Effect of Fungal Infection and Temperature on Physiological Changes in Banana Fruit

An attempt was made to investigate which factor temperature or fungal infection, has a greater effect in accelerating physiological processes such as respiration and ethylene evolution in fruit. For this study the effect of infection with Trichothecium roseum, Alternaria alternata and Diplodia natalensis on banana fruit, at 14°C and 20°C was examined. The results show that the development of rot caused by different fungi and the physiological changes in the infected fruit may depend not only on the effect of temperature on fungi (as in vitro) but also on fruit ripening, or on processes connected with fruit ripening, which may accelerate fungal development.     

猕猴桃果实采后生理研究进展

猕猴桃（Actinidia chinensis Planch.）属于呼吸跃变型果实,采后易软化腐烂,不耐贮藏,如何延长猕猴桃果实贮藏期限已成为猕猴桃产业发展壮大亟待解决的问题。猕猴桃果实采后生理变化强烈影响果实的贮藏期限和果实品质,特别是呼吸作用、乙烯合成及其信号转导系统和果实软化等,并且它们与猕猴桃贮藏保鲜技术的研发与应用密切相关。本文重点从这3个方面就国内外相关研究进展进行综述,并讨论它们对猕猴桃耐贮性的影响,以期为猕猴桃耐贮新品种的培育和贮藏保鲜技术的研发提供理论依据。     

挥发性香气物质和乙烯生产在"湖景蜜露"桃果实发育过程中的变化

以"湖景蜜露"水蜜桃(Prunus persica L.)为试材,检测了果实从未成熟到成熟发育过程中乙烯生成、呼吸速率及挥发性香气性物质的变化;同时对果实大小、果皮色泽、果肉硬度、可溶性固形物、可滴定酸进行了测定;对与果实乙烯产生密切相关的1-氨基环丙烷-1-羧酸(ACC)含量、ACC合成酶活性、ACC氧化酶活性也进行了测定.结果表明,随果实成熟度的增加,果实大小、果皮L*值、可溶性固形物含量增加,而果实硬度、果皮h°值、可滴定酸含量减少.在未成熟的果实中,C6的醛类(反式-2-己烯醛)和醇类(顺式-3-己烯醇)是主要的成分;乙烯生成量很低;呼吸速率较高.到跃变阶段C6～C12的内酯类物质明显增加,尤其是γ和δ-内酯类成为果实主要的香气挥发性物质.推测果实乙烯、呼吸作用等基本的生理变化可能调节着内酯类物质的生成.在乙烯跃变上升时果肉中ACC氧化酶的活性下降,ACC含量和ACC合成酶活力的变化与乙烯生成量变化的趋势一致.根据以上结果可以认为桃果实主要的香气挥发性物质的形成与乙烯、呼吸跃变的开始密切相关.香气物质形成速率动态变化可能是桃果实发育过程中成熟度的另一个生理学指标.     

Stimulation of Fruit Ethylene Production by Wounding and by Botrytis cinerea and Geotrichum candidum Infection in Normal and Non-Ripening Tomatoes

Inoculations with both Botrytis cinerea and Geotrichum candidum stimulated ethylene evolution in the pre-climacteric normal tomato fruit and the non-ripening nor mutant which did not show any rise in ethylene when uninfected. In the post-climacteric normal fruits, new peaks in ethylene production were formed. The rise in ethylene evolution in all types of infected fruits has already been detected during the incubation period of the disease. Ethylene peaks were detected earlier and were higher in fruits infected with B. cinerea than with G. candidum, coinciding with the faster rate of growth of the former. Mechanical wounding also stimulated ethylene synthesis by the non-ripening fruits, production being directly proportional to wound dimension. Considerably higher rates of ethylene were recorded for infected fruits than for mechanically-injured fruits in which wound dimensions were similar to those of lesion development. Applying aminoxyacetic acid at the site of inoculation inhibited ethylene production by 55–60 % in the normal fruits and by about 80 % in the nor mutant fruits. A similar pathway of ethylene synthesis was suggested for normally ripening tomato fruit and non-ripening infected tissues.     

