Short‐term anoxia treatment maintains tissue energy levels and membrane integrity and inhibits browning of harvested litchi fruit

Pericarp browning is the main limitation to the postharvest storage, handling and marketing life of litchi fruit. Pre‐storage treatment with pure N₂ gas is potentially effective in reducing skin browning and maintaining eating quality of litchis. To better understand inhibition of pericarp browning by a short period of anoxia, adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) levels, adenylate energy charge and membrane permeability were investigated. Litchi fruit were exposed to pure N₂ gas for 6 h and then kept in closed but vented containers for 6 days in the dark at 25 °C and 95–100% relative humidity. Changes in the mentioned fruit physiology and biochemistry parameters and in browning index were measured. ATP concentration and adenylate energy charge decreased rapidly and membrane permeability (relative leakage) increased gradually during storage. Fruit exposed to N₂ gas exhibited higher concentrations of ATP, ADP and AMP and adenylate energy charge levels, and lower levels of browning index and membrane permeability, compared to control (non‐N₂‐treated) fruit. Greater differences in ATP and ADP concentrations and adenylate energy charge levels of pericarp tissues between N₂‐treated and control fruit were more manifest after 4 and 6 days of storage, in association with significant differences at the 5% level in the pericarp browning index. It is suggested that pre‐storage anoxia treatment maintains membrane integrity of pericarp tissues, with high ATP and ADP concentrations and high adenylate energy charge levels. Thus, the loss of cellular compartmentalization (mixing of enzymes and substrates) that leads to enzymatic browning of litchi fruit pericarp is delayed. Copyright © 2007 Society of Chemical Industry     

Effects of O2 and CO2 concentrations on physiology and quality of litchi fruit in storage

Litchi (Litchi chinensis Sonn. cv Heiye) fruit were stored in air, modified atmosphere packaging (MAP) and controlled atmospheres (CA) at 3 °C to determine the effects of different O₂ and CO₂ atmospheres on physiology, quality and decay during the storage periods. The results indicated that CA conditions were more effective in reducing total phenol content, delaying anthocyanidin decomposition, preventing pericarp browning, and decreasing fruit decay in comparison with MAP treatment. Polyphenol oxidase (PPO), peroxidase (POD), anthocyanin and total phenols were involved in cellular browning. High O₂ treatment significantly limited ethanol production of litchi flesh in the early period of storage. The fruit stored in CA conditions for 42 days maintained good quality without any off-flavour.     

壳聚糖对荔枝果皮渗漏性及贮藏时间的影响

通过分析经壳聚糖涂膜处理的荔枝果实在贮藏期间果皮细胞的渗漏率的变化,研究果皮细胞电解质渗漏性与果实贮藏性能之间的关系。在研究的29、22、15和8℃4个温度范围内,随贮藏温度下降,果皮渗漏率降低,果实贮藏时间延长。在贮藏时间范围内,渗漏率表现为先是升高至峰值,然后下降。渗漏率开始发生下降后,果实失去商品价值。并且渗漏率峰值出现得越晚,果实贮藏时间越长。渗漏率(X)与褐变指数(Y)呈显著的正相关(R=0.9506),其回归方程为Y=0.0683X-1.0039。壳聚糖涂膜处理明显抑制荔枝果皮细胞的渗漏率,果实贮藏时间较长。     

荔枝果皮变色机理的研究

白蜡荔枝研究表明：荔枝果皮花色苷在510nm有最大吸收峰,随着氢离子浓度的降低,花色苷的红色变浅或光谱吸收峰变低,吸收峰消失的临界氢离子浓度为1×10-3．5mol／L。但氢离子浓度在1×10-2～10-3mol／L范围内对多酚化合物的影响不明显。新鲜荔枝在室温放置两天半,果皮外表全部变色。荔枝果皮变色主要由3方面原因引起：1）果皮可滴定酸降低,H＋离子浓度逐渐降低并小于临界值而引起花色苷颜色由深变浅,最终失去红色,它是荔枝初期可逆性变色的重要原因。2）花色苷氧化破坏。3）酶促多酚褐变。     

