Germ Tube Elongation Research Articles

Alternaria Spots in Tomato Leaves Differently Delayed by Four Plant Essential Oil Vapours

Alternaria leaf spot disease has been a concern during a tomato production in greenhouse. In vitro antifungal activities of vapours of four plant essential oils, cinnamon oil, fennel oil, origanum oil and thyme oil, were investigated during in vitro conidial germination and mycelial growth of Alternaria alternata causing the tomato leaf spots to find eco-friendly alternatives for chemical fungicides. The four plant essential oils showed different antifungal activities against in vitro conidial germination of A. alternata in dose-dependent manners, and cinnamon oil vapour was most effective to suppress the conidial germination. The four plant essential oils showed similar antifungal activities against the in vitro mycelial growth of A. alternata in dosedependent manners, but low doses of thyme oil vapour slightly increased in vitro mycelial growth of A. alternata. Necrotic lesions on the A. alternata-inoculated tomato leaves were reduced differently depending on kinds and concentrations of plant essential oils. Delayed conidial germination and germ-tube elongation of A. alternata were found on the tomato leaves treated with cinnamon oil and origanum oil vapours at 6 hpi. These results suggest that volatiles from cinnamon oil and origanum oil can be provided as alternatives to manage Alternaria leaf spot during the tomato production eco-friendly.

Antifungal Efficacy of Plant Oils Containing Thymol and Carvacrol in Controlling Botrytis cinerea, the Causal Agent of Grape (Vitis vinifera) GrayMould

The efficacy of a large number of plant extracts and essential oils in controlling plant diseases has been proven worldwide. Botrytis cinerea has attacked a wide host range causing severe loss in the field and at storage. In this study the antifungal efficacy of essential oils of three medicinal plants namely wild marjoram (Zataria multifolia), wild savory (perennial) (Satureja mutica) and savory (annual) (Satureja hortensis) possessing these compounds at three concentrations: 50, 100 and 200 ppm were evaluated in controlling the mycelial growth, spore germination and germ tube elongation of B. cinerea. All treatments except savory (annual) essential oil at 50 ppm showed significant differences with the control in inhibiting the mycelial growth as well as spore germination and germ tube elongation of B. cinerea (p=0.01). It was also noted that wild marjoram at 100 ppm and wild savory (perennial) essential oils at 200 ppm could completely (100%) inhibit the growth of the fungus. Essential oils from wild marjoram and wild savory had higher antifungal activity than annual savory. Their suitable formulations could be prepared and used as safe alternatives for controlling moulds of horticultural products during storage.
 The Agriculturists 2018; 16(2) 15-24

Biocontrol of Penicillium digitatum on Postharvest Citrus Fruits by Pseudomonas fluorescens

The effectiveness of the bacteria antagonist Pseudomonas fluorescens to control green mold caused by Penicillium digitatum on oranges (Citrus sinensis Osbeck, cv. Jincheng) and the possible modes of action were evaluated. Whether in vitro or in vivo, treatments with cell-free autoclaved cultures or culture filtrate had limited capacity to suppress P. digitatum, while P. digitatum was significantly inhibited by bacterial fluid (P. fluorescens in the nutrient broth liquid medium) and bacterial suspension (P. fluorescens in sterile distilled water) with living cells. There was a positive relationship between the concentration of P. fluorescens in bacterial suspension and its biological efficacy. In addition, P. fluorescens was effective when applied preventatively but not when applied curatively. In the inoculated wounds, the population of P. fluorescens was an approximately 28- and 34-fold increase after being incubated at 20°C for 8 d and at 4°C for 16 d, respectively, and P. digitatum could effectively stimulate the growth and reproduction of P. fluorescens. Moreover, P. fluorescens was able to inhibit spore germination and germ tube elongation of P. digitatum as well as induce resistance on citrus peel by increasing the chitinase (CHI) activity and advancing the activities peaks of β-1,3-glucanase (GLU), peroxidase (POD), and phenylalanine ammonia lyase (PAL). All of these results support the potential application of P. fluorescens against green mold on postharvest citrus.

