Phenylpropanoid metabolism in relation to peel browning development of cold-stored 'Nanguo' pears.

Cold-stored 'Nanguo' pears are susceptible to peel browning during subsequent shelf life. In this study, 'Nanguo' pears were cold-stored for different periods to elucidate the metabolism of phenylpropanoid accompanying browning. Changes in phenolics and flavonoids and the crucial enzyme activity and related gene expression involved in the phenylpropanoid pathway were monitored. It was found that the fruit that underwent long-term storage showed peel browning symptoms prior to softening, and the symptom got worse with increasing shelf life. Meanwhile, the accumulation of reactive oxygen species (ROS) and the decrease of ROS scavenging ability were noted. The content of phenolics and flavonoids and the activity and expression of shikimate dehydrogenase (SKDH), phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), and 4-coumarate-CoA ligase (4CL) involved in the phenylpropanoid pathway decreased with prolonged storage. Correlation analysis revealed that browning was positively correlated with ROS accumulation, and the content of phenolics and flavonoids directly affected ROS scavenging ability. In addition, the decrease in phenolics and flavonoids might be owing to the reduced activity of SKDH, PAL, and 4CL and the down-regulated expression of PuPAL and Pu4CL. Collectively, this study indicated that the metabolism of phenylpropanoid is associated with the browning response induced by low-temperature stress.

24-epibrassinolide alleviates postharvest yellowing of broccoli via improving its antioxidant capacity.

Postharvest crop yellowing is a major concern in the broccoli industry. The effect and underlying mechanisms of 24-epibrassinolide (EBR) treatment on yellowing in postharvest broccoli were investigated. Treatment with 2 µM EBR markedly inhibited the increase of the yellowing index and L* values, causing higher retention of the metric hue angle and chlorophyll content compared to the control. Treatment also alleviated oxidative damage by preventing the accumulation of malondialdehyde and superoxide anion (O2•-). The ascorbic acid content of broccoli reached its lowest value at the end of its shelf life, whereas that of the treated sample was obviously higher than the control. Moreover, treated broccoli exhibited higher superoxide dismutase, ascorbate peroxidase, and phenylalanine ammonia-lyase activities. Multivariate statistical analysis further demonstrated the effective enhancement of EBR treatment on antioxidant enzymes. These results indicate that exogenous application of EBR ameliorates postharvest yellowing by improving the antioxidant capacity of broccoli.

Phenolics Profile and Antioxidant Activity Analysis of Kiwi Berry (Actinidia arguta) Flesh and Peel Extracts From Four Regions in China.

The kiwi berry (Actinidia arguta) has been widely studied because of its rich phenolic, flavonoid, and vitamin C contents. Numerous reports have demonstrated that fruit peels contain higher phenolic content and antioxidant activity than that of flesh. In this study, the phytochemical content and antioxidant activities of peel and flesh extracts of six kiwi berries were analyzed from four regions (namely, Dandong, Benxi, Taian, and Tonghua) in China. The antioxidant activity was determined using the peroxyl radical scavenging capacity (PSC) and cellular antioxidant activity (CAA) assays. The phenolic, flavonoid, and vitamin C contents of kiwi berry peel were 10.77, 13.09, and 10.38 times richer than that of kiwi berry flesh, respectively. In addition, the PSC and CAA values of kiwi berry peel were higher than those of kiwi berry flesh. The analysis of the separation and contents of phenolics were performed by the high-performance liquid chromatography (HPLC)-diode-array detectormass spectrometry/mass (DAD-MS/MS) system, and the results illustrated that protocatechuic acid, caffeic acid, chlorogenic acid, and quinic acid were the major phenolic compounds. In conclusion, this study indicated that kiwi berry peel contains a rich source of antioxidants. These data are of great significance for the full development and utilization of kiwi berries in these four regions of China to produce nutraceutical and functional foods.

Glycine betaine treatment alleviates loss of aroma-related esters in cold-stored 'Nanguo' pears by regulating the lipoxygenase pathway.

