Latency of Infection in Unripe Avocados by Colletotrichum gloeosporioides is Not Due to Inadequate Enzyme Potential

Colletotrichum gloeosporioides produced exo-pectin lyase and protease in a) liquid cultures with incorporated washed cell wall material from unripe or ripe avocado and b) autoclaved immature fruit. The activity of exo-pectin lyase and protease produced in liquid cultures incorporating washed cell walls from immature fruits was almost the same as when washed cell walls from ripe fruits were incorporated. Ripe fruit tissue rotted by the fungus contained exo-pectin lyase, endo-polygalacturonase (endo-PG) and protease. The endo-PG was found to be endogenous to avocado fruit, and had a pH optimum of 5.5. The pH optima of exo-pectin lyase and protease were 8.5 and 7.5 respectively in all three enzyme preparations. All these enzyme preparations completely macerated avocado fruit tissue discs in vitro in less than 3 h of incubation but not potato tuber discs. Neither immature nor ripe fruit contained substances, proteinaceous or otherwise, which could inhibit the exo-pectin lyase or protease activity of these preparations. The results indicated that C. gloeosporioides possesses sufficient enzyme potential to invade cell walls of unripe fruit and that the fruit tissue does not have a mechanism to inactivate such enzymes.     

The Effect of Nanostructured Chitosan and Chitosan‐thyme Essential Oil Coatings on Colletotrichum gloeosporioides Growth in vitro and on cv Hass Avocado and Fruit Quality

In this study, nanostructured edible coatings based on chitosan nanoparticles (CSNPs) and chitosan‐thyme essential oil nanoparticles (CSTEO‐NPs) were characterized and evaluated on in vitro growth of Colletotrichum gloeosporioides and on inoculated avocado fruit cv. Hass, to evaluate their fungicidal activity and the effect on fruit quality. From TEM and particle size distribution characterization, the size of nanoparticles increased after thyme essential oil incorporation. Overall a synergistic effect between the chitosan and thyme essential oil (TEO) was observed. For in vitro evaluation, incorporation of this essential oil to CSNPs improved the control of C. gloeosporioides as there was a complete growth inhibition. CSNPs with concentrations of TEO at 3 and 5% had a fungicidal effect. The coating formulation with 55% CSTEO‐NPs notably reduced the incidence of C. gloeosporioides on avocado cv. Hass by up to 60%. Also, at the end of the 8‐day storage period, CSTEO‐NPs incorporation into the coating did not affect the quality of avocado; moreover, fruit firmness was better maintained than untreated fruit.     

Identification and Characterization of Colletotrichum spp. affecting Fruit after Harvest in Brazil

Colletotrichum spp. cause anthracnose in various fruits post‐harvest and are a particularly important problem in tropical and subtropical fruits. The disease in fruits of avocado, guava, papaya, mango and passion fruit has been reported to be caused by C. gloeosporioides, and in banana by C. musae. In subtropical and temperate crops such apple, grape, peach and kiwi, the disease is caused by C. acutatum. The variation in pathogenic, morphological, cultural and molecular characteristics of Brazilian isolates of Colletotrichum acutatum Simmonds and isolates from post‐harvest decays of avocado, banana, guava, papaya, mango and passion fruit was evaluated. The fruits were inoculated with mycelium of C. acutatum, Colletotrichum spp. and C. musae on a disc of potato dextrose agar. The morphological, cultural and molecular characteristics studied were conidia morphology, colony growth at different temperatures, colony coloration and PCR with primers CaInt2 and ITS4 for C. acutatum and CgInt and ITS4 for C. gloeosporioides. C. acutatum was pathogenic to avocado, guava, papaya, mango and passion fruit, but it was not pathogenic to banana. The morphological, cultural and molecular studies indicated that the avocado, papaya, mango and passion fruit isolates were C. gloeosporioides. The natural guava isolate was identified as C. acutatum, which had not been found previously to produce anthracnose symptoms on guava in Brazil.     