The role of ethylene and cell wall modifying enzymes in raspberry (Rubus idaeus) fruit ripening

This study focuses on four raspberry ( Rubus idaeus ) genotypes from two different genetic backgrounds: cvs Glen Prosen and Glen Clova, bred at the Scottish Crop Research Institute (SCRI) and genotypes bred at Horticulture Research International (HRI), East Malling (EM), EM 4997 and EM 5007. The ripe fruit of each genotype pair were characterised subjectively by raspberry breeders as relatively firm or soft, respectively. Different stages of fruit development from each genotype were used to quantify fruit firmness, rates of ethylene evolution and ripening rate. Penetrometry data confirmed suspected firmness differences. Firmness correlated with rates of ethylene evolution. Rates of ethylene production also correlated with receptacle size. Storage of green fruits in 20 μl l⁻¹ ethylene reduced fruit firmness, enhanced respiration rate and colour (anthocyanin) development and stimulated the development of cell wall hydrolase activities. However, during natural ripening in the field, fruit respiration rate declined, which indicates a non-climacteric ripening pattern. In drupelets, the activities of polygalacturonase (PG), pectin methylesterase (PME), C x -cellulase (C x ) and β -galactosidase ( β -gal.) increased substantially as ripening progressed. More detailed studies with ripe fruit of cv. Glen Clova indicated major isoforms of PG at pIs 3.3, 8.6 and 10.1; of PME at pIs 7.2, 8.5, 8.7, 8.8; of C x at pI 2.4; and of β -gal. at pIs 6.3 and 6.7.     

In vitro growth and ripening of strawberry fruit in the presence of ACC, STS or propylene

Strawberry ( Fragaria ananassa Duch.) fruit exhibit limited capacity for continued development following harvest. This problem can be circumvented by maintaining harvested strawberry fruit in solutions containing sucrose and a bactericide. In this study, we investigated the respiratory and ethylene production kinetics and ethylene responsiveness in strawberry fruit harvested immature and ripened in vitro in the presence of propylene. The effects of 1-amino-cyclopropane-1-carboxylic acid (ACC) and silver thiosulfate (STS) alone and in combination were also examined. Respiration and ethylene patterns of fruit harvested green and developed in vitro declined with maturation and ripening, as did those of field-grown fruit harvested at different stages of ripeness. Exposure of detached green strawberry fruit to 5000 μl litre^-1 propylene failed to stimulate respiration or ethylene production, but advanced pigmentation changes and fresh-weight gain significantly. Excised fruit provided with 1 mol^-3 ACC exhibited increased ethylene production, enhanced fresh-weight gain, and accelerated anthocyanin accumulation, but showed no change in respiration. The developmental response of harvested strawberry fruit to propylene or ACC was dependent on fruit maturity at harvest, with white fruit exhibiting greater insensitivity compared with green fruit. Silver thiosulfate (0.5 mol^-3) applied alone or in combination with ACC failed to delay ripening in excised strawberry fruit. These experiments demonstrate that ripening in detached strawberry fruit can be modified by ethylene only in green fruit that are provided with a carbohydrate source. Ethylene, when applied exogenously as ACC or propylene to green fruit, can slightly increase fruit growth and the rate of colour development.     

Effect of Abscisic Acid on Banana Fruit Ripening in Relation to the Role of Ethylene

