Quality assurance of postharvest grapes against Botrytis cinerea by terbinafine

Worldwide,fruit is an indispensable treasure house of nutrition for human beings,occupying a vital position of human diet.Postharvest fruit storage requires efficient antifungal agents to control Botrytis cinerea,which is a vital postharvest disease affecting fruit and leading to enormous losses.However,with the enormous abuse of existing antifungal drugs,the problem of drug-resistant fungi is imminent,making the controlling diseases caused by pathogenic fungi even more challenging.Drug repurposing is an efficient alternative method,we evaluated a well-known antifungal chemical,terbinafine,against the agricultural pathogen,B.cinerea in vitro,as a result,terbinafine showed strong anti-fungal activity.Furthermore,the in vivo antifungal activity of terbinafine was evaluated,the results showed that terbinafine could reduce the decay area on grapes.Terbinafine could disrupt the cell membrane integrity,increase cell membrane permeability,and eventual cell death of B.cinerea.In addition,terbinafine reduced decay incidence,and weight loss and maintained the soluble solids,titratable acidity,ascorbic acid,total phenolic,and malondialdehyde content during the storage period of grapes.Overall,terbinafine could be an antifungal preservative for postharvest table grapes fresh-keeping.

Identification of Resistance to Pathogenesis Related Protein GmPR1L in Tobacco Botrytis cinerea Infection

Soybean(Glycine max(Linn.)Merr.)annual leguminous crop is cultivated all over the world.The occurrence of diseases has a great impact on the yield and quality of soybean.In this study,based on the RNA-seq of soybean variety M18,a complete CDS(Coding sequence)GmPR1L of the pathogenesis-related protein 1 family was obtained,which has the ability to resist fungal diseases.The overexpression vector and interference expression vector were transferred into tobacco NC89,and the resistance of transgenic tobacco(Nicotiana tabacum L.)to Botrytis cinerea infection was identified.The results show that:Compared with the control,the activities of related defense enzymes SOD(Superoxide dismutase),POD(Peroxidase),PAL(L-phenylalanine ammonia-lyase)and PPO(Polyphenol oxidase)in the over-expressed transgenic tobacco OEA1 and OEA2 increased to different degrees,and increased significantly at different infection time points.The activities of defense enzymes in the interfering strains IEA1 and IEA2 were significantly lower than those in the control strains.The results of resistance level identification showed that the disease spot rate of OEA1 was significantly lower than that of the control line,and the disease spot rate of OEA2 was significantly lower than that of the control line.The plaque rate of the interfering expression line IEA1-IEA2 was significantly higher than that of the control line.It is preliminarily believed that the process related protein GmPR1L can improve the resistance of tobacco to B.cinerea.

Baseline Sensitivity of Botrytis cinerea from Liaoning Province to Boscalid

[Objective] This study aimed to avoid and delay the generation of resis- tance in Botrytis cinerea to boscalid. [Method] The sensitivity of Botrytis cinerea to boscalid was tested by the mycelial growth rate method. The distribution of sensitivity variation of B. cinerea to boscalid in different regions was cleared, and the sen- sitivity baseline of B. cinerea from Liaoning Province to boscalid was established. [Result] The ECso values of B. cinerea strains from Liaoning Province ranged from 0.080 0 to 7.787 2μg/ml, and the highest ECho value was 97.34 times higher than the minimum. The average EC50 value （1.973μg/ml） of the 158 strains was treated as the baseline sensitivity of B. cinerea from Liaoning Province to boscalid. [Conclu- sion] It is essential to conduct a risk assessment of drug resistance in B. cinerea to boscalid.