Effect of pure oxygen atmosphere on antioxidant enzyme and antioxidant activity of harvested litchi fruit during storage

The effects of pure oxygen on pericarp browning, reactive oxygen species (ROS) metabolism, antioxidant enzyme and antioxidant activity of harvested litchi fruit were investigated. Application of pure oxygen significantly prevented pericarp browning and delayed the increase in membrane permeability of litchi fruit during storage. Litchi fruit exposed to pure oxygen showed a lower level of lipid peroxides, compared to control fruit, with the delay in the increases of both H₂O₂ content and superoxide production rate. Furthermore, it was found that the treatment with pure oxygen induced the activities of superoxide dismutase (SOD), ascorbated peroxidase (APX) and catalase (CAT), which could be beneficial in scavenging of H₂O₂ and superoxide and alleviating lipid peroxidation. In addition, antioxidant ability (reducing power and free-radical scavenging activity against DPPH radical, superoxide anions and hydroxyl radical) of methanol extracts from litchi fruit pericarp declined gradually, with decreasing contents of anthocyanins and phenolic compounds, as storage time of the fruit progressed. There was a linear relationship between the contents of either anthocyanins or phenolic compounds and antioxidant ability or free radical scavenging activity. Treatment with pure oxygen markedly increased antioxidant ability, which was related to higher levels of anthocyanins and phenolic compounds, compared with those of control fruit. It is suggested that enhanced antioxidant activity and antioxidant enzyme induced by pure oxygen may contribute to alleviating lipid peroxidation and maintenance of membrane integrity, which reduced decompartmentation of enzymes and substrates, resulting in enzymatic browning.     

Role of anthocyanin degradation in litchi pericarp browning

Pericarp browning is the main problem of post-harvest litchi fruit, resulting in an accelerated shelf life and reduced commercial value of the fruit. Underhill and Critchley (1994). Anthocyanin decolorisation and its role in lychee pericarp browning. Australian Journal of Experimental Agriculture, 34, 115-122 found that there was not an obvious change in the content of anthocyanins when the fruit browned. This work was conducted with a view to explaining this unexpected observation. Litchi pericarp browning index increased while the content of anthocyanins decreased with storage time when 0.1 M HCl was used as the extract solution instead of acidic methanol. The visible spectum of the anthocyanin extract, at a range of 400–600 nm and pH values of 1.0, 3.0 and 5.0, were recorded, with an absorbance peak of about 510 nm. The colour of the extract depended on the pH values and the half-degradation constants for anthocyanins at pHs 1.0, 3.0 and 5.0 were, respectively, 29, 15.3 and 10.5 days, as calculated from the kinetics of the degradation. Compared with the anthocyanin extract, anthocyanidin is more vulnerable, with a half-degradation of about 5.3 min at pH 5.0. Furthermore, the product from the anthocyanidin degradation had a similar structure to catechol (a good substrate for polyphenol oxidase), which, in turn, could accelerate enzymatic browning reaction by the enzyme polyphenol oxidase. In addition, an anthocyanase, catalyzing anthocyanin hydrolysis and producing anthocyanidin was extracted from litchi fruit pericarp. High activity of the enzyme was observed in the pericarp. Thus, it is suggested that anthocyanase might contribute to the browning of litchi pericarp involved in the anthocyanase-anthocyanin-PPO reaction.     

采后荔枝果实安全保鲜技术研究进展

荔枝果实在采后贮藏和运输中极易发生病原菌侵染、褐变及腐败而导致品质劣变。目前常用的物理保鲜方法成本高,而人工合成化学杀菌剂处理可能会有一定残留且病原菌容易产生抗药性,故使用无毒害、可生物降解、对环境无污染的新型安全保鲜技术是目前的研究热点。对国内外壳寡糖涂膜保鲜技术、植物提取物保鲜技术和生物拮抗菌保鲜技术等安全保鲜技术在采后荔枝果实保鲜应用的研究进展进行了综述,旨在为维持采后荔枝果实品质,延长荔枝果实贮藏期提供技术参考。     