Abstract The role of abscisic acid (ABA) in banana fruit ripening was examined with the ethylene binding inhibitor, 1-methylcyclopropene (1-MCP). ABA (0, 10⁻⁵, 10⁻⁴, or 10⁻³ mol/L) was applied by vacuum infiltration into fruit. 1-MCP (1 μL/L) was applied by injecting a measured volume of stock gas into sealed glass jars containing fruit. Fruit ripening, as judged by ethylene evolution and respiration associated with color change and softening, was accelerated by 10⁻⁴ or 10⁻³ mol/L ABA. ABA at 10⁻⁵ mol/L had no effect. The acceleration of ripening by ABA was greater at 10⁻³ mol/L than at 10⁻⁴ mol/L. ABA-induced acceleration of banana fruit ripening was not observed in 1-MCP treated fruit, especially when ABA was applied after exposure to 1-MCP. Thus, ABA's promotion of ripening in intact banana fruit is at least partially mediated by ethylene. Exposure of ABA-treated fruit to 0.1 μL/L ethylene for 24 h resulted in increased ethylene production and respiration, and associated skin color change and fruit softening. Control fruit (no ABA) was unresponsive to similar ethylene treatments. The data suggest that ABA facilitates initiation and progress in the sequence of ethylene-mediated ripening events, possibly by enhancing the sensitivity to ethylene. Received 29 January 1999; accepted 16 January 2000     

采前二氧化氯处理对‘海沃德’猕猴桃的防腐保鲜效果

研究了采前二氧化氯（ClO2）处理对采后‘海沃德’猕猴桃果实在冷藏条件下的防腐保鲜效果。分别用浓度（有效成分）为0、20、40、60、80mg·L-1的ClO2溶液,在采收前对果实进行喷布处理,在采收后冷藏过程中定期测定相关生理指标,并在贮藏末期（120d）观察统计其腐烂指数。结果表明,浓度60mg·L-1的ClO2采前处理可有效清除猕猴桃果实表面菌落,并能显著延缓冷藏期间果实硬度的下降、抑制可溶性固型物的上升、降低猕猴桃呼吸速率、乙烯释放速率、失重和腐烂指数,提高过氧化物酶（POD）和苯丙氨酸解氨酶（PAL）的活性。通过比较,确定采前ClO2处理的适宜浓度为60mg·L-1。该浓度的ClO2处理对‘海沃德’猕猴桃具有明显的防腐保鲜效果,可有效延长贮藏期。     

Energy conservation and dissipation in mitochondria isolated from developing tomato fruit of ethylene-defective mutants failing normal ripening: the effect of ethephon,a chemical precursor of ethylene

Alternative oxidase (AOX) and uncoupling protein (UCP) are present simultaneously in tomato fruit mitochondria. In a previous work, it has been shown that protein expression and activity of these two energy-dissipating systems exhibit large variations during tomato fruit development and ripening on the vine. It has been suggested that AOX and UCP could be responsible for the respiration increase at the end of ripening and that the cytochrome pathway could be implicated in the climacteric respiratory burst before the onset of ripening. In this study, the use of tomato mutants that fail normal ripening because of deficiencies in ethylene perception or production as well as the treatment of one selected mutant with a chemical precursor of ethylene have revealed that the bioenergetics of tomato fruit development and ripening is under the control of this plant hormone. Indeed, the evolution pattern of bioenergetic features changes with the type of mutation and with the introduction of ethylene into an ethylene-synthesis-deficient tomato fruit mutant during its induced ripening.     

Involvement of wound and climacteric ethylene in ripening avocado discs

Avocado (Persea americana Mill. cv Hass) discs (3 mm thick) ripened in approximately 72 hours when maintained in a flow of moist air and resembled ripe fruit in texture and taste. Ethylene evolution by discs of early and midseason fruit was characterized by two distinct components, viz. wound ethylene, peaking at approximately 18 hours, and climacteric ethylene, rising to a peak at approximately 72 hours. A commensurate respiratory stimulation accompanied each ethylene peak. Aminoethoxyvinyl glycine (AVG) given consecutively, at once and at 24 hours following disc preparation, prevented wound and climacteric respiration peaks, virtually all ethylene production, and ripening. When AVG was administered for the first 24 hours only, respiratory stimulation and softening (ripening) were retarded by at least a day. When AVG was added solely after the first 24 hours, ripening proceeded as in untreated discs, although climacteric ethylene and respiration were diminished. Propylene given together with AVG led to ripening under all circumstances. 2,5-Norbornadiene given continuously stimulated wound ethylene production, and it inhibited climacteric ethylene evolution, the augmentation of ethylene-forming enzyme activity normally associated with climacteric ethylene, and ripening. 2,5-Norbornadiene given at 24 hours fully inhibited ripening. When intact fruit were pulsed with ethylene for 24 hours before discs were prepared therefrom, the respiration rate, ethylene-forming enzyme activity buildup, and rate of ethylene production were all subsequently enhanced. The evidence suggests that ethylene is involved in all phases of disc ripening. In this view, wound ethylene in discs accelerates events that normally take place over an extended period throughout the lag phase in intact fruit, and climacteric ethylene serves the same ripening function in discs and intact fruit alike.     

