Non-Chemical Treatments for the Pre- and Post-Harvest Elicitation of Defense Mechanisms in the Fungi-Avocado Pathosystem.

The greatest challenge for the avocado (Persea americana Miller) industry is to maintain the quality of the fruit to meet consumer requirements. Anthracnose is considered the most important disease in this industry, and it is caused by different species of the genus Colletotrichum, although other pathogens can be equally important. The defense mechanisms that fruit naturally uses can be triggered in response to the attack of pathogenic microorganisms and also by the application of exogenous elicitors in the form of GRAS compounds. The elicitors are recognized by receptors called PRRs, which are proteins located on the avocado fruit cell surface that have high affinity and specificity for PAMPs, MAMPs, and DAMPs. The activation of defense-signaling pathways depends on ethylene, salicylic, and jasmonic acids, and it occurs hours or days after PTI activation. These defense mechanisms aim to drive the pathogen to death. The application of essential oils, antagonists, volatile compounds, chitosan and silicon has been documented in vitro and on avocado fruit, showing some of them to have elicitor and fungicidal effects that are reflected in the postharvest quality of the fruit and a lower incidence of diseases. The main focus of these studies has been on anthracnose diseases. This review presents the most relevant advances in the use of natural compounds with antifungal and elicitor effects in plant tissues.

Sodium hydrosulfide priming improves the response of photosynthesis to overnight frost and day high light in avocado (Persea americana Mill, cv. 'Hass').

Radiation frost events, which have become more common in the Mediterranean Basin in recent years, inflict extensive damage to tropical/subtropical fruit crops. During radiation frost, sub-zero temperatures are encountered in the dark, followed by high light during the subsequent clear-sky day. One of the key processes affected by these conditions is photosynthesis, which, when significantly inhibited, leads to the enhanced accumulation of reactive oxygen species (ROS) and damage. The use of 'chemical priming' treatments that induce plants' endogenous stress responses is a possible strategy to improve their coping with stress conditions. Herein, we studied the effects of priming with sodium hydrosulfide (NaHS), a donor of hydrogen sulfide (H2 S), on the response of photosynthesis to overnight frost and day high-light conditions in 'Hass' avocado (Persea americana Mill). We found that priming with a single foliar application of NaHS had positive effects on the response of grafted 'Hass' plants. Primed plants exhibited significantly reduced inhibition of CO2 assimilation, a lower accumulation of hydrogen peroxide as well as lower photoinhibition, as compared to untreated plants. The ability to maintain a high CO2 assimilation capacity after the frost was attained on the background of considerable inhibition in stomatal conductance. Thus, it was likely related to the lower accumulation of ROS and photodamage observed in primed 'Hass' plants. This work contributes toward the understanding of the response of photosynthesis in a subtropical crop species to frost conditions and provides a prospect for chemical priming as a potential practice in orchards during cold winters.

Identification and characterization of a new Bacillus atrophaeus strain B5 as biocontrol agent of postharvest anthracnose disease in soursop (Annona muricata) and avocado (Persea americana).

Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous fruit, including soursop and avocado. The use of fungicides to maintain the high quality of fruit creates a potential health risk. One alternative to this problem is the biological control, which has been applied successfully during postharvest. The Bacillus species are one of the most studied biological agents against postharvest pathogens because accomplish their biocontrol performance by producing a variety of metabolites. In this study, we evaluated the activity of metabolites contained in the cell free supernatant, obtained from Bacillus strain B5 culture, against micelial growth and spore germination of two virulent strains of C. gloeosporioides isolated from soursop and avocado. On the basis of 16S rDNA gene sequence analysis, this strain was identified as Bacillus atrophaeus. A preventive treatment using cell free supernatant, reduced severity and incidence of anthracnose disease on harvested soursop and avocado fruit. B. atrophaeus strain B5 harbors genes involved in the production of antibiotics such as surfactin, bacillomycin and iturin, which could be contributing to the efficiency of the preventive treatment during postharvest. The antagonistic role of metabolites contained in the cell free supernatant against anthracnose disease, provide a new approach by which to attack this problem and can help reduce the use of chemical pesticides, environmental pollution, leading to the safer fruit preservation.

