Oxidative Stress Associated with Chilling Injury in Immature Fruit: Postharvest Technological and Biotechnological Solutions

Immature, vegetable-like fruits are produced by crops of great economic importance, including cucumbers, zucchini, eggplants and bell peppers, among others. Because of their high respiration rates, associated with high rates of dehydration and metabolism, and their susceptibility to chilling injury (CI), vegetable fruits are highly perishable commodities, requiring particular storage conditions to avoid postharvest losses. This review focuses on the oxidative stress that affects the postharvest quality of vegetable fruits under chilling storage. We define the physiological and biochemical factors that are associated with the oxidative stress and the development of CI symptoms in these commodities, and discuss the different physical, chemical and biotechnological approaches that have been proposed to reduce oxidative stress while enhancing the chilling tolerance of vegetable fruits

Involvement of ethylene biosynthesis and signalling in fruit set and early fruit development in zucchini squash (Cucurbita pepo L.)

Background We have identified a kind of parthenocarpy in zucchini squash which is associated with an incomplete andromonoecy, i.e. a partial conversion of female into bisexual flowers. Given that andromonoecy in this and other cucurbit species is caused by a reduction of ethylene production in the female flower, the associated parthenocarpic development of the fruit suggested the involvement of ethylene in fruit set and early fruit development.Results We have compared the production of ethylene as well as the expression of 13 ethylene biosynthesis and signalling genes in pollinated and unpollinated ovaries/fruits of two cultivars, one of which is parthenocarpic (Cavili), while the other is non-parthenocarpic (Tosca). In the latter, unpollinated ovaries show an induction of ethylene biosynthesis and ethylene signal transduction pathway genes three days after anthesis, which is concomitant with the initiation of fruit abortion and senescence. Fruit set and early fruit development in pollinated flowers of both cultivars and unpollinated flowers of Cavili is coupled with low ethylene biosynthesis and signalling, which would also explain the partial andromonoecy in the parthenocarpic genotype. The reduction of ethylene production in the ovary cosegregates with parthenocarpy and partial andromonoecy in the selfing progeny of Cavili. Moreover, the induction of ethylene in anthesis (by ethephon treatments) reduced the percentage of bisexual parthenocarpic flowers in Cavili, while the inhibition of ethylene biosynthesis or response (by AVG and STS treatments) induces not only andromonoecy but also the parthenocarpic development of the fruit in both cultivars.Conclusions Results demonstrate that a reduction of ethylene production or signalling in the zucchini flower is able to induce fruit set and early fruit development, and therefore that ethylene is actively involved in fruit set and early fruit development. Auxin and TIBA treatments, inducing fruit set and early fruit development in this species, also inhibit ethylene production and the expression of ethylene biosynthesis and response genes. A model is presented that discusses the crosstalk between ethylene and auxin in the control of fruit set and early fruit development in zucchini squash.This work was supported by grants AGL2008-05619-C02-02/ALI and AGL2011-30568-C02-02/ALI, partly funded by ERDF (European Regional Development Fund) and by the Spanish Ministry of Science and Innovation, and grant CVI-02617, funded by ERDF and by the Consejería de Innovación, Ciencia y Empresa, Junta de Andalucía, Spain. C.M. and Z.M. acknowledge FPU program scholarships from MEC, Spain. S.M. is funded by grant PTA2011-479-I from the Spanish Ministry of Science and Innovation

Genetic and Pre- and Postharvest Factors Influencing the Content of Antioxidants in Cucurbit Crops