Role of endogenous and exogenous phenolics in litchi anthocyanin degradation caused by polyphenol oxidase

The degradation of anthocyanins and/or the oxidation of phenolics caused by polyphenol oxidase (PPO) results in an enzymatic browning reaction of fruits and vegetables. This work was conducted with a view to explaining the unexpected observation that litchi (Litchi chinensis Sonn.) PPO did not directly oxidise litchi anthocyanins. PPO and anthocyanin from litchi fruit pericarp were extracted and purified, respectively, and then the anthocyanin degradation by PPO in the presence of (−)-epicatechin (endogenous PPO substrate), and catechol and gallic acid (exogenous PPO substrates) were analysed comparatively. The results showed that catechol was the most effective in litchi anthocyanin degradation, followed by (−)-epicatechin and gallic acid, but no significant differences existed between catechol and (−)-epicatechin. The study suggested that litchi PPO directly oxidised (−)-epicatechin; then oxidative products of (−)-epicatechin in turn catalysed litchi anthocyanin degradation, and finally resulted in the browning reaction, which can account for pericarp browning of postharvest litchi fruit.     

INHIBITORY EFFECT OF ANTHOCYANIN EXTRACT FROM SEED COAT OF BLACK BEAN ON PERICARP BROWNING AND LIPID PEROXIDATION OF LITCHI FRUIT DURING STORAGE

ABSTRACT

Anthocyanins were extracted from seed coats of black beans (Glycine max[L.]) and the inhibitory effects of anthocyanin extract on pericarp browning and lipid peroxidation of litchi fruit were investigated. Litchi fruit were infiltrated for 3 min with 0 (control) or 50 mg/L of anthocyanin extract at a reduced pressure of 53 kPa, then packed in 0.03 mm thick polyethylene bags, and finally stored at 28C for 6 days. Changes in browning index, contents of anthocyanins and total phenol, peroxidase (POD) activity, levels of relative leakage rate and lipid peroxidation, α,α‐diphenyl‐β‐picrylhydrazy (DPPH) radical scavenging activity and reducing power were evaluated. Application of anthocyanin extract from black bean delayed pericarp browning of litchi fruit during storage, which was associated with reduced POD activity and higher contents of anthocyanins and total phenol. Moreover, the anthocyanin extract was found to have a direct inhibition on the POD activity in vitro. Furthermore, application of the anthocyanin extract apparently reduced lipid peroxidation and relatively maintained membrane integrity, which may account for browning inhibition to an extent. Finally, higher DPPH radical scavenging activity and reducing power of the fruits treated with the anthocyanin extract than control fruits possibly benefited in scavenging free radicals and reducing lipid peroxidation. It is, therefore, suggested that inhibited POD activity and reduced lipid peroxidation by the anthocyanin extract from seed coats of black beans were responsible for the inhibition of pericarp browning of litchi fruits.

PRACTICAL APPLICATIONS

Lipid peroxidation is a major cause of quality deterioration of postharvest fruits and vegetables. Some synthetic antioxidants are beneficial in inhibiting lipid peroxidation. However, considering that synthetic antioxidants, such as butylhydroxyanisole and butylated hydroxytoluene (BHT), have potential toxicity, the use of natural extracts to extend the shelf life of postharvest fruits and vegetables is a tendency. In our previous study, it has been found that anthocyanin extract from litchi pericarp has stronger antioxidant activities or free radical scavenging activities than BHT and ascorbic acid ( Duan et al. 2007 ). In this study, the application of anthocyanin extract from seed coat of black soybean showed reduced lipid peroxidation and pericarp browning. It could be used potentially as a postharvest technology for reducing or replacing the use of other chemicals, but it requires further investigation.     