Maintaining postharvest quality of cold stored 'Hass' avocados by altering the fatty acids content and composition with the use of natural volatile compounds - methyl jasmonate and methyl salicylate.

BACKGROUND: Low temperatures are often used to reduce metabolic processes and extend the storage life of fruit; however, in the case of avocado, a temperature below 3 °C will often result in the development of physiological disorders associated with chilling injury. The objective of this study was to investigate the ability of methyl jasmonate (MeJA) and methyl salicylate (MeSA) vapours to alleviate chilling injury in 'Hass' avocado fruit kept at 2 °C for 21 days followed by 6-7 days of shelf-life at 20 °C, simulating supply chain conditions. RESULTS: The incidence and severity of chilling injury were significantly reduced in MeJA- and MeSA-exposed fruit, especially at 100 µmol L-1 . The mechanism involved improved membrane integrity via alteration of the fatty acid content and composition, down-regulation of LOX gene expression and reduced activity of lipoxygenase. CONCLUSION: MeJA and MeSA have the potential for being used with 'Hass' avocado fruit shipped at low temperature to reduce its susceptibility to chilling injury. © 2017 Society of Chemical Industry.

Investigation into the role of endogenous abscisic acid during ripening of imported avocado cv. Hass.

BACKGROUND: The importance of ethylene in avocado ripening has been extensively studied. In contrast, little is known about the possible role of abscisic acid (ABA). The present work studied the effect of 1-methylcyclopropene (1-MCP) (0.3 µL L-1 ), e+® Ethylene Remover and the combination thereof on the quality of imported avocado cv. Hass fruit stored for 7 days at 12 °C. Ethylene production, respiration, firmness, colour, heptose (C7) sugars and ABA concentrations in mesocarp tissue were measured throughout storage. RESULTS: Treatment with e+® Ethylene Remover reduced ethylene production, respiration rate and physiological ripening compared with controls. Fruit treated with 1-MCP + e+® Ethylene Remover and, to a lesser extent 1-MCP alone, had the lowest ethylene production and respiration rate and hence the best quality. Major sugars measured in mesocarp tissue were mannoheptulose and perseitol, and their content was not correlated with ripening parameters. Mesocarp ABA concentration, as determined by mass spectrometry, increased as fruit ripened and was negatively correlated with fruit firmness. CONCLUSIONS: Results suggest a relationship between ABA and ethylene metabolism since blocking ethylene, and to a larger extent blocking and removing ethylene, resulted in lower ABA concentrations. Whether ABA influences avocado fruit ripening needs to be determined in future research. © 2017 Society of Chemical Industry.

Inhibitory effect of Persea cordata Mez. (pau-andrade) bark extracts against Clostridium perfringens causing gangrenous mastitis

Introduction: Gangrenous mastitis is a special clinical presentation of mastitis in cattle and small ruminants. The bark of the tree Persea cordata Mez. is used in Brazilian ethnoveterinary medicine to treat wounds in farm animals. Objectives: Examine in vitro antimicrobial action of apolar fractions of P. cordata bark against a wild strain of C. perfringens isolated from the udder of a cow with gangrenous mastitis, and against a reference strain. Methods: A milk sample was collected from the udder, aliquots were diluted and Gram-stained smears were performed. The aliquots were inoculated in broth and planted in blood agar, and then incubated in anaerobiosis at 37oC / 24h. Biochemical identification was based on bacterial isolation. In vitro inhibitory activity of apolar fractions of P. cordata was evaluated by agar diffusion and MIC (minimum inhibitory concentration) using the agar dilution method. Results: In both tests the plant extracts displayed significant in vitro inhibitory activity against the clinical and reference strains of C. perfringens assayed. Conclusion: The study is the first demonstration of the inhibitory effect of P. cordata on C. perfringens, due to its antimicrobial properties, which serves as evidence supporting its folk use. The extracts could be used as coadjuvants in the treatment of gangrenous mastitis(AU)

