Current challenges in postharvest biology of fruit ripening

This paper reviews the recent advances in the understanding of the fruit ripening process and describes future challenges. Fruit ripening is a complex developmental process which is orchestrated by the expression of ripening-related genes under the control of a network of signaling pathways. In climacteric fruit components responsible for the production of climacteric ethylene have been identified. Less progress has been made on non-climacteric fruit. Great advances have been made in the characterization of transcription factors (ERFs, RIN, etc...) that induce gene expression through the binding to their promoters. Genetic resources, genome sequencing and “omics” tools have been developed bringing a huge amount of data that will help to draw together an integrative network of regulatory and signaling pathways responsible for triggering and coordinating the ripening process. The discovery that some ripening events are controlled at the epigenetic level and, not in relation with the DNA sequences, opens novel perspectives

Identification of biomarkers associated to ‘Gala’ apples ripening and postharvest quality

Apple is, sociocultural and economically, on of the most important species in the world, and stands out for its high storage potential. However, the monitoring of factors that could result in fruit quality modifications during postharvest is essential to ensure the acceptability and for the development of new storage technologies in order to increase fruit shelf life. Approaches focused on molecular biology, such as RT-qPCR have been used to better understand the mechanisms involved in fruit development and maturation. In this study the use of RT-qPCR to monitoring apple quality during ripening and development in different postharvest conditions such as room temperature, cold storage with or without control of atmosphere and the 1-methylcyclopropene usage were proposed. The potential of genes involved in ethylene biosynthesis and response, cell wall modification and degradation, sugar and aroma metabolisms for employment as biomarkers of fruit development and quality were evaluated. Thus MdEXP4 is highlighted as biomarker for development and MdACO1, MdPG1, MdAF1, MdAF3 and MdAAT2 as potential biomarkers for ripening. MdACO1 and MdPG1 appear as suitable markers for quality, conservation technologies and storage time in apples. This work suggests that the study of gene expression by RT-qPCR may be an alternative for a better fruit characterization during the development and postharvest period.Apple is, sociocultural and economically, on of the most important species in the world, and stands out for its high storage potential. However, the monitoring of factors that could result in fruit quality modifications during postharvest is essential to ensure the acceptability and for the development of new storage technologies in order to increase fruit shelf life. Approaches focused on molecular biology, such as RT-qPCR have been used to better understand the mechanisms involved in fruit development and maturation. In this study the use of RT-qPCR to monitoring apple quality during ripening and development in different postharvest conditions such as room temperature, cold storage with or without control of atmosphere and the 1-methylcyclopropene usage were proposed. The potential of genes involved in ethylene biosynthesis and response, cell wall modification and degradation, sugar and aroma metabolisms for employment as biomarkers of fruit development and quality were evaluated. Thus MdEXP4 is highlighted as biomarker for development and MdACO1, MdPG1, MdAF1, MdAF3 and MdAAT2 as potential biomarkers for ripening. MdACO1 and MdPG1 appear as suitable markers for quality, conservation technologies and storage time in apples. This work suggests that the study of gene expression by RT-qPCR may be an alternative for a better fruit characterization during the development and postharvest period

Characteristics of the tomato chromoplast revealed by proteomic analysis

Chromoplasts are non-photosynthetic specialized plastids that are important in ripening tomato fruit (Solanum lycopersicum) since, among other functions, they are the site of accumulation of coloured compounds. Analysis of the proteome of red fruit chromoplasts revealed the presence of 988 proteins corresponding to 802 Arabidopsis unigenes, among which 209 had not been listed so far in plastidial databanks. These data revealed several features of the chromoplast. Proteins of lipid metabolism and trafficking were well represented, including all the proteins of the lipoxygenase pathway required for the synthesis of lipid-derived aroma volatiles. Proteins involved in starch synthesis co-existed with several starch-degrading proteins and starch excess proteins. Chromoplasts lacked proteins of the chlorophyll biosynthesis branch and contained proteins involved in chlorophyll degradation. None of the proteins involved in the thylakoid transport machinery were discovered. Surprisingly, chromoplasts contain the entire set of Calvin cycle proteins including Rubisco, as well as the oxidative pentose phosphate pathway (OxPPP). The present proteomic analysis, combined with available physiological data, provides new insights into the metabolic characteristics of the tomato chromoplast and enriches our knowledge of non-photosynthetic plastids

UV-C radiation modifies the ripening and accumulation of ethylene response factor (ERF) transcripts in tomato fruit

