Transcriptome Analysis of Pre-Storage 1-MCP and High CO2-Treated 'Madoka' Peach Fruit Explains the Reduction in Chilling Injury and Improvement of Storage Period by Delaying Ripening.
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ABSTRACT: Cold storage of peach fruit at low temperatures may induce chilling injury (CI). Pre-storage 1-MCP and high CO2 treatments were reported among the methods to ameliorate CI and reduce softening of peach fruit. However, molecular data indicating the changes associated with pre-storage 1-MCP and high CO2 treatments during cold storage of peach fruit are insufficient. In this study, a comparative analysis of the difference in gene expression and physico-chemical properties of fruit at commercial harvest vs. stored fruit for 12 days at 0 °C (cold-stored (CS), pre-storage 1-MCP+CS, and pre-storage high CO2+CS) were used to evaluate the variation among treatments. Several genes were differentially expressed in 1-MCP+CS- and CO2+CS-treated fruits as compared to CS. Moreover, the physico-chemical and sensory data indicated that 1-MCP+CS and CO2+CS suppressed CI and delayed ripening than the CS, which could lead to a longer storage period. We also identified the list of genes that were expressed commonly and exclusively in the fruit treated by 1-MCP+CS and CO2+CS and compared them to the fruit quality parameters. An attempt was also made to identify and categorize genes related to softening, physiological changes, and other ripening-related changes. Furthermore, the transcript levels of 12 selected representative genes from the differentially expressed genes (DEGs) in the transcriptome analysis were confirmed via quantitative real-time PCR (qRT-PCR). These results add information on the molecular mechanisms of the pre-storage treatments during cold storage of peach fruit. Understanding the genetic response of susceptible cultivars such as 'Madoka' to CI-reducing pre-storage treatments would help breeders release CI-resistant cultivars and could help postharvest technologists to develop more CI-reducing technologies.
Project description:Storage at the putative chilling threshold temperature (CTT) to avoid chilling injury still limits postharvest handling of tropical fruit like banana in that ripening may occur at the CTT. To determine whether chilling injury (CI) symptoms would develop in mature green (MG) banana fruit if the CTT exposure was extended by inhibiting ethylene action and thus ripening, 1-methylcyclopropene (1-MCP) was applied. Individual 'fingers' from multiple 'clusters' of MG bananas were either immersed in water or 50 μg L-1 1-MCP (a.i.) solution and each treatment was divided into three subgroups for storage at 5.0°C (severe CI), 13.0°C (mild CI), or 14.0°C (CTT) ± 0.1°C. 1-MCP delayed ripening in terms of color change for 10 days for fruit stored at the CTT. Ethylene production by fruit at 5.0°C remained around 0.04 ng kg-1 s-1 with no obvious increase during 31-day storage. Ethylene production at 14.0°C (-1-MCP/+1-MCP) increased on Day 33 while increasing on Day 38 for 13.0°C fruit without 1-MCP and on Day 39 for fruit with 1-MCP. Peak climacteric ethylene occurred on Days 44 and 39 for 13.0 and 14.0°C fruit without 1-MCP, respectively, and on Days 59 and 51 for 13.0°C and 14.0°C 1-MCP-treated fruit, respectively. As hypothesized, longer exposure of MG banana fruit to the CTT of 14.0°C without onset of ripening as was allowed by prior 1-MCP treatment allowed CI to develop at that normally non-chilling temperature. Vascular browning was the first visual and most sensitive CI symptom in the experiment and was observed on Day 4 at 5.0°C, Day 10 at 13.0°C, Day 19 at 14.0°C without 1-MCP, and on Day 28 at 14.0°C with 1-MCP. Using a 1-MCP pre-treatment to remove the influence of ethylene from bananas stored at 13°C or 14°C also resulted in slight reduction in vascular browning severity. In conclusion, a putative safe temperature may become a CI temperature if the shelf-life-limiting factor is removed, allowing longer exposure. Chilling at the CTT caused relatively mild injury on fruit, and vascular browning is a sensitive indicator of CI status, while the light-adapted quantum yield of photosystem II [Y(II)] could be a non-destructive indicator of early CI stress in MG banana. Fruit at 13.0/14.0°C developed CI symptoms slightly later with 1-MCP than without 1-MCP. This suggests that ethylene might be involved in early CI symptom development.