Introducción: la mastitis gangrenosa es una presentación clínica especial de mastitis en el ganado y pequeños rumiantes. Persea cordata Mez. , es un árbol conocido en la etnoveterinaria brasileña, cuya corteza se utiliza en la curación de heridas en animales de granja. Objetivos: investigar el efecto antimicrobiano in vitro de fracciones apolares de la corteza de P. cordata contra una cepa salvaje de C. perfringens, aislada de la ubre de una vaca con mastitis gangrenosa y una cepa de referencia. Métodos: Se recogió una muestra de leche de la ubre, se diluyeron alícuotas, y se realizaron frotis teñidos por Gram. Las alícuotas fueron inoculadas en caldo y sembradas en agar sangre, y posteriormente incubados en anaerobiosis a 37 oC/24h. La identificación bioquímica fue realizada a partir del aislamiento bacteriano. La actividad inhibitoria in vitro de las fracciones apolares de P. cordata fue evaluada utilizando la técnica de difusión en agar y la CMI (concentración minima inhibitoria) mediante el método de dilución en agar. Resultados: los extractos de la planta, en ambas pruebas, presentaron significativa actividad inhibitoria in vitro contra las cepas clínica y de referencia de C. perfringens ensayadas. Conclusión: se concluyó que, por primera vez, se demuestra un efecto inhibitorio de P. cordata sobre C. perfringens, reforzando el uso popular, debido a sus propiedades antimicrobianas. Los extractos podrán ser utilizados como coadyuvantes en el tratamiento de la mastitis gangrenosa(AU)

Development of Metal-Organic Framework for Gaseous Plant Hormone Encapsulation To Manage Ripening of Climacteric Produce.

Controlled ripening of climacteric fruits, such as bananas and avocados, is a critical step to provide consumers with high-quality products while reducing postharvest losses. Prior to ripening, these fruits can be stored for an extended period of time but are usually not suitable for consumption. However, once ripening is initiated, they undergo irreversible changes that lead to rapid quality loss and decay if not consumed within a short window of time. Therefore, technologies to slow the ripening process after its onset or to stimulate ripening immediately before consumption are in high demand. In this study, we developed a solid porous metal-organic framework (MOF) to encapsulate gaseous ethylene for subsequent release. We evaluated the feasibility of this technology for on-demand stimulated ripening of bananas and avocados. Copper terephthalate (CuTPA) MOF was synthesized via a solvothermal method and loaded with ethylene gas. Its crystalline structure and chemical composition were characterized by X-ray diffraction crystallography, porosity by N2 and ethylene isotherms, and morphology by electron microscopy. The MOF loaded with ethylene (MOF-ethylene) was placed inside sealed containers with preclimacteric bananas and avocados and stored at 16 °C. The headspace gas composition and fruit color and texture were monitored periodically. Results showed that this CuTPA MOF is highly porous, with a total pore volume of 0.39 cm(3)/g. A 50 mg portion of MOF-ethylene can absorb and release up to 654 µL/L of ethylene in a 4 L container. MOF-ethylene significantly accelerated the ripening-related color and firmness changes of treated bananas and avocados. This result suggests that MOF-ethylene technology could be used for postharvest application to stimulate ripening just before the point of consumption.

Combined Treatments Reduce Chilling Injury and Maintain Fruit Quality in Avocado Fruit during Cold Quarantine.