Tomato Resistance to Botrytis cinerea Induced by Chitosan

[Objective] The aim was to explore the effect of tomato＇s resistance to Botrytis cinerea induced by exogenous Chitosan.[Method] The leaf spraying method was used to determine the induced resistance effect of Chitosan to tomato B.cinerea and the chlorophyll content,soluble protein,soluble sugar,proline influence,the activities of peroxidase（POD）,malondialdehyde（MDA）and the Proline content in leaves of tomato.[Result] The induced resistance of Chitosan reached 58.26% at 14th d after treatment;Compared with the B.cinerea stress group,the chlorophyll content in leaves of tomato seedlings of B.cinerea stress ＋ Chitosan group had increased by 34.63%,while that of soluble protein content,soluble sugar content,Pro content and POD activity was 5.30%,10.83%,16.21% and 16.88%,respectively（except for the MDA content,which was decreased by 16.54%）.[Conclusion] Exogenous Chitosan could improve photosynthetic efficiency and protect enzymes activity to improve the resistance of tomato seedlings to B.cinerea.

Inhibitory Effects of Original Nano-Cu_2O Drug and Nano-Cu_2O Suspension on Snake Melon Botrytis cinerea

The drug-containing culture medium method for the test of toxicity was adopted to compare inhibitive effects of original nano-Cu2O drug and nano-Cu2O suspension, and nano-Cu2O drug has better inhibitive effects on snake melon Botry- tis cinerea than original nano-Cu2O drug with the same mass concentration, and inhibitory effects are positively correlated with concentration. Correlation coefficients of the toxicity regression equation are 0.892 2 and 0.996 1, effective concentration EC50 of original nano-Cu2O drug and that of nano-Cu2O suspension are 3 948.9 and 167.9 mg/kg. Original nano-Cu2O drug has an inhibitive effect on snake melon Botrytis cinerea, but the inhibition of nano-Cu2O suspension is more obvious.

Effects of Chitosan and Organosilicon on Antioxidant Enzyme Activity of Tomato Seedlings Diseased with Botrytis cinerea

[Objective] The aim was to explore effects of chitosan and organosilicon on tomato resistance against Botrytis cinerea. [Method] With leaf spraying method adopted, four groups were set in the test, including control group, groups treated with Botrytis cinerea, with chitosan and Botrytis cinerea, and with chitosan, organosilicon, and Botrytis cinerea, in order to study on effects of chitosan and organosilicon on antioxidant enzyme activities of tomato. [Result] Antioxidant enzyme activities of tomato seedlings were improved in groups with chitosan, organosilicon, and Botrytis cinerea and with chitosan and Botrytis cinerea compared with the group treated with Botrytis cinerea. On the 6th d, activities of PAL, SOD, POD, PPO and CAT in groups with chitosan and Botrytis cinerea, and with chitosan, organosilicon, and Botrytis cinerea increased by 27.36%, 52.07%, 43.55%, 82.02% and 71.82%, and 18.91%, 30.22%, 57.14%, 38.09% and 53.64%, respectively. [Conclusion] Antioxidant enzyme activities of tomato seedlings could be improved by chitosan and organosilicon.

Antifungal Activity of Botanical Extracts against Botrytis cinerea and Alternaria solani

[Objective] The antifungal activity of the extracts from,Atractylodes macracephal Koidz and Pulsatilla chinensis Bunge Regel,against Botrytis cinerea and Alternaria solani were studied under the condition of laboratory,in order to develop and utilize these two plants.[Method] The mycelium growth rate test was applied to measure the antifungal activities of extracts against fungi.[Result] the extracts of all the two plants showed strong antifungal activity against the target pathogenic fungi,especially the antifungal activity of the extract from Pulsatilla chinensis Bunge Regel was stronger and more stable.The inhibition rate to the mycelium growth of Botrytis cinerea was 80.25%.At the same concentration,the extract from Atractylodes macracephal Koidz showed little inhibition to Botrytis cinerea and Alternaria solani.The petrolelum ether extract of Atractylodes macracephal Koidz showed stronger antifungal activities and the EC50 was 5.31 mg/ml,and the n-butanol extract of Pulsatilla chinensis Bunge Regel showed stronger antifungal activities and the EC50 was 2.93 mg/ml.[Conclusion] The extracts from Pulsatilla chinensis Bunge Regel showed the stronger antifungal activity against Botrytis cinerea and Alternaria solani.