Project description:We investigated the effects of at-harvest maturity of 'YuLu' peach fruit on soluble sugar metabolism and their relationship with chilling injury susceptibility. Peaches were sorted into four maturity groups at harvest by I AD (index of the absorbance difference between 670 and 720 nm) then stored at 5 °C for 28 days. Fruit quality parameters, flesh browning index, malondialdehyde (MDA) content, soluble sugar content, gene expression, and enzyme activities associated with sucrose metabolism were measured. The more mature fruit groups had significantly (p < 0.05) lower firmness, higher soluble solid content, a* values of background color, sorbitol and sucrose content at harvest. During the cold storage, the higher flesh browning index in the mature groups (M3 and M4) maybe due to the double stress of senescence and chilling injury because there was concomitant sharp increase in MDA content. However, the most immature at-harvest group (M1) had the significantly (p < 0.05) higher MDA content after 14 days of cold storage, and a flesh browning index significantly (p < 0.05) higher than the M2 group (the next more mature group), late in the storage period. Moreover, the M1 group had lower sucrose content at postharvest and higher activities and transcript levels of sucrose degrading enzymes and lower levels of sucrose synthesis enzymes, which was responsible for the lower sucrose levels than M2 group during storage. It was concluded that the more immature peach fruit with lower sucrose content, have a higher chilling susceptibility than more mature fruit.
Project description:Consumers' choices are mainly based on fruit external characteristics such as the final size, weight, and shape. The majority of edible fruit are by tree fruit species, among which peach is the genomic and genetic reference for Prunus. In this research, we used a peach with a slow ripening (SR) phenotype, identified in the Fantasia (FAN) nectarine, associated with misregulation of genes involved in mesocarp identity and showing a reduction of final fruit size. By investigating the ploidy level, we observed a progressive increase in endoreduplication in mesocarp, which occurred in the late phases of FAN fruit development, but not in SR fruit. During fruit growth, we also detected that genes involved in endoreduplication were differentially modulated in FAN compared to SR. The differential transcriptional outputs were consistent with different chromatin states at loci of endoreduplication genes. The impaired expression of genes controlling cell cycle and endocycle as well as those claimed to play a role in fruit tissue identity result in the small final size of SR fruit.
Project description:Melatonin has been reported to alleviate chilling symptoms in postharvest peach fruit during cold storage, however, the mechanism involved is largely unknown. To better understand its role in chilling tolerance, here we investigated the effects of melatonin on oxidative damage in peach fruit subjected to chilling after harvest. Chilling injury of peaches was dramatically reduced by melatonin treatment. Melatonin induced hydrogen peroxide (H2O2) content at the early stage of storage but inhibited its accumulation thereafter. Meanwhile, melatonin also up-regulated the expression of genes involved in antioxidant responses in peaches. In addition, compared to the control fruit, peaches treated with melatonin displayed higher transcript abundance of ascorbic acid (AsA) biosynthetic genes and consequently increased the AsA content. Our results suggested that in response to melatonin during chilling, the high H2O2 level in the treated peaches at the initial time of storage, may work as a signaling molecule to induce protective mechanisms via up-regulating the expression of antioxidative genes and increasing AsA content. On the other hand, after the transient increase in the treated peaches, H2O2 was efficiently removed because of the activated antioxidant systems, which was associated with the higher chilling tolerance induced by melatonin.
Project description:Peach (Prunus persica) is a climacteric fruit with a relatively short shelf life due to its fast ripening or softening process. Here, we report the association of gene families encoding ethylene insensitive-3 like (EIL) and ethylene response factor (ERF) with fruit ripening in peach. In total, 3 PpEILs and 12 PpERFs were highly expressed in fruit, with the majority showing a peak of expression at different stages. All three EILs could activate ethylene biosynthesis genes PpACS1 and PpACO1. One out of the 12 PpERFs, termed PpERF.E2, is a homolog of ripening-associated ERFs in tomato, with a consistently high expression throughout fruit development and an ability to activate PpACS1 and PpACO1. Additionally, four subgroup F PpERFs harboring the EAR repressive motif were able to repress the PpACO1 promoter but could also activate the PpACS1 promoter. Promoter deletion assay revealed that PpEILs and PpERFs could participate in transcriptional regulation of PpACS1 through either direct or indirect interaction with various cis-elements. Taken together, these results suggested that all three PpEILs and PpERF.E2 are candidates involved in ethylene biosynthesis, and EAR motif-containing PpERFs may function as activator or repressor of ethylene biosynthesis genes in peach. Our study provides an insight into the roles of EILs and ERFs in the fruit ripening process.
Project description:BackgroundFruit ripening in Prunus persica melting varieties involves several physiological changes that have a direct impact on the fruit organoleptic quality and storage potential. By studying the proteomic differences between the mesocarp of mature and ripe fruit, it would be possible to highlight critical molecular processes involved in the fruit ripening.ResultsTo accomplish this goal, the proteome from mature and ripe fruit was assessed from the variety O'Henry through shotgun proteomics using 1D-gel (PAGE-SDS) as fractionation method followed by LC/MS-MS analysis. Data from the 131,435 spectra could be matched to 2740 proteins, using the peach genome reference v1. After data pre-treatment, 1663 proteins could be used for comparison with datasets assessed using transcriptomic approaches and for quantitative protein accumulation analysis. Close to 26% of the genes that code for the proteins assessed displayed higher expression at ripe fruit compared to other fruit developmental stages, based on published transcriptomic data. Differential accumulation analysis between mature and ripe fruit revealed that 15% of the proteins identified were modulated by the ripening process, with glycogen and isocitrate metabolism, and protein localization overrepresented in mature fruit, as well as cell wall modification in ripe fruit. Potential biomarkers for the ripening process, due to their differential accumulation and gene expression pattern, included a pectin methylesterase inhibitor, a gibbellerin 2-beta-dioxygenase, an omega-6 fatty acid desaturase, a homeobox-leucine zipper protein and an ACC oxidase. Transcription factors enriched in NAC and Myb protein domains would target preferentially the genes encoding proteins more abundant in mature and ripe fruit, respectively.ConclusionsShotgun proteomics is an unbiased approach to get deeper into the proteome allowing to detect differences in protein abundance between samples. This technique provided a resolution so that individual gene products could be identified. Many proteins likely involved in cell wall and sugar metabolism, aroma and color, change their abundance during the transition from mature to ripe fruit.
