Juan Pablo Zoffoli

Effects of continuous 0.3 ppm ozone exposure on decay development and physiological responses of peaches and table grapes in cold storage

Continuous ozone exposure at 0.3 ppm (v/v) (US-OSHA Threshold Limit Value for short term exposure) inhibited aerial mycelial growth and sporulation on ‘Elegant Lady’ peaches wound inoculated with Monilinia fructicola, Botrytis cinerea, Mucor piriformis ,o rPenicillium expansum and stored for 4 weeks at 5 °C and 90% relative humidity (RH). Aerial growth and sporulation, however, resumed afterward in ambient atmospheres. Ozone exposure did not significantly reduce the incidence and severity of decay caused by these fungi with the exception of brown rot. Gray mold nesting among ‘Thompson Seedless’ table grapes was completely inhibited under 0.3 ppm ozone when fruit were stored for 7 weeks at 5 °C. Gray mold incidence, however, was not significantly reduced in spray inoculated fruit. Continuous ozone exposure at 0.3 ppm increased water loss after 5 weeks of storage at 5 °C and 90% RH in ‘Zee Lady’ peaches but not after 4 weeks of storage in ‘Flame Seedless’ grapes. Respiration and ethylene production rates of ‘O’Henry’ peaches were not affected by previous exposure to 0.3 ppm ozone. In every test, no phytotoxic injuries of fruit tissues were observed in ozonated or ambient atmosphere treatments.

Temperature, length of cold storage and maturity influence the ripening rate of ethylene-preconditioned kiwifruit

Abstract The effect of temperature, length of cold storage and maturity on the ripening of ethylene-preconditioned (100 μl l −1 for 12 or 24 h) kiwifruit was investigated. Low (0 o C) temperatures at any point prior to, during or after ethylene preconditioning significantly delayed softening and soluble solids concentration (SSC) accumulation compared to higher temperatures (i.e. 20 o C). Freshly-harvested kiwifruit responded to ethylene-preconditioning (100 μl l −1 at 0°C for 24 h) by softening faster than control fruit even if harvested 5 weeks after commercial maturity. In contrast, kiwifruit harvested at commercial maturity and stored at 0°C softened faster than the control only if preconditioned with ethylene during the first 2 weeks of storage. Kiwifruit had high respiration rates 1 day after being transferred from 0 to 20°C, but respiration dropped to near base-line levels by day 2. Fruit stored at 0°C always respired faster upon transfer to 20°C than did freshly-harvested fruit and preconditioning with ethylene increased the initial rate of respiration of freshly-harvested fruit but had less of an effect on stored fruit. Ethylene preconditioning did not significantly hasten the climacteric rise in respiration or ethylene production of either freshly-harvested or stored kiwifruit. The climacteric rise of individual kiwifruit began only after fruit softened to ≤7 N.

Comparison of the total phenolic content, total anthocyanin content and antioxidant activity of polyphenol-rich fruits grown in Chile

In the last 10 years, interest in research on polyphenol-rich fruit species has increased due to the potential health benefits of these species, mainly attributed to their high anthocyanin content and antioxidant activity. Six polyphenol-rich fruit species (blueberries, raspberries, blackberries, strawberries, pomegranates and maqui berries) were harvested at the same maturity stage during the same growing season and were compared according to their total phenolic (Folin-Ciocalteu method) and total anthocyanin (pH differential method) contents and antioxidant activity using ferric-reducing antioxidant power (FRAP assay) and 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging capacity methods. With the results of this study, the polyphenolic status of the main polyphenol-rich fruit species that are grown in Chile were compared, and maqui berry showed the highest total phenolic (14.6 g gallic acid equivalent kg -1 of fresh weight [g GAE kg -1 FW]), total anthocyanin (9.3 g cyanidin-3-glu kg -1 of fresh weight [g cy-3-glu kg -1 FW]) contents, and antioxidant activity (152.0 mmol Fe 2+ kg -1 of fresh weight [mmol Fe 2+ kg -1 FW] and 1.5 mg of fresh weight [FW]) with significant differences from the other fruit species that were analyzed. Nevertheless, bioavailability studies to test the benefits of the species’ dietary antioxidants should be performed in order to establish scientific evidence in this area. Durante los ultimos 10 anos, el interes en la investigacion de los frutos ricos en polifenoles ha incrementado debido a sus potenciales efectos beneficiosos para la salud atribuidos principalmente a contenidos altos de antocianinas y una alta actividad antioxidante. Seis especies de frutos ricos en polifenoles (arandanos, frambuesas, moras, frutillas, granadas y maquis) fueron cosechados en el mismo estado de madurez durante la misma temporada de produccion y comparadas de acuerdo a sus contenidos de fenoles totales (metodo Folin Ciocalteu), antocianinas totales (metodo pH diferencial) y actividad antioxidante determinada por los metodos de FRAP (poder antioxidante para reducir el hierro ferrico) y del radical DPPH (2,2-diphenylpicrylhydrazil). Los resultados de este estudio permiten comparar el estatus polifenolico de los principales frutos ricos en polifenoles que crecen en Chile, donde el maqui presenta el mayor contenido de fenoles totales (14,6 g EAG kg -1 pf), antocianinas totales (9,3 g ci-3-glu kg -1 pf), y actividad antioxidante (152,0 mmol E Fe +2 kg -1 and 1,5 mg pf) con diferencias significativas con las otras especies de frutos analizadas. Sin embargo, se deben realizar estudios de biodisponibilidad para probar los efectos beneficos sobre la salud humana de los antioxidantes de esta especie para poder establecer evidencia cientifica en esta area.

