquosos de plantas espontâneas no controle de doenças pós-colheita do morango

2016 ◽  
Vol 11 (1) ◽  
pp. 57
Author(s):  
Gabriela Silva Moura ◽  
Jonas Marcelo Jaski ◽  
Gilmar Franzener
Keyword(s):  
Antifungal Activity ◽  
Botrytis Cinerea ◽  
Gray Mold ◽  
Alternative Methods ◽  
Wild Plants ◽  
Rhizopus Nigricans ◽  
Post Harvest ◽  
Control Water

<p>A cultura do morangueiro é severamente acometida por várias doenças, dentre elas o mofo cinzento, causado por <em>Botrytis cinerea</em> é considerada a doença mais severa na pós-colheita. Visando reduzir o uso de fungicidas sintéticos, vem sendo realizadas pesquisas propondo a utilização de métodos alternativos de controle de patógenos pós-colheita envolvendo a utilização de extratos vegetais, uso de biofungicidas e óleos essenciais. Assim, o presente trabalho teve como objetivo avaliar o potencial de diferentes extratos de própolis e plantas espontâneas no controle de podridão pós-colheita causada pelo fungo <em>B. cinerea</em> em morangos. Para avaliar a atividade antifúngica direta dos extratos etanólico de própolis e extratos aquosos de plantas espontâneas sobre <em>B. cinerea,</em> foi realizado o experimento <em>in vitro</em>, utilizando-se os tratamentos própolis verde 0,5%; própolis verde 2,5%; própolis marrom 0,5%; própolis marrom 2,5%; língua-de-vaca 10%; assa-peixe 10%; rubim 10%; tansagem 10%; testemunha (água). As medições do diâmetro das colônias foram iniciadas 48, 72 e 96 horas após a instalação do experimento. No experimento <em>in vivo </em>os frutos foram imersos nos tratamentos descritos acima. Após cinco dias avaliou-se a incidência e severidade da doença mofo cinzento e das doenças pós-colheita como antracnose e podridão de Rhizopus que apareceram no experimento. Utilizou-se o delineamento experimental inteiramente casualizado (DIC) com quatro e cinco repetições para o ensaio <em>in vitro</em> e <em>in vivo,</em> respectivamente<em>.</em> Os resultados mostram que os extratos etanólicos de própolis verde e marrom a 2,5% apresentaram <em>in vitro</em> e <em>in vivo </em>atividade antifúngica a <em>B. cinerea</em> e <em>Rhizopus nigricans,</em> respectivamente.</p><p align="center"><strong><em>Potential of propolis extracts and extracts etanol spontaneous plants aqueous in control of diseases of strawberry post-harvest</em></strong><strong><em></em></strong></p><p><strong>Abstract</strong><strong>: </strong>The strawberry crop is severely affected by various diseases, including gray mold, caused by <em>Botrytis cinerea</em> is considered the most severe disease in post-harvest fruit. To reduce the use of synthetic fungicides, has been carried out research proposing the use of alternative methods of control postharvest pathogens involving the use of plant extracts, use of biofungicides, essential oils among others. Thus, this study aimed to evaluate the potential of different propolis extracts and wild plants in the control of post-harvest rot caused by the fungus <em>Botrytis cinerea</em> in strawberries. To evaluate the direct antifungal activity of ethanolic extracts of propolis and aqueous extracts of wild plants of B. cinerea, the in vitro experiment was performed, using treatments propolis 0.5%; propolis 2.5%; brown propolis 0.5%; brown propolis 2.5%; control (water + alcohol 2%); cow tongue 10%; assa-fish 10%; rubim 10%; tansagem 10%; control (water). Measurements of the diameter of the colonies were started 48, 72 and 96 hours after installation of the experiment. Conducted the in vivo experiment in which the fruits of strawberry plants were immersed in the treatments described above. After five days we evaluated the incidence and severity of gray mold disease and post-harvest diseases such as anthracnose and Rhizopus rot appearing in the experiment. We used a completely randomized design (CRD) with four and five replicates for the in vitro assay and in vivo, respectively. The results show that ethanol extracts of green and brown propolis 2.5% presented in vitro and in vivo antifungal activity to <em>B. cinerea</em> and <em>Rhizopus nigricans</em> respectively.</p>

scholarly journals Molecular and Biochemical Characterization of Laboratory and Field Mutants of Botrytis cinerea Resistant to Fludioxonil