Project description:BackgroundCold storage induces chilling injury (CI) disorders in peach fruit (woolliness/mealiness, flesh browning and reddening/bleeding) manifested when ripened at shelf life. To gain insight into the mechanisms underlying CI, we analyzed the transcriptome of 'Oded' (high tolerant) and 'Hermoza' (relatively tolerant to woolliness, but sensitive to browning and bleeding) peach cultivars at pre-symptomatic stages. The expression profiles were compared and validated with two previously analyzed pools (high and low sensitive to woolliness) from the Pop-DG population. The four fruit types cover a wide range of sensitivity to CI. The four fruit types were also investigated with the ROSMETER that provides information on the specificity of the transcriptomic response to oxidative stress.ResultsWe identified quantitative differences in a subset of core cold responsive genes that correlated with sensitivity or tolerance to CI at harvest and during cold storage, and also subsets of genes correlating specifically with high sensitivity to woolliness and browning. Functional analysis indicated that elevated levels, at harvest and during cold storage, of genes related to antioxidant systems and the biosynthesis of metabolites with antioxidant activity correlates with tolerance. Consistent with these results, ROSMETER analysis revealed oxidative stress in 'Hermoza' and the progeny pools, but not in the cold resistant 'Oded'. By contrast, cold storage induced, in sensitivity to woolliness dependant manner, a gene expression program involving the biosynthesis of secondary cell wall and pectins. Furthermore, our results indicated that while ethylene is related to CI tolerance, differential auxin subcellular accumulation and signaling may play a role in determining chilling sensitivity/tolerance. In addition, sugar partitioning and demand during cold storage may also play a role in the tolerance/sensitive mechanism. The analysis also indicates that vesicle trafficking, membrane dynamics and cytoskeleton organization could have a role in the tolerance/sensitive mechanism. In the case of browning, our results suggest that elevated acetaldehyde related genes together with the core cold responses may increase sensitivity to browning in shelf life.ConclusionsOur data suggest that in sensitive fruit a cold response program is activated and regulated by auxin distribution and ethylene and these hormones have a role in sensitivity to CI even before fruit are cold stored.
Project description:Storage at low temperatures is a common practice to prolong postharvest life of fruit and vegetables with a minimal negative impact on human/environmental health. Storage at low temperatures, however, can be restricted due to produce susceptibility to non-freezing chilling temperatures, when injuries such as physiological disorders and decays may result in unmarketable produce. We have investigated tomato fruit response to postharvest chilling stress in a recombinant inbred line (RIL) population developed from a cross between a chilling-sensitive cultivated tomato (Solanum lycopersicum L.) breeding line and a chilling-tolerant inbred accession of the tomato wild species S. pimpinellifolium L. Screening of the fruit of 148 RILs under cold storage (1.5°C) indicated presence of significant variations in chilling tolerance, manifested by varying degrees of fruit injury. Two extremely contrasting groups of RILs were identified, chilling-tolerant and chilling-sensitive RILs. The RILs in the two groups were further investigated under chilling stress conditions, and several physiological parameters, including weight loss, chlorophyll fluorescence parameters Fv/Fm, and Performance Index (PI), were determined to be efficient markers for identifying response to chilling stress in postharvest fruit. The Fv/Fm values reflected the physiological damages endured by the fruit after cold storage, and PI was a sensitive marker for early changes in photosystem II function. These two parameters were early indicators of chilling response before occurrence of visible chilling injuries. Antioxidant activities and ascorbic acid content were significantly higher in the chilling-tolerant than the chilling-sensitive lines. Further, the expression of C-repeat/DREB binding factors (CBFs) genes swiftly changed within 1-hr of fruit exposure to the chilling temperature, and the SlCBF1 transcript level was generally higher in the chilling-tolerant than chilling-sensitive lines after 2-hr exposure to the low temperature. This research demonstrates the presence of potential genetic variation in fruit chilling tolerance in the tomato RIL population. Further investigation of the RIL population is underway to better understand the genetic, physiological, and biochemical mechanisms involved in postharvest fruit chilling tolerance in tomato.