POLYPHENOL CONTENT AND ANTIOXIDANT ACTIVITY OF MAQUI ( Aristoteliachilensis [MOLINA] STUNTZ) DURING FRUIT DEVELOPMENT AND MATURATION INCENTRAL CHILE

Maqui ( Aristotelia chilensis [Molina] Stuntz, Elaeocarpaceae) is a Chilean native species which produces small berries that are mainly collected from the wild. The health benefits of maqui fruit are attributed to their high polyphenol content as well as their wide variety of anthocyanins and flavonols. One of the main factors that affect the polyphenol content in fruit is the maturity stage at harvest. The objective of this study was to determine total phenol and total anthocyanin content and antioxidant activity (by ferric reducing ability of plasma [FRAP] assay) of maqui fruits harvested at different fruit maturity stages from two wild populations located in Central Chile. Each maturity stage was determined by days from fruit set, berry size, and soluble solids. Total phenol content declined while total anthocyanin content increased from the green to light red stage. Nevertheless, both total phenol and anthocyanin content increased from the light red to dark purple stage. The highest anthocyanin content and antioxidant activity was found in the late maturity stage (dark purple). The results show that ripening in maqui fruit can be expected with 1100 growing degree-days (91 d after fruit set) in Central Chile. At this moment of harvest, fruits with 18-19° Brix have the highest anthocyanin content and antioxidant activity (FRAP). This study constitutes the first advances in the understanding of maqui fruit ripening and corresponding antioxidant activity.

Identification and Characterization of Botrytis Blossom Blight of Japanese Plums Caused by Botrytis cinerea and B. prunorum sp. nov. in Chile

Blossom blight is a destructive disease of plums (Prunus salicina) when humid and temperate weather conditions occur in Chile. Disease incidence ranging from 4 to 53% has been observed. Symptoms include light brown petal necrosis, starting as light brown mottles or V-shaped necrosis at the margins of the petals, progressing to the stamen and pistils. In this study, the etiology of blossom blight of plums was determined. High- and low-sporulating isolates of Botrytis were obtained consistently from blighted blossoms and apparently healthy flowers of plums. Based on colony morphology, conidial production and molecular phylogenetic analysis, these high- and low-sporulating isolates were identified as B. cinerea and B. prunorum sp. nov., respectively. Phylogenetic analysis of the genes glyceraldehyde 3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) grouped B. prunorum isolates in a single cluster, distantly from B. cinerea and other Botrytis species. The phylogenetic analysis of necrosis and ethylene-inducing protein (NEP1 and NEP2) genes corroborated these results. Analysis of the internal transcribed spacer region and large-subunit (26S) ribosomal DNA and detection of Boty and Flipper transposable elements, were not useful to differentiate between these Botrytis species. Both species were pathogenic on plum flowers and the fruit of plums, apples, and kiwifruits. However, B. prunorum was less virulent than B. cinerea. These pathogens were re-isolated from inoculated and diseased tissues; thus, Kochs postulates were fulfilled, confirming its role in blossom blight of plums. B. cinerea was predominant, suggesting that B. prunorum may play a secondary role in the epidemiology of blossom blight in plums in Chile. This study clearly demonstrated that the etiology of blossom blight of plums is caused by B. cinerea and B. prunorum, which constitute a species complex living in sympatry on plums and possibly on other stone fruit trees.

Evolución del daño por insolación de manzanas 'Granny Smith' durante el almacenaje refrigerado

)to reduce scald (the physiological disorder), and an equal number of non-treated apples were used as controls. The concentration of conjugated trienes (CTs) was determined and compared between the apparently healthy and damaged side of the same fruits. Sunscald, characterized by brown skin tissue, developed on the sunburnt side of the fruit, and superfi cial and senescent scald symptoms developed on the healthy side of the apple. A signifi cant and negative relationship between sunscald and scald were obtained in fruit treated with DPA. A low concentration of CT 281 was found on the sunburnt side of apples treated with DPA and 1-MCP. Application of 1-MCP and DPA controlled scald but not sunscald. DPA treatment resulted in better control of scald after 6 months of storage on fruit with moderate sunburn damage at the time of harvest.