Glycine betaine (GB) is known to alleviate chilling injury in many fruit species. Therefore, we studied how GB affects the biosynthesis of esters in 'Nanguo' pears. Based on the kinds of esters, total esters, and the quantity of the main esters, it was evident that aroma losses were alleviated by GB treatment. In addition, unsaturated fatty acids contents (linoleic and linolenic acid) and the activities of lipoxygenase (LOX) and alcohol acyltransferase (AAT) enzymes were also increased. Meanwhile, comparing with the control fruit, the genes directly involved in ester synthesis were up-regulated in the GB-treated fruit. In addition, an increase in the activities and gene expression of antioxidant enzymes was observed in the treated samples. Thus, GB treatment promotes the synthesis of esters by regulating the LOX pathway and increasing antioxidant capacity, thereby effectively improving the quality of esters in cold-stored fruit.

Transcriptome analysis of postharvest blueberries (Vaccinium corymbosum 'Duke') in response to cold stress.

BACKGROUND: Blueberry (Vaccinium spp.) is a small berry with high economic value. Although cold storage can extend the storage time of blueberry to more than 60 days, it leads to chilling injury (CI) displaying as pedicle pits; and the samples of 0 °C-30 days was the critical point of CI. However, little is known about the mechanism and the molecular basis response to cold stress in blueberry have not been explained definitely. To comprehensively reveal the CI mechanisms in response to cold stress, we performed high-throughput RNA Seq analysis to investigate the gene regulation network in 0d (control) and 30d chilled blueberry. At the same time, the pitting and decay rate, electrolyte leakage (EL), malondialdehyde (MDA) proline content and GSH content were measured. RESULTS: Two cDNA libraries from 0d (control) and 30d chilled samples were constructed and sequenced, generating a total of 35,060 unigenes with an N50 length of 1348 bp. Of these, 1852 were differentially expressed, with 1167 upregulated and 685 downregulated. Forty-five cold-induced transcription factor (TF) families containing 1023 TFs were identified. The DEGs indicated biological processes such as stress responses; cell wall metabolism; abscisic acid, gibberellin, membrane lipid, energy metabolism, cellular components, and molecular functions were significantly responsed to cold storage. The transcriptional level of 40 DEGs were verified by qRT-PCR. CONCLUSIONS: The postharvest cold storage leads serious CI in blueberry, which substantially decreases the quality, storability and consumer acceptance. The MDA content, proline content, EL increased and the GSH content decreased in this chilled process. The biological processes such as stress responses, hormone metabolic processes were significantly affected by CI. Overall, the results obtained here are valuable for preventing CI under cold storage and could help to perfect the lack of the genetic information of non-model plant species.

Potential of jasmonic acid (JA) in accelerating postharvest yellowing of broccoli by promoting its chlorophyll degradation.

Chlorophyll degradation is the main reason for postharvest yellowing of broccoli. To uncover the role of jasmonic acid (JA) on the degradation of chlorophyll, broccoli flowers were treated with exogenous methyl jasmonate (MeJA) and diethyldithiocarbamic acid (DIECA). We found a surge of endogenous JA content with the yellowing process, and a significant correlation between JA and chlorophyll content. MeJA treatments led to increased endogenous JA, increased allene oxide cyclase (AOC) activity, and enhanced expression of JA synthesis genes. MeJA caused a stronger reduction in the maximum quantum yield (Fv/Fm), fluorescence decline ratio (Rfd), and total chlorophyll content, advanced the peak of pheide a oxygenase (PAO) activity, and up-regulated the expression of chlorophyll degradation genes. The DIECA treatment resulted in lower endogenous levels of JA, and AOC and 12-oxo-phytodienoic acid reductase (OPR) activity. This study revealed that the potential role of JA on broccoli yellowing is to promote the chlorophyll degradation.

Exogenous glycine betaine treatment alleviates low temperature-induced pericarp browning of 'Nanguo' pears by regulating antioxidant enzymes and proline metabolism.

The effect of glycine betaine (GB) on chilling injury (CI)-induced pericarp browning in 'Nanguo' pears was investigated during shelf life at 20 °C after storage at 0 °C for 120 d. GB treatment alleviated the severity of browning in 'Nanguo' pears as represented by lower browning index (BI) and browning incidence. Membrane lipid peroxidation in GB-treated fruit was lower than that in the control, and membrane integrity was maintained in good condition. The activities and expression of ascorbate peroxidase (APX), catalase (CAT), and superoxide dismutase (SOD) were higher in GB-treated fruit than in control fruit. Furthermore, significantly higher proline content, proline synthesis key enzyme activities, and gene expression were observed in the treated fruit, including ornithine d-aminotransferase (OAT) and Δ1-pyrroline-5-carbox-ylate synthetase (P5CS), which were consistent with the browning tendency. In a nutshell, GB treatment can effectively alleviate pericarp browning of cold-stored 'Nanguo' pears by regulating antioxidant enzymes and proline metabolism.

