Role of Antifungal Gallotannins,Resorcinols and Chitinases in the Constitutive Defence of Immature Mango (Mangifera indica L.) against Colletotrichum gloeosporioides

Conidia of Colletotrichum gloeosporioides germinate and form infection hyphae on inoculated, immature mango but remain quiescent until fruit ripening. Antifungal resorcinols have previously been implicated for quiescence of C. gloesoporioides and Alternaria alternata on mango. This study revealed the presence of a mixture of several gallotannins with glycosidic linkages, including 1,2,3,4,6‐penta‐O‐galloyl‐β‐D‐glucopyranose, with significant antifungal activity in the unripe mango fruit peel. Gallotannin antifungal activity was greater in a cultivar resistant (295.8 mm² inhibition) to anthracnose than in a susceptible (148.4 mm² inhibition) cultivar. In both, the activity decreased with ripening but the decrease was 10% less in the resistant cultivar. Three recorcinols, 5‐pentadecylresorcinol, 5‐(12‐cis‐heptadecenyl)resorcinol, AR 21 and another resorcinol derivative were present in the unripe fruit peel and all declined during ripening, more significantly the 5‐(12‐cis‐heptadecenyl)resorcinol and AR 21. Mango latex, when drained out, separates into an oily and aqueous phase. The aqueous phase showed significant chitinase activity and the ability to digest conidia of C. gloeosporioides. The oily phase has previously been reported to contain resorcinols. Draining fruits of latex soon after harvest resulted in greater incidence and severity of anthracnose at ripe stage. Chitinase activity was less in the peel of fruits from which latex was drained. The evidence suggests that the resistance of unripe mango to C. gloeosporioides is because of an elaborate constitutive defence system comprising antifungal resorcinols, gallotannins and chitinases.     

A method for pre-harvest assessment of latent infections in fruits

A method was developed for the assessment of latent infections of Alternaria alternata in mango fruits. Fruit discs were sampled and assayed for fungal contamination under suitable growth conditions. By this method it was possible to evaluate the percentage of infections and the relative surface infected by Alternaria in mango fruits before harvest. The percentage of infections increased during the growing season but fungicide treatments prevented this increase. A significant positive correlation was found between the relative surface infected by latent Alternaria at harvest time and the incidence of black spots that developed on the fruit during storage.     

Utilization of Caffeic Acid Phenethyl Ester to Control Alternaria alternata Rot in Tomato (Lycopersicon esculentum Mill.) Fruit

Chemical fungicides that are related with resistant strains develop negative effects on human health and environment. Propolis is a resinous substance collected by bees with positive effects on human health and inhibitory activity against Alternaria alternata. Caffeic acid phenethyl ester (CAPE) is a component of the propolis. The objective of this experiment was to test the effect of CAPE on fungi infecting tomato fruit using as a model the pathosystem A. alternata‐tomato. CAPE was chemically synthesized in our laboratory and analysed with nuclear magnetic resonance spectroscopy. Different concentrations (0, 16, 32, 48, 64, 80, 90 and 100 μm ) of CAPE were tested on A. alternata growing in vitro. For the in vivo experiment, red ripe tomato fruit was inoculated with A. alternata and untreated or treated with 1, 50 and 100 μm of CAPE. After that, the fruit was stored at 25°C for up to 20 days. Colony size (CS) was recorded in vitro. In tomato fruits, the severity of infection (SI), respiration rate (RR), ethylene production (EP), pH, total soluble solids (TSS), weight loss (WL) and titratable acidity (TA) were evaluated during the storage time. CAPE melting point and spectral data probed to be the right molecule. In vitro, 64 and 100 μm of CAPE reduced CS by 30%. In vivo, 50 and 100 μm of CAPE reduced SI higher than the fungicide Captan^® with no effects on RR, EP, WL, pH, TSS and TA. It was concluded that CAPE controls A. alternata infection better than a commercial fungicide without negative effects on tomato fruit ripening and fruit quality.     