Cucurbitaceae is one of the most economically important plant families, and includes some worldwide cultivated species like cucumber, melons, and squashes, and some regionally cultivated and feral species that contribute to the human diet. For centuries, cucurbits have been appreciated because of their nutritional value and, in traditional medicine, because of their ability to alleviate certain ailments. Several studies have demonstrated the remarkable contents of valuable compounds in cucurbits, including antioxidants such as polyphenols, flavonoids, and carotenoids, but also tannins and terpenoids, which are abundant. This antioxidant power is beneficial for human health, but also in facing plant diseases and abiotic stresses. This review brings together data on the antioxidant properties of cucurbit species, addressing the genetic and pre- and postharvest factors that regulate the antioxidant content in different plant organs. Environmental conditions, management, storage, and pre- and postharvest treatments influencing the biosynthesis and activity of antioxidants, together with the biodiversity of this family, are determinant in improving the antioxidant potential of this group of species. Plant breeding, as well as the development of innovative biotechnological approaches, is also leading to new possibilities for exploiting cucurbits as functional products

Effect of Ethylene-Insensitive Mutation etr2b on Postharvest Chilling Injury in Zucchini Fruit

Zucchini is a vegetable fruit that is very susceptible to postharvest chilling injury, and fruit ethylene production is correlated with chilling injury sensitivity, such that the more tolerant the cultivar, the lower is its ethylene production. It is expected that zucchini fruit with reduced sensitivity to ethylene would have a higher chilling injury tolerance. In this study, we compared the postharvest fruit quality of wild type and ethylene-insensitive mutant etr2b, in which a mutation was identified in the coding region of the ethylene receptor gene CpETR2B. Flowers from homozygous WT (wt/wt), mutant plants in homozygous (etr2b/etr2b) and heterozygous (wt/etr2b) were hand-pollinated, and all fruits were harvested with the same length, at about 8 days after pollination. After harvesting, fruit of each genotype was randomly divided in 3 batches of 12 fruits each (four replications with three fruits each), and then stored at 4 °C and 95% RH. At 0, 7, and 14 days after cold storage, each batch was used to assess ethylene production, respiration rate, weight and firmness loss, chilling injury, and oxidative stress metabolites. The results showed a lower chilling injury associated with lower cold-induced ethylene production in the mutant fruit, in comparison with the WT fruit. These data demonstrated that the ethylene-insensitive etr2b mutant fruit was more tolerant to chilling injury, confirming that basal ethylene in the still undamaged fruit could function as a modulator of post-harvest chilling injury. Moreover, the higher chilling tolerance of the etr2b mutant fruit was not associated with MDA content, but was concomitant with a reduction in the accumulation of hydrogen peroxide in the refrigerated mutant fruit

Individual Shrink Wrapping of Zucchini Fruit Improves Postharvest Chilling Tolerance Associated with a Reduction in Ethylene Production and Oxidative Stress Metabolites

We have studied the effect of individual shrink wrapping (ISW) on the postharvest performance of refrigerated fruit from two zucchini cultivars that differ in their sensitivity to cold storage: Sinatra (more sensitive) and Natura (more tolerant). The fruit was individually shrink wrapped before storing at 4°C for 0, 7 and 14 days. Quality parameters, ethylene and CO2 productions, ethylene gene expression, and oxidative stress metabolites were assessed in shrink wrapped and non-wrapped fruit after conditioning the fruit for 6 hours at 20°C. ISW decreased significantly the postharvest deterioration of chilled zucchini in both cultivars. Weight loss was reduced to less than 1%, pitting symptoms were completely absent in ISW fruit at 7 days, and were less than 25% those of control fruits at 14 days of cold storage, and firmness loss was significantly reduced in the cultivar Sinatra. These enhancements in quality of ISW fruit were associated with a significant reduction in cold-induced ethylene production, in the respiration rate, and in the level of oxidative stress metabolites such as hydrogen peroxide and malonyldialdehyde (MDA). A detailed expression analysis of ethylene biosynthesis, perception and signaling genes demonstrated a downregulation of CpACS1 and CpACO1 genes in response to ISW, two genes that are upregulated by cold storage. However, the expression patterns of six other ethylene biosynthesis genes (CpACS2 to CpACS7) and five ethylene signal transduction pathway genes (CpCTR1, CpETR1, CpERS1, CpEIN3.1 and CpEN3.2), suggest that they do not play a major role in response to cold storage and ISW packaging. In conclusion, ISW zucchini packaging resulted in improved tolerance to chilling concomitantly with a reduction in oxidative stress, respiration rate and ethylene production, as well as in the expression of ethylene biosynthesis genes, but not of those involved in ethylene perception and sensitivity.This work was supported by grants AGL2011-30568-C02/ALI from the Spanish Ministry of Science and Innovation, and AGR1423 from the Consejería de Economía, Innovación y Ciencia, Junta de Andalucía, Spain. Z.M. acknowledges FPU program scholarships from MEC, Spain. S.M. is funded by grant PTA2011-479-I from the Spanish Ministry of Science and Innovation