Responses of strawberry fruit to 1-Methylcyclopropene (1-MCP) and ethylene

1-Methylcyclopropene (1-MCP), a competitive inhibitorof ethylene action, binds to the ethylene receptor toregulate tissue responses to ethylene. In this work,we investigated the effects of 1-MCP and exogenousethylene on ripening, respiration rate, ionicconductivity and peroxidase activity in strawberryfruit. Strawberry fruit can ripen without exogenousethylene treatment, but exogenous ethylene inducessecondary ripening processes. Results indicated thatstimulation of respiration by ethylene wasdose-dependent. Fruit colour development and softeningwere slightly accelerated by ethylene, but changes insoluble solid content were not. 1-MCP may/may notaffect the respiratory rise induced by exogenousethylene dependent on fruit maturity. Cycloheximide(CHI) reduced the ethylene-induced respiratoryincrease. Combinations of 1-MCP and CHI reducedrespiration more than CHI alone. 1-MCP and CHI did notinfluence the primary respiratory change in nonethylene-treated fruit. This indicates that ethyleneinduced respiratory increase may involve an ethylenereceptor in early harvested fruit, but not in laterharvested fruit. Exogenous ethylene stimulatedrespiration by regulating new respiratory enzyme(s)synthesis in strawberry fruit. Ethylene induced anionic leakage increase, and this was positivelycorrelated to fruit water loss and peroxidaseactivity. These results suggest that non-climactericfruit, such as strawberry, may have different ethylenereceptor(s) and/or ethylene receptor(s) may havedifferent regulatory functions. It may be thesecondary effect of ethylene to stimulate respirationin strawberry. Non-climacteric fruit ripening may berelated to the development of active oxygen species(AOS) induced by postharvest stress.     

Effect of a Protein Polygalacturonidase Inhibitor from Apple Fruit Tissue on the Enzyme Released by Fungal Pathogens

A protein polygalacturonidase inhibitor isolated from fruit of the apple cultivars Antonovka and Mantuanskoe differently affects the polygalacturonidases of different phytopathogenic fungi. Three groups of fungi were recognized according to the sensitivity of their polygalacturonidases to the inhibitory effect. Storage of apples after harvesting is accompanied by changes in the inhibitor activity, and the time pattern of these changes depends on the cultivar. An increase in the inhibitor activity occurs concurrently with the elevation in ethylene release characteristic of the stage of elevated respiration (a climacteric increase). The data suggest that a decrease in the apple fruit resistance to microbes at the end of the storage period is related, along with other reasons, to a change in the activity of the protein polygalacturonidase inhibitor.     

Role of Brassinosteroids, Ethylene, Abscisic Acid, and Indole-3-Acetic Acid in Mango Fruit Ripening

