Relation between catalase activity and fungal survival in lemon fruit infected by Phytophthora citrophthora

In lemon fruit inoculated with Phytophthora citrophthora, a continuous increase in H₂O₂ during the incubation and rot development periods was found at the infection site; seven to ten days after inoculation of the fungus, the mycelium and fruit cells were dead. The suggestion is made that the increase in H₂O₂ may be related to the decrease in catalase activity and be responsible for the death of the fungus and fruit cells.The effect of different concentrations of peroxide on the fungus in-vitro was tested.Contribution from the Volcani Center, Agricultural Research Organisation, Bet Dagan, Israel. 1974 Series, No. 104-E. Division of fruit and Vegetable Storage, Bet Dagan 50 200, Israel/     

Brown spot disease of citrus caused by Phaeoisariopsis sp.

Brown spot disease of Citrus spp. was shown to be caused by Phaeoisariopsis sp. It severely affects all varieties of sweet orange, tangerine, grapefruit, lime and rough lemon, smooth lemon being relatively resistant. Fruits and leaves are much more susceptible than stems on which symptoms are rare. On the young fruit circular, slightly sunken, brown necrotic lesions are often associated with a surrounding ring of raised epicarp, giving the fruit a blistered appearance. Lesions on older fruits are usually flat and brown and surrounded by yellow haloes. Premature abscission, especially of young fruits, is common. Leaf symptoms start as greenish yellow patches and a fully formed leaf spot consists of light brown or greyish centre which is surrounded by a dark brown margin bordered by a yellow halo. Generalised foliar chlorosis, caused by coalescence of several lesions, culminates in premature defoliation. During wet weather, centres of both fruit and foliar lesions sporulate and become black. Stem lesions, dark brown and mostly occurring as extensions of petiole lesions, may coalesce causing stem die-back or resulting in the formation of corky internodal lesions. In artificial culture, the fungus sporulated only on media containing an extract of citrus leaf or fruit peel. The characteristics of the fungus mostly based on examination of structures formed on naturally infected parts of the plant, are described. This is probably the first report of a Phaeoisariopsis sp. on citrus.     

Oviposition behaviour and larval development of Anastrepha fraterculus from Argentina in citrus

Citrus peel physicochemical attributes are considered the main components conferring partial or even total resistance to fruit fly (Diptera: Tephritidae) infestation. Fruit fly females adapt their ovipositional strategies to overcome such resistance. Here, we explored the effects of citrus species (Rutaceae) on the ovipositional behaviour of the South American fruit fly, Anastrepha fraterculus (Wiedemann), and on its immature development. Particularly, we investigated the effects of (1) citrus species on oviposition behaviour and immature development, (2) citrus species on oviposition preference and on the location of the eggs at different depth in the citrus peel, and (3) harvest season and post‐harvest storage time on oviposition behaviour and immature development in lemon. Citrus species influenced ovipositional behaviour and affected survival of immature stages. Females laid eggs in lemon [Citrus limon (L.) Burm.], orange [Citrus sinensis (L.) Osbeck], and grapefruit (Citrus paradisi Macfadyen). In orange and lemon, larvae were found dead close to the oviposition areas, suggesting chemically mediated resistance mechanisms. Under choice conditions, females preferred grapefruit over lemon and bigger clutches were found in the layers where embryonic development is favoured. Unsuitability of lemon as a medium to complete development was neither affected by harvest season nor by storage time of the fruit after harvest. The physical and chemical characteristics of the peel were distinctive to each citrus species and may have affected the specific levels of resistance of these citrus species to infestation by A. fraterculus.     