Population fluctuations and interspecific competition between Tephritid flies attacking fruit crops in the New Valley oases,Egypt

Population fluctuations of the Mediterranean fruit fly Ceratitis capitata (Wiedemann) and the peach fly Bactrocera zonata (Saunders) were monitored with lure trap collections in three provinces in the New Valley oases, Western Desert, Egypt. Results showed marked temporal differences in peak trap catches of the two flies in the selected sites all over the entire studied areas. One annual peak of C. capitata was recorded during both October 2005 and February 2006 and coincided with the ripening period of citrus trees in Kharga oases. However, two annual peaks were recorded during June and September 2005 in Bodkholow province and coincided with the ripening period of apple and mango. On the other hand, two annual peaks of B. zonata were recorded in Kharga oases throughout May and September and coincided with the ripening periods of apricot, mango and guava. One annual peak only was recorded in the round up of September and/or October in both Moot and Bodkholow in Dakhla oases and coincided with the ripening period of mango, guava and citrus. The occurrence of C. capitata was very limited in comparison with B. zonata. Population fluctuations of the two pests in the studied sites were significantly different. The ability of the traps used in capturing both C. capitata and B. zonata indicated that the yellow sticky trap was more effective in capturing C. capitata. However, the Abdel-Kawi trap was significantly efficient at trapping B. zonata. The occurrence of B. zontata in high numbers all over the study period compared to C. capitata is considered as good proof that this invading fly may be considered as a vigorous competitive tephritid fly to the native fly C. capitata.     

Mango stem end rot pathogens - Fruit infection by endophytic colonisation of the inflorescence and pedicel

The stem end rot pathogens of mango (Mangifera indica), (Dothiorella dominicana, Dothiorella mangiferae, Lasiodiplodia theobromae (Syn. Diplodia natalensis Phomopsis mangiferae, Cytosphaera mangiferae, Pestalotiopsis sp. and Dothiorella‘long’), as well as other fungi (including Alternaria alternata), were found to occur endophytically in the stem tissue of mango trees prior to inflorescence emergence. On samples from trees with a record of low stem end rot levels, colonisation did not extend into the most recently produced flush of stem tissue. At a site with a history of high stem end rot levels, sequential monitoring of inflorescence tissue between flowering and harvest by plating out small (c. 8 mm³) tissue pieces revealed, that at least some of the pathogens - Dothiorella spp., P. mangiferae, Pestalotiopsis sp. and C. mangiferae gradually colonised the inflorescence, reaching the pedicel tissue of young fruit - 8 wk after flowering. Subsequently, detection frequency of the pathogens in inflorescence tissue declined, possibly because of interference from copper residues (from field sprays) accumulating on tissue samples. The detection frequency of A. alternata also increased as Dothiorella spp. declined, however these changes could not be attributed to antagonistic interactions between the two fungi. Using larger tissue pieces (1–2 mm thick transverse sections, or a square of tissue 25 mm²× 3 mm thick) in isolations, endophytic colonisation by Dothiorella spp. and P. mangiferae was detected in stem, inflorescence and pedicel tissues of mature-fruit-specimens from two different sites, one unsprayed, and the other regularly sprayed with copper. The fungi were detected more frequently in the samples from unsprayed trees. Fruit from the sprayed orchard subsequently developed a high level of stem end rot caused by D. dominicana, while a lower level of stem end rot developed in unsprayed fruit, possibly because the latter fruit were also extensively diseased by anthracnose (Colletotrichum gloeosporioides Penz.). Endophytic colonisation of inflorescence and pedicel tissue was found to be a primary route of infection for fruit which develop stem end rot during ripening.     

Physical postharvest treatments in the control of stem‐end rot of mango

Botryosphaeria dothidea is the major pathogen of mango in Brazil, causing stem‐end rot, which causes significant losses during transportation and storage. The current strategy to control this particular disease using synthetic fungicides has been ineffective, leaving residues in the fruit. The objective of the research was to study the effect of physical treatments, with hot water rinse brushing (HWRB) and ultraviolet C irradiation (UV‐C), individually and in combination, to control stem‐end rot of mango. Physicochemical parameters, respiration and resistance induction of the fruit were also analysed. The in vitro trials demonstrated that B. dothidea is a thermoresistant fungus. The individual treatments with HWRB at 65°C for 15 s and 2.5 kJ/m² of UV‐C presented the best results, showing less symptoms of the disease during 18 days of storage. The combination of HWRB with UV‐C did not improve the control of the disease when compared to the treatments applied individually. The physicochemical parameters and the consumer acceptance evaluation showed that both physical treatments preserved the appearance of the fruit and delayed the ripening–senescence process. The induction of defence‐related enzymes revealed that induced resistance was an important mechanism involved in the control of stem‐end rot of mango.     

