Dehydrin from citrus, which confers in vitro dehydration and freezing protection activity, is constitutive and highly expressed in the flavedo of fruit but responsive to cold and water stress in leaves

A cDNA encoding a dehydrin was isolated from the flavedo of the chilling-sensitive Fortune mandarin fruit (Citrus clementina Hort. Ex Tanaka x Citrus reticulata Blanco) and designed as Crcor15. The predicted CrCOR15 protein is a K2S member of a closely related dehydrin family from Citrus, since it contains two tandem repeats of the unusual Citrus K-segment and one S-segment (serine cluster) at an unusual C-terminal position. Crcor15 mRNA is consistently and highly expressed in the flavedo during fruit development and maturation. The relative abundance of Crcor15 mRNA in the flavedo was estimated to be higher than 1% of total RNA. The high mRNA level remained unchanged during fruit storage at chilling (2 degrees C) and nonchilling (12 degrees C) temperatures, and it was depressed by a conditioning treatment (3 days at 37 degrees C) that induced chilling tolerance. Therefore, the expression of Crcor15 appears not to be related to the acquisition of chilling tolerance in mandarin fruits. However, Crcor15, which was barely detected in unstressed mandarin leaves, was rapidly induced in response to both low temperature and water stress. COR15 protein was expressed in Escherichia coli, and the purified protein conferred in vitro protection against freezing and dehydration inactivation. The potential role of Citrus COR15 is discussed.     

Active oxygen detoxifying enzymes and phenylalanine ammonia-lyase in the ethylene-induced chilling tolerance in citrus fruit

The effects of applying ethylene (2 microL x L(-)(1)) during cold storage of Fortune mandarins on the development of chilling-induced peel damage and on changes in the activities of the enzymes of the antioxidant system, superoxide dismutase, catalase (CAT), ascorbate peroxidase, guaiacol peroxidase, and glutathione reductase, and on phenylalanine ammonia-lyase (PAL) have been investigated. Chilling damage was reduced by applying ethylene during fruit storage at 1.5 degrees C. PAL activity increased in response to cold stress and was higher in fruit held under ethylene than under air during the whole storage period, whereas CAT was temporarily higher in ethylene-treated fruit. In contrast, the activities of the other enzymes were not increased by ethylene. The global results suggest that the ethylene-induced chilling tolerance in Fortune mandarins might be due to increased PAL and CAT activities.     

Phenylalanine ammonia-lyase as related to ethylene in the development of chilling symptoms during cold storage of citrus fruits.

Low-temperature, nonfreezing, storage induces pitting and necrosis in the flavedo tissue of chilling susceptible citrus fruits. In this study the role of ethylene and phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) in the cold-induced citrus peel damage has been investigated. It has been shown that increasing PAL activity by applying ethylene at a nonchilling temperature did not cause fruit damage or reduce the incidence of this peel disorder when fruits were subsequently held at a chilling temperature (2 degrees C). The cold-induced peel damage was enhanced by applying inhibitors of PAL activity and ethylene synthesis and action. These results indicate that the induction of PAL and ethylene during fruit cold storage, but not before, plays a role in reducing the development of chilling symptoms. The cold-induced PAL activity was reduced by inhibitors of ethylene production, but inhibitors of ethylene action exerted little effect on the activation of this enzyme. Therefore, the activation of PAL may be dependent on ethylene but also an independent cold signal apparently related to the cold-induced peel damage.     

Variation in sugar levels and invertase activity in mature fruit representing a broad spectrum of Cucumis melo genotypes

Sugar accumulation is a very important physiological process that determines dessert-melon fruit quality. Considerable variation in the sugar content and composition in the mature flesh of Cucumis melo L. fruits was observed among 56 genotypes which represent the wide range of morphological and horticultural types found in this species. Sucrose accumulation was observed not only among 'dessert melons' of the inodorus and cantalupensis types, but also in representatives of other subgroups of C. melo, including some of the accessions from agrestis and conomon groups. Among the genotypes that accumulate sucrose, the levels of this sugar, and not of the hexoses, were correlated with the total sugar concentration. Hexose levels were correlated with total sugar levels only among low-sugar genotypes that did not accumulate sucrose. Activities of the sucrose hydrolyzing enzymes acid invertase (EC 3.2.1.26) and alkaline invertase (EC 3.2.1.27) were measured in the mature fruit flesh and the results of this survey support the hypothesis that low acid invertase activity is a prerequisite for sucrose accumulation. The sucrose accumulating, high sugar genotypes had less-acidic flesh pH values (pH> 6) than the low-sugar genotypes. The latter were characterized by a broader range of pH values, including acidic (pH< 5.5) fruit flesh.     