Inhibitory of grey mold on green pepper and winter jujube by chlorine dioxide (ClO2) fumigation and its mechanisms

As a powerful sanitizer, chlorine dioxide (ClO2) has mainly focused on control of bacteria, but little is known about its antifungal effects, especially on postharvest fungi found in vegetable and fruit. This study investigated the effect of ClO2 on Botrytis cinerea growth in vitro and on its disease severity in green pepper and winter jujube. Result showed that ClO2 fumigation reduced disease development of grey mold caused by B. cinerea on green pepper and winter jujube, and the efficiency was positively correlated with ClO2 concentrations. ClO2 treatment directly inhibited mycelial growth, spore germination and germ tube elongation of B. cinerea. To further survey the mechanisms by which ClO2 treatment inhibited hypha growth of B. cinerea, we observed the mycelial morphology and the integrity of the plasma membrane; the results showed that ClO2 treatment resulting in B. cinerea mycelial morphology was strongly affected, and leading to the loss of plasma membrane integrity, causing the cellular leakage. These data showed that ClO2 was effective for controlling grey mold caused by B. cinerea on green pepper and winter jujube, and the antifungal activity of ClO2 could be attributed to the disruption of cell membrane of the fungal pathogen.

The modes of action of epsilon-polylysine (ε-PL) against Botrytis cinerea in jujube fruit

Epsilon-polylysine (ε-PL) is a natural antimicrobial and has been widely used as a food preservative. Previous studies showed ε-PL being effective for control of foodborne bacteria; however, little is known about its control effects on postharvest pathogenic fungi and the possible modes of action. Here, we found that ε-PL could significantly inhibit mycelial growth, spore germination and germ tube elongation of Botrytis cinerea in vitro, as well as effectively control gray mold in harvested jujube fruit. Further investigation indicates that the modes of action of ε-PL against B. cinerea include (i) stimulating the accumulation of intracellular reactive oxygen species (ROS) and reducing the expression of pathogenesis-related genes of the pathogen, (ii) resulting in the leakage of soluble carbohydrates and nucleic acids, as well as the damage of plasma membrane integrity of the fungal cells, (iii) inducing the expression of respiratory burst oxidase homolog (RBOH) genes in ε-PL-treated jujube fruit.

Potential Impact of Populations Drift on Botrytis Occurrence and Resistance to Multi- and Single-Site Fungicides in Florida Southern Highbush Blueberry Fields.

Incidence of blossom blight and Botrytis fruit rot (BFR), caused by Botrytis cinerea, on two southern highbush blueberry cultivars was evaluated in several blueberry fields grown in the vicinity (BB-Str(+)) or not (BB-Str(-)) of strawberry fields in central Florida. Blossom blight and BFR incidence were higher in BB-Str(+) fields in 2014 and significantly higher in 2015 compared to BB-Str(-) fields. In total, 613 B. cinerea isolates (i.e., 181 and 432 isolates from BB-Str(-) and BB-Str(+) fields, respectively) were collected. The isolates were evaluated for sensitivity to eight single-site and one multisite fungicides using a spore germination and a germ tube elongation assay. Overall, 5, 15, 24, 28, 54, and 93% of isolates collected from BB-Str(-) were resistant to penthiopyrad, cyprodinil, boscalid, fenhexamid, pyraclostrobin, and thiophanate-methyl, respectively. Respective resistance frequencies in BB-Str(+) isolates were 10, 30, 65, 66, 89, and 99%. Resistance frequencies for all fungicides were always higher in BB-Str(+) fields compared to BB-Str(-) fields. Isolates exhibiting resistance to six or five fungicides simultaneously were predominant (50 to 70%) in blueberry fields regardless if they were grown in the vicinity of strawberry fields or not. Among 308 and 305 B. cinerea isolates tested in 2014 and 2015, 41.8 and 47.1%, respectively, showed reduced sensitivity to the multisite fungicide captan. The lower label rate of captan applied preventively did not control isolates with reduced sensitivity on detached blueberry fruit. These findings suggest a potential population flow between strawberry and blueberry fields that may impact blossom blight and gray mold development in blueberry fields. The relatively lower fungicide input applied to blueberry fields compared with strawberry fields seems to be sufficient to select for resistance and multiple-resistant phenotypes in B. cinerea populations in blueberry.

