2,4-二硝基苯酚对采后龙眼果皮脂氧合酶活性和膜脂脂肪酸组分的影响

以'福眼'龙眼(Dimocarpus longan Lour.'Fuyan')果实为材料,研究呼吸解偶联剂2,4-二硝基苯酚(DNP)对采后果皮脂氧合酶(LOX)活性、膜脂脂肪酸组分和细胞膜透性的影响及其与果皮褐变的关系.结果表明:DNP处理导致龙眼果皮细胞膜透性、LOX活性和褐变指数增加,膜脂脂肪酸组分中的棕榈酸(C16:0)、硬脂酸(C18:0)等饱和 脂肪酸的组分增加,亚油酸(C18:2)、亚麻酸(C18:3)和花生一烯酸(C20:1)等不饱和脂肪酸的组分下降,脂肪酸不饱和指数和脂肪酸不饱和度下降.因此认为,DNP促进了龙眼果实果皮褐变可能是由于提高了LOX活性,促进了膜脂不饱和脂肪酸的降解而引起膜系统完整性受损,最终导致细胞膜结构的破坏,使酚酶与酚类物质接触而引起酚类物质氧化的结果.     

拟茎点霉侵染对采后龙眼果皮LOX活性和膜脂脂肪酸组分的影响

用拟茎点霉(Phomopsis longanae Chi)侵染福眼龙眼(Dimocarpus longan Lour. ‘Fuyan’)果实,研究龙眼果皮脂氧合酶(LOX)活性、膜脂脂肪酸组分和细胞膜透性的变化及其与果皮褐变的关系。结果表明：拟茎点霉侵染导致龙眼果皮褐变指数、LOX活性和细胞膜透性增加,不饱和脂肪酸组分[亚油酸(C18∶2)、亚麻酸(C18∶3)和花生一烯酸(C20∶1)]下降而饱和脂肪酸组分[棕榈酸(C16∶0)和硬脂酸(C18∶0)]增加,脂肪酸不饱和指数(IUFA)和脂肪酸不饱和度下降。拟茎点霉侵染提高龙眼果皮LOX活性和加速膜脂不饱和脂肪酸的降解,从而破坏细胞膜系统的完整性,导致膜系统区室化功能丧失,使多酚氧化酶与酚类物质接触,引起酚类物质的酶促氧化和黑褐色高聚物形成,导致龙眼果皮褐变。     

炭疽病菌侵染对荔枝果实生理生化变化的影响

本研究测定了荔枝果实人工接种炭疽病菌后呼吸速率、乙烯释放量的变化和果皮氧化、过氧化作用以及与酚类代谢有关的几种酶活性的变化。结果表明,接种炭疽病菌的荔枝果实呼吸速率和乙烯释放量显著增高,果皮活性氧(O₂^·)产生速率和丙二醛(MDA)含量显著增加,超氧化物歧化酶(SOD)活性显著降低,过氧化物酶(POD)、多酚氧化酶(PPO)和苯丙氨酸解氨酶(PAL)活性显著增高。说明炭疽病菌的侵染可导致荔枝果实呼吸速率和乙烯释放量的增高,加速荔枝果皮氧化和过氧化进程,并诱导荔枝果皮PPO、POD、PAL活性增高,是加速采收后荔枝果实衰老、褐变、腐烂的一个重要原因。     