Identification of an ETR1-homologue from mango fruit expressing during fruit ripening and wounding

We have isolated a mango (Mangifera indica L.) cDNA homologue of the ethylene receptor gene ETR-1, referred to as METR1, which codes for a polypeptide of 802 amino acids with a predicted molecular weight of 89 kDa. The amino acid sequence is highly homologous (over 80 percnt;) to ETRs from other fruits. Genomic Southern blot analysis indicates that two or more ETR homologues exist in mango. RNA blot analysis revealed that the level of METR1 mRNA in the mesocarp increased during fruit ripening. In addition, it was found that the METR1 mRNA increases transiently during wounding of the tissue. This is the first report of an ETR homologue showing an induction during fruit ripening and wounding.     

5-(12-heptadecenyl)-resorcinol,the major component of the antifungal activity in the peel of mango fruit

A mixture of5-(12-cis-heptadecenyl)- and 5-pentadecyl-resorcinol was isolated from the peel of mango fruit; it acts as a preformed agent against Alternaria alternata, a fungus responsible for the black spot disease of mango fruits in Israel. The chemical characterization was done by ¹HNMR, ¹³CNMR and MS.     

Effect of Fungal Infection and Temperature on Physiological Changes in Banana Fruit

An attempt was made to investigate which factor temperature or fungal infection, has a greater effect in accelerating physiological processes such as respiration and ethylene evolution in fruit. For this study the effect of infection with Trichothecium roseum, Alternaria alternata and Diplodia natalensis on banana fruit, at 14°C and 20°C was examined. The results show that the development of rot caused by different fungi and the physiological changes in the infected fruit may depend not only on the effect of temperature on fungi (as in vitro) but also on fruit ripening, or on processes connected with fruit ripening, which may accelerate fungal development.     

Aflatoxins in sunflower seeds: Influence of Alternaria alternata on aflatoxin production by Aspergillus parasiticus

The aim of the present work was to determine the influence of Alternaria alternata upon aflatoxin production by Aspergillus parasiticus.A mixture of spores of both strains was inoculated in sunflower seeds at 0,90 a_w, and incubated for 42 days at 28 °C ±1.The cultures were observed and analyzed every 7 days to determine the infection level of the seeds and the production of aflatoxins. Results showed that when the seeds were inoculated only with Aspergillus parasiticus, 100% were infected from the 7th day.When Aspergillus parasiticus and Alternaria alternata were simultaneously inoculated the infection level of the seeds was 100% for Aspergillus parasiticus following 7 days of inoculation and 0% for Alternaria alternata. After the 14th day of inoculation there was no significant difference in the infection percentage of both strains (approximately 80% of each one). As far as toxin production is concerned a remarkable decrease was observed when seeds were inoculated with both strains simultaneously.In accordance to the results, Alternaria alternata would not compete with Aspergillus parasiticus in colonization of seeds but would either degrade the aflatoxins by Aspergillus parasiticus or compete for aflatoxin biosynthesis precursors. Alternaria alternata could also secrete some substance that specifically inhibits aflatoxin synthesis.     

Effect of storage conditions on virulence of Fusarium avenaceum and Alternaria alternata on apple fruits

To overcome difficulty in phytopathogenic fungi control during storage of apple fruits, the effect of different storage conditions on the occurrence and development of Fusarium avenaceum and Alternaria alternata infections on apple cultivar “Cripps Pink” was investigated during and after storage. Inhibitory effects of wild oregano essential oil on apple fruit rots caused by F. avenaceum and A. alternata were also tested as possible rot control measure. Artificially inoculated apple fruits were kept in cold storage with normal (NA) and controlled (CA) atmosphere for 95 days and at room temperature only. The obtained results indicated that different storage conditions significantly affect necrosis development on apple fruits caused by F. avenaceum and A. alternata after storage, as well as during shelf life.     