The importance of avocado (Persea americana Mill.) fruits anthracnose and factors influencing the disease in Mana district,south-western Ethiopia

Anthracnose disease surveys were conducted in 25 farmers’ orchards, wholesaler and retailer shops in south-west Ethiopia. In addition, harvesting and postharvest practices, and storage conditions influencing disease development were studied with observation and questionnaire. The assessment results indicated significant variation among farmers’ orchards with the highest incidence (84.0 ± 16.7%) and severity index (26.0 ± 5.4%). Anthracnose damage of fruit was higher at retailers (76.7 ± 20.8%) than in the wholesalers shop (56.7 ± 32.5%). The total number of isolates identified was 249 and Colletotrichum gloeosporioides was the predominant pathogen proved by pathogenecity test. Among the major factors, harvesting avocado fruits with children (88%) and climbing on the tree (72%) resulted in fruit dropping that caused substantial injury and bruise. Generally, anthracnose caused by C. gloeosporioides of avocado fruit was prevalent in producer orchards that aggravated by traditional harvest and postharvest practices coupled to inadequate transportation and storage facilities at wholesaler and retailer shops with subsequent decay and loss of avocado fruits.     

Involvement of Epicatechin in Cultivar Susceptibility of Avocado Fruits to Colletotrichum gloeosporioides after Harvest1

Avocado cultivars were defined as susceptible and resistant to Colletotrichum gloeosporioides depending upon the length of the incubation period of the disease after fruit softening. In the susceptible cultivars Fuerte, Horshim, Vurtz, Rincon, and Benik, epicatechin concentration of the peel decreased to 60-130 μg.g^?1, fr. wt. at fruit softening and symptoms appeared on the same or one day later. In the resistant cultivars Hass, Nabal, Netaim and Pinkerton, epicatechin concentration was still 632–1740 μg.g^?1 fr. wt. when fruit softening and symptoms appeared only 4-10 days later. When susceptible Fuerte fruits became soft the concentration of the antifungal compound 1-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15 diene, had decreased to 120 μg.g^?1 fr. wt. and symptoms appeared. In resistant Hass fruits, the antifungal diene was still 238 μg.g^?1 fr. wt. at fruit softening; and it had further decreased to 159 μg.g^?1 fr. wt. when symptoms appeared, four days later. A modified atmosphere and 0.2 M CaCl₂ infiltration both delayed softening of Fuerte fruits; but symptom appearance on these fruits was related to diene decrease and not to fruit softening. The results are discussed in relation to the hypothesis that the susceptibility of avocado cultivars to post-harvest decay by C. gloeosporioides is related to the degradation of the antifungal diene, catalyzed by avocado lipoxygenase, the activity of which is regulated by the decline of its inhibitor epicatechin.     

Molecular characterization and pathogenicity of Colletotrichum sp. from guava

Guava (Psidium guajava) fruit is vulnerable to postharvest diseases, such as anthracnose. In the present study, molecular characterisation and pathogenicity of Colletotrichum associated with antharcnose disease of guava fruit were conducted. From anthracnose lesion of guava, 20 isolates were successfully recovered. Based on colony colours, conidia, appressoria and presence or absence of setae, and ITS regions and ß-tubulin gene sequences, the isolates were identified as Colletotrichum gloeosporioides. Phylogenetic analysis based on combined data-sets using neighbour-joining method showed that C. gloeosporioides isolates did not group with C. gloeosporioides epitype strain, and thus the isolates were referred to as C. gloeosporioides species complex or C. gloeosporioides sensu lato. Pathogenicity tests using wounded treatment showed that C. gloeosporioides isolates from guava were pathogenic causing anthracnose on the fruits. The present study showed that C. gloeosporioides sensu lato is the most common species causing antharcnose disease of guava fruit.     

Gibberellin-like Activity in the Developing Avocado Fruit

Gibberellin-like activity in avocado (Persea americana) fruit extracts was measured by the barley endosperm bioassay. Fruit tissues were anlyzed separately during fruit growth. The level of the activity found was very high in the endosperm and in the seed coats, but decreased in the latter during fruit growth. No measurable gibberellin-like activity was detected in the mesocarp or in the embryo. It is assumed that the seed coats are a site of production of gibberellin-like substances in the avocado fruit.     