Quarantine treatment enables export of avocado fruit (Persea americana) to parts of the world that enforce quarantine against fruit fly. The recommended cold-based quarantine treatment (storage at 1.1°C for 14 days) was studied with two commercial avocado cultivars 'Hass' and 'Ettinger' for 2 years. Chilling injuries (CIs) are prevalent in the avocado fruit after cold-quarantine treatment. Hence, we examined the effect of integrating several treatments: modified atmosphere (MA; fruit covered with perforated polyethylene bags), methyl jasmonate (MJ; fruit dipped in 2.5 µM MJ for Hass or 10 µM MJ for Ettinger for 30 s), 1-methylcyclopropene (1-MCP; fruit treated with 300 ppb 1-MCP for 18 h) and low-temperature conditioning (LTC; a gradual decrease in temperature over 3 days) on CI reduction during cold quarantine. Avocado fruit stored at 1°C suffered from severe CI, lipid peroxidation, and increased expression of chilling-responsive genes of fruit peel. The combined therapeutic treatments alleviated CI in cold-quarantined fruit to the level in fruit stored at commercial temperature (5°C). A successful therapeutic treatment was developed to protect 'Hass' and 'Ettinger' avocado fruit during cold quarantine against fruit fly, while maintaining fruit quality. Subsequently, treated fruit stored at 1°C had a longer shelf life and less decay than the fruit stored at 5°C. This therapeutic treatment could potentially enable the export of avocado fruit to all quarantine-enforcing countries. Similar methods might be applicable to other types of fruit that require cold quarantine.

The effect of ultrasound on particle size, color, viscosity and polyphenol oxidase activity of diluted avocado puree.

The effect of ultrasound treatment on particle size, color, viscosity, polyphenol oxidase (PPO) activity and microstructure in diluted avocado puree was investigated. The treatments were carried out at 20 kHz (375 W/cm(2)) for 0-10 min. The surface mean diameter (D[3,2]) was reduced to 13.44 µm from an original value of 52.31 µm by ultrasound after 1 min. A higher L(∗) value, ΔE value and lower a(∗) value was observed in ultrasound treated samples. The avocado puree dilution followed pseudoplastic flow behavior, and the viscosity of diluted avocado puree (at 100 s(-1)) after ultrasound treatment for 1 min was 6.0 and 74.4 times higher than the control samples for dilution levels of 1:2 and 1:9, respectively. PPO activity greatly increased under all treatment conditions. A maximum increase of 25.1%, 36.9% and 187.8% in PPO activity was found in samples with dilution ratios of 1:2, 1:5 and 1:9, respectively. The increase in viscosity and measured PPO activity might be related to the decrease in particle size. The microscopy images further confirmed that ultrasound treatment induced disruption of avocado puree structure.

Avocado roots treated with salicylic acid produce phenol-2,4-bis (1,1-dimethylethyl), a compound with antifungal activity.

We demonstrated the ability of salicylic acid (SA) to induce a compound in avocado roots that strengthens their defense against Phytophthora cinnamomi. The SA content of avocado roots, before and after the application of exogenous SA, was determined by High-Performance Liquid Chromatography (HPLC). After 4h of SA feeding, the endogenous level in the roots increased to 223 µg g(-1) FW, which was 15 times the amount found in control roots. The methanolic extract obtained from SA-treated avocado roots inhibited the radial growth of P. cinnamomi. A thin layer chromatographic bioassay with the methanolic extract and spores of Aspergillus showed a distinct inhibition zone. The compound responsible for the inhibition was identified as phenol-2,4-bis (1,1-dimethylethyl) by gas chromatography and mass spectrometry. At a concentration of 100 µg/mL, the substance reduced germinative tube length in Aspergillus and radial growth of P. cinnamomi. A commercial preparation of phenol-2,4-bis (1,1-dimethylethyl) caused the same effects on mycelium morphology and radial growth as our isolate, confirming the presence of this compound in the root extracts. This is the first report of the induction of this compound in plants by SA, and the results suggest that it plays an important role in the defense response of avocado.

Integrating transient heterogeneity of non-photochemical quenching in shade-grown heterobaric leaves of avocado (Persea americana L.): responses to CO2 concentration, stomatal occlusion, dehydration and relative humidity.