Carbohydrate metabolism as related to high-temperature conditioning and peel disorders occurring during storage of citrus fruit

The aim of this research was to understand the involvement of the carbohydrate metabolism in physiological disorders occurring during the postharvest storage of citrus fruit. These disorders, manifested in the rind, depreciate fruit quality and often originate important losses. There has been increasing interest in the use of nonharmful treatments, such as high-temperature conditioning, to avoid citrus peel damage during fruit storage at low temperature in chilling-sensitive cultivars, but their influence in postharvest disorders occurring at nonchilling temperatures and the mechanisms related to them are poorly understood. The data obtained showed that heat conditioning (3 days/37 degrees C) increases the chilling tolerance of cv. Navelate fruit and favored sucrose, but not hexoses, accumulation and its maintenance after the fruit was transferred to low temperature. This effect was related to heat-induced increase in the activities of the sucrose-synthesizing enzymes sucrose phosphate synthase (SPS) and sucrose synthase (SS). Furthermore, sucrose levels and the activities of both enzymes were higher in cv. Pinalate oranges, a chilling-tolerant spontaneous abscisic acid deficient mutant of Navelate. In contrast, carbohydrates appeared not to be involved in the susceptibility of oranges to rind staining, a physiological disorder different from chilling injury, which mainly occurred at a nonchilling temperature (12 degrees C) and was not reduced by heat conditioning. The effect of low temperature in SS and SPS activities was less than that of high temperature, which might be related to the lower changes occurring in sucrose during fruit storage at 2 degrees C.     

Methyl jasmonate reduces chilling injury and maintains postharvest quality of mango fruit

Exposure of mango (Mangifera indica cv. Tommy Atkins) fruit to methyl jasmonate (MJ) vapors (10(-)(4) M) for 24 h at 25 degrees C reduced chilling injury during subsequent storage for 21 days at 7 degrees C and after 5 days of shelf life at 20 degrees C. The chilling tolerance induced by MJ was positively correlated with the reduction in the percent ion leakage of mango tissue. The overall quality of MJ-treated fruit was also better than that of control fruit. MJ treatment increased the total soluble solids but did not affect titratable acidity or pH. MJ also did not change the normal climacteric rise in respiration, water loss, and softening rates. The efficacy of MJ to reduce chilling injury and decay of mango could be related to the tolerance induced at low temperature. It was concluded that MJ treatment may prevent chilling injury symptoms of mango without altering the ripening process.     

NO处理对采后映霜红桃果实冷害及呼吸作用的影响

为探究一氧化氮(NO)处理对桃果实冷藏期间冷害及呼吸作用的影响,本研究以映霜红桃果实为试验材料,采用不同浓度NO(0、5、10、15 μL·L^-1)处理,检测桃在4℃贮藏期间冷害症状、冷害指数、硬度、呼吸速率、呼吸途径关键酶的变化。结果表明,10 μL·L^-1NO处理延缓了桃冷害的发生,显著降低了冷害指数、呼吸速率(P<0.05),维持了桃果实的硬度,使果实正常软化后熟。同时,10 μL·L^-1NO处理通过抑制桃果实糖酵解、三羧酸(TCA)循环、细胞色素氧化酶途径关键酶—葡萄糖磷酸异构酶(GPI)、琥珀酸脱氢酶(SDH)和细胞色素氧化酶(COX)的活性(P<0.05),抑制了果实呼吸速率,延缓了果实衰老。10 μL·L^-1 NO处理通过提高磷酸戊糖途径关键酶—葡萄糖-6-磷酸脱氢酶(G-6-PDH)和6-磷酸葡萄糖酸脱氢酶(6-PGDH)的联合活性及抗氰呼吸途径关键酶—交替氧化酶(AOX)的活性,增强了桃果实的抗冷性,减轻了桃果实的冷害。本研究结果为NO在桃果实贮藏保鲜中的应用提供了理论依据。     