The antagonistic effect and mechanisms of Bacillus amyloliquefaciens DGA14 against anthracnose in mango cv. ‘Carabao’

Bacillus amyloliquefaciens strain DGA14 was tested for in vitro antagonism towards Colletotrichum gloeosporioides, a causal pathogen of anthracnose in mango cv. ‘Carabao’. DGA14 produced extracellular metabolites in solid and liquid media that suppressed the growth of C. gloeosporioides. The cells of DGA14 were often observed adjacent to the pathogen so affecting its spore germination and mycelium development. DGA14 colonised mango fruit 48 h after artificial inoculation and persisted 14 days after storage at 18–20°C. On fruit surfaces, DGA14 attached and produced dents to spores of C. gloeosporioides. Dipping mangoes in aqueous cell suspension (10⁸ mL L^?1) of DGA14 significantly decreased the incidence of anthracnose as compared to untreated fruit.     

Penetration by Botryosphaeriaceae species in avocado,guava and persimmon fruit during postharvest

Botryosphaeriaceae species have a wide host range and a worldwide distribution. These fungal species can colonize several plant organs, such as the trunk, leaves and fruit. Some Botryosphaeriaceae species cause important diseases on persimmon, avocado and guava fruit. However, there is a lack of information regarding the mechanisms of penetration by Botryosphaeriaceae species on these tropical and subtropical fruits. This study aimed to better understand the mechanisms involved in fungal penetration, host specificity and aggressiveness of Botryosphaeria dothidea, Lasiodiplodia pseudotheobromae and Neofusicoccum parvum on avocado (Persea americana), guava (Psidium guajava) and persimmon (Diospyros kaki) fruit. Scanning electron microscopy (SEM) image analysis showed that in avocado fruit, the three studied Botryosphaeriaceae species penetrated through lenticels. In guava fruit, penetration through stomata was verified for Botryosphaeria dothidea and Neofusicoccum parvum. In persimmon fruit, an appressoria-like structure was observed for B. dothidea, which suggests direct penetration. Disease incidence in wounded fruit was 24% higher than in non-wounded fruit. L. pseudotheobromae and N. parvum showed differences in aggressiveness in guava fruit. The longest incubation period was observed for N. parvum inoculated on guava, with an average of 4.5 days, and the shortest incubation period was verified for B. dothidea inoculated on avocado, with an average of 2.8 days. The area under the disease progress curve (AUDPC) did not differ between Botryosphaeriaceae species on avocado, whereas on guava and persimmon fruit, the AUDPC was lower for B. dothidea. The information regarding penetration mechanisms and aggressiveness is important to improve postharvest disease control strategies.     

A <Emphasis Type="Italic">Colletotrichum gloeosporioides</Emphasis>-induced esterase gene of nonclimacteric pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis>) fruit during ripening plays a role in resistance against fungal infection

Ripe fruits of pepper (Capsicum annuum) are resistant to the anthracnose fungus, Colletotrichum gloeosporioides, whereas unripe-mature fruits are susceptible. A pepper esterase gene (PepEST) that is highly expressed during an incompatible interaction between the ripe fruit of pepper and C. gloeosporioides was previously cloned. Deduced amino acid sequence of PepEST cDNA showed homology to both esterases and lipases, and contained -HGGGF- and -GXSXG- motifs and a catalytic triad. Inhibition of PepEST activity by a specific inhibitor of serine hydrolase demonstrated that a serine residue is critical for the enzyme activity. Expression of PepEST gene was fruit-specific in response to C. gloeosporioides inoculation, and up-regulated by wounding or jasmonic acid treatment during ripening. PepEST mRNA and protein was differentially accumulated in ripe vs. unripe fruit from 24 h after inoculation when C. gloeosporioides isinvading into fruits. Immunochemical examination revealed that PepEST accumulation was localized inepidermal and cortical cell layers in infected ripe fruit, but rarely even in epidermal cells in infected unripe one. Over-expression of PepEST in transgenic Arabidopsis plants caused restriction of Alternaria brassicicola colonization by inhibition of spore production, resulting in enhanced resistance against A.brassicicola. These results suggest that PepEST is involved in the resistance of ripe fruit against C.gloeosporioides infection.^†These authors contributed equally to the work     

Systemic Infection of Avocado,Persea americana,by Raffaelea lauricola,Does Not Progress Into Fruit Pulp or Seed