Antifungal activity of proteolytic fraction (P1G10) from (Vasconcellea cundinamarcensis) latex inhibit cell growth and cell wall integrity in Botrytis cinerea

The aim of this study was to determine the antifungal activity of the proteolytic fraction P1G10 from Vasconcellea cundinamarcencis (ex-Carica candamarcensis) against Botrytis cinerea, the causative agent of pre- and postharvest damaging disease in fruit and vegetables. The survival of B. cinerea at different concentrations of P1G10 showed that 1 mg/mL inhibited 50% of mycelium growth after 72 h incubation. The kinetic of growth inhibition fits the Weibull distribution function, and the data was confirmed by the IC50 survival assay. The study shows that P1G10 inhibits conidia germination and germ tube elongation of B. cinerea relative to untreated conidia. Hypersensitivity to cell wall-perturbing agents (Calcofluor white and Congo red) was observed in mycelium cells treated with P1G10. In addition, P1G10 exhibited inhibitory effect on the adhesion of conidia, provoked alterations in membrane integrity and induced production of reactive oxygen species accompanied by cellular damage. Our results highlight the effect of P1G10 on mycelium growth, cell wall alterations, membrane integrity and adhesion. P1G10 emerges as promising antifungal to control disease causing agents in the food agroindustry.

Antagonistic Activity against Dirty Panicle Rice Fungal Pathogens and Plant Growth-Promoting Activity of Bacillus amyloliquefaciens BAS23.

Bacterial strain BAS23 was isolated from rice field soil and identified as Bacillus amyloliquefaciens. Based on dual culture method results, the bacterium BAS23 exhibited potent in vitro inhibitory activity on mycelial growth against a broad range of dirty panicle fungal pathogens of rice (Curvularia lunata, Fusarium semitectum and Helminthosporium oryzae). Cell-free culture of BAS23 displayed a significant effect on germ tube elongation and mycelial growth. The highest dry weight reduction (%) values of C. lunata, H. oryzae and F. semitectum were 92.7%, 75.7% and 68.9%, respectively. Analysis of electrospray ionization-mass spectrometry (ESI-MS) and 1H nuclear magnetic resonance (NMR) spectroscopy revealed that the lipopeptides were iturin A with a C14 side chain (C14 iturinic acid) and with a C15 side chain (C15 iturinic acid), which were produced by BAS23 when it was cultured in nutrient broth (NB) for 72 h at 30°C.BAS23, the efficient antagonistic bacterium, also possessed in vitromultipletraits for plant growth promotionand improved rice seedling growth. The results indicated that B.amyloliquefaciens BAS23 represents a useful option either for biocontrol or as a plant growth-promoting agent.

Direct antifungal activity of tiadinil, a systemic acquired resistance inducer, and thymol formulations on Stagonosporopsis citrulli and control of watermelon gummy stem blight