Evaluation of commercial essential oil samples on the growth of postharvest pathogen Monilinia fructicola (G. Winter) Honey

Aim: To assess the effect of several commercial essential oils samples Australian lemon myrtle (Backhousia citriodora), cinnamon bark (Cinnamomum zeylanicum), oregano (Origanum vulgare), thyme oil (Thymus vulgaris), clove bud (Eugenia caryophyllata), valerian (Valeriana officinalis) and Australian tea tree oil (Melaleuca alternifolia) on mycelium growth and spore germination of Monilinia fructicola. The effectiveness of lemon myrtle essential oil as a fumigant for the control of brown rot in nectarines was evaluated. Methods and Results: Monilinia fructicola exhibited a different level of sensitivity to each tested essential oil with results suggesting that the essential oils provide excellent control of the pathogen with respect to mycelium growth and spore germination at very low concentrations, whereas for others higher concentrations are needed to reduce significant fungal growth. In vivo application of lemon myrtle essential oil effectively reduced the incidence of M. fructicola on noninoculated fruit. Fumigation of nectarines following inoculation did not reduce the incidence of brown rot in comparison with the inoculated control treatment. No evidence of phytotoxicity on the fruit was recorded. Conclusions: Lemon myrtle essential oil exhibited the strongest antifungal activity against M. fructicola, in vitro and to a lesser extent, under in vivo conditions. Significance and Impact of the Study: The results demonstrate that lemon myrtle essential oil, in particular, has potential as an antifungal agent to control M. fructicola.     

Control of green mold and sour rot of stored lemon by biofumigation with Muscodor albus

Control of postharvest lemon diseases by biofumigation with the volatile-producing fungus Muscodor albus was investigated. In vitro exposure to M. albus volatile compounds for 3 days killed Penicillium digitatum and Geotrichum citri-aurantii, causes of green mold and sour rot of lemons, respectively. Lemons were wound-inoculated with P. digitatum and placed in closed 11-L plastic boxes with rye grain cultures of M. albus at ambient temperature. There was no contact between the fungus and the fruit. Biofumigation for 24–72 h controlled green mold significantly, even when treatment began 24 h after inoculation. Effectiveness was related to the amount of M. albus present. In tests conducted inside a 11.7-m³ degreening room with 5 ppm ethylene at 20 °C, green mold incidence on lemons was reduced on average from 89.8 to 26.2% after exposure to M. albus for 48 h. Ethylene accelerates color development in harvested citrus fruit. M. albus had no effect on color development. Biofumigation in small boxes immediately after inoculation controlled sour rot, but was ineffective if applied 24 h later. G. citri-aurantii may be less sensitive to the volatile compounds than P. digitatum or escapes exposure within the fruit rind. Biofumigation with M. albus could control decay effectively in storage rooms or shipping packages.     

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.     

Construction and characterization of a cloned probe for the detection of Phoma tracheiphila in plant tissues

Summary The construction and characterization of the recombinant clone, pPhoB25 is described. This clone contains an highly repeated sequence of DNA from the fungus Phoma tracheiphila, a pathogen of the lemon. The specificity of the probe was determined by molecular hybridization with the DNA of the host plant and with those of several fungi which can be found associated with the lemon plants. The clone was used successfully as a specific and sensitive probe for the detection of the pathogen in infected plant tissues by dotblot hybridization under laboratory and field conditions.     

Temperature, soluble solids and pH effect on Alicyclobacillus acidoterrestris viability in lemon juice concentrate

Alicyclobacillus acidoterrestris is a thermoacidophilic, non-pathogenic, spore-forming bacterium detected in spoiled commercial pasteurized fruit juice. Apple, white grape and tomato are particularly susceptible. A. acidoterrestris spores are resistant to lemon juice pasteurization (2 min at 82°C), and they can germinate and grow causing spoilage. This contamination is characterized by a medicinal or disinfectant smell attributed to guaiacol (o-dihydroxybenzene) production and other taint chemicals. The aim of this work was to study the influence of temperature (82, 86, 92 and 95 °C), total soluble solids (SS) (6.20, 9.8, 50 and 68°Brix) and pH (2.28, 2.45, 2.80, 3.25, 3.5) on decimal reduction time (D) of the A. acidoterrestris in clarified and non-clarified concentrated lemon juice. Once D-value was determined, the resistance of A. acidoterrestris at the assayed temperatures was confirmed. SS and pH influence spore viability, because spore resistance increases with higher SS (50°Brix 22 min 82 °C–68°Brix 28 min 82 °C) and pH values (pH 2.28, 17 min–pH 4.00, 22 min). Bacterial growth was lower in clarified lemon juice, 26 min at 82 °C, than in non-clarified lemon juice, 51 min at 82 °C. Temperature was the parameter that had the greatest influence on the D value.     