Rapid ripening of mango fruit limits its distribution to distant markets. To better understand and perhaps manipulate this process, we investigated the role of plant hormones in modulating climacteric ripening of ??Kensington Pride?? mango fruits. Changes in endogenous levels of brassinosteroids (BRs), abscisic acid (ABA), indole-3-acetic acid (IAA), and ethylene and the respiration rate, pulp firmness, and skin color were determined at 2-day intervals during an 8-day ripening period at ambient temperature (21?±?1°C). We also investigated the effects of exogenously applied epibrassinolide (Epi-BL), (+)-cis, trans-abscisic acid (ABA), and an inhibitor of ABA biosynthesis, nordihydroguaiaretic acid (NDGA), on fruit-ripening parameters such as respiration, ethylene production, fruit softening, and color. Climacteric ethylene production and the respiration peak occurred on the fourth day of ripening. Castasterone and brassinolide were present in only trace amounts in fruit pulp throughout the ripening period. However, the exogenous application of Epi-BL (45 and 60?ng?g^?1 FW) advanced the onset of the climacteric peaks of ethylene production and respiration rate by 2 and 1?day, respectively, and accelerated fruit color development and softening during the fruit-ripening period. The endogenous level of ABA rose during the climacteric rise stage on the second day of ripening and peaked on the fourth day of ripening. Exogenous ABA promoted fruit color development and softening during ripening compared with the control and the trend was reversed in NDGA-treated fruit. The endogenous IAA level in the fruit pulp was higher during the preclimacteric minimum stage and declined during the climacteric and postclimacteric stages. We speculate that higher levels of endogenous IAA in fruit pulp during the preclimacteric stage and the accumulation of ABA prior to the climacteric stage might switch on ethylene production that triggers fruit ripening. Whilst exogenous Epi-BL promoted fruit ripening, endogenous measurements suggest that changes in BRs levels are unlikely to modulate mango fruit ripening.     

Respiration and protein synthesis in nongrowing cultured pear fruit cells in response to ethylene and modified atmospheres: a model system for fruits postharvest

The respiration of pear fruit (Pyrus communis L. Passe Crassane) cells was monitored after subculture into an auxin-free, mannitol-enriched medium in which the cells remained viable but did not grow. Respiration rates were affected by the presence or absence of sucrose in the medium even though the cells retained reserves of sucrose and starch. Provided the medium contained respirable carbohydrate, exposure to ethylene (1-10 microliters per liter) increased the respiration rate with some acceleration of cell death. In the range from 10 to 2% oxygen by volume, the respiration rate of the cells decreased with oxygen concentration resulting in some prolongation of cell life. Thus, in their responses to ethylene and modified atmospheres, the cells reflected the behavior of harvested fruits. Having defined conditions under which respiration rate could be varied without apparent influence on the quiescent state of the cells, we sought a connection between maintenance respiration and protein turnover. Relative rates of protein synthesis were assessed by measuring ribosome distribution between monosomes and polysomes. In general, the higher the respiration rate the higher the proportion of polysomes supporting the thesis that protein turnover is a variable component of maintenance metabolism. Protein turnover in cells incubated in the presence or absence of sucrose was measured as retained α-amino-³H following a pulse of ³H₂O. Turnover was shown to be a quantitatively important component of the maintenance budget and to be more rapid in cells in media supplemented with sucrose through the chase period. The experiments illustrate that cultured cells may be used to explore aspects of the maintenance metabolism of resting or senescent cells that are not amenable to study in bulky fruit tissues.     

Ripening in papaya fruit is altered by ACC oxidase cosuppression

Papaya (Carica papaya) is a very important crop in many tropical countries but it is highly susceptible to parasitic diseases, physiological disorders, mechanical damage and fruit overripening. Here we report a study on ACC oxidase cosuppression and its effects on papaya fruit ripening. Papaya ACC oxidase was isolated using PCR and embriogenic cells transformed by biolistic using the CaMV 35S promoter to drive the expression of the PCR fragment in sense orientation. Fifty transgenic lines were recovered and 20 of those were grown under field conditions. Southern analysis showed incorporation of the transgene in different copy numbers in the papaya genome. Fruits were evaluated in terms of texture (firmness), colour development, respiration and ethylene production. A sharp reduction in ethylene and CO2 production was detected, whereas softening and colour development of the peel were also altered. Overall, transgenic fruits showed a delay in ripening rate. A reduction in mRNA level for ACC oxidase in transgenic fruit was clearly detectable by northern blot. More studies are necessary before this technology can be used to extend the shelf life of papaya fruit.     