O2和CO2配比对气调贮藏梨采后褐变及相关理化因子的影响

以采后'丰水'梨果实为材料,在乐扣气调试验箱中研究了O2和CO2配比对果实褐变率、多酚氧化酶(PPO)和过氧化物酶(POD)活性、丙二醛(MDA)和总酚含量的影响,以探讨适宜减轻梨气藏褐变的气体成份.结果表明:在整个贮藏过程(150 d)中,'丰水'梨果肉未发生褐变.从贮藏60 d开始,气调处理和冷藏对照果实的果皮均出现褐变,气调处理在贮藏120 d之前对果皮褐变的影响不显著,而在贮藏120～150 d内可显著减轻果皮的褐变、抑制果皮PPO和POD活性及降低总酚含量.与冷藏对照相比,气调处理可推迟果心褐变的时间,且(8%～10%)O2+3% CO2处理可完全抑制果心的褐变;气调处理亦可降低果心PPO活性、减少总酚及MDA含量;(8%～10%)O2+1% CO2处理能够显著提高果心的POD活性,而(8%～10%)O2+3% CO2处理对果实POD活性的影响不显著.可见,气调贮藏主要是通过降低'丰水'梨果皮PPO、POD活性及总酚含量来减轻组织的褐变,并以(8%～10%)O2+3% CO2处理对果实褐变因子的控制效果较理想.     

采后枸杞鲜果褐变与其活性氧代谢的关系

以采后'宁杞3号'枸杞鲜果为试材,研究其在(4±1)℃和(10±1)℃贮藏过程中果实褐变与活性氧代谢的关系.结果表明:枸杞果实褐变指数在两种贮藏温度下的贮藏初期(0～10 d)增加幅度均较小(分别为17.6%和5.1%),(10±1)℃下的果实从第15天显著增大(64.1%),而(4±1)℃下从第20天显著增大(67.9%).(4±1)℃贮藏果实的超氧歧化酶(SOD)活性和还原型抗坏血酸(AsA)含量在采后贮藏的前10 d增大,之后降低,贮藏后期又增大;超氧阴离子(O2-)产量在采后0～5 d降低,而后逐渐增大;过氧化氢(H2O2)和丙二醛(MDA)含量在采后0～5 d增大.之后前者降低又缓慢升高.与(10±1)℃贮藏果实相比,(4±1)℃下贮藏果实O2-产量显著降低,H2O2含量在贮藏第5天和第25天也显著降低,MDA含量在贮藏5～20 d显著降低;SOD活性在贮藏5～10 d显著提高.过氧化氢酶(cAT)活性和AsA含量明显提高;多酚氧化酶(PPO)活性在贮藏15～25 d显著下降,总酚含量在贮藏5～20 d显著增加.研究发现,枸杞果实褐变可能是细胞活性氧代谢失调,使PPO与酚类物质接触,导致酚类物质氧化的结果;(4±1)℃低温贮藏能通过调整鲜果活性氧代谢来有效延缓褐变进程.     

香蕉低温酶促褐变

7℃低温导致香蕉果皮变褐,褐变程度可用提取液的OD_(450)值表示。随低温处理时间的延长,OD_(450)增大;还原性物质抗坏血酸、谷胱甘肽含量迅速下降;多酚氧化酶底物多巴胺先有增加,接着降低。 低温下,多酚氧化酶(PPO)、过氧化物酶(POD)和过氧化氢酶(CAT)活性较常温低,PPO和POD的游离态酶活性增高。此外,H_2O_2处理加速果皮变褐,刺激PPO及POD活性;棓酸丙酯处理起相反的效应。故认为香蕉低温褐变是低温损伤、酶促褐变引起的一个复杂过程。     

臭氧水对荔枝采后若干生理生化指标的影响

用不同浓度臭氧水对荔枝果实进行采后处理,测定其花色素苷、类黄酮、可滴定酸和糖含量以及多酚氧化酶和过氧化物酶活性。结果显示,处理后荔枝果皮中花色素苷、类黄酮含量下降速度低于对照,PPO活性低于对照,POD活性高于对照,而果汁中可滴定酸及糖含量与对照相比无显著变化。表明一定浓度臭氧水对荔枝果实保色护色有一定效果,并有防褐变作用。     