Ultraviolet-C (UV-C) radiation is used as a postharvest treatment to prolong the shelf life of fruit. However, this stressful process may also affect ethylene production and, consequently, the expression of genes encoding ethylene response factors (ERFs). To test this hypothesis, MicroTom tomatoes harvested at the breaker stage were subjected to: 1 – application of 3.7 kJ m-2 UV-C radiation, 2 – application of 2 µLL-1 1-methylcyclopropene (1-MCP) followed by UV-C radiation; and 3 – without 1-MCP or UV-C (control treatment). After treatment all fruit were stored for 12 d at 21 +/- 2C° and 80 +/- 5% relative humidity (RH). Although UV-C radiation increased ACC oxidase transcripts and stimulated ethylene production, the ripening evolution was delayed. Fruit treated with UV-C showed lower accumulation of lycopene, b-carotene, lutein + zeaxanthin and d-tocopherol; but retained higher levels of chlorogenic acid, r-coumaric acid and quercetin after 6 d. Additionally, UV-C treated fruit had higher contents of polyamines (putrescine and spermidine). Among the 14 ERFs studied, 11 (Sl-ERF A.1, Sl-ERF A.3,Sl-ERF B.1, Sl-ERF B.2, Sl-ERF B.3, Sl-ERF C.6, Sl-ERF D.1, Sl-ERF D.3, Sl-ERF E.1, Sl-ERF F.5, Sl-ERF G.2) exhibited increased transcript accumulation, 2 ERFs (Sl-ERF E.2 and Sl-ERF E.4) showed decreased transcript accumulation and only 1 ERF (Sl-ERF E.3) was not significantly affected by UV-C treatment. As expected, the transcript profiles of 1-MCP and/or UV-C-treated tomatoes demonstrate that ethylene plays an important role in the expression of ERFs. The delay in fruit ripening may be caused by the activation of ERFs that could act as regulators of metabolic pathways during ripening. However, this hypothesis needs to be better tested. In conclusion, a relationship has been established between UV-C treatment and ripening delay, correlated to changes in 13 ERF transcripts evaluated during postharvest treatment

Genome-wide analysis of the AP2/ERF superfamily in apple and transcriptional evidence of ERF involvement in scab pathogenesis

The APETALA2 (AP2)/ETHYLENE RESPONSE FACTOR (ERF) superfamily of transcriptional regulators is involved in several growth, development and stress responses processes in higher plants. Currently, the available information on the biological roles of AP2/ERF genes is derived from Arabidopsis thaliana. In the present work, we have investigated genomic and transcriptional aspects of AP2/ERF genes in the economically important perennial species, Malus ×domestica. We have identified 259 sequences containing at least one ERF domain in apple genome. The vast majority of the putative proteins display predicted nuclear localization, compatible with a biological role in transcription regulation. The AP2 and ERF families are greatly expanded in apple. Whole-genome analyses in other plant species have identified a single genomic sequence with divergent ERF, whereas in apple seven soloists are present. In the apple genome, the most noteworthy expansion occurred in sub-groups V, VIII and IX of the ERF family. Expression profiling analyses have revealed the association of ripening-involved ERF genes to scab (Venturia inequalis) pathogenesis in the susceptible Gala cultivar, indicating that gene expansion processes were accompanied by functional divergence. The presented analyses of AP2/ERF genes in apple provide evidences of shared ethylene-mediated signaling pathways in ripening and disease responses

Putative role of cytokinin in differential ethylene response of twolines of antisense ACC oxidase cantaloupe melons

Two transgenic lines of ‘Cantaloupe’ melon derived from the same wild type genotype were previouslygenerated using ACC oxidase antisense constructs from melon (pMEL1AS) and apple (pAP4AS). Both linesyielded fruit with reduced ethylene production and low ACC oxidase (ACCO) expression. ACCO antisensefruit also exhibited lower expression of ACC synthase genes, ACCS1 and ACCS3, indicating that these genesare positively regulated by ethylene and participate in the autocatalytic ethylene production process. Incontrast, a higher expression of ACCS5 was observed in antisense lines when compared to the wild typeindicating a negative feedback regulation of ACCS5 by ethylene. Fruit of both transformed lines exhibiteddelayed ripening and reduction in ester volatile production but differed in their response to exogenousethylene supply. While postharvest ethylene application fully restored the ripening process in pMEL1ASmelon, it only restored flesh softening of pAP4AS melon but not rind color change or aroma volatile pro-duction. Up-regulation of lipoxygenase pathway associated genes (hydroxyperoxide lyase, lipoxygenase,and alcohol acyl transferases 1, 3 and 4) occurred in ethylene-treated pMEL1AS fruit but not in pAP4ASmelons. Polygalacturonase1 gene transcript accumulation increased in pMEL1AS and pAP4AS fruit uponethylene supply. Zeatin and zeatin riboside content of roots and fruit (rind and flesh) of pAP4AS plantswere 5-fold higher than the wild type and pMEL1AS counterparts. Higher relative transcript accumula-tion of a gene involved in the cytokinin synthesis and a gene involved in cytokinin response were alsofound in the roots and fruit of pAP4AS. In addition, polyamines, which are known to reduce sensitiv-ity to ethylene, remained unchanged in all fruit. Collectively the results suggest a putative role for theincreased endogenous cytokinin content in counteracting ethylene action in some aspects of the fruitripening process