ATP-regulation of antioxidant properties and phenolics in litchi fruit during browning and pathogen infection process

The impact of energy level on antioxidant properties in relation to pericarp browning and loss of disease resistance of litchi fruit was investigated. Litchi fruits were vacuum-infiltrated with distiled water (control), 1 mM adenosine triphosphate (ATP) and 0.5 mM 2,4-dinitrophenol (DNP) under 75 kPa for 3 min before being inoculated with Peronophythora litchi or not. ATP-treated fruits presented higher activities of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX). Higher activities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging, reducing power and contents of phenolic compounds were also observed in ATP-treated fruit during storage. In contrast, DNP treatment lowered the enzymes activities, scavenging ability and the contents of phenolic compounds. Higher levels of antioxidant enzymatic system and non-enzymatic system were observed in P. litchii-inoculated fruits than those in non-inoculated fruits. Application of ATP and DNP exhibited a similar change patterns and effects in inoculated fruits. When related to previously reported ATP levels, the results suggested that ATP levels could regulate the antioxidant system. Sufficient available energy is crucial for inhibiting browning and preventing the loss of disease resistance in harvested litchi fruit.     

Two Endogenous Substrates for Polyphenoloxidase in Pericarp Tissues of Postharvest Rambutan Fruit

Abstract: The catalytic oxidation of phenolic substrates by polyphenoloxidase (PPO) causes pericarp browning of postharvest rambutan fruit. In the present study, PPO and its endogenous substrates were extracted from rambutan pericarp tissues (RPT). The substrate extracts were sequentially partitioned with ethyl acetate and n-butanol. The analysis of total phenolic content showed that the most phenolic compounds were distributed in ethyl acetate fraction. By high-performance liquid chromatography (HPLC), (−)-epicatechin (EC) and proanthocyanidin A2 (PA2) were identified from this fraction. After reacting with rambutan PPO, EC turned brown rapidly within 10 min, indicating that it was a significant endogenous substrate. Although PA2 could also be oxidized by the PPO, it turned brown very slowly. In addition, because EC and PA2 were continually catalyzed into browning products by PPO during storage of the fruit at 4 and 25 °C, their contents in RPT gradually declined with the extended storage time. It was further observed that both substrate contents in rambutan fruit storing at 25 °C decreased more rapidly than that storing at 4 °C, suggesting that low temperature inhibited the catalytic oxidation of substrates so as to slow down pericarp browning. Practical Application: Pericarp browning is a serious problem to storage and transport of harvested rambutan fruit. A generally accepted opinion on the browning mechanism is the oxidation of phenolic substrates by PPO. Ascertaining PPO substrates will effectively help us to control enzymatic reaction by chemical methods so as to delay or even prevent pericarp browning of harvested rambutan fruit.     

脱硫对熏硫‘黑叶’荔枝采后品质及果肉亚硫酸盐代谢的影响

本实验研究了脱硫对熏硫‘黑叶’荔枝果实采后品质、亚硫酸盐残留量和果肉中亚硫酸盐代谢的影响。测定对照（施保克）、熏硫和脱硫处理的果实4 ℃贮藏期间的色度值、褐变指数、腐烂率、亚硫酸盐残留量及果肉中亚硫酸盐氧化酶（sulfite oxidase,SO）、腺苷-5’-磷酰硫酸还原酶（adenosine 5’-phosphosulfatezoline reductase,APR）、亚硫酸盐还原酶（sulfite reductase,SiR）、丝氨酸酰基转移酶（serine acetyltransferase,SAT）和乙酰丝氨酸裂解酶（O-acetylserine (thiol) lyase,OAS-TL）活力和基因表达。结果表明：贮藏期间脱硫果实较熏硫果实提前转红;脱硫和熏硫荔枝的果皮褐变指数和腐烂率均较对照组显著降低;相比熏硫,脱硫使果皮水分质量分数降低和相对电导率升高的幅度更大,抑制呼吸的效果则相近;脱硫使果皮亚硫酸盐含量大幅降低,并使得果肉亚硫酸盐含量在贮藏16 d后降至接近对照组的水平;贮藏期间除APR外,熏硫和脱硫后果肉中其他4 种酶的活力均呈现上调,即贮藏过程中处理组SO、SAT活力分别较同时期对照组上调2.0、8.6～26.7 倍,而脱硫果肉SAT活力仅在贮藏0 d较对照组上调11.1 倍,之后迅速下降,贮藏32 d后与对照组无显著差异;熏硫和脱硫果肉中5 个基因的表达则均在贮藏中后期总体高于对照。上述结果表明,脱硫可加快熏硫荔枝果皮恢复红色,获得与熏硫相当的保鲜效果,大幅降低了熏硫荔枝的硫残留,保障其食用安全。脱硫和熏硫荔枝果肉中SO和SAT活力的上调表明,SO酶促氧化反应是荔枝果肉亚硫酸盐降解中占主导和最稳定的方式,SAT和SiR主导的还原途径则可能作为氧化途径的补充。     