不同灰霉病抗性苹果果实中酚类物质代谢特征

以‘秦冠’、‘富士’、‘金冠’苹果果实为材料,通过对损伤接种灰葡萄孢菌(Botrytis cinerea Pers)后果肉组织酚类代谢主要产物和相关酶活性变化的分析测定,揭示苹果采后酚类物质代谢与灰霉病抗性的关系,为苹果灰霉病抗性鉴定和筛选抗灰霉病苹果资源提供理论指导。结果表明:(1)接种灰葡萄孢菌后,3个苹果品种的果实灰霉病发病率和病斑直径大小均为‘秦冠’‘富士’‘金冠’,而且3个品种间的发病率和病斑直径均差异显著,各品种对灰霉病的抗性由强到弱依次为‘秦冠’‘富士’‘金冠’。(2)抗病品种‘秦冠’果肉组织中类黄酮、木质素含量及苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)活性均显著高于感病品种‘富士’和‘金冠’,但总酚含量为‘秦冠’‘金冠’‘富士’,且3品种间总酚含量差异显著。研究表明,抗病苹果品种通过调节果肉内酚类物质代谢,增强次生代谢能力,其中类黄酮和木质素含量的增加强化了果实的抗性反应,进而提高对灰霉病的抗性,但总酚含量与植物抗病性关系不大。     

香蕉芒果果皮中酶活性与炭疽病菌潜伏侵染的关系

在树上正生长发育的香蕉果实,随着果实长大,果皮中过氧化物酶(POD)和多酚氧化酶(PPO)活性不断升高。采后的香蕉和芒果果实的果皮中,POD和PPO活性变化与呼吸代谢相关。苯丙氨酸解氨酶(PAL)主要分布在果皮中,并随着果实成熟度的提高,其活性呈极显著下降趋势。感病果实的果皮中三种酶活性均比健康果皮高。说明果皮中三种酶活性的变化与炭疽菌的潜伏侵染有关。     

马铃薯块茎发芽过程中酚类物质含量及其相关酶活性的变化

马铃薯块茎中含有促进发芽的酚类物质 ,块茎发芽过程中 ,酚类物质含量增加 ,多酚氧化酶 (PPO)活性降低 ,过氧化物酶 (POD)活性在萌发初期升高 ,以后随着芽的生长又下降     

PRO-LONG涂膜对采后贮藏荔枝果实色泽和酶活性变化的影响

本文以荔枝品种“槐枝”为材料,以1．5％和2．5％Pro-long涂膜处理果实,研究Pro-long涂膜对果后贮于4℃的荔枝果皮色泽变化和有关酶活性的变化。对照和处理的HunterL值和b值均随贮藏时间的延长而降低,处理的比对照的下降慢。采后贮藏开始21d内,对照和处理的HuntCra值变化很小,保持相对稳定,之后的贮藏时间内有较显著的下降,说明贮藏初期果实的红色特性变化较少,之后变化很明显,这与果皮花色素苷、类黄酮、总酚的变化是相对应的。在贮藏后期,处理的HuntCra值下降明显较对照慢。Hunter值的变化与在贮藏过程中果实外观逐渐变成暗红和揭变以及在果实的来后根变中起着重要作用的多酚氧化酶和部分地涉及褐变过程的过氧化物酶的变化相对应。处理的过氧化物酶活性增加较对照慢,而且,与对照比较,多酚氧化酶的峰值稍稍延迟了。未发现l‘5％和2．5％处理间有明显差别。因此,Pro-long涂胶对荔枝的应用可以部分地降低多酚氧化酶和过氧化物酶,影响果皮花色素苷的降解、类黄酮和总酚的变化,延迟了褐变过程的发生。     

Salicylic acid and methyl jasmonate improve chilling tolerance in cold-stored lemon fruit (Citrus limon)