This study evaluated the direct antifungal activity of tiadinil [N-(3-chloro-4-methylphenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide], a systemic acquired resistance (SAR) inducer and two formulations of thymol (thymol I and thymol II) against Stagonosporopsis citrulli, the causal agent of gummy stem blight (GSB) disease of watermelon. Tiadinil, thymol I and thymol II completely inhibited the mycelial growth of S. citrulli in vitro at ≥ 100 ppm. Conidial germination and germ tube elongation were completely inhibited by tiadinil at ≥ 2000 ppm and by thymol-based formulations at ≥ 100 ppm. A single foliar application of tiadinil at ≥ 10 ppm or a single application of thymol I and II at ≥ 1 ppm, 48 h before or after pathogen inoculation, significantly reduced disease severity of watermelon seedlings inoculated with 105/ml conidial suspension of S. citrulli, compared to respective nontreated controls. Plants treated with foliar application of tiadinil at ≥ 1000 ppm before pathogen inoculation had significantly lower disease severity than plants that received an equivalent drench application. The efficacy of foliar application of tiadinil was affected by concentration and frequency of application. The study suggests direct antifungal activity of tiadinil, indicating a new mode of action of tiadinil against GSB disease of watermelon. The study also demonstrated direct antifungal action of thymol, a formulated active compound of essential oils, against S. citrulli and GSB disease of watermelon.

Efficacy of citronella essential oil for the management of chilli anthracnose

The potential of citronella essential oil for the management of chilli anthracnose caused by Colletotrichum acutatum was investigated. In in vitro tests, citronella essential oil inhibited mycelial growth at 0.25, 1.25 and 2.5 μL citronella/mL water, reduced conidial germination, and inhibited germ tube elongation at 1.25 μL/mL. Citronella essential oil applied as a protective or curative treatment to chilli cv. Django fruits reduced anthracnose infection. Efficacy of citronella essential oil was substantially effective to chemical fungicides and relatively superior to a biofungicide in both protective and curative tests. However, citronella essential oil at 2.5 μL/mL was phytotoxic to chilli fruits. Therefore, the most effective rate of citronella essential oil was at 1.5 μL/mL with inhibition of pathogen growth, reduction of anthracnose symptoms, and no observable phytotoxic response on chilli fruits. Citronella essential oil may be a viable alternative to chemical fungicides for the management of chilli anthracnose.

Inhibitory effects of methyl thujate on mycelial growth of Botrytis cinerea and possible mechanisms

Methyl thujate, a monoterpenoid substance, was effective in controlling postharvest gray mold caused by Botrytis cinerea on apple fruit, during application in aqueous solution. Methyl thujate strongly inhibited spore germination, germ tube elongation and mycelial spreading of B. cinerea in the culture medium, and the inhibition on fruit decay was closely correlated with methyl thujate concentrations. By means of propidium iodide and fluorescein diacetate staining, loss of membrane integrity and cell viability in B. cinerea was detected after methyl thujate treatment. Furthermore, methyl thujate led to cytoplasmic accumulation of reactive oxygen species, leakage of cellular constituents and variations in cell wall component deposition in B. cinerea hyphae, as the genes encoded subunits for NADPH oxidase were synergistically upregulated, whereas the genes involved in cell wall modeling and virulence responded differentially. Taken together, these data suggest that the mechanisms by which methyl thujate decreased fruit decay may be directly attributed to the induced ROS burst, breakdown of cell membrane integrity and loss of cytoplasmic contents from the hyphae.

Toxicity and biochemical action of the antibiotic fungicide tetramycin on Colletotrichum scovillei

Tetramycin, a novel polyene macrolide antibiotic, has strong activity against a broad spectrum of fungi and may have potential uses in future agricultural applications. Thus, the antifungal activity and biochemical action of tetramycin on Colletotrichum scovillei were investigated in this study. The experimental results indicated that tetramycin had strong inhibitory activity against the mycelial growth, spore germination and germ tube elongation of C. scovillei. The baseline sensitivity curves were unimodal, with mean EC50 values of 1.98 ± 0.078 μg/mL and 0.003 ± 0.005 μg/mL for mycelial growth and spore germination inhibition, respectively. Tetramycin also inhibited the germination of spores and formation of appressoria. After tetramycin treatment, the edge of the mycelial diaphragm showed protuberances, with decreased offshoots at the top. Additionally, disruption of the membrane was detected through an increase in membrane permeability, leakage of sugars and a reduction in the ergosterol content. Tetramycin effectively controlled C. scovillei on detached pepper fruits. These results will contribute to our evaluation of the potential of tetramycin for successful management of pepper anthracnose and to our understanding of the possible biochemical action of tetramycin against C. scovillei.