Effect of nitric oxide on pericarp browning of harvested longan fruit in relation to phenolic metabolism

The effects of nitric oxide (NO) on enzymatic browning of harvested longan fruit in relation to phenolic metabolisms were investigated. Fruits were dipped for 5 min in 1 mM sodium nitroprusside (SNP), a nitric oxide donor, then packed in 0.03 mm thick polyethylene bags, and finally stored for 6 days at 28 °C. Changes in pericarp browning and pulp breakdown were evaluated, while total phenol content, activities of phenolic-associated enzymes, polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase (PAL), and concentrations of total soluble solids, titratable acidity and ascorbic acid were measured. SNP treatment delayed pericarp browning, inhibited activities of PPO, POD and PAL and maintained a high total phenol content of longan fruit during storage. Furthermore, NO showed a significant inhibition of the in vitro activities of PPO and POD, indicating that the beneficial effect of NO was direct. Moreover, application of NO resulted in a lower pulp breakdown and maintained relatively high levels of total soluble solids and ascorbic acid.     

Effect of hexanal vapour on longan fruit decay,quality and phenolic metabolism during cold storage

The effects of postharvest treatment with hexanal vapour on longan fruit decay, quality, hexanal residue, phenolic compound content, and polyphenoloxidase (PPO) and peroxidase (POD) activities were studied during storage at 5 °C for 30 days. Hexanal exposure for 2 h at 900 μL L^?1 before cold storage reduced the percentage of fruit with decay and was deemed the optimum treatment. Hexanal exposure resulted in a pericarp that was more reddish brown and less intense in colour. Hexanal residue in the pericarp and aril of fumigated fruit was several fold higher than that of nonfumigated fruit, although levels were low at the end of cold storage. Electrolyte leakage of pericarp increased during 5 °C storage and was further increased by hexanal exposure. Hexanal reduced pericarp phenolic content, and increased PPO and POD activities. Overall, use of hexanal vapour reduced postharvest disease of longan fruit but increased the likelihood of pericarp browning.     

荔枝保藏中的褐变研究进展

本文根据荔枝果实的结构与生理特点，综述了荔枝保藏中果皮褐变的原因，着重从水分、多酚氧化酶及花色素苷三方面进行了评述，并阐述了褐变产生的机制及控制的途径。     

采收期对冷藏橄榄果实贮藏期间冷害的影响

针对‘檀香’橄榄果实冷藏期间易发生冷害现象,本实验研究了7 个采收期（Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ、Ⅶ）对（2±1）℃、相对湿度85%～90%条件下冷藏橄榄果实冷害的影响。定期测定贮藏期间橄榄果实冷害指数、果皮褐变指数、果实好果率和质量损失率、果实呼吸强度和细胞膜相对渗透率、果实表面色度角、果皮叶绿素含量、果肉可溶性固形物、可滴定酸、可溶性总糖和还原糖含量的变化。结果表明：与采收期Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅵ、Ⅶ相比,采收期Ⅴ能保持冷藏橄榄果实较低的冷害指数、果皮褐变指数、果实质量损失率和较高的好果率,降低果实呼吸强度和呼吸峰值,延缓果实细胞膜相对渗透率升高,保持中后期较高的果实表面色度角、果皮叶绿素含量、果肉可溶性固形物、可滴定酸、可溶性总糖和还原糖含量。因此认为,采收期Ⅴ可作为减轻冷藏‘檀香’橄榄果实冷害发生的适宜采收成熟度。     