Biological Characteristics of Ginseng Botrytis cinerea Pers

[ Objective ] The study systematically studied the biological characteristics of ginseng Botrytis cinerea Pers. [ Method ] The pure pathogenic fungus was isolated from ginseng B. cinerea collected in the field by tissue segregation and purification cultivation. Subsequently, using PDA medium plate culture method, the effect of various culture conditions on mycelium growth and sporulation of ginseng B. cinerea was detected. [ Result] The optimum temperature for mycelium growth and sporulation of B. cinerea was 25 ℃. The appropriate temperature for conidia germination ranged from 20 to 25 ℃. And the optimum pH value for mycelium growth and conidia germination was 6.0. The optimum carbon source was sucrose, followed by glucose and fructose. The optimum nitrogen source was peptone, fol- lowed by beef extract, yeast extract, alanine and ammonium nitrate. Among the media, the growth of mycelium cultured on PDA medium was the fastest with the production of gray mycelium and dense colonies. Lethal temperatures for sclerotia, mycelium and conidia were 60, 55 and 50 ℃, respectively. [ Conclusion] The study provided the scientific basis for the research on the incidence law of B. cinerea and its control.

Hot air treatment activates defense responses and induces resistance against Botrytis cinerea in strawberry fruit

The effect of hot air（HA, 45°C, 3.5 h） treatment on reducing gray mold caused by Botrytis cinerea in strawberry fruit and the possible mechanisms were investigated. The results showed that HA treatment significantly reduced lesion diameter and enhanced activities of chitinase（CHI）, β-1,3-glucanase and phenylalanine ammonia-lyase（PAL） in strawberry fruit. Total phenolic contents were also increased by HA treatment. The activities of antioxidant enzymes including superoxide dismutase（SOD）, catalase（CAT） and ascorbate peroxidase（APX） were higher in HA treated strawberry fruit than those in control. Expression of three defense related genes such as CAT, CCR-1 allele and PLA6 was greatly induced in HA treated strawberry fruit with or without inoculation by B. cinerea. In addition, the in vitro experiment showed that HA treatment inhibited spore germination and tube growth of B. cinerea. These results suggested that HA treatment directly activated disease resistance against B. cinerea in strawberry fruit without priming response and directly inhibiting growth of B. cinerea.

The Herbicidal Activity of Mutant Isolates from Botrytis cinerea

Fifteen mutant isolates were obtained by ultraviolet mutation from parent isolate Botrytis cinerea BC-4. Among them three mutant isolates, BC4-1, BC4-2, and BC4-15, showed strong herbicidal activity. BC4-1 showed maximum herbicidal activity for inhibition of germination and growth of Digitaria sanguinalis L. and Arnaranthus retroflexus L. The results also showed that herbicidal activity was influenced by differing pH of PD media, with pH value of 4.0 being the optimum. The crude toxin was extracted using chloroform, petroleum ether, and ethyl acetate, respectively, and the ethyl acetate extracts showed the strongest inhibitory activity on the germination and growth of D. sanguinalis L. and A. retroflexus L. Using HPLC, one fraction with an absorption peak at 271 nm was separated from the crude toxin. This fraction could strongly inhibit the growth of D. sanguinalis L. at a concentration of 100 mg L^-1 and could completely inhibit the seed germination of D. sanguinalis L. and A. retroflexus L. at a concentration of 50 mg L^-1 .