Inhibitory effect and possible mechanism of a Pseudomonas strain QBA5 against gray mold on tomato leaves and fruits caused by Botrytis cinerea.

The fungal pathogen Botrytis cinerea causes gray mold disease on various hosts, which results in serious economic losses. Over the past several decades, many kinds of fungicides have been used to successfully control the disease. Meanwhile, the uses of fungicides lead to environmental pollution as well as a potential threat to the human health by the chemical residues in tomato fruit. Also, the gray mold disease is difficult to control with fungicides. Therefore, exploring alternative measures such as biological controls could be the best choice to control the disease and alleviate damages caused by fungicides. In this study, we isolated and identified a novel Pseudomonas strain termed as QBA5 from healthy tomato plant based on the morphological, biochemical characteristics and molecular detection. The antifungal activity assays revealed that, in the presence of QBA5, conidia germination, germ tube elongation and mycelial growth of B. cinerea were significantly inhibited. Most importantly, QBA5 exerted a significant preventive effectiveness against gray mold on tomato fruits and plants. The possible mechanism of QBA5 involved in the inhibition of B. cinerea was investigated. It revealed that the conidia plasma membrane of B. cinerea was severely damaged by QBA5. Further, four different antifungal compounds in the supernatant of QBA5 were separated by preparative high performance liquid chromatography (PHPLC). Overall, the data indicate that there is a considerable potential for QBA5 to reduce the damage caused by gray mold disease on tomato.

Biocontrol Activity of Volatile-Producing Bacillus megaterium and Pseudomonas protegens Against Aspergillus and Penicillium spp. Predominant in Stored Rice Grains: Study II

In our previous studies, Bacillus megaterium KU143, Microbacterium testaceum KU313, and Pseudomonas protegens AS15 have been shown to be antagonistic to Aspergillus flavus in stored rice grains. In this study, the biocontrol activities of these strains were evaluated against Aspergillus candidus, Aspergillus fumigatus, Penicillium fellutanum, and Penicillium islandicum, which are predominant in stored rice grains. In vitro and in vivo antifungal activities of the bacterial strains were evaluated against the fungi on media and rice grains, respectively. The antifungal activities of the volatiles produced by the strains against fungal development and population were also tested using I-plates. In in vitro tests, the strains produced secondary metabolites capable of reducing conidial germination, germ-tube elongation, and mycelial growth of all the tested fungi. In in vivo tests, the strains significantly inhibited the fungal growth in rice grains. Additionally, in I-plate tests, strains KU143 and AS15 produced volatiles that significantly inhibited not only mycelial growth, sporulation, and conidial germination of the fungi on media but also fungal populations on rice grains. GC-MS analysis of the volatiles by strains KU143 and AS15 identified 12 and 17 compounds, respectively. Among these, the antifungal compound, 5-methyl-2-phenyl-1H-indole, was produced by strain KU143 and the antimicrobial compounds, 2-butyl 1-octanal, dimethyl disulfide, 2-isopropyl-5-methyl-1-heptanol, and 4-trifluoroacetoxyhexadecane, were produced by strain AS15. These results suggest that the tested strains producing extracellular metabolites and/or volatiles may have a broad spectrum of antifungal activities against the grain fungi. In particular, B. megaterium KU143 and P. protegens AS15 may be potential biocontrol agents against Aspergillus and Penicillium spp. during rice grain storage.