Long-lived shade leaves of avocado had extremely low rates of photosynthesis. Gas exchange measurements of photosynthesis were of limited use, so we resorted to Chl fluorescence imaging (CFI) and spot measurements to evaluate photosynthetic electron transport rates (ETRs) and non-photochemical quenching (NPQ). Imaging revealed a remarkable transient heterogeneity of NPQ during photosynthetic induction in these hypostomatous, heterobaric leaves, but was adequately integrated by spot measurements, despite long-lasting artifacts from repeated saturating flashes during assays. Major veins (mid-vein, first- and second-order veins) defined areas of more static large-scale heterogeneous NPQ, with more dynamic small-scale heterogeneity most strongly expressed in mesophyll cells between third- and fourth-order veins. Both responded to external CO2 concentration ([CO2]), occlusion of stomata with Vaseline™, leaf dehydration and relative humidity (RH). We interpreted these responses in terms of independent behavior of stomata in adjacent areoles that was largely expressed through CO2-limited photosynthesis. Heterogeneity was most pronounced and prolonged in the absence of net CO2 fixation in 100 p.p.m. [CO2] when respiratory and photorespiratory CO2 cycling constrained the inferred ETR to ~75% of values in 400 or 700 p.p.m. [CO2]. Likewise, sustained higher NPQ under Vaseline™, after dehydration or at low RH, also restricted ETR to ~75% of control values. Low NPQ in chloroplast-containing cells adjacent to major veins but remote from stomata suggested internal sources of high [CO2] in these tissues.

Field evaluation of systemic imidacloprid for the management of avocado thrips and avocado lace bug in California avocado groves.

BACKGROUND: The efficacy of systemic applications of imidacloprid for the management of avocado thrips and avocado lace bug was determined in field trials. Following insecticide treatment by chemigation, leaves of appropriate age for each insect were sampled over a 6 month period and used for bioassays. Imidacloprid residues were measured by ELISA in leaves used for bioassays to determine concentrations of insecticide that were toxic to both pests. RESULTS: The uptake of imidacloprid into treated trees was extremely slow, peaking in the current year's leaf flush at only 8 ng cm(-2) leaf tissue after 15 weeks. Avocado thrips mortality in bioassays with young flush leaves, the preferred feeding substrate for this insect, was minimal, indicating that imidacloprid concentrations were below threshold levels needed for effective control. Residues present in older leaves, which are preferred by the avocado lace bug, were higher than in young flush leaves, and provided good control of this pest. Probit analysis of bioassay data showed that the avocado lace bug (LC(50) = 6.1 ng imidacloprid cm(-2) leaf tissue) was more susceptible to imidacloprid than the avocado thrips (LC(50) = 73 ng imidacloprid cm(-2) leaf tissue). CONCLUSIONS: In spite of the slow uptake of imidacloprid into avocado trees, the levels of imidacloprid would be sufficient to control avocado lace bug infestations. In contrast, the slow uptake would be problematic for avocado thrips control because inadequate levels of insecticide accumulate in new flush foliage and would allow avocado thrips populations to build to levels that would subsequently damage developing avocado fruit.

Electric signalling in fruit trees in response to water applications and light-darkness conditions.

A fundamental property of all living organisms is the generation and conduction of electrochemical impulses throughout their different tissues and organs, resulting from abiotic and biotic changes in environmental conditions. In plants and animals, signal transmission can occur over long and short distances, and it can correspond to intra- and inter-cellular communication mechanisms that determine the physiological behaviour of the organism. Rapid plant and animal responses to environmental changes are associated with electrical excitability and signalling. The same molecules and pathways are used to drive physiological responses, which are characterized by movement (physical displacement) in animals and by continuous growth in plants. In the field of environmental plant electrophysiology, automatic and continuous measurements of electrical potential differences (DeltaEP) between plant tissues can be effectively used to study information transport mechanisms and physiological responses that result from external stimuli on plants. A critical mass of data on electrical behaviour in higher plants has accumulated in the last 5 years, establishing plant neurobiology as the most recent discipline of plant science. In this work, electrical potential differences were monitored continuously using Ag/AgCl microelectrodes, which were inserted 15mm deep into sapwood at various positions in the trunks of several fruit-bearing trees. Electrodes were referenced to an unpolarisable Ag/AgCl microelectrode, which was installed 5cm deep in the soil. Systematic patterns of DeltaEP during day-night cycles and at different conditions of soil water availability are discussed as alternative tools to assess early plant stress conditions. This research relates to the adaptive response of trees to soil water availability and light-darkness cycles.