茉莉酸甲酯处理对采后龙眼果皮褐变的影响

本实验以10、50、100μmol/L MeJA处理‘储良’龙眼(Dimocarpus longan Lour.)果实,研究其对(20±1)℃、85%相对湿度贮藏条件下的龙眼果皮褐变的影响。结果表明：与对照组相比,MeJA处理可有效降低‘储良’龙眼采后果皮褐变指数、果皮细胞膜透性和果肉自溶指数,而且MeJA浓度越高,抑制效果越明显。与对照组相比,100μmol/L MeJA处理龙眼果实能够保持较高的果皮总酚和类黄酮含量,降低果皮多酚氧化酶活力,有效提高果皮过氧化物酶和苯丙氨酸解氨酶活力;另一方面,MeJA处理可以有效降低超氧阴离子自由基产生速率以及H₂O₂含量,有效增强抗氧化酶(超氧化物歧化酶、过氧化氢酶)活力以及1,1-二苯基-2-三硝基苯肼自由基清除能力,提高果皮抗氧化能力。综上,100μmol/L MeJA处理可抑制龙眼果实采后部分酚类物质代谢,增强其抗氧化能力,从而延缓龙眼采后果皮褐变的发生,延长贮藏保鲜期。     

EFFECT OF OXYGEN CONCENTRATION ON THE BIOCHEMICAL AND CHEMICAL CHANGES OF STORED LONGAN FRUIT

ABSTRACT

Longan fruits were stored for 6 days in atmosphere of 5, 21 (air) or 60% O₂ (balance N₂) at 28C and 90–95% relative humidity to examine effects of low and high O₂ concentration on enzymatic browning and quality attributes of the fruit. Changes in pericarp browning, pulp breakdown, disease development, total phenol content, activities of phenol metabolism‐associated enzymes, relative leakage rate, α,α‐diphenyl‐β‐picrylhydrazy (DPPH) radical scavenging activity, and contents of total soluble solids, titratable acidity and ascorbic acid were evaluated. Storage of fruit in a 5% O₂ atmosphere markedly delayed pericarp browning in association with maintenance of high total phenolic content and reduced activities of polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase. Moreover, the fruit stored in a 5% O₂ atmosphere exhibited a lower relative leakage rate and higher DPPH radical scavenging activity than fruit stored in air. This presumably was beneficial in maintaining compartmentation of enzymes and substrates, and thus, reducing pericarp browning. Pulp breakdown and disease development were also reduced by exposure to a 5% oxygenatmosphere. On the contrary, exposure of longan fruit to a 60% O₂ atmosphere accelerated pericarp browning, pulp breakdown and decay development. PPO and POD activities and relative leakage rate were similar for control and 60% O₂‐treated fruit after 4 and 6 days of storage. Furthermore, treatment with 60% O₂ significantly decreased the phenolic content and DPPH scavenging activity of fruit. In addition, exposure to 5 or 60% O₂ resulted in a higher level of total soluble solids, but a lower level of ascorbic acid of longan fruit flesh. In conclusion, exposure to a 5% O₂ atmosphere showed great potential to reduce pericarp browning and extend shelf life of longan fruit.

PRACTICAL APPLICATIONS

Pericarp browning and pulp breakdown are the major causes of deterioration in postharvest longan. Conventional controlled atmosphere with low O₂ and high CO₂ is effective in maintaining quality and extending shelf life of fruits and vegetables, including inhibition of tissue browning. In this study, 5%‐controlled atmosphere reduced significantly pericarp browning, pulp breakdown and rot development. It could potentially be useful as a postharvest technology of longan fruit for reducing or replacing the use of chemicals such as SO₂ and fungicides, but it requires further investigation.     

枇杷果实酶促褐变控制研究

枇杷果实在贮藏及加工过程中极易产生酶促褐变。本文采用分光光度法,研究了几种因素对枇杷果实多酚氧化酶活性的影响。结果表明:多酚氧化酶的最适 pH 为 6.3,抗坏血酸、L-半胱氨酸、柠檬酸、草酸、柠檬酸-抗坏血酸混合物可明显抑制该酶活性。