Plant Disease ◽  
2016 ◽  
Vol 100 (7) ◽  
pp. 1414-1423 ◽  
Author(s):  
Weichao Ren ◽  
Wenyong Shao ◽  
Xu Han ◽  
Mingguo Zhou ◽  
Changjun Chen
Keyword(s):  
Botrytis Cinerea ◽  
Biochemical Characterization ◽  
Resistance Mechanism ◽  
Gray Mold ◽  
Terminal Region ◽  
Excellent Activity ◽  
Resistant Mutants ◽  
First Time

Botrytis cinerea is a filamentous phytopathogen with a high risk of developing resistance to fungicides. The phenylpyrrole fungicide fludioxonil has been reported to have excellent activity against B. cinerea and increasingly has been applied to control gray mold in China. In this study, molecular and biochemical characteristics of laboratory and field mutants of B. cinerea resistant to fludioxonil has been investigated. During 2012 to 2014, B. cinerea isolates collected from Jiangsu and Shandong Provinces in China were tested in vitro for sensitivity to fungicides commonly used to suppress gray mold of cucumber and tomato. Among the 75 isolates collected from cucumber in 2013, two were highly resistant (HR) to fludioxonil. Of the 308 isolates collected from tomato in 2014, four were fludioxonil-HR. This was the first time that B. cinerea isolates HR to fludioxonil had been detected in the field. Six fludioxonil-resistant mutants were obtained in the laboratory by selection on fungicide-amended media. These mutants exhibited stable resistance to fludioxonil, as indicated by resistance factor values that ranged from 34.38 to >10,000. In comparison with fludioxonil-sensitive isolates of B. cinerea, all field and laboratory mutants showed reduced fitness, as defined by mycelial growth, sporulation, virulence, and sensitivity to osmotic stress. When treated with fludioxonil at 1 μg/ml, sensitive isolates showed increased glycerol contents in mycelium and expression levels of Bchog1, while levels in field and laboratory HR mutants increased only slightly. Sequences of the Bos1 gene of field and laboratory fludioxonil-HR mutants showed that mutations in field mutants were located in the histidine kinase, adenylyl cyclase, methyl-accepting chemotaxis protein, and phosphatase (HAMP) domains of the N-terminal region, whereas mutations in the laboratory mutants were distributed in HAMP domains or in the HATPase_c domain of the C-terminal region. These results will enhance our understanding of the resistance mechanism of B. cinerea to fludioxonil.

scholarly journals Characterization of a Dye-Decolorizing Peroxidase from Irpex lacteus Expressed in Escherichia coli: An Enzyme with Wide Substrate Specificity Able to Transform Lignosulfonates

2021 ◽  
Vol 7 (5) ◽  
pp. 325
Author(s):  
Laura Isabel de de Eugenio ◽  
Rosa Peces-Pérez ◽  
Dolores Linde ◽  
Alicia Prieto ◽  
Jorge Barriuso ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Inclusion Bodies ◽  
Veratryl Alcohol ◽  
Dye Decolorization ◽  
Irpex Lacteus ◽  
Type D ◽  
Lignin Peroxidases

A dye-decolorizing peroxidase (DyP) from Irpex lacteus was cloned and heterologously expressed as inclusion bodies in Escherichia coli. The protein was purified in one chromatographic step after its in vitro activation. It was active on ABTS, 2,6-dimethoxyphenol (DMP), and anthraquinoid and azo dyes as reported for other fungal DyPs, but it was also able to oxidize Mn2+ (as manganese peroxidases and versatile peroxidases) and veratryl alcohol (VA) (as lignin peroxidases and versatile peroxidases). This corroborated that I. lacteus DyPs are the only enzymes able to oxidize high redox potential dyes, VA and Mn+2. Phylogenetic analysis grouped this enzyme with other type D-DyPs from basidiomycetes. In addition to its interest for dye decolorization, the results of the transformation of softwood and hardwood lignosulfonates suggest a putative biological role of this enzyme in the degradation of phenolic lignin.

scholarly journals Detrimental Role of Heat Shock Protein 60&70 and Toll-like Receptor 4 in Skeletal Muscle Ischaemia In Vitro

2013 ◽  
Vol 57 (5) ◽  
pp. 77S
Author(s):  
Ali Navi ◽  
Rebekah Yu ◽  
Xu Shi-Wen ◽  
Sidney Shaw ◽  
George Hamilton ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Heat Shock Protein 60 ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Muscle Ischaemia

scholarly journals Identification and characterization of molecular interactions between mortalin/mtHsp70 and HSP60