Chilling injury (CI) is associated with the degradation of membrane integrity which can be aligned to phenolic oxidation activated by polyphenol oxidase (PPO) and peroxidase (POD), enzymes responsible for tissue browning. Phenylalanine ammonia-lyase (PAL) is a further enzyme prominent in the phenolic metabolism that is involved in acclimation against chilling stress. It was hypothesized that treatment with methyl jasmonate (MJ) and salicylic acid (SA) may enhance chilling tolerance in lemon fruit by increasing the synthesis of total phenolics and PAL by activating the key enzyme regulating the shikimic acid pathway whilst inhibiting the activity of POD and PPO. Lemon fruit were treated with 10 μM MJ, 2 mM SA or 10 μM MJ plus 2 mM SA, waxed, stored at −0.5, 2 or 4.5 °C for up to 28 days plus 7 days at 23 °C. Membrane integrity was studied by investigating membrane permeability and the degree of membrane lipid peroxidation in lemon flavedo following cold storage. The 10 μM MJ plus 2 mM SA treatment was most effective in enhancing chilling tolerance of lemon fruit, significantly reducing chilling-induced membrane permeability and membrane lipid peroxidation of lemon flavedo tissue. This treatment also increased total phenolics and PAL activity in such tissue while inhibiting POD activity, the latter possibly contributing to the delay of CI manifestation. PPO activity was found to be a poor biochemical marker of CI. Treatment with 10 μM MJ plus 2 mM SA resulted in an alteration of the phenolic metabolism, enhancing chilling tolerance, possibly through increased production of total phenolics and the activation of PAL and inhibition of POD.     

Characterisation of phenol oxidase and peroxidase from maize silk

Silk of some maize genotypes contains a high level of phenolics that undergo enzymatic oxidation to form quinones, which condense among themselves or with proteins to form brown pigments. Two phenolic oxidizing enzymes, peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; EC 1.10.3.1), from maize (Zea mays L.) silk were characterised with respect to their preferred substrate, different isoforms and specific effectors. One browning silk sample with high, and two non‐browning samples with low phenolic content were investigated. Although POD oxidizes a wide range of phenolic substrates in vitro, its activity rate was independent of silk phenolic content. PPO activity, detected with o‐diphenolic substrates, was abundant only in browning silk, and low or absent in non‐browning silk. Pollination increased POD but not PPO activity. Isoelectric‐focusing (IEF) and specific staining for POD and PPO showed a high degree of polymorphism that varied with silk origin. The IEF pattern of POD revealed a number of anionic and several cationic isoenzymes, with the most pronounced having neutral pI 7 and a basic isoform with pI 10. Detected isoforms of PPO were anionic, except for one neutral form found only in browning silk, and occupied positions different from those of POD. Different inhibitory effects of NaN₃, EDTA, KCN, and L‐cysteine, as well as different impacts of a variety of cations on the oxidation of chlorogenic acid, mediated by PPO or POD, were detected. The findings are discussed in terms of a possible roles of these enzymes in defence and pollination.     

裂果易发性不同的荔枝品种果皮中细胞壁代谢酶活性的比较

“糯米糍”荔枝裂果率极显著高于“淮枝”,前者果皮中的果胶酶、纤维素酶和果胶甲酯酶的活性高于后者,其中以果胶酶活性差异最明显,其次是纤维素酶,果胶甲酯酶差异最小;“糯米糍”细胞壁结合型的过氧化物酶(POD)和多酚氧化酶(PP0)活性明显高于“淮枝”,而水溶性POD和PP0的活性则两个品种间无明显差异。据此认为,果皮细胞壁水解酶活性以及细胞壁结合型的POD和PPO的活性高的荔枝品种,其裂果率也高。文章对细胞壁代谢相关酶类在果皮抗裂性形成中的作用进行了讨论。     

Cloning and Expression Analysis of Litchi (Litchi Chinensis Sonn.) Polyphenol Oxidase Gene and Relationship with Postharvest Pericarp Browning