套黄、白袋凯特芒果采后果实糖代谢的比较研究

为探究果实生长期间套不同颜色纸袋对芒果采后果实软鼻病和贮藏特性的影响,本试验以套黄、白袋凯特芒果(Mangifera indica L.cv. Keitt)为试验材料,比较研究了两种套袋果实采后软鼻病发病率、硬度及糖代谢的差异。结果表明,在常温贮藏下(10 d),套黄袋芒果采后果实的软鼻病发病率显著低于套白袋芒果(P<0.05),而且采后果实果肉的淀粉酶、酸性转化酶(AI)、中性转化酶(NI)和蔗糖磷酸合成酶(SPS)活性均显著低于套白袋芒果(P<0.05),因而缓解了果实淀粉含量的下降速率和蔗糖积累的速率,延缓了果实的软化成熟进程,提高了采后果实的耐贮性。另外,套袋影响采后芒果果肉的蔗糖糖代谢可能与果实软鼻病的病情发展相关。本研究为选取合理有效的芒果套袋材料提供了一定的参考依据。     

Antioxidant enzymes and DPPH-radical scavenging activity in chilled and heat-shocked rice (Oryza sativa L.) seedlings radicles.

Chilling whole rice seedlings at 5 degrees C significantly increased the time needed to recover linear growth and reduced the subsequent linear rate of radicle growth. Subjecting nonchilled seedlings to a 45 degrees C heat shock for up to 20 min did not alter subsequent growth, whereas a 3 min heat shock was optimal in reducing growth inhibition caused by 2 days of chilling. The activity of five antioxidant enzymes [superoxide dismutase (EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11), glutathione reductase (GR; EC 1.6.4.2), and guaiacol peroxidase (GPX; EC 1.11.1.7)] and DPPH (1,1-diphenyl-2-picrylhydrazyl)-radical scavenging activity were measured in heat-shocked and/or chilled radicles. Heat shock slightly increased the activity of CAT, APX, and GR and suppressed the increase of GR and GPX activity during recovery from chilling. Increased CAT, APX, GR, and DPPH-radical scavenging activity and protection of CAT activity during chilling appear to be correlated with heat shock-induced chilling tolerance.     

Simultaneous transgenic suppression of LePG and LeExp1 influences fruit texture and juice viscosity in a fresh market tomato variety

Tomatoes are grown for fresh consumption or for processing of the fruit. Some ripening-associated processes of the fruit can either contribute to or degrade attributes associated with both fresh and processing quality. For example, cell wall disassembly is associated with loss of fresh fruit firmness as well as with loss of processed tomato product viscosity. Several enzymes contribute to cell wall polysaccharide disassembly. Polygalacturonase (PG, poly[1,4-alpha-d-galactouronide] glucanohydrolase, EC 3.2.1.15) is among the most abundant polysaccharide hydrolases in ripening tomato fruit and is the major contributor to pectin depolymerization. Expansin (LeExp1) is also abundant in ripening fruit and is proposed to contribute to cell wall disassembly by nonhydrolytic activity, possibly by increasing substrate accessibility to other enzymes. Suppression of either LePG or LeExp1 expression alone results in altered softening and/or shelf life characteristics. To test whether simultaneous suppression of both LePG and LeExp1 expression influences fruit texture in additive or synergistic ways, transgenic Lycopersicon esculentum var. Ailsa Craig lines with reduced expression of either LePG or LeExp1 were crossed. Fruits from the third generation of progeny, homozygous for both transgenic constructs, were analyzed for firmness and other quality traits during ripening on or off the vine. In field-grown transgenic tomato fruit, suppression of LeExp1 or LePG alone did not significantly increase fruit firmness. However, fruits suppressed for both LePG and LeExp1 expression were significantly firmer throughout ripening and were less susceptible to deterioration during long-term storage. Juice prepared from the transgenic tomato fruit with reduced LePG and LeExp1 expression was more viscous than juice prepared from control fruit.     