First Report of Diaporthe novem Causing Postharvest Rot of Kiwifruit During Controlled Atmosphere Storage in Chile

Chile is considered the third major exporter of kiwifruits (Actinidia deliciosa (A. Chev.) C. F. Liang & A. R. Ferguson) worldwide after Italy and New Zealand (1). The genus Diaporthe Nitschke (anamorph: genus Phomopsis) has been reported as causing postharvest rot in kiwifruit (4). During the current study, 1,400 fruits arbitrarily collected from seven controlled atmosphere (CA) rooms after 90 days of storage conditions (2% O2, 5% CO2) determined that 21.5% of the fruit were affected by decay and 0.86% developed symptoms different than those caused by Botrytis cinerea, the main postharvest pathogen associated to kiwifruit. Symptoms were soft rot with brown skin that started at the stem-end and in severe cases affected the entire fruit. Internally, affected fruit showed browning and watery tissues. Twelve affected fruits were surface disinfested (75% ethanol) and small pieces of internal rotten tissues were placed on acidified potato dextrose agar (APDA) for 7 days at 20°C. Twelve isolates were obtained, and four of them were identified morphologically and molecularly as Diaporthe ambigua, a species that has been previously described causing rot in stored kiwifruits in Chile (2). However, eight other flat, white to grayish colonies with sparse dirty-white aerial mycelium at the edge of the dish were obtained (3). Black pycnidia contained unicellular, hyaline, biguttulate, oval to cylindrical alpha conidia, with obtuse ends of (7.9) 6.7 (5.3) × (2.9) 2.5 (2.1) μm (n = 30). These isolates were tentatively identified as a Diaporthe sp. The species identification was determined by sequencing comparison of the internal transcribed spacer (ITS1-5.8S-ITS2) region of the rDNA (GenBank Accession Nos. KJ210020 to 24, KJ210027, and KJ210033) and a portion of beta-tubulin (BT) (KJ210034 to 38, KJ210041, and KJ210047) using primers ITS4-ITS5 and Bt2a-Bt2b, respectively. BLAST analyses showed 99 to 100% identity with D. novem J.M. Santos, Vrandecic & A.J.L Phillips reference ex-type (KC343156 and KC344124 for ITS and BT, respectively) (3). Eighteen mature kiwifruits cv. Hayward were inoculated using a sterile cork borer on the surface of the fruit and placing 5-mm agar plugs with mycelial of D. novem (DN-1-KF). An equal number of fruits treated with sterile agar plugs were used as negative controls. After 30 days at 0°C under CA, all inoculated fruit showed rot symptoms with lesions 7.8 to 16.4 mm in diameter. The same D. novem isolate was inoculated with 30 μl of a conidial suspension (106 conidia/ml) on the surface of 18 ripe kiwifruits that were previously wounded and non-wounded as described above. An equal number of wounded and non-wounded fruits, treated with 30 μl sterile water, were used as negative controls. All inoculated wounded fruits developed rot symptoms with necrotic lesions of 14.1 to 20.2 mm of diameter after 14 days at 25°C. Inoculated non-wounded and negative control fruits remained symptomless. Kochs postulates were fulfilled by re-isolating D. novem only from the symptomatic fruits. To our knowledge, this is the first report of rot caused by D. novem on kiwifruit during cold storage in Chile and worldwide. Therefore, both Diaporthe species appears to be associated to Diaporthe rot of kiwifruit in Chile. References: (1) Belrose, Inc. World Kiwifruit Review. Belrose, Inc. Publishers, Pullman, WA, 2012. (2) J. Auger et al. Plant Dis. 97:843, 2013. (3) R. Gomes et al. Persoonia 31:1, 2013. (4) L. Luongo et al. J. Plant Pathol. 93:205, 2011.

Pruning effects on vegetative growth and fruit quality of 'Bing'/'Gisela®5' and 'Bing'/'Gisela®6' sweet cherry trees (Prunus avium)