Effects of different LED light wavelengths on the resistance of tomato against Botrytis cinerea and the corresponding physiological mechanisms

New types of light-emitting diode （LED） sources were applied to irradiate Botrytis cinerea mycelium and tomato leaves that were inoculated with B. cinerea to assess the effect of different LED light wavelengths on the infection of tomato with B. cinerea, to determine the optimum light wavelengths to control B. cinerea, and to explore the mechanism of LED influ- ence on the development of gray mold. The results showed that purple light and blue light irradiation significantly inhibited the growth of B. cinerea mycelium, and the inhibition rates were 22.3 and 15.16%, respectively, and purple light exhibited a better inhibitory effect than blue light. The lesion development of B. cinerea on tomato leaves was significantly inhibited upon irradiation with red and purple light with inhibition rates of 32.08 and 36.74%, respectively. Irradiation with red light inhibited the oxidative burst of superoxide anion （O2-） that was caused by infection with B. cinerea, and red light regulated the H2O2 content in the tomato leaf, which increased and rapidly returned to a lower level. In addition, red light irradiation improved the activity of superoxide dismutase （SOD）, catalase （CAT） and peroxidase （POD） in tomato leaves. However, purple light irradiation did not make tomato leaves exhibit this kind of physiological response. Therefore, red light and purple light can suppress gray mold in tomatoes, but the disease suppression mechanisms of these two types of LED light are different. In general, red light suppresses gray mold primarily by regulating the tomato defense mechanism for disease, whereas the suppression of tomato gray mold by purple light can be partially explained by the photo-inhibition of B. cinerea.

Plastic bronchitis associated with Botrytis cinerea infection in a child:A case report

BACKGROUND Plastic bronchitis(PB)frequently occurs in children after the surgical repair of congenital cardiac defects or in the presence of inflammatory or allergic diseases of the lung.Accurate epidemiological data of this condition are still lacking.CASE SUMMARY A 5-year-old boy,with a clear medical history,presented to our hospital with persistent cough and pneumonia with segmental atelectasis on chest computerized tomography.He showed no significant improvement after 1 wk of amoxicillin-clavulanate potassium treatment.Bronchial casts were extracted using flexible bronchoscopy.Pathological examination of the dendritic cast confirmed the diagnosis of type I PB.Botrytis cinerea was detected by next-generation sequencing of the bronchoalveolar lavage fluid.After the removal of the airway obstruction and fluconazole treatment,the patient recovered and was discharged 14 d after admission without the recurrence of cough.CONCLUSION Botrytis cinerea pneumonia should be considered in children with PB who still have prolonged cough and atelectasis after a regular course of antibiotic therapy.Flexible bronchoscopy and etiological examination should be performed in a timely manner to determine the diagnosis,clear the airway obstruction,and target etiological treatment.

Occurrence of Boscalid Resistance in Botrytis cinerea from Greenhouse Cucumbers in China

Gray mold caused by Botrytis cinerea is a major disease of greenhouse cucumbers and its control is largely dependent on the succinate dehydrogenase inhibitor boscalid.In this study,gray mold-infected cucumbers were collected from greenhouses located in 11 towns in which boscalid had been used,and 163 B.cinerea strains were isolated by single-spore method.The EC50 value effective concentration that could inhibit conidia germination by 50%compared with the control for sensitive isolates ranged from 0.045μg/mL to 50.148μg/mL in the assays.Additionally,101 isolates were resistant to boscalid,and the resistance frequency ranged from 33.33%to 86.67%in these towns.Most of the resistant isolates showed a low or moderately resistant phenotype,with the resistance frequency of moderately resistant isolates being as high as 80%.The SdhB gene harbored a single mutation corresponding to His272.The fitness evaluation showed that the hyphal growth,sporulation,germinability and pathogenicity were different among the resistant and sensitive isolates.The results suggested that dual resistance to boscalid in B.cinerea populations could result in failure to control gray mold.