Patulin and aflatoxin in brown rot lesion of apple fruits and their regulation

Aspergillus flavus, A. niger, Penicillium expansum and Rhizopus stolonifer were the most frequently isolated fungi from healthy apple fruits. Alternaria alternata was the most common organism of rotten apple fruits, followed by A. niger, A. flavus, P. expansum and R. stolonifer. The prevalent type of decay, brown rot lesion, is caused by R. stolonifer followed by A. flavus, A. niger, A. alternata and P. expansum. Sodium hypochlorite had good curative properties against fruit rots. The main natural mycotoxins produced in rotten apple were patulin and aflatoxins. The optimum temperature for patulin production by P. expansum was 15 °C after 15 days. Complete inhibition of patulin formation was attained using 0.2% lemon oil and > 90% inhibition using 0.05% lemon and 0.2% orange oils. Also significant inhibition (> 90%) of aflatoxin production was observed with 0.2% lemon oil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Induction of serotonin biosynthesis is uncoupled from the coordinated induction of tryptophan biosynthesis in pepper fruits (<Emphasis Type="Italic">Capsicum annuum</Emphasis>) upon pathogen infection

It has been suggested that serotonin biosynthesis is regulated by tryptophan decarboxylase (TDC) in plants. To determine if TDC plays a rate-limiting role in serotonin biosynthesis, two TDC genes, PepTDC1 and PepTDC2, were cloned from pepper (Capsicum annuum L.) fruits infected with anthracnose fungus and their expression was then examined in various organs, including fruit that had been treated with the fungus or various chemicals. PepTDC1 expression was highly induced in pepper fruits after treatment with fungus and ethylene, while PepTDC2 was constitutively expressed at low levels in all pepper tissues. Additionally, predominant induction of PepTDC1 mRNA and TDC enzyme activity was detected in the unripe-green fruit, but not in the ripe-red fruit upon pathogen infection. Higher expression of TDC in unripe-green fruit was closely associated with increased levels of tryptamine, serotonin, and serotonin derivatives. However, unlike the enhanced serotonin synthesis, tryptophan levels responded unchanged when challenged with the pathogen in both the unripe-green fruit and the ripe-red fruit. Expression of two key tryptophan biosynthetic genes, anthranilate synthase (ASα) and tryptophan synthase (TSβ), remained unchanged in response to treatment. Also, anthranilate synthase enzyme activity remained steady regardless of pathogen infection. Taken together, these results suggest that the synthesis of serotonin was regulated by the induction of TDC without a simultaneous increase in tryptophan levels in pepper fruits.     

A thaumatin-like gene in nonclimacteric pepper fruits used as molecular marker in probing disease resistance,ripening, and sugar accumulation

During pepper (Capsicum annuum) fruit ripening, the ripe fruit interaction with the anthracnose fungus, Colletotrichum gloeosporioides, is generally incompatible. However, the unripe fruit can interact compatibly with the fungus. A gene, designated PepTLP (for pepper thaumatin-like protein), was isolated and characterized by using mRNA differential display. The PepTLP gene encodes a protein homologous to other thaumatin-like proteins and contains 16 conserved cysteine residues and the consensus pattern of thaumatin. PepTLP gene expression is developmentally regulated during ripening. The accumulation of PepTLP mRNA and PepTLP protein in the incompatible interaction was higher than that in the compatible one. Furthermore, PepTLP gene expression was stimulated by both jasmonic acid treatment and wounding during ripening, but by wounding only in the unripe fruit. Immunolocalization studies showed that it is localized to the intercellular spaces among cortical cells. The expression of the PepTLP gene upon fungal infection was a rise from the early-breaker fruit. The development of anthracnose became significantly prevented with beginning of fruit ripening, and the sum total of sugar accumulation increased. The results suggest that the PepTLP gene can be used as a molecular marker in probing for disease resistance, ripening, and sugar accumulation in nonclimacteric pepper fruits.     