Abstract M. Villasante, S. Godoy, J.P. Zoffoli, and M. Ayala. 2012. Pruning effects on growth and fruit quality of ‘Bing’/‘Gisela®5’ and ‘Bing’/‘Gisela®6’ sweet cherry trees ( Prunus avium ). Cien. Inv. Agr. 39(1): 117-126. Annual pruning is one of the most efficient ways to regulate crop load and renew fruiting wood in highly productive sweet cherry ( Prunus avium L.) combinations. Although Chilean growers did not previously prune cherry trees of more vigorous combinations, in recent years, the adoption of more dwarfing rootstocks and self-fertile cultivars has led to the inclusion of annual pruning as a practice in modern orchards. At first, this alteration in orchard management practices was not considered by growers, and thus, many of the initially established cherry orchards were not pruned as intensively as they should have been. As a consequence, many trees showed a reduction in fruit quality after 4 or 5 years of being planted, as they became overcropped and, consequently, registered reductions in their vegetative growth. There are only a few studies related to the effect of corrective pruning on dwarfing combinations that display an imbalance between reproductive and vegetative growth due to a reduction in the leaf area to fruit ratio of the tree. For this reason, the objective of this research was to study the effect of pruning in an orchard consisting of the dwarfing combinations ‘Bing’/‘Gisela®5’ (‘Bing’/‘GI®5’) and ‘Bing’/‘Gisela®6’ (‘Bing’/‘GI®6’), which shown a reduction in vigor, fruit quality and yield. Trees of both combinations were treated with a medium intensity pruning in late winter (early September). Several vegetative (shoot length, leaf area of spurs and shoots, trunk cross sectional area) and reproductive (total yield per tree, fruit growth and quality) parameters were evaluated after pruning. One of the most important effects of pruning for both combinations was an increase in the total current season shoot (CSS) growth, which was 112.5 and 125.6% for ‘Bing’/‘GI®5’ and ‘Bing’/‘GI®6’, respectively. Additionally, the average shoot length increased by 820.0 and 325.4% for ‘Bing’/‘GI®5’ and ‘Bing’/‘GI®6’, respectively. Furthermore, CSSs developed a higher leaf number in the pruned trees. There was no change in leaf number for reproductive spurs, but these had bigger leaves in the pruned trees, demonstrating increased total leaf area per spur. Additionally, pruning allowed crop load regulation and increased fruit size by 8.5 and 6.1% for ‘Bing’/‘GI®5’ and ‘Bing’/‘GI®6’, respectively. However, fruits from pruned trees showed a higher susceptibility to mechanical damage compared with unpruned trees of both combinations.

Identification and Characterization of Diaporthe ambigua, D. australafricana, D. novem, and D. rudis Causing a Postharvest Fruit Rot in Kiwifruit

Diaporthe spp. are important plant pathogens causing wood cankers, blight, dieback, and fruit rot in a wide range of hosts. During surveys conducted during the 2013 and 2014 seasons, a postharvest rot in Hayward kiwifruit (Actinidia deliciosa) was observed in Chile. In order to identify the species of Diaporthe associated with this fruit rot, symptomatic fruit were collected from seven kiwifruit packinghouses located between San Francisco de Mostazal and Curicó (central Chile). Twenty-four isolates of Diaporthe spp. were identified from infected fruit based on morphological and cultural characters and analyses of nucleotides sequences of three loci, including the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2), a partial sequences of the β-tubulin, and translation elongation factor 1-α genes. The Diaporthe spp. identified were Diaporthe ambigua, D. australafricana, D. novem, and D. rudis. Multilocus phylogenetic analysis revealed that Chilean isolates were grouped in separate clades with their correspondent ex-types species. All species of Diaporthe were pathogenic on wounded kiwifruit after 30 days at 0°C under normal and controlled-atmosphere (2% O2 and 5% CO2) storage and they were sensitive to benomyl, pyraclostrobin, and tebuconazole fungicides. D. ambigua isolates were the most virulent based on the lesion length measured in inoculated Hayward and Jintao kiwifruit. These findings confirm D. ambigua, D. australafricana, D. novem, and D. rudis as the causal agents of kiwifruit rot during cold storage in Chile. The specie D. actinidiae, a common of Diaporthe sp. found associated with kiwifruit rot, was not identified in the present study.

Postharvest Control of Gray Mold on Blueberry Based on Critical Growth Stages and Infection Risk Estimations

Gray mold (Botrytis cinerea) is an important postharvest disease of blueberries (Vaccinium corymbosum) in Chile, favored by the long (>15 days) transportation to reach the international markets. The aims of this research were to study the critical blueberry growth stages for postharvest gray mold control and to determine the infection risks on the basis of weather conditions. The critical stages for gray mold control were studied on blueberry ‘Brigitta’ and ‘Duke’ in two planting localities. Differential fungicide applications (0.5 g/L fenhexamid), performed between the early pink bud stage and mature fruit stage, showed that the best control of postharvest gray mold was obtained when fungicides were applied between the first blue fruit and mature fruit stages. The infection risks for B. cinerea infection were defined as >6 h of wetness and temperatures between 14 and 25 °C. This algorithm to estimate the infection risks was studied in blueberry ‘Brigitta’, ‘Duke’ and ‘Liberty’ in four planting localities. A significant correlation between the infection risk and gray mold prevalence in stored fruit was obtained (r = 0.96, P < 0.0001), suggesting that this algorithm could be used to optimize fungicide applications, but field validation remains to be determined. In conclusion, the mature fruit stage appears as the most critical stage for postharvest gray mold control if weather conditions, defined by this algorithm, occur.