An ABAGS-Like Metabolite of Botrytis cinerea Isolate BC4 and Its Inhibitory Activity to Seed Germination of Weeds

Aimed at finding out natural compounds to kill weeds, a plant pathogenic fungus, Botrytis cinerea isolate BC4, was chosen as a source. A significantly polar metabolite, which was different in chromatographic behavior from ABAGE-like metabolite, was isolated from cultural filtrate of the fungus by column chromatography on silica gel, preparative LC, TLC and HPLC. The structure of the metabolite was determined by HPLC-ESI MS, g.l.c, IR, ^1H NMR and hydrolysis. The results showed that this metabolite was quite similar with ABAGS. The bioassay showed the metabolite had inhibition to seed germination of both broadleaf weeds [Amaranthus retroflerus L. and Chenopodium album L.] and gramineae weeds [Digitaria sanguinalis L. Scop and Echinochloa crusgalii L. Beauv]. It also had inhibitory activity to the seedling growth of broadleaf weeds. The bioassay, using Amaranthus retroflexus L. as an indicator, showed that the lowest concentration of the metabolite to inhibit root＋sprout growth of A. retroflexus was 0.6 μM, and the concentration for 50% inhibition was 1.3μM.

An ABAGE-Like Metabolite of Botrytis cinerea Isolate BC4 Inhibited the Growth of Hypocotyls and Roots of Amaranthus retroflexus Seedlings

A metabolite, which had an inhibitory effect on plant growth, was isolated from cultural filtrate of Botrytis cinerea isolate BC4 by column chromatography on silica gel and preparative HPLC. Its structure was determined from HPLC-ESI MS, GC, IR, ^1H NMR and ^13C NMR spectral data, as well as chemical hydrolysis. The results showed that this metabolite was quite similar to abscisic acid-β-D-glucopyranosyl ester（ABAGE） in structure. The inhibitory effect of the ABAGE-like metabolite on plant growth was investigated using a weed Amaranthus retroflexus L. as a bioassay material. The results showed that it inhibited hypocotyls and roots growth of A. retroflexus （Amaranthus retroflexus L.） seedlings when its concentration was over 0.5 μM. The concentrations for 50% inhibition of hypocotyls and roots growth of A. retroflexus seedlings were 2.8 and 1.4 μM, respectively.

Tomato SlPti5 plays a regulative role in the plant immune response against Botrytis cinerea through modulation of ROS system and hormone pathways

While SlPti5 has been shown to play a crucial role in the regulation of antagonistic genes in Solanum lycopersicum and Arabidopsis against pathogen infection,there have been no comprehensive studies on the effects of SlPti5 on the regulatory response mechanism of reactive oxygen species(ROS) system and hormone pathways during growth and disease resistance of tomato plants.Here,we investigated the function of SlPti5 in the defense response of tomato against Botrytis cinerea utilizing a virus-induced gene silencing(VIGS)-based system.Expression profile analysis showed that SlPti5 was significantly induced upon B.cinerea infection,with high expression levels in the leaves and fruit of tomato.VIGS-based silencing of SlPti5 inhibited early vegetative growth,increased the plant’s susceptibility to infection,promoted the development of ROS,affected the expression of genes involved in the ROS scavenging system,and attenuated the expression of genes associated with pathogenesis and the ethylene/jasmonic acid signaling pathways.In sum,our data demonstrated that SlPti5 stimulates the immune response of tomato plant to Botrytis cinerea infection by involving the ethylene(ET)-and jasmonic acid(JA)-mediated pathways and modulating the expression of some key pathogenesis-related(PR) genes.

Control Effect of Bacillus cereus CGMCC4348 against Botrytis cinerea and Its Mechanism

[ Objectivel The paper was to study antifungal activity and control mechanism of Bacillus cereus CGMCCA348 against Botrytis cinerea, and further find new methods against the disease. [Method] By quantitative bioassay, in vitro experiment and pot test, the antagonistic protein was extracted with ammonium sul- fate precipitation fractionation to determine the control efficiency. ECso of B. cereus CGMCC4348 was 6. 19 mg/L. Average controlling effect of B. cereus CGM- CC4348 on B. cinerea was 75.8%, equal to the control group which was treated by 50 mg/L pyrimethanil. Bacillus cereus CGMCCA348 had good inhibitory effect against B. cinerea. [ Conclusion] The paper provides scientific basis for green control technology against solanaceous vegetable diseases.