Volatile cues used by the parasitoid, Aphytis melinus, for host location: California red scale revisited

Aphytis melinus DeBach (Hymenoptera: Aphelinidae) is a biological control agent of diaspidid scale insects. The parasitoid has a narrow host range but its hosts are polyphagous. We determined the source of volatile cues the wasp uses to locate its few host species when those hosts occur on more than one host plant species. We addressed four questions in regard to the use of volatile cues in host location of California red scale, Aonidiella aurantii (Maskell) (Homoptera: Diaspididae): (1) Does A. melinus use volatile cues to assist in host location? (2) Are these cues innately recognized or learned? (3) Are cues produced by female California red scale, or from other sources? (4) Are the cues specific to the host or host plant? These questions were tested through the use of a Y-tube olfactometer. Female A. melinus used volatile cues to orient toward both infested and uninfested host plant material. Wasps learned these cues by associating odors from the host plant with host presence. They had no innate preferences for scale insect or host plant volatile stimuli. Contrary to previous studies, we found no evidence of orientation toward the female-produced sex pheromone of California red scale, nor to volatile cues from the attacked host stage. Wasps given experience with scale insects growing on lemon fruit subsequently oriented toward lemon and orange fruit and leaves. The scale species with which the wasp was given experience did not affect this preference. Wasps given experience with California red scale growing on squash did not orient toward infested lemon fruit. The host ranges of the parasitoid and its hosts are used to explain the adaptive value for the evolution of learned rather than fixed responses to cues used in foraging behavior.     

Acid fast-green staining as a cytological indicator of chromosomal activity in interphase nuclei of lemon fruit explants

Summary

The bodies along the chromatin strands as well as the nucleoli of interphase nuclei of freshly excised lemon fruit tissue showed a far stronger positive staining reaction with acid fast-green than was observed previously with alkaline fast-green-stained material. The enlarged interphase nucleoli of 48-hour-old explants behaved similarly. Evidence indicates that the apparent absence of the fast-green staining bodies from the chromatin strands of interphase nuclei having enlarged nucleoli is a cytological indication of chromosomal activity in lemon fruit explants responding to new environmental situations following excision from the fruit.     

Freckle disease of banana in Hawaii caused by Phyllostictina musarum (Cke.) Petr

‘Freckle’ (‘black-spot’ disease) of bananas is common on leaves and fruit of Dwarf Cavendish and other varieties in Hawaii, especially after rainy periods. On fruit, symptoms may appear 2–4 weeks after the bunch has opened, and become more severe as maturity is approached. The disease is usually confined to older leaves on affected plants. Freckled tissue contains numerous pycnidia of Phyllostictina musarum and disease was experimentally induced by inoculating leaves and fruit with conidia of this fungus. This appears to be the first record of successful inoculation with P. musarum. Conidia of P. musarum germinate after 3–6 h in a film of water on banana peel, appressoria being formed after 18–30 h. Penetration of the epidermis occurs 24–96 h after inoculation, and is brought about by an infection hypha which grows from the appressorium. The progressive increase in severity of freckle as fruit matures is due to repeated infection by further conidia of P. musarum, rather than to enlargement of original infections. Some banana clones, including Gros Michel, appear to be resistant to the fungus Dispersal of P. musarum conidia immediately after discharge from the pycnidium is chiefly by rainwater and dew. Secondary infections contribute greatly to the total number of infections. Conidium dispersal by water often results in the development of characteristic patterns of spotting, chiefly in the form of streaks or circular areas, coinciding with the directions of movement of rainwater and dew. Large numbers of conidia of P. musarum are washed on to fruit in rainwater and dew running from diseased, overhead leaves.     

