The Expression of Key Ethylene and Anthocyanin Biosynthetic Genes of 'Honeycrisp' Apples Subjected to the Combined Use of Reflective Groundcovers and Aminoethoxyvinylglycine in the Mid-Atlantic US.

The decreased profitability of important apple cultivars, such as 'Honeycrisp', results from the poor red skin coloration and high fruit drop in the mid-Atlantic US. Apple red skin coloration is determined by the anthocyanin concentration. Reflective groundcovers promote red skin coloration, whereas aminoethoxyvinylglycine (AVG) decreases the ethylene production and fruit drop, thus reducing the coloration. Although our previous study showed that combinations of these practices impact the fruit quality and color, research is lacking regarding their effects at the gene and metabolite levels. In this work, for two years, we compared the differences in the internal ethylene concentration (IEC), red skin coloration, fruit drop, transcript accumulation of key ethylene and anthocyanin biosynthetic pathway-related genes, and total anthocyanin concentration of 'Honeycrisp' apples. The fruit was treated with combinations of reflective groundcover (Extenday) and AVG (130 mg L-1) and was assessed throughout ripening. Extenday-only-treated fruit displayed the highest upregulation of ethylene and anthocyanin biosynthetic-related genes and of total anthocyanins, exceeding 50% blush, while boosting the IEC. In contrast, AVG significantly decreased the expression of key ethylene and anthocyanin biosynthetic-related genes and total anthocyanins, thus preventing apples from reaching 50% blush, while also decreasing the IEC and fruit drop. The combination of Extenday x AVG fine-tuned the transcript accumulation of ethylene and anthocyanin biosynthetic-related genes as well as the total anthocyanins, allowing the 'Honeycrisp' fruit to exceed 50% blush, while increasing the IEC moderately and reducing the fruit drop (as compared to Extenday-only and control), thus enhancing the fruit economic value.

Changes in ethylene and sugar metabolism regulate flavonoid composition in climacteric and non-climacteric plums during postharvest storage.

Plums are rich in flavonoids, key contributors to fruit coloration and putative health benefits. We studied the impact of changes in ethylene and sugars in flavonoid metabolism-related pathways of the climacteric Santa Rosa and its non-climacteric mutant Sweet Miriam, throughout the postharvest period. Fruits were harvested at optimal maturity, subjected to ethylene treatments, and evaluated during storage. We examined transcript profiles of structural and regulatory genes of flavonoid-related pathways and their associated metabolites in skin and flesh, integrated with multivariate analyses of ethylene and sugar metabolism. Ethylene treatments were positively correlated with anthocyanin and negatively correlated with flavonol and flavan-3-ol metabolism. Sucrose and galactose were positively associated with anthocyanin concentration, while sorbitol, fructose, glucose and minor sugars were correlated with flavonol and flavan-3-ol metabolism. Our results support the notion that ethylene is playing key roles in shifting plum fruit flavonoid profiles, which are also associated with changes in fruit sugars.

Sensory, physicochemical and volatile compound analysis of short and long shelf-life melon (Cucumis melo L.) genotypes at harvest and after postharvest storage.

Flavor is a key attribute defining melon fruit quality and driving consumer preferences. We characterized and compared fruit ripening patterns (ethylene, respiration), physicochemical properties (rind/flesh color, firmness, soluble solids, acidity), aroma volatiles, and flavor-related sensory attributes in seven melon genotypes differing in shelf life capacity. Fruits were evaluated at optimal maturity and after storage for six days at 5 °C plus one day at room temperature. Total volatile content increased after storage in all genotypes, with esters being dominant. Shorter shelf-life genotypes, displaying a sharper climacteric phase, correlated with fruity/floral/sweet flavor-related descriptors, and with esters, sulfur-containing compounds and a terpenoid. Longer shelf-life types were associated with firmness, green and grassy aroma/flavor and aldehydes. Multivariate regression identified key volatiles that predict flavor sensory perception, which could accelerate breeding of longer shelf-life melons with improved flavor characteristics.

Ethylene Response of Plum ACC Synthase 1 (ACS1) Promoter is Mediated through the Binding Site of Abscisic Acid Insensitive 5 (ABI5).