Polyphenol oxidase (PPO) plays a key role in the postharvest pericarp browning of litchi fruit, but its underlying mechanism remains unclear. In this study, we cloned the litchi PPO gene (LcPPO, {"type":"entrez-nucleotide","attrs":{"text":"JF926153","term_id":"350601663","term_text":"JF926153"}}JF926153), and described its expression patterns. The LcPPO cDNA sequence was 2120 bps in length with an open reading frame (ORF) of 1800 bps. The ORF encoded a polypeptide with 599 amino acid residues, sharing high similarities with other plant PPO. The DNA sequence of the ORF contained a 215-bp intron. After carrying out quantitative RT-PCR, we proved that the LcPPO expression was tissue-specific, exhibiting the highest level in the flower and leaf. In the pericarp of newly-harvested litchi fruits, the LcPPO expression level was relatively high compared with developing fruits. Regardless of the litchi cultivar and treatment conditions, the LcPPO expression level and the PPO activity in pericarp of postharvest fruits exhibited the similar variations. When the fruits were stored at room temperature without packaging, all the pericarp browning index, PPO activity and the LcPPO expression level of litchi pericarps were reaching the highest in Nandaowuhe (the most rapid browning cultivar), but the lowest in Ziniangxi (the slowest browning cultivar) within 2 d postharvest. Preserving the fruits of Feizixiao in 0.2-μm plastic bag at room temperature would decrease the rate of pericarp water loss, delay the pericarp browning, and also cause the reduction of the pericarp PPO activity and LcPPO expression level within 3 d postharvest. In addition, postharvest storage of Feizixiao fruit stored at 4°C delayed the pericarp browning while decreasing the pericarp PPO activity and LcPPO expression level within 2 d after harvest. Thus, we concluded that the up-regulation of LcPPO expression in pericarp at early stage of postharvest storage likely enhanced the PPO activity and further accelerated the postharvest pericarp browning of litchi fruit.     

Improved methods for the extraction of polyphenol oxidase from d'anjou pears

The conditions for extracting polyphenol oxidase (PPO, monophenol monooxygenase, EC 1.14.18.1) from d'Anjou pears have been studied. Water extracts of pear PPO contained artefacts which were present as additional bands on polyacrylamide-gel electrophoresis. Buffer extracts of an acetone powder did not remove sufficient endogenous phenolics to prevent browning of the extract. The following phenolic absorbents, arranged in order of increasing efficiency, reduced the formation of artefacts in extracts of PPO: PVPP, Amberlite XAD-4, Bio-Rad AG 1-X8, and Bio-Rad AG 2-X8. Greatest activity was extracted within a pH range of 5.6–5.9. Anion exchange resins were particularly effective in removing phenolics. XAD-4, AG 1-X8, or AG 2-X8 did not adsorb PPO and reduced the electrophoretically separable bands of PPO activity from 11 in water extracts to 3. The properties of the crude PPO were also studied.     

The Changes of Pigments,Phenolics Contents and Activities of Polyphenol Oxidase and Phenylalanine Ammonia-Lyase in Pericarp of Postharvest Litchi Fruit

A common litchi cultivar “Huaizhi” was used for the experiment to understand the changes of pigment, phenolics content and activities of two enzymes involing in phenolics metabolism in pericarp during storage at room temperature and relative humidity of 73–79%. The maturation of "Huaizhi" fruit was 80% when harvested. The contents of carotenoid, anthocynin and activities of phenylalanine ammonia-lyase, polyphenol oxidase were increased, and the content of total phenolics was kept at its initial level in the first 2 days of storage. It indicated that the active synthesis of pigments and phenolics still continued with the progressive ripening. A decline of above parameters and content of flavonoid ware observed during the senescence of fruit. The relative contents of anthocynin, flavonoid, phenolics and activity of phenylalanine ammonialyase were 90%, 59%,71% and 46% in the day of 7 compared with that when harvested, respectively. Less change of anthocynin content was found in browning pericarp. The relation between phenolics, anthocynin and activities of two enzymes during repining and senescence of litchi fruit was discussed.