First steps towards biological control of the pear gall midge (Contarinia pyrivora) with the insect pathogenic fungus Metarhizium brunneum

Gall midges are important pests in many crops. In fruit, they are difficult to control due to their life cycle, which takes place partially within the fruit. Here, we provide the first successful laboratory experiment to infect pear gall midge (Contarinia pyrivora) with the insect pathogenic fungus Metarhizium brunneum. We developed a procedure for sampling larvae, maintaining them in the laboratory and subjecting them to the fungus. We demonstrated that dipping larvae in a fungus suspension or adding a fungus suspension to the soil result in significant fungus induced mortality of the pear gall midge. An immune response in treated larvae was recorded proving that there was a real pathogenesis. Finally, we discuss next steps and a strategy for field experiments.     

Spatial and Temporal Variation in Chaetanaphothrips orchidii Moulton (Thysanoptera: Thripidae) Population and Its Damage on Lemon

Chaetanaphothrips orchidii Moulton has recently been detected in lemon (Citrus limon) orchards in northwest Argentina, causing high levels of damage on fruits. Severe damage results in the rejection of fruit for export, which must then be sold in the industry. However, the restrictions imposed by the citrus industry on insecticide residues sometimes also result in fruit rejection. Here, we studied the ecology and behavior of C. orchidii in order to propose a pest management strategy that could meet both export and industry demands. Seasonal occurrence and canopy distribution of C. orchidii in lemon orchards were evaluated, and field experimental manipulations of thrips populations were performed to analyze how the length (45, 100, 130, and 200 days) and timing (January, February, or March) of C. orchidii activity related with fruit damage. Lemons harvested during summer showed lower infestation levels (～0.64 individual per fruit) than those harvested in winter (～1.88 individuals per fruit). Higher proportions of damaged fruits were recorded in the lower part of the tree. Changes in the population levels of C. orchidii were closely associated with fruit phenology. The longer the lemon fruits were exposed to the thrips, the higher was the damage. However, the time of infestations did not affect fruit damage. Our data provide a first step towards understanding the factors that determine the severity of fruit damage caused by C. orchidii in northwest Argentina.     

Engineering d‐limonene synthase down‐regulation in orange fruit induces resistance against the fungus Phyllosticta citricarpa through enhanced accumulation of monoterpene alcohols and activation of defence

Terpene volatiles play an important role in the interactions between specialized pathogens and fruits. Citrus black spot (CBS), caused by the fungus Phyllosticta citricarpa, is associated with crop losses in different citrus‐growing areas worldwide. The pathogen may infect the fruit for 20–24 weeks after petal fall, but the typical hard spot symptoms appear when the fruit have almost reached maturity, caused by fungal colonization and the induction of cell lysis around essential oil cavities. d ‐Limonene represents approximately 95% of the total oil gland content in mature orange fruit. Herein, we investigated whether orange fruit with reduced d ‐limonene content in peel oil glands via an antisense (AS) approach may affect fruit interaction with P. citricarpa relative to empty vector (EV) controls. AS fruit showed enhanced resistance to the fungus relative to EV fruit. Because of the reduced d ‐limonene content, an over‐accumulation of linalool and other monoterpene alcohols was found in AS relative to EV fruit. A global gene expression analysis at 2 h and 8 days after inoculation with P. citricarpa revealed the activation of defence responses in AS fruit via the up‐regulation of different pathogenesis‐related (PR) protein genes, probably as a result of enhanced constitutive accumulation of linalool and other alcohols. When assayed in vitro and in vivo, monoterpene alcohols at the concentrations present in AS fruit showed strong antifungal activity. We show here that terpene engineering in fruit peels could be a promising method for the development of new strategies to obtain resistance to fruit diseases.     

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.