The enzyme 1-amino-cyclopropane-1-carboxylic acid synthase (ACS) participates in the ethylene biosynthesis pathways and it is tightly regulated transcriptionally and post-translationally. Notwithstanding its major role in climacteric fruit ripening, the transcriptional regulation of ACS during ripening is not fully understood. We studied fruit ripening in two Japanese plum cultivars, the climacteric Santa Rosa (SR) and its non-climacteric bud sport mutant, Sweet Miriam (SM). As the two cultivars show considerable difference in ACS expression, they provide a good system for the study of the transcriptional regulation of the gene. To investigate the differential transcriptional regulation of ACS1 genes in the SR and SM, their promoter region, which showed only minor sequence differences, was isolated and used to identify the binding of transcription factors interacting with specific ACS1 cis-acting elements. Three transcription factors (TFs), abscisic acid-insensitive 5 (ABI5), GLABRA 2 (GL2), and TCP2, showed specific binding to the ACS1 promoter. Synthetic DNA fragments containing multiple cis-acting elements of these TFs fused to ß-glucuronidase (GUS), showed the ABI5 binding site mediated ethylene and abscisic acid (ABA) responses of the promoter. While TCP2 and GL2 showed constant and similar expression levels in SM and SR fruit during ripening, ABI5 expression in SM fruits was lower than in SR fruits during advanced fruit ripening states. Overall, the work demonstrates the complex transcriptional regulation of ACS1.

Hormone balance in a climacteric plum fruit and its non-climacteric bud mutant during ripening.

Hormone balance plays a crucial role in the control of fruit ripening. We characterized and compared hormone balance in two Japanese plum cultivars (Prunus salicina Lindl.), namely Santa Rosa, a climacteric type, and Sweet Miriam, its non-climacteric bud-sport mutant. We assessed hormonal changes in gene expression associated with hormone biosynthesis, perception and signaling during ripening on-the tree and throughout postharvest storage and in response to ethylene treatments. Non-climacteric fruit displayed lower ethylene levels than climacteric fruit at all stages and lower auxin levels during the initiation of ripening on-the-tree and during most of post-harvest storage. Moreover, 1-MCP-induced ethylene decrease also resulted in low auxin contents in Santa Rosa, supporting the role of auxin in climacteric fruit ripening. The differences in auxin contents between Santa Rosa and Sweet Miriam fruit could be the consequence of different routed auxin biosynthesis pathways as indicated by the significant negative correlations between clusters of auxin metabolism-associated genes. Ethylene induced increased ABA levels throughout postharvest storage in both ripening types. Overall, ripening of Santa Rosa and Sweet Miriam fruit are characterized by distinct hormone accumulation pathways and interactions.

Sugar metabolism reprogramming in a non-climacteric bud mutant of a climacteric plum fruit during development on the tree.

We investigated sugar metabolism in leaves and fruits of two Japanese plum (Prunus salicina Lindl.) cultivars, the climacteric Santa Rosa and its bud sport mutant the non-climacteric Sweet Miriam, during development on the tree. We previously characterized differences between the two cultivars. Here, we identified key sugar metabolic pathways. Pearson coefficient correlations of metabolomics and transcriptomic data and weighted gene co-expression network analysis (WGCNA) of RNA sequencing (RNA-Seq) data allowed the identification of 11 key sugar metabolism-associated genes: sucrose synthase, sucrose phosphate synthase, cytosolic invertase, vacuolar invertase, invertase inhibitor, α-galactosidase, ß-galactosidase, galactokinase, trehalase, galactinol synthase, and raffinose synthase. These pathways were further assessed and validated through the biochemical characterization of the gene products and with metabolite analysis. Our results demonstrated the reprogramming of sugar metabolism in both leaves and fruits in the non-climacteric plum, which displayed a shift towards increased sorbitol synthesis. Climacteric and non-climacteric fruits showed differences in their UDP-galactose metabolism towards the production of galactose and raffinose, respectively. The higher content of galactinol, myo-inositol, raffinose, and trehalose in the non-climacteric fruits could improve the ability of the fruits to cope with the oxidative processes associated with fruit ripening. Overall, our results support a relationship between sugar metabolism, ethylene, and ripening behavior.

Non-climacteric ripening and sorbitol homeostasis in plum fruits.

During ripening fruits undergo several physiological and biochemical modifications that influence quality-related properties, such as texture, color, aroma and taste. We studied the differences in ethylene and sugar metabolism between two genetically related Japanese plum cultivars with contrasting ripening behaviors. 'Santa Rosa' (SR) behaved as a typical climacteric fruit, while the bud sport mutant 'Sweet Miriam' (SM) displayed a non-climacteric ripening pattern. SM fruit displayed a delayed ripening that lasted 120 days longer than that of the climacteric fruit. At the full-ripe stage, both cultivars reached similar final size and weight but the non-climacteric fruits were firmer than the climacteric fruits. Fully ripe non-climacteric plum fruits, showed an accumulation of sorbitol that was 2.5 times higher than that of climacteric fruits, and the increase in sorbitol were also paralleled to an increase in sucrose catabolism. These changes were highly correlated with decreased activity and expression of NAD(+)-dependent sorbitol dehydrogenase and sorbitol oxidase and increased sorbitol-6-phosphate dehydrogenase activity, suggesting an enhanced sorbitol synthesis in non-climacteric fruits.