In vitro effects of glycyrrhetinic acid and hyaluronic acid on the growth of vulvovaginal Candida albicans and other yeasts

Aims. The present study aimed to test the in vitro activity against Candida albicans and non-albicans strains of 18-β glycyrrhetinic acid (18-β GA) and hyaluronic acid (HA), both alone and in combination. This antimicrobial activity was assessed using the National Committee for Clinical Laboratory Standards (NCCLS) method on Candida strains that were isolated from patients with recurrent vulvovaginal candidiasis (RVVC). Results. Our results demonstrate that the anti-Candida activity is independent from antifungal susceptibility level and the fact that the growth inhibition is stronger at acidic pH level makes the two drugs a promising biological alternative for the topical treatment of vulvovaginal candidiasis (VVC) and RVVC. Conclusions. Furthermore, the reduction of both budding cells formation and germ tube elongation, on mammalian cell monolayers, may explain the observed growth inhibition and suggest a decreased virulence, respectively.

English

Chemical analysis of aqueous extract of Ziziphus mucronata Willd. was determined by liquid chromatography–mass spectrometry (LC-MS) analysis. Among the 11 compounds found, catechin, rutin (quercetin 3-O-rutinoside), delphinidin-3-glucoside, isoquercetin (hyperoside) and quercitrin (quercetine3,7-O-L-dirhanmopyranoside) were identified as the major phenolics components in this aqueous plant extract. To elute the target compounds, the fractionation of crude extract was carried out on solid phase extraction (SPE) columns. The different fractions (from FZ1 to FZ5) obtained after fractionation were evaluated in vitro against economically important foliar fungal pathogens of peanut, including Cercospora arachidicola, Phaeoisariopsis personata and Puccinia arachidis. The treatments with Z. mucronata fractions were compared with negative control (water) and standard solutions of cathechin and rutin (1 mg/mL). All the fractions recorded an inhibitory effect, firstly on conidial germination and germ tube elongation, secondly on disease evolution on peanut leaves previously inoculated by fungi; the level of efficiency of inhibition varied from 40.55 (FZ1 against C. arachidicola) to 57.14% (FZ2 and FZ3 against P. arachidis). Then, spores of P. arachidis seemed to be more sensitive to the treatment.   Key words: Fungal pathogens, peanut, plant extract, phenolics, Ziziphus mucronata Willd.

Sensitivity of Colletotrichum acutatum to six fungicides and reduction in incidence and severity of chili anthracnose using pyraclostrobin

Anthracnose, caused by Colletotrichum acutatum, is one of the most destructive diseases of chili worldwide. Fungicides including picoxystrobin, azoxystrobin, difenoconazole, thiophanate-methyl and mancozeb have been used to manage anthracnose diseases for many years. Pyraclostrobin was introduced in China in 2016 to control chili anthracnose but has not seen widespread use. In this study, 40 isolates of C. acutatum were examined for their corresponding sensitivities to these fungicides using mycelial growth and spore germination assays. In greenhouse and field trials, the duration of protection of chili fruit from infection provided by pyraclostrobin against C. acutatum was determined. In vitro experiments showed that pyraclostrobin and azoxystrobin were inhibitory to both mycelial growth and spore germination, while mancozeb was only effective in inhibiting spore germination, and difenoconazole and thiophanate-methyl were effective in inhibiting mycelial growth. The results obtained suggest that these fungicides inhibited C. acutatum. It was observed that germ tube elongation and sporulation were reduced by pyraclostrobin. Pyraclostrobin provided protective activity for up to 96 h before inoculation, as well as curative activity up to 72 h after inoculation. In field trials, pyraclostrobin was effective in reducing the incidence and severity of anthracnose, and enhanced the marketable yield of chili. These results demonstrate that pyraclostrobin is a useful fungicide for the management of chili anthracnose.