不同变种甜瓜糖分积累及蔗糖代谢酶活性动态变化

为了解不同类型甜瓜糖分积累及糖代谢特点,选用厚皮甜瓜品种X228、普通甜瓜品种B154及越瓜品种H227为材料,定期取样测定果实成熟过程中的葡萄糖、果糖、蔗糖含量和蔗糖代谢相关酶活性,研究不同变种甜瓜果实发育过程中糖分积累及相关酶动态变化差异。结果表明,授粉15 d至果实成熟期间,3个甜瓜品种的果实葡萄糖与果糖含量的变化均较小,品种间差异不显著。3个甜瓜品种果实蔗糖含量存在显著性差异,其中H227果实几乎无蔗糖积累,葡萄糖和果糖是果实的主要糖组分;B154和X228果实蔗糖含量随着果实发育而快速增加,蔗糖积累存在明显的转折点,蔗糖是B154和X228这2个品种成熟果实中最主要的糖组分,且果实蔗糖含量提高的同时蔗糖磷酸合成酶(SPS)活性上升、酸性转化酶(AI)活性降低,蔗糖合成酶(SS)合成方向的活性与蔗糖含量关系不显著。根据蔗糖含量的差异,可将甜瓜分为蔗糖积累型和低蔗糖积累型两类,前者果实蔗糖含量的上升被认为是SPS活性上升与转化酶(特别是AI)活性下降共同作用的结果,后者果实内极低的蔗糖含量被认为是SPS活性较低导致的。本研究结果为甜瓜种质资源创新利用和甜瓜果实糖分积累调控研究奠定了理论基础。     

Loss of ripening capacity of pawpaw fruit with extended cold storage

The fruit ripening traits of pawpaw [ Asimina triloba (L.) Dunal] were examined after harvest and after cold storage at -2, 2, 4, and 6 degrees C for up to 12 weeks. Generally, fruits stored at 2-4 degrees C for 4 weeks ripened normally, but those stored at -2 degrees C did not ripen normally, those stored at 6 degrees C were overripe, and by 6-8 weeks those stored at 2-4 degrees C had a lower respiration rate and ethylene production, lower firmness, and lower pH than fruit cold-stored for 4 weeks or less. These changes, and the occasional development of brown discoloration in the pulp once the fruits were moved back to room temperature, were evidence of chilling injury by 6 weeks. After harvest and through 4 weeks of cold storage, the main volatile compounds produced by fruit were methyl and ethyl octanoates and hexanoates. Volatile production significantly increased >5-fold in fruit ripened for 72 h after harvest or after removal from up to 4 weeks of cold storage. Fruit cold-stored for 6 weeks or more produced fewer total volatiles and esters but increased levels of such off-flavor compounds as ethyl acetate, ethyl propionate, and hexanoic and decanoic acids. Alcohol acyltransferase (AAT) activity declined in cold-stored fruit but was not correlated with either total volatile production or total ester production. Alcohol dehydrogenase activity did not change during ripening after harvest or cold storage. Lipoxygenase activity was highest just after harvest or after 2 weeks of cold storage, but was low by 4 weeks. Thus, ripening of pawpaw fruit seems to be limited to 4 weeks at 2-4 degrees C with loss of ability to continue ripening and chilling injury symptoms evident at colder temperatures and after longer periods of cold storage.     

Metabolism of phenolic compounds during loquat fruit development.

Phenolic compounds in loquat fruit were identified as 5-caffeoylquinic acid (chlorogenic acid), neochlorogenic acid, hydroxybenzoic acid, 5-p-feruloylquinic acid, protocatechuic acid, 4-caffeoylquinic acid, epicatechin, o-coumaric acid, ferulic acid, and p-coumaric acid. Neochlorogenic acid was found to be dominant in the early stages of loquat fruit development. Both the concentrations and types of phenolic compounds were high in young fruit but then decreased steadily during growth. However, the concentration of chlorogenic acid increased during ripening and became predominant in ripe fruit. The large rise in chlorogenic acid concentration appears to be a characteristic of loquat fruit ripening. In all of the cultivars tested, the types of phenolic compounds were similar but the total phenolic content varied from 81.8 to 173.8 mg/100 g of fresh pulp. In the biosynthetic pathway of chlorogenic acid, the enzyme activities of phenylalanine ammonia-lyase (PAL), 4-coumarate:CoA ligase (CL), and hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase (CQT) were high at the early stage of growth, diminished to low levels approximately 3 weeks prior to harvest, but then rose to a peak at 1 week before harvest. The changes of these enzyme activities seemed to be associated with variations in chlorogenic acid concentration during development, maturation, and ripening of loquat fruit.     