Phytochrome B regulates jasmonic acid-mediated defense response against Botrytis cinerea in Arabidopsis

The phytochrome B mediated light signaling integrates with various phytohormone signalings to control plant immune response.However,it is still unclear whether phyB-mediated light signaling has an effect on the biosynthesis of jasmonate during plant defense response against Botrytis cinerea.In this study,we demonstrated that phyB-mediated light signaling has a role in this process.Initially,we confirmed that phyb plants were obviously less resistant to B.cinerea while phyB overexpressing plants showed significantly enhanced resistance.We also found that the expression of numerous JA biosynthesis genes was promoted upon treatment with red or white light when compared to that of darkness,and that this promotion is dependent on phyB.Consistent with the gene expression results,phyb plants accumulated reduced pool of JA-lle,indicating that phyB-mediated light signaling indeed increased JA biosynthesis.Further genetic analysis showed that light-mediated JAZ9 degradation and phyB-enhanced resistance were dependent on the receptor COI1,and that pifl/3/4/5(pi/q)can largely rescue the severe symptom of phyb.Taken together,our study demonstrates that phyB may participate in plant defense against B.cinerea through the modulation of the biosynthesis of JA.

In field control of Botrytis cinerea by synergistic action of a fungicide and organic sanitizer

A new Integrated Pest Management program based on the combination of synthetic pesticide with a GRAS （generally recognized as safe）-classified sanitizer for the control of Botrytis cinerea in field conditions was described. The aim behind this research was to determine whether the use of this mixture would enhance the efficiency of pesticides while decreasing the recommended dose. Naturally infected tomato plants, grown in the greenhouse, were treated with two commonly used fungicides SWITCH （Syngenta, Switzerland） and SIGNUM （BASF, Germany） each alone or combined with a commercially available organic sanitizer PERACLEAN？5 （Evonik Industries, Germany）. A total of 27 treatments were tested consisting of three concentrations of synthetic fungicide （×1, ×1/2 and ×1/4 of the recommended dose） either applied separately or combined with three concentrations of the tested sanitizer （0.5, 1 and 1.5％）. The control efficacy achieved by the fungicides applied alone ranged between 0 and 66.7％ while all fungicide-sanitizer mixtures resulted in up to 70％ control of grey mould. The treatment that provides the maximum control of B. cinerea was the result mixture of ×1/4 of the recommended concentration of SWITCH （15 g L-1） with 0.5％ of PERACLEAN？5. This combination suppressed 85％ of grey mold infections while decreasing the usually used amount of this pesticide by 75％, reducing therefore the well known negative impacts of chemical pesticides on environment and consumers health.

Enhanced resistance to Botrytis cinerea and Rhizoctonia solani in transgenic broccoli with a Trichoderma viride endochitinase gene

A endochitinase gene（Tch） from the fungus Trichoderma viride was introduced into broccoli（Brassica oleracea var. italica） by Agrobacterium-mediated transformation. Sixty-eight putative transformants were obtained and the presence of the Tch gene was confirmed by both PCR and Southern blot analysis. RT-PCR analysis showed an accumulation of the transcript encoding the endochitinase protein in the transgenic plants. Using real-time quantitative PCR, the expression profiling of endochitinase gene was analyzed. Primary transformants and selfed progeny were examined for expression of the endochitinase using a fluorometric assay and for their resistance to the pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The endochitinase activities in T0 in vitro plants, T0 mature plants and T1 mature plants were correlated with leaf lesions, and the transgenic line T618 had high endochitinse activities of 102.68, 114.53 and 120.27 nmol L–1 MU min–1 mg–1 protein in the three kinds of plants, respectively. The endochitinase activity showed a positive correlation with the resistance to the pathogens. Most transgenic T0 broccoli had increased resistance to the pathogens of B. cinerea and R. solani in leaf assays and this resistance was confirmed to be inheritable. These findings suggested that expression of the Tch gene from T. viride could enhance resistance to pathogenic fungi in Brassica species.