2005 ◽  
Vol 391 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Renu Wadhwa ◽  
Syuichi Takano ◽  
Kamaljit Kaur ◽  
Satoshi Aida ◽  
Tomoko Yaguchi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Molecular Interactions ◽  
Heat Shock Protein 60 ◽  
Hsp60 Expression ◽  
Mitochondrial Hsp70 ◽  
Shock Proteins ◽  
First Time

Mortalin/mtHsp70 (mitochondrial Hsp70) and HSP60 (heat-shock protein 60) are heat-shock proteins that reside in multiple subcellular compartments, with mitochondria being the predominant one. In the present study, we demonstrate that the two proteins interact both in vivo and in vitro, and that the N-terminal region of mortalin is involved in these interactions. Suppression of HSP60 expression by shRNA (short hairpin RNA) plasmids caused the growth arrest of cancer cells similar to that obtained by suppression of mortalin expression by ribozymes. An overexpression of mortalin, but not of HSP60, extended the in vitro lifespan of normal fibroblasts (TIG-1). Taken together, this study for the first time delineates: (i) molecular interactions of HSP60 with mortalin; (ii) their co- and exclusive localizations in vivo; (iii) their involvement in tumorigenesis; and (iv) their functional distinction in pathways involved in senescence.

scholarly journals AVALIAÇÃO DO REAL POTENCIAL INIBIDOR DE EXTRATOS ETANÓLICOS DE Ottonia martiana SOBRE Cylindrocladium spathulatum E Botrytis cinerea

FLORESTA ◽  
2013 ◽  
Vol 43 (2) ◽  
pp. 225
Author(s):  
Miriam Machado Cunico ◽  
Celso Garcia Auer ◽  
Marlon Wesley Machado Cunico ◽  
Obdulio Gomes Miguel ◽  
Patricio Peralta Zamora ◽  
...  
Keyword(s):  
Factorial Design ◽  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Black Spot ◽  
Gray Mold ◽  
Causal Agent ◽  
In Vitro Testing ◽  
Ethanolic Extracts ◽  
Antifungal Potential

 Extratos etanólicos de anestesia, Ottonia martiana Miq., foram reavaliados quanto à inibição do crescimento micelial dos fungos Cylindrocladium spathulatum (pinta-preta da erva-mate) e Botrytis cinerea (mofo-cinzento do eucalipto), por meio do planejamento fatorial. A ocorrência de decomposição de bioativos no processo de autoclavagem também foi investigada, por meio de teste de eficiência de extratos filtrados (filtro Millipore) e esterilizados (autoclave) no controle dos fitopatógenos, nas concentrações de 1, 10, 100 e 1000 ppm. Os extratos etanólicos filtrado e esterilizado inibiram o crescimento micelial dos fungos e foram mais ativos frente a B. cinerea.O extrato filtrado exibiu maior potencial antifúngico que o extrato esterilizado. O processo de esterilização por autoclavagem causou pequena decomposição dos bioativos presentes no extrato de anestesia.Palavras-chave: Anestesia; mofo-cinzento; pinta-preta. Abstract Fungitoxic potential of ethanolic extracts of anestesia in the control of phytopathogenic diseases. The antifungal potential of anestesia, Ottonia martiana Miq. was reassessed by factorial design, in vitro testing of fungal mycelial growth compared to the pathogenic isolates Cylindrocladium spathulatum, causal agent of black spot onyerba mate, and Botrytis cinerea causal agent of gray-mold on eucalypts. Occurrence of decomposition of bioactive of the autoclaving process was investigated using foliar detached test compared to the pathogens (1000 ppm). Ethanolic extracts - EBEtOH (filtered and autoclaved) inhibited the mycelial growth of C. spathulatum and B. cinerea (1000 ppm) and were more pronounced against B. cinerea (43.6 % and 68.9 %). EBEtOH filtered (0.22 µm) presented higher activity than EBEtOH autoclaved (C. spathulatum: 52.8 % and 43.6 %, B. cinerea: 68.9 % and 43.6 %), suggesting little decomposition ofbioactive after autoclaving. EBEtOH filtrate presented potential inhibition of 28 % in eucalypt leaves against B. cinerea.  Keywords: Ottonia martiana; black spot; gray-mold.

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