Catalase in the heat-induced chilling tolerance of cold-stored hybrid Fortune mandarin fruits

Hybrid Fortune mandarins developed chilling injury (CI) upon cold storage, unless the fruits were conditioned at 37 degrees C for 3 days before they were held at low temperature. This heat treatment induced 2.5-, 1.2-, and 1.4-fold increases in the activities of catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD), respectively, and reduced the activity of glutathione reductase (GR). The differences in the activities afforded by the heat treatment were, in general, maintained during cold storage. However, SOD levels in nonconditioned Fortune fruits exhibiting CI were similar to those of conditioned fruits stored for 0 or 6 weeks at 2 degrees C. No difference between APX activity in the conditioned and nonconditioned fruits stored for 6 weeks at 2 degrees C was found. The data indicate that CAT may be a major antioxidant enzyme operating in the heat-induced chilling tolerance of cold-stored Fortune mandarin fruits.     

Changes in oxygen-scavenging systems and membrane lipid peroxidation during maturation and ripening in blackberry

Maturation and ripening of blackberry (Rubus sp.) fruit was accompanied by decreased activities of oxygen-scavenging enzymes [superoxide dismutase (EC 1.15.1.1), glutathione-peroxidase (EC 1.11.1.9), catalase (EC 1.11.1.6)] and enzymes in the ascorbate-glutathione cycle [ascorbate peroxidase (EC 1.11.1.11), monodehydroascorbate reductase (EC 1.6.5.4), dehydroascorbate reductase (EC 1.8.5.1), and glutathione reductase (EC 1.6.4.2)]. Nonenzyme components in the ascorbate-glutathione cycle such as ascorbate (AsA), dehydroascorbate (DHAsA), glutathione (GSH), and oxidized glutathione (GSSG) and the ratios of AsA/DHAsA, GSH/GSSG were also decreased. These decreases in antioxidant capacity were correlated with increases in the ratios of saturated to unsaturated fatty acid of polar lipids and free sterols to phospholipids, thus contributing to decreased fluidity, enhanced lipid peroxidation, and membrane deterioration, which may be associated with ripening and senescence in blackberry fruit.     

设施油桃果实发育过程中有机酸代谢的研究

以设施栽培"超红珠"油桃为试材,测定了果实发育过程有机酸含量及相关代谢酶——柠檬酸合酶(CS)、苹果酸酶(ME)、苹果酸脱氢酶(MDH)的活性,并对果实中有机酸积累及酶活性的关系进行了分析。研究结果表明:随油桃果实发育,其有机酸含量呈先升高后降低趋势,于盛花后49d达到最高值;苹果酸含量与总有机酸含量变化趋势相似;柠檬酸含量呈逐渐升高趋势,至果实成熟有所降低;果实成熟期苹果酸与柠檬酸含量相近。有机酸相关代谢酶CS活性变化与柠檬酸含量相关性不大;果实发育后期ME活性逐渐升高,促进了苹果酸的降解和转化;MDH活性变化与苹果酸含量显著相关。因此抑制果实发育前期MDH的活性、促进果实发育后期ME的活性可以降低果实酸含量,提高糖酸比。     

High CO2 atmosphere modulating the phenolic response associated with cell adhesion and hardening of Annona cherimola fruit stored at chilling temperature

Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5.) activity, tanning ability, and polyphenols levels were measured in cherimoya (Annona cherimola Mill.) fruit treated with 20% CO(2) + 20% O(2) + 60% N(2) for 1, 3, or 6 days during chilling temperature (6 degrees C) storage. The residual effect of CO(2) after transfer to air was also studied. These observations were correlated with texture and cellular characteristics, visualized by cryo-SEM. Tanning ability and the early increase in tannin polyphenols induced by chilling temperature were reduced by CO(2) treatment. Conversely, high CO(2) atmosphere enhanced the nontannin polyphenol fraction as compared with fruit stored in air. Lignin accumulation and PAL activation observed in untreated fruit after prolonged storage at chilling temperature were prevented by high CO(2). Moreover, the restraining effect on lignification was less effective when the CO(2) treatment was prolonged for 6 days. In addition, fruits held at these conditions had greater firmness and the histological characterization of the separation between cells was similar to that in untreated fruits. We conclude that CO(2) treatment modulates the phenolic response that seems to regulate the strength of cell adhesion and so to prevent hardening caused by chilling temperature storage.     

脱落酸对果实成熟的生理效应与机制

本文概述了果实成熟过程中脱落酸含量的变化和生理效应．分析了脱落酸促进果实成熟的细胞分子生物学基础及其可能的分子机制。