Changes in Membrane Lipid Metabolism Accompany Pitting in Blueberry During Refrigeration and Subsequent Storage at Room Temperature.

Low-temperature storage is the primary postharvest method employed to maintain fruit quality and commercial value. However, pitting can develop during refrigeration, especially during the shelf life. In this study, a membrane lipidomic approach was employed to analyze the potential relationship between pitting and membrane lipid metabolism during post-cold-storage shelf life. We also determined the changes in ultrastructure and water distribution by low-field nuclear magnetic resonance (LF-NMR) and assessed the permeability of membrane, membrane lipid peroxidation, proline and malondialdehyde contents, and the activity and gene expression of phospholipase D and lipoxygenase, which are involved in membrane lipid metabolism. The results indicated that the changes in blueberry phospholipids during storage could be caused by cold stress. Furthermore, dehydration is a manifestation of chilling injury. Finally, the significant increase in electrolyte leakage, content of malondialdehyde and proline, and activity of phospholipase D and lipoxygenase in chilled blueberry also indicated that membrane lipid metabolism plays an important role in cold stress response.

Chlorophyll degradation and carotenoid biosynthetic pathways: Gene expression and pigment content in broccoli during yellowing.

Broccoli undergoes yellowing in unfavorable conditions, thereby diminishing the sensory quality and commodity value. This study aimed to investigate systematically cellular and/or biomolecular changes involved in broccoli yellowing by analyzing changes in microstructural integrity, pigment content, and gene expression. On day-5 of storage at 20 °C, the buds turned yellow without blooming and showed structural damage; ultrastructural analysis revealed plastid transformation and abnormal chloroplast development. Genes regulating pigment content and chloroplast structure directly were identified. More specifically, BoCAO and BoNYC1 regulated chlorophyll turnover, affecting chlorophyll a and b contents. Changes in the ß-cryptoxanthin content were influenced by the combined action of up- (BoHYD) and downstream (BoZEP) genes. BoZEP and BoVDE were activated after cold-temperature induction. High BoHO1 expression delayed yellowing at low temperature, inducing BoZEP expression. Color intensity correlated significantly with the chlorophyll b, ß-cryptoxanthin, and ß-carotene contents, which were associated with increased yellowing of plant tissues.

GABA application improves the mitochondrial antioxidant system and reduces peel browning in 'Nanguo' pears after removal from cold storage.

Although refrigeration is commonly used in the storage of 'Nanguo' pears (Pyrus ussuriensis Maxim.), long-term refrigeration can result in browning of the pericarp. In this study, we aimed to determine how γ-aminobutyrate (GABA) affects the mitochondrial oxidation defense system in 'Nanguo' pears and how it might be used to prevent post-refrigeration peel browning. We found that fruit treated with GABA browned slower, and had lower browning indices and reactive oxygen and malondialdehyde content; increased peroxidase, superoxide dismutase, alternating oxidase, and catalase enzyme activities; and heightened enzyme-related gene expression. The mitochondria of GABA-treated fruit also showed less damage following cold storage, and there were decreases in mitochondrial membrane permeability transition pore concentrations. Furthermore, we detected an increase in the endogenous GABA content of fruits following GABA treatment. These observations indicate that, by regulating the mitochondrial oxidative defense system and maintaining mitochondrial structure, GABA is effective in terms of reducing peel browning.

Low temperature conditioning alleviates loss of aroma-related esters of 'Nanguo' pears by regulation of ethylene signal transduction.

The optimum harvest time for 'Nanguo' pears lasts for only 20 days. Refrigeration is a common technique used to extend the fruit supply. However, with low-temperature storage, the proper aroma of the pear is lost when it matures during shelf-life at room temperature. In this study, we investigated the ability of low-temperature conditioning (LTC) to regulate aroma esters of 'Nanguo' pears, and identified the key genes in the ethylene signal transduction pathway based on RNA-seq analysis. LTC caused higher ethylene production and effectively alleviated the loss of aroma-related esters in 'Nanguo' pears during refrigeration and during the subsequent shelf-life at room temperature. Furthermore, the expression levels of PuERS1, PuEIN4, PuEIN2, and PuERF were increased under LTC treatment. Thus, we speculate that the alleviation effect of LTC on fruit aroma esters is closely related to ethylene signal transduction, including up-regulated expression of PuERS1, PuEIN4, PuEIN2, and PuERF.