Effect of environmental factors on tenuazonic acid production by Alternaria alternata on soybean-based media

Aims: To determine the effects of water activity (a_W; 0·995–0·90), temperature (5, 18, 25 and 30°C), time of incubation (7–35 days) and their interactions on tenuazonic acid (TA) production on 2% soybean‐based agar by two Alternaria alternata strains isolated from soybean in Argentina. Methods and Results: TA production by two isolates of A. alternata was examined under interacting conditions of a_W, temperature and time of incubation on 2% soybean‐based agar. Maximum TA production was obtained for both strains at 0·98 a_W, but at 30 and 25°C for the strains for RC 21and RC 39, respectively. The toxin concentration varied considerably depending on a_W, temperature, incubation time and strain interactions. TA was produced over the temperature range from 5 to 30°C and a_W range from 0·92 to 0·995, however at 5 and 18°C little TA was produced at a_W below 0·94. Contour maps were developed from these data to identify areas where conditions indicate a significant risk for TA accumulation. Conclusions: The optimum and marginal conditions for TA production by A. alternata on soybean‐based agar were identified. The results indicated that TA production by A. alternata is favoured by different temperatures in different strains. Significance and Impact of the Study: Data obtained provide very useful information for predicting the possible risk factors for TA contamination of soybean as the a_W and temperature range used in this study simulate those occurring during grain ripening. The knowledge of TA production under marginal or sub‐optimal temperature and a_W conditions for growth are relevant as improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality.     

Role of Brassinosteroids, Ethylene, Abscisic Acid, and Indole-3-Acetic Acid in Mango Fruit Ripening

Rapid ripening of mango fruit limits its distribution to distant markets. To better understand and perhaps manipulate this process, we investigated the role of plant hormones in modulating climacteric ripening of ??Kensington Pride?? mango fruits. Changes in endogenous levels of brassinosteroids (BRs), abscisic acid (ABA), indole-3-acetic acid (IAA), and ethylene and the respiration rate, pulp firmness, and skin color were determined at 2-day intervals during an 8-day ripening period at ambient temperature (21?±?1°C). We also investigated the effects of exogenously applied epibrassinolide (Epi-BL), (+)-cis, trans-abscisic acid (ABA), and an inhibitor of ABA biosynthesis, nordihydroguaiaretic acid (NDGA), on fruit-ripening parameters such as respiration, ethylene production, fruit softening, and color. Climacteric ethylene production and the respiration peak occurred on the fourth day of ripening. Castasterone and brassinolide were present in only trace amounts in fruit pulp throughout the ripening period. However, the exogenous application of Epi-BL (45 and 60?ng?g^?1 FW) advanced the onset of the climacteric peaks of ethylene production and respiration rate by 2 and 1?day, respectively, and accelerated fruit color development and softening during the fruit-ripening period. The endogenous level of ABA rose during the climacteric rise stage on the second day of ripening and peaked on the fourth day of ripening. Exogenous ABA promoted fruit color development and softening during ripening compared with the control and the trend was reversed in NDGA-treated fruit. The endogenous IAA level in the fruit pulp was higher during the preclimacteric minimum stage and declined during the climacteric and postclimacteric stages. We speculate that higher levels of endogenous IAA in fruit pulp during the preclimacteric stage and the accumulation of ABA prior to the climacteric stage might switch on ethylene production that triggers fruit ripening. Whilst exogenous Epi-BL promoted fruit ripening, endogenous measurements suggest that changes in BRs levels are unlikely to modulate mango fruit ripening.     

Occurrence and Latent Infection of Alternaria Rot of Pingguoli Pear (Pyrus bretschneideri Rehd. cv. Pingguoli) Fruits in Gansu, China

Alternaria rot of Pingguoli pear occurred after latent infection. Fruit surfaces were asymptomatic within 60 days storage under cold condition (0°C, RH 85–90%), but black‐grey hyphae could be seen in the lenticels or calyx tube of Pingguoli pear after 90 days of storage. The tissue collapsed and resulted in visible black spots as the hyphae spread over the fruit. Average incidence of Alternaria rot of fruits from an orchard in Gansu was 28.86% at 100 days of storage. The main fungus isolated from the Alternaria rot on stored Pingguoli pear was identified as Alternaria alternata (Fr. : Fr.) Keissl. This pathogen was able to initially infect the fruit via two pathways during the growing season, and then remain in a latent state. The fungus first colonized the styles at the full‐blossoming stage, and then grew into the carpel cavities progressively after 50 days from petal fall. The percentage latent infection of A. alternata was up to 45% in the carpel cavity until the harvest time. The fungus also attacked fruit surfaces and remained latent in the fruit peel during fruit development. The percentage of A. alternata latent infection at the calyx end, middle part and stem end of the fruit peel was 40%, 24% and 42.8%, respectively, at harvest time.     