Análisis molecular de los cromosomas A y B de Crepis capillaris

Tesis Univ. de Granada. Departamento de Genétic

Oxidative Stress Associated with Chilling Injury in Immature Fruit: Postharvest Technological and Biotechnological Solutions

Immature, vegetable-like fruits are produced by crops of great economic importance, including cucumbers, zucchini, eggplants and bell peppers, among others. Because of their high respiration rates, associated with high rates of dehydration and metabolism, and their susceptibility to chilling injury (CI), vegetable fruits are highly perishable commodities, requiring particular storage conditions to avoid postharvest losses. This review focuses on the oxidative stress that affects the postharvest quality of vegetable fruits under chilling storage. We define the physiological and biochemical factors that are associated with the oxidative stress and the development of CI symptoms in these commodities, and discuss the different physical, chemical and biotechnological approaches that have been proposed to reduce oxidative stress while enhancing the chilling tolerance of vegetable fruits

Exogenous γ-aminobutyric acid treatment improves the cold tolerance of zucchini fruit during postharvest storage

This work examines the effect of a treatment with 1 mM of γ-aminobutyric acid (GABA) on zucchini fruit during postharvest cold storage. Specifically, the effect of GABA on postharvest quality was measured, as well as its implication in the GABA shunt and other related metabolic pathways. The treatments were performed in Sinatra, a variety of zucchini highly sensitive to low-temperature storage. The application of GABA improved the quality of zucchini fruit stored at 4 °C, with a reduction of chilling-injury index, weight loss, and cell death, as well as a lower rate of electrolyte leakage. GABA content was significantly higher in the treated fruit than in the control fruit at all times analyzed. At the end of the storage period, GABA-treated fruit had higher contents of both proline and putrescine. The catabolism of this polyamine was not affected by exogenous GABA. Also, over the long term, the treatment induced the GABA shunt by increasing the activities of the enzymes GABA transaminase (GABA-T) and glutamate decarboxylase (GAD). GABA-treated fruit contained higher levels of fumarate and malate than did non-treated fruit, as well as higher ATP and NADH contents. These results imply that the GABA shunt is involved in providing metabolites to produce energy, reduce power, and help the fruit to cope with cold stress over the long term.</p

The Ethylene Biosynthesis Gene CitACS4 Regulates Monoecy/Andromonoecy in Watermelon (Citrullus lanatus).

Monoecious and andromonoecious cultivars of watermelon are characterised by the production of male and female flower or male and hermaphrodite flowers, respectively. The segregation analysis in the offspring of crosses between monoecious and andromonoecious lines has demonstrated that this trait is controlled by a single gene pair, being the monoecious allele M semi-dominant to the andromonoecious allele A. The two studied F1 hybrids (MA) had a predominantly monoecious phenotype since both produced not only female flowers, but also bisexual flowers with incomplete stamens, and hermaphrodite flowers with pollen. Given that in other cucurbit species andromonoecy is conferred by mutations in the ethylene biosynthesis genes CmACS7, CsACS2 and CpACS27A we have cloned and characterised CitACS4, the watermelon gene showing the highest similarity with the formers. CitACS4 encoded for a type ACS type III enzyme that is predominantly expressed in pistillate flowers of watermelon. In the andromonoecious line we have detected a missense mutation in a very conserved residue of CitACS4 (C364W) that cosegregates with the andromonoecious phenotype in two independent F2 populations, concomitantly with a reduction in ethylene production in the floral buds that will develop as hermaphrodite flowers. The gene does not however co-segregates with other sex expression traits regulated by ethylene in this species, including pistillate flowering transition and the number of pistillate flowers per plant. These data indicate that CitAC4 is likely to be involved in the biosynthesis of the ethylene required for stamen arrest during the development of female flowers. The C364W mutation would reduce the production of ethylene in pistillate floral buds, promoting the conversion of female into hermaphrodite flowers, and therefore of monoecy into andromonoecy