Effects of methyl jasmonate and melatonin treatments on the sensory quality and bioactive compounds of harvested broccoli.

Harvested broccoli is prone to decline in quality with regard to its appearance and nutrition. In this study, freshly harvested broccoli was treated with methyl jasmonate (MeJA) and melatonin (MT) and stored at 20 °C and the changes in sensory qualities and bioactive compounds were analyzed. The control samples began yellowing on day 2, whereas MeJA and MT treatments delayed the yellowing by 2 and 4 days, respectively. Upon yellowing, sweetness and bitterness of control samples increased sharply, accompanied by the accumulation of bioactive compounds, except for sulforaphane; however, no significant change in volatile components was detected. When the samples started losing their green color, MeJA alleviated the bitterness while increasing the sweetness and sulforaphane content. The bitterness, astringency, umami level, and the content of sulfurous volatiles improved significantly in the MT-treated samples. Moreover, these samples showed high antioxidant activity; the protective effect on VC and carotenoids was extremely significant.

Proteomic analysis of peel browning of 'Nanguo' pears after low-temperature storage.

BACKGROUND: Postharvest ripening of the 'Nanguo' pear (Pyrus ussuriensis Maxim.) can be impeded by low-temperature storage. However, pears after long-term refrigeration are prone to peel browning when returned to room temperature conditions. This study investigated the browning mechanism of 'Nanguo' pear stored at a low temperature by analysing the differentially expressed proteins between healthy fruit and fruit with peel browning. RESULTS: The results showed that 181 proteins underwent statistically significant changes. A categorisation of the disparately accumulated proteins was performed using gene ontology annotation. The results showed that the 'metabolic process', 'cellular process', 'catalytic activity', and 'binding' proteins were the most affected after low-temperature storage. Further analysis revealed that the differentially expressed proteins, which are related to peel browning, are primarily involved in the phenylpropanoid pathway, linoleic acid pathways, fatty acid biosynthesis pathway, glutathione metabolism pathway, photosynthesis pathway, oxidative phosphorylation pathway, and glycolysis pathway. CONCLUSION: This study reveals that there are variations in key proteins in 'Nanguo' pear after low-temperature storage, and the identification of these proteins will be valuable in future functional genomics studies, as well as provide protein resources that can be used in the efforts to improve pear quality. © 2016 Society of Chemical Industry.

Changes in energy metabolism accompanying pitting in blueberries stored at low temperature.

Low-temperature storage and transport of blueberries is widely practiced in commercial blueberry production. In this research, the storage life of blueberries was extended at low temperature, but fruit stored for 30 d at 0°C pitted after 2d at room-temperature. Fruit cellular structure and physiological parameters accompanying pitting in blueberries were changed. The objective of this research was to characterise properties of energy metabolism accompanying pitting in blueberries during storage, including adenosine phosphates and mitochondrial enzymes involved in stress responses. Physiological and metabolic disorders, changes in cell ultrastructure, energy content and ATPase enzyme activity were observed in pitting blueberries. Energy shortages and increased activity of phenylalanine ammonia lyase (PAL) and lipoxygenase (LOX) were observed in fruit kept at shelf life. The results suggested that sufficient available energy status and a stable enzymatic system in blueberries collectively contribute to improve chilling tolerance, thereby alleviating pitting and maintaining quality of blueberry fruit in long-term cold storage.

Rhodamine B induces long nucleoplasmic bridges and other nuclear anomalies in Allium cepa root tip cells.

The cytogenetic toxicity of rhodamine B on root tip cells of Allium cepa was investigated. A. cepa were cultured in water (negative control), 10 ppm methyl methanesulfonate (positive control), and three concentrations of rhodamine B (200, 100, and 50 ppm) for 7 days. Rhodamine B inhibited mitotic activity; increased nuclear anomalies, including micronuclei, nuclear buds, and bridged nuclei; and induced oxidative stress in A. cepa root tissues. Furthermore, a substantial amount of long nucleoplasmic bridges were entangled together, and some nuclei were simultaneously linked to several other nuclei and to nuclear buds with nucleoplasmic bridges in rhodamine B-treated cells. In conclusion, rhodamine B induced cytogenetic effects in A. cepa root tip cells, which suggests that the A. cepa root is an ideal model system for detecting cellular interactions.