Peroxisomal thiolase mRNA is induced during mango fruit ripening

Fruit ripening is a complex, developmentally regulated process. A series of genes have been isolated from various ripening fruits encoding enzymes mainly involved in ethylene and cell wall metabolism. In order to aid our understanding of the molecular basis of this process in a tropical fruit, a cDNA library was prepared from ripe mango (Mangifera indica L. cv. Manila). By differential screening with RNA poly(A)⁺ from unripe and ripe mesocarp a number of cDNAs expressing only in ripe fruit have been isolated. This paper reports the characterization of one such cDNA (pTHMF 1) from M. indica which codes for a protein highly homologous to cucumber, rat and human peroxisomal thiolase (EC 2.3.1.16), the catalyst for the last step in the -oxidation pathway.The cDNA for the peroxisomal mango thiolase is 1305 bp in length and codes for a protein of 432 amino acids with a predicted molecular mass of 45 532 Da. Mango thiolase is highly homologous to cucumber thiolase (80%), the only other plant thiolase whose cloning has been reported, and to rat and human thiolases (55% and 55% respectively).It is shown by northern analysis that during fruit ripening THMF 1 is up-regulated. A similar pattern of expression was detected in tomato fruit. Wounding and pathogen infection do not appear to affect THMF 1 expression. The possible involvement of thiolase in fatty acid metabolism during fruit ripening will be discussed. To our knowledge this is the first report cloning of a plant gene involved in fatty acid metabolism showing an induction during fruit ripening.     

Modulation of mango ripening by chemicals: physiological and biochemical aspects

During ripening, fleshy fruits undergo textural changes that lead to loss of tissue firmness and consequent softening due to cell wall dismantling carried out by different and specifically expressed enzymes. The effect of various chemical treatments on the ripening of mango fruit (Mangifera indica) was investigated at physiological and biochemical level. Based on changes in respiration, firmness, pH, total soluble sugar and a cell wall degrading enzyme pectate lyase (PEL) activity, treatment with 1-methylcyclopropene (1-MCP), silver nitrate (AgNO₃), gibberlic acid (GA₃), sodium metabisulphite (SMS) and ascorbic acid led to delaying of ripening process while those of ethrel and calcium chloride (CaCl₂) enhanced the process. PEL of mango was found to be inhibited by certain metabolites present in dialysed ammonium sulphate enzyme extract as well as EDTA. Mango PEL activity exhibited an absolute requirement for Ca²⁺ and an optimum pH of 8.5.     

Effects of 1-methylcyclopropene alone and in combination with polyethylene bags on the postharvest life of mango fruit

Experiments were conducted to determine how 1‐methylcyclopropene (1‐MCP) treatments influence ethylene‐stimulated ripening of harvested mango cv. Zihua fruit at 20°C. The ripening response of fungicide (prochloraz) treated fruit was characterised following various 1‐MCP treatments in sealed jars followed by storage in polyethylene bags and/or subsequent ethephon (ethylene) exposure. Exposure of fruit to increasing concentrations of 1‐MCP for 12 h resulted in the reduced softening of produce when subsequently held in air for 7 days after ethephon treatment. Application levels of between 1 and 100 μl litre^?1 1‐MCP had increasing impact, while 200 μl litre^?1 1‐MCP apparently began to approach response saturation. Exposure of fruit to 50 or 100 μl litre^?1 concentrations of 1‐MCP for periods from 1 to 24 h subsequently resulted in reduced softening of produce when held in air for 7 days after ethephon treatment. Increasing periods of exposure from 1 to 12 h had increasing impact, while exposure times greater that 12 h appeared to reach saturation. In the absence of ethephon‐stimulation, the natural ripening of mangoes held in polyethylene bags was delayed by prior exposure to 100 μl litre^?1 1‐MCP for 12 h. Extended holding of 1‐MCP treated and non‐1‐MCP treated control fruit in polyethyene bags encouraged physiological and pathological deterioration. Following exposure to 100 μl litre^?1 1‐MCP for 12 h, mango fruit held for 10 days in polyethylene bags showed a delay in the onset of ripening relative to bagged but non‐1‐MCP treated control fruit. Treatment with 1‐MCP allowed storage of mango fruit in plastic bags at 20°C for 30 days. Observations suggest that 1‐MCP treatments do not adversely influence the quality of the post‐storage ethephon‐ripened fruit. Thus, application of 1‐MCP in combination with the use of polyethylene bags can extend the postharvest life of mango fruit at ambient temperature. Treatments that extend postharvest life are important in developing countries, such as China, where the cold chain infrastructure is often lacking.