Avocado, Persea americana, is an important fruit crop in the tropics and warm subtropics. Laurel wilt, caused by Raffaelea lauricola, is a systemic vascular wilt of avocado that spread recently to Florida, an important producing state in the USA. As fruit and seed of avocado produced in Florida are sold in other states and countries where this crop is produced, there is concern that commerce in these commodities might spread this disease. Potted, fruit‐bearing trees were artificially inoculated with R. lauricola, and plants were systemically colonized by the fungus. In no instance did infection progress further than the hilum (87 total fruit), as determined by re‐isolation of R. lauricola on a semi‐selective medium or its detection, with qPCR and high fidelity PCR, of diagnostic small subunit (SSU) 18s rDNA. Thus, it would apparently be safe to propagate avocado with seed from trees affected by this disease. Pedicels/peduncles and hila associated with these fruit were colonized by the pathogen. The latter tissues would be associated with/attached to marketed fruit, but they do not harbour the pathogen’s ambrosia beetle vector, Xyleborus glabratus. Thus, commerce in avocado fruit appears to be a negligible risk for expanding the geographic range of laurel wilt.     

Dieback of Passion Fruit in Surinam

In Surinam, the commercial cultivation of the yellow passion fruit (Passiflora edulis f. flavicarpa) is difficult due to the occurrence of dieback. Symptoms referred to as dieback include a decrease in elongation of the shoot end internodes after a period of normal growth leading to wilting and death of the shoots. Fruits from plants showing dieback symptoms are much smaller than those from healthy plants. From shoots with dieback symptoms, three fungi were isolated including Colletotrichum gloeosporioides. However, inoculation experiments with these fungi on shoots of vigorously growing plants were negative, even after wound inoculation. It appeared that plants with dieback symptoms had a poorly developed root system, From these roots Fusarium solani was isolated, which appeared to be highly pathogenic to roots of the yellow passion fruit. After inoculation of the roots of 3-month-old plants, roots became infected and the aerial plant parts showed typical dieback symptoms. Plants with their root system reduced either by inoculating with F. solani or by clipping, and subsequently inoculated with C. gloeosporioides on the aerial parts 2 weeks later, showed dieback symptoms and infection by C. gloeosporioides in shoots with these symptoms. Thus, a badly functioning root system, for example caused by infection of F. solani leads to dieback and predisposes plants to infection by C. gloeosporioides. The latter fungus itself is not a primary pathogen of shoots of the yellow passion fruit in Surinam.     

The antagonistic action of Trichoderma harzianum strain DGA01 against anthracnose-causing pathogen in mango cv. ‘Carabao’

The mechanisms of control and efficacy of Trichoderma harzianum strain DGA01 against anthracnose-causing pathogen Colletotrichum gloeosporioides in mango cv. ‘Carabao’ were examined. The action of DGA01 towards C. gloeosporioides was mycoparasitism and production of metabolites. DGA01 parasitised the pathogen by coiling its mycelia and spores on both artificial media and mango fruit surfaces. DGA01 was a parasitic necrotroph capable of killing C. gloeosporioides in 14 days of coexistence in artificial media. Dipping fruit in conidial suspension (10⁶?mL?L^?1) of DGA01 significantly decreased the incidence of anthracnose as compared to untreated fruit. Reduction in anthracnose severity was 87.90% showing high antagonistic potential of DGA01 in vivo.     

First Report of Anthracnose Disease on Jatropha curcas Caused by Colletotrichum gloeosporioides in Korea

In the summers of 2010 and 2011, an anthracnose disease was observed on the Jatropha curcas L. grown at the research field of Gyeongsangnam‐do Agricultural Research and Extension Services, South Korea. The symptoms included the appearance of dark brown spots on the leaf and fruit and the mummification of the fruit. The causal fungus formed grey to dark grey colony on potato dextrose agar. Conidia were single celled, ovoid or oblong, and 8–15 × 3–5 μm in size while seta was dark brown, cone‐shaped and 25–46 × 2–6 μm in size. The optimum temperature for growth was approximately 30°C. On the basis of mycological characteristics, pathogenicity test and molecular identification using internal transcribed spacer rDNA sequence, the fungus was identified as Colletotrichum gloeosporioides. To our knowledge, this is the first report of an anthracnose caused by C. gloeosporioides on J. curcas plant in Korea.     