Fungitoxicity of organic extracts ofOcimum basilicumon growth and morphogenesis ofBipolarisspecies (teleomorphCochliobolus)

This study aimed at evaluating the inhibitory effects of various organic extracts of Ocimum basilicum against some species of Bipolaris and Cochliobolus with GC-MS and HPLC analysis. The ethyl acetate extract consisted of methyl cinnamate as the most abundant component, while butylated hydroxytoluene was the major component in the methanol extract. Pyrogallol and chlorogenic acid were major phenolic compounds in the ethyl acetate and methanol extracts, respectively. Complete growth inhibition of all fungi except Cochliobolus australiensis was observed by ethyl acetate extract, and on Bipolaris hawaiensis, Bipolaris spicifera and Cochliobolus cynodontis by methanol extract. Spore germination was completely inhibited for Bipolaris hawaiensis by ethyl acetate extract. Scanning electron microscopic observations revealed that the organic extracts cause considerable morphological changes of the fungal hyphae such as mycelial asymmetry, hyphal swelling, sunken, curling, distorted and broken hyphae. The ethyl acetate and methanol extracts of O. basilicum can result in an effective suppression of mycelial growth, spore germination and germ tube elongation of Bipolaris and Cochliobolus species. The organic extracts of O. basilicum are potential and promising natural tools for controlling Bipolaris and Cochliobolus species, economically important plant and human fungal pathogens.

The oligosaccharyl transferase subunit STT3 mediates fungal development and is required for virulence in Verticillium dahliae.

Verticillium dahliae is the most overwhelming plant pathogen, causing Verticillium wilt in a number of economic crops. The molecular mechanism is still unclear and identification of the genes involved in the pathogenicity or virulence of this fungus would benefit to uncover such mechanism. STT3 is a catalytic subunit of the multi-subunit oligosaccharyl transferase (OST) and plays an essential role in glycoprotein modification. Here, we characterized STT3 gene (VDAG_03232.1) of V. dahliae to explore its regulatory role in the development and virulence by deletion and complementation of this gene, as well as its silence in transgenic plants. The expression of the STT3 gene increased at the stage of conidia germination and reached its peak level with germ tube formation and elongation. We generated the knockout mutants (ΔSTT3) using protoplast transformation. Mycelial growth, sporulation ability and glycoprotein secretion were impaired when ΔSTT3 mutants were grown on media supplemented with different carbon sources. Moreover, ΔSTT3 mutants exhibited distinctly decreased germination ratio and reduction in virulence compared with the wild type (Vd wt) and complementary (ΔSTT3-C) strains. We also generated transgenic Nicotiana benthamiana (Trans-1 and -2) plants by expressing dsRNA against the STT3 gene. Transgenic plants showed significant reduction in the disease index and fungal biomass resulting in elevated resistance to V. dahliae compared with the wild-type plants when inoculated with Vd wt. Our results indicated that STT3 mediates the full virulence through the regulation in fungal development, hyphal growth, glycoprotein secretion of V. dahliae and merits further study as a potential RNAi target to control this fungus.

Microencapsulation of clove essential oil improves its antifungal activity against Penicillium digitatum in vitro and green mould on Navel oranges

ABSTRACT​Relatively high levels of clove essential oil (CEO) necessary for in vivo inhibition of postharvest pathogens imply that its antifungal activity should be reinforced. Hence, this study aimed to determine whether microencapsulation technology enhances the antifungal activity of CEO against Penicillium digitatum, the causal agent of citrus green mould, in vitro and on Navel oranges. Microencapsulation of CEO resulted in a respective reduction of minimum inhibitory concentration and minimum fungicidal concentration from 500 μg/ml to 125 μg/ml and 250 μg/ml, indicating the enhancement of fungistatic and fungicidal activity with respect to that of the pure oil. CEO microcapsules had the best control of lesion diameter and decay development (0.79 mm, 8.64%) in Navel oranges after 5 days’ storage at 25°C, followed by pure oil treatment (4.2 mm, 55.56%). In the vapour phase, CEO microcapsules also exhibited the highest ability to control green mould of Navel oranges. Furthermore, CEO microcapsules showed stronger suppressive activity of spore germination and germ tube elongation of P. digitatum than pure oil. These results indicate that the antifungal activity of CEO was considerably enhanced when it was converted into a microcapsule.