Evaluation of ethylene production by ten Mediterranean carnation cultivars and their response to ethylene exposure

Vase life is one of the most important characters of postharvest cut flower quality. The onset of ethylene production and the amount of ethylene produced by flowers vary with the carnation cultivar, and thus influence their vase life. In the present study, differences in ethylene production and the response to exogenous ethylene among carnation cultivars were evaluated. Ten different cultivars: �Baltico�, �Domingo�, �Exotica�, �Famosa�, �Kiro�, �Madame Augier�, �Master�, �Mundo�, Pilar� and �Reina�were studied. Five flowers for each cultivar were exposed for 8 hours to 1 ìL L�1 exogenous ethylene concentration. Ethylene production, fresh weight and water uptake was measured daily throughout the experiments. The 10 different cultivars studied showed clear differences in vase life, ethylene production, onset time in ethylene production and response to exogenous ethylene. The shortest vase life was for �Exotica� flowers which was only 11.6 days, while �Baltico� and �Pilar� lasted 2.5-3 times longer than �Exotica�. Most of the investigated cultivars showed notable increases in the amount of ethylene. However, �Baltico� and �Pilar� flowers produced only a trace amount of ethylene and had the longest vase life. Results showed that cultivars with a long vase life (�Baltico� and �Pilar�) display high ethylene responsiveness and, in contrast, cultivars with a short vase life (�Exotica� and �Mundo�) present low responsiveness. The decline in fresh weight of cut flowers observed in the last phase of their vase life occurred earlier in short-lived cultivars than in the longer-lived ones.La vida util de la flor cortada es uno de los caracteres mas importantes de evaluacion de la calidad en poscosecha. El momento y cantidad de etileno producido por la flor de clavel varia entre cultivares y eso influye en la vida util de la flor. El objetivo de este trabajo fue evaluar diferencias en la produccion de etileno y su repuesta al etileno exogeno en diez cultivares de clavel (�eBaltico�f, �eDomingo�f, �eExotica�f, �eFamosa�f, �eKiro�f, �eMadame Augier�f, �eMaster�f, �eMundo�f, Pilar�f y �eReina�f). Se expusieron durante 8 h cinco flores de cada cultivar a una concentracion de 1 �ÊL L.1 de etileno exogeno. Se midio la produccion de etileno, peso fresco y absorcion de agua diariamente. Los 10 cultivares estudiados presentaron diferencias claras en la vida util de la flor, tiempo y produccion de etileno y respuesta al etileno. �eExotica�f fue el cultivar con vida util mas corta (11,6 dias), mientras �eBaltico�f y �ePilar�f presentaron una vida util 2,5- 3 veces mayor que �eExotica�f. La mayoria de los cultivares produjeron una notable cantidad de etileno, sin embargo �eBaltico�f y �ePilar�f, con la vida util mas larga, produjeron solo cantidades trazas de etileno. Estos cultivares de vida util larga (�eBaltico�f y �ePilar�f) mostraron alta sensibilidad al etileno, sin embargo, los de vida util corta (�eExotica�f y �eMundo�f) presentaron baja sensibilidad. La reduccion del peso fresco observado en la primera fase de la vida util de la flor, ocurre antes en los cultivares con una vida util corta que larga