Respiratory Patterns in Fruit of Pineapple, Ananas comosus, Detached at Different Stages of Development

The postharvest respiratory drifts for six stages of development of pineapple fruit (Ananas comosus cv. Cayenne) ranging from dry flower to senescence are presented. Based upon these data, pineapple is a non-climacteric fruit. Pineapple does produce ethylene gas hut when levels ranging from 0.01 to 1000 μl/l were applied to stage 4 fruits (fruit just at the start of ripening) no respiration or chemical changes were induced which could he interpreted as affecting the ripening processes. A decrease in the oxygen concentration (to 2.5 per cent) resulted in a decrease in the respiration rate. An increased carbon dioxide concentration (up to 10 per cent) had not detectable effect on the respiration rate.     

Effect of proteinaceous polygalacturonase inhibitors from apple seed tissue on an enzyme isolated from phytopathogenic fungi

A protein polygalacturonidase inhibitor isolated from fruit of the apple varieties Antonovka and Mantuanskoe differently affects the polygalacturonidases of different phytopathogenic fungi. Three groups of fungi were recognized by the sensitivity of their polygalacturonidases to the inhibitory effect. Storage of apples after harvesting is accompanied by changes in the inhibitor activity, and the time pattern of these changes depends on the variety. An increase in the inhibitor activity occurs concurrently with the elevation in ethylene release characteristic of the stage of elevated respiration (a climacteric increase). The data suggest that a decrease in the apple fruit resistance to microbes at the end of the storage period is related, along with other reasons, to a change in the activity of the protein polygalacturonidase inhibitor.     

The role of ethylene in the prevention of chilling injury in nectarines

Woolliness is a chilling injury phenomenon occurring in nectarines held at low temperatures for extended periods. It is a disorder marked by altered cell wall metabolism during ripening leading to a dry, woolly texture in the fruit. Two treatments were found to alleviate this disorder. One was holding the fruits for 2 days at 20 °C before 0 °C storage (delayed storage) and the second was having ethylene present during cold storage (ethylene). Immediately stored fruit (control) had 88 percnt; woolliness while 7 percnt; of delayed storage and 15 percnt; of ethylene fruit showed woolliness. The severity of the injury in individual fruits was closely related to inhibition of ethylene evolution. Woolly fruit had higher levels of 1-aminocyclopropane-1-carboxylic acid (ACC) and less 1-aminocyclopropane-1-carboxylic acid oxidase (ACO, EC 1.4.3) activity than healthy fruit. It is suggested that ethylene is essential for promoting the proper sequence of cell wall hydrolysis necessary for normal fruit softening. This is in contrast to chilling injury in other fruits, whereby ethylene is often a sign of incipient damage. Respiration was also found to be associated with chilling injury, in that fruit with woolliness had a depressed respiration.     

Physiology and firmness determination of ripening tomato fruit

Tomato ( Lycopersicon esculentum Mill.) genotypes varying in intrinsic firmness were examined to determine the quantitative relationships between polygalacturonase (EC 3.2.1.15) activity, firmness and other ripening parameters including rate (days from mature-green to full red) and intensity (rate of ethylene production at climacteric peak) of ripening. Texture, respiration and ethylene production were monitored in the immature-green through the red (ripe) stages of development. Polygalacturonase activity was measured by direct assay of salt-extractable wall protein or by monitoring the release of pectins from isolated, enzymically active wall. In all fruit, polygalacturonase activity was highly correlated with pericarp softening, but only moderately correlated with softening of whole fruit (r = 0.920 and 0.757, respectively). Polygalacturonase activity was positively correlated with cell-wall autolytic activity in pink (r = 0.969) and red (r = 0.900) fruit. Firmer genotypes exhibited lower rates of respiration and ethylene production during ripening. Polygalacturonase activity in isolates prepared from fruit at the climacteric peak was positively correlated with ethylene production and respiration, and negatively correlated with days to ripening (r = 0.929, 0.805, and -0.791, respectively). The data demonstrate the importance of selecting the appropriate method of firmness determination and are consistent with the hypothesis that pectin fragments released by polygalacturonase contribute to the production of autocatalytic (system II) ethylene.     

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。