Cellulase gene expression in ripening avocado fruit: The accumulation of cellulase mRNA and protein as demonstrated by cDNA hybridization and immunodetection

Summary A cDNA library was constructed from poly(A)⁺RNA of ripe avocado fruit. Colony hybridization identified a number of ripening specific clones of which one, pAV5, was shown to be specific for cellulase. Hybrid selection with pAV5 provided a message from ripe fruit that on in vitro translation yielded a polypeptide of 53kD, comigrating with purified avocado cellulase on SDS polyacrylamide gel electrophoresis. The translation product was selectively immunoprecipitated by antiserum to purified avocado cellulase. Immunoblotting of unripe and ripe avocado fruit extracts following SDS-PAGE showed a plentiful immunoreactive polypeptide in ripe fruit, and essentially none in unripe fruit. Hybridization of pAV5 to poly(A)⁺-RNA from unripe and ripe avocado fruit demonstrated that there is at least a 50-fold increase in the cellulase message concentration during ripening. Thus, the expression of cellulase enzyme activity during ripening is regulated by the appearance of mRNA coding for cellulase rather than by either translational or post-translational control mechanisms.Abbreviations poly(A)⁺ polyadenylated - DS sodium dodecyl sulfate - D kilodalton - bp base pairs Supported by Research Grant GM 19807 from the United States Public Health Service and by additional funds from the University of California Research Council.     

Response of sucrose metabolizing enzyme activity to CPPU and p-CPA treatments in excised discs of muskmelon

We investigated the effects of 1-(2-chloro-4-pyridyl)-3-phenylurea(CPPU) and para-chlorophenoxyacetic acid(p-CPA) treatments on the sucrose metabolism-relatedenzymeactivities in excised mesocarp discs of muskmelon fruit at different growthstages. Both a CPPU and a p-CPA treatment applied to discsprepared at 5 and at 20 days after anthesis (DAA) increased acid invertase (AI)activity and neutral invertase (NI) activity, but neither treatment affectedthese activities in the discs prepared at 45 DAA. Both plant growth substancesincreased the activity of sucrose phosphate synthase (SPS) in the discs at 5and20 DAA, but neither affected it in the 45 DAA discs. The sucrose synthase (SS)activity was markedly increased by p-CPA treatment in the20 and 45 DAA discs, but was not affected at 5 DAA. CPPU treatment did notactivate SS of discs throughout the growth stage.     

Physiological characterisation of Colletotrichum gloeosporioides,the incitant of anthracnose disease of noni in India

Ten isolates of Colletotrichum gloeosporioides were collected from different noni growing areas of Tamil Nadu, Karnataka and Kerala in India and their pathogenicity was proved under glass house conditions. Effect of different pH levels, temperature, light intensity and media were tested against the growth of C. gloeosporioides under in vitro. The results indicated that the growth of C. gloeosporioides was maximum in pH range of 6.50–7.00 and temperature range of 25–30°C. Exposure of the fungus to alternate cycles of 12 h light and 12 h darkness resulted in the maximum mycelial growth of C. gloeosporioides compared to the 24 h exposure to continuous light and 24 h exposure to continuous dark. Among the different media tested, host leaf extract medium supported significantly the maximum growth of all the 10 isolates of C. gloeosporioides followed by potato dextrose agar. Further, the strains were found to vary morphologically between the isolates under the study.     

The Influences of Epicuticular Wax Disruption and Cutinase Resistance on Penetration of Tomatoes by Colletotrichum gloeosporioides

Conidia of Colletotrichum gloeosporioides germinated on green and ripe tomato fruit with intact epicuticular wax, and formed penetration pegs below melanized appressoria. If the delicate layer of epicuticular wax was distupted by abrasion or removed by solvents before inoculation, apparent increased diffusion of fruit substances into the inoculum stimulated fungal growth, hyphal anastomosis and the production of penetration pegs from hyaline appressoria. This was followed by cutlcle erosion centred on the penetration pegs in green fruit allowing sec-ondary growth of infection hyphae. Due to the development of cutinase resistance when the cuticle became yellow at ripening, no cuticle erosion occurred at penetrations on ripe fruit Since cuticle erosion followed penetration of the cutinase-susceptible cuticle and since penetration peg formation was not hindered by the cutinase cuticle, the process of primary penetration is regarded as mechanical.