采后果蔬对乙烯受体抑制剂的响应及贮运保鲜技术的研究

近几年来 ,随着重氮环戊二烯和环丙烯类等乙烯受体抑制剂的发现 ,为控制乙烯敏感型的果蔬采后成熟、衰老提供新的技术手段。从乙烯受体抑制剂的特性、作用特点以及可能作用机理等方面概述了采后果蔬对乙烯受体抑制剂的响应和应用乙烯受体抑制剂延长采后果蔬贮运保鲜的技术。     

Differential Suppression of Ethylene Biosynthesis and Receptor Genes in ‘Golden Delicious’ Apple by Preharvest and Postharvest 1-MCP Treatments

Harvista?, a sprayable formulation of 1-methylcyclopropene (1-MCP), has recently been developed for preharvest use on horticultural products, whereas SmartFresh? is a widely used 1-MCP treatment for products after harvest. The effects of Harvista? on apple fruit ripening when sprayed at different maturities and on expression patterns of ethylene biosynthesis and receptor genes during storage have been investigated. Harvista? applied to on-tree maturing apple fruit at an average starch pattern index of 2.5 resulted in a higher at-harvest firmness value compared with those treated at a starch pattern index of 1.5 and 3.5. This indicates that the timing of the Harvista? application is critical. An application of Harvista? led to better postharvest fruit firmness retention as well as reduced ethylene production. In addition, both preharvest and postharvest 1-MCP treatments resulted in contrasting responses in the expression patterns of two ethylene biosynthesis genes and in differentially suppressing effects on four ethylene receptor genes. Furthermore, the combined application of Harvista? + SmartFresh? resulted in greater fruit firmness retention and longer ethylene suppression. The expression profiles of these genes during on-tree fruit maturation prior to Harvista? application were also characterized. Different regulation patterns of receptor genes could contribute to differential effects by 1-MCP treatments. The potential roles of Harvista? to manipulate the ripening process as well as the molecular mechanism influencing 1-MCP treatment efficacy are discussed.     

Growth inhibition of pollen tubes by blocking the aromatic or branched-chain amino acid pathways of biosynthesis

A number of organic molecules that appear to block the ethylene receptor have been discovered recently. For example, on irradiation with visible light, diazocyclopentadiene (DACP), gives rise to some potent but as yet unidentified inhibitor compounds. Some synthetic cyclopropenes have been shown to bind to the ethylene receptor and prevent the physiological action of ethylene for extended periods. Cyclopropene (CP). 1-methylcyclopropene (1-MCP) and 3,3-dimethylcyclopropene (3,3-DMCP) have been shown to prevent ethylene effects in a number of plants. As low a concentration as 0.5 nl l⁻¹ of 1-MCP is sufficient to protect carnation ( Dianthus caryophyllus ) flowers for several days against ethylene, and 0.7 nl l⁻¹ 1-MCP or CP will prevent the ripening of banana ( Musa sapientum ) for 12 days at 24°C. Some plant organs require higher concentrations of these inhibitors. Complete inhibition of ethylene effects in pea seedlings requires treatment with 40 n1 1⁻¹ of 1-MCP. These novel inhibitors appear to be suitable for many commercial applications including extending the vase life of cut flowers and the display life of potted plants. Since 1-MCP apparently is non-toxic at concentrations that are active, it may in future be available for regulating the ripening of fruits and preventing the deleterious effects of ethylene in vegetables.     

Compounds Interacting with the Ethylene Receptor in Plants

Abstract: Some of the compounds binding to the ethylene receptor induce an ethylene response, but others prevent it. The compounds preventing an ethylene response have been developed into a means for protecting plants against ethylene and extending the life of some plant material. 1-Methylcyclopropene (1-MCP), a compound now commercially available under the names EthylBloc and SmartFresh™, is currently being used on flowers, fruit and vegetables with great success. In ethylene sensitive flowers, among other responses, it prevents senescence and abscission of plant organs; in fruit and vegetables it slows down the ripening process. Other similar compounds are now being developed for a range of methods of application.     

Responses of banana fruit to treatment with 1-methylcyclopropene

Experiments were conducted to determine levels of 1-methylcyclopropene (1-MCP) exposure needed to prevent ethylene-stimulated banana fruit ripening, characterise responses of ethylene-treated fruit to subsequent treatment with 1-MCP, and to test effects of subsequent ethylene treatment on 1-MCP-treated fruit softening. Fruit softening was measured at 20°C and 90% relative humidity. One hour exposure at 20°C to 1000 nl 1-MCP/l essentially eliminated ethylene-stimulated ripening effects. Exposure for 12 h at 20°C to just 50 nl 1-MCP/l was similarly effective. Fruit ripening initiated by ethylene treatment could also be delayed with subsequent 1-MCP treatment. However, 1-MCP treatment only slowed down ripening of ethylene-treated fruit when applied at 1 day after ethylene and was ineffective when applied 3 or 5 days after ethylene treatment. The ripening response of fruit treated with 1-MCP and subsequently treated with ethylene varied with interval time between 1-MCP and ethylene treatments. As time increased, the response of 1-MCP-treated fruit to ethylene was enhanced. Responses to 0.1, 1, 10 or 100 µl ethylene/l concentrations were similar. Enzyme kinetic analysis applied to 1-MCP effects on ethylene-induced softening of banana fruit suggested that 1-MCP inhibition is by noncompetitive antagonism of ethylene binding.     

Control of ethylene activity in various plant systems by structural analogues of 1-methylcyclopropene

Two structural analogues of 1-methylcyclopropene (1-MCP), 1-ethylcyclopropene (1-ECP) and 1-propylcyclopropene (1-PCP) were found to inhibit ethylene action and thereby the responses to ethylene in various plant systems. When applied prior to exposure to ethylene, the analogues considerably delayed ethylene-induced ripening of avocado and tomato fruits, delayed citrus leaf explants abscission and reversed ethylene-induced swelling and inhibition of elongation in etiolated pea plants. The analogues exerted their effect in a concentration-depended manner, at a range of several parts per million. Of the two analogues, 1-ECP was found in all cases more potent than 1-PCP but less potent then the mother compound 1-MCP. It is proposed that the analogues inhibit ethylene action by competing for the sites of binding on the ethylene receptor, similar to the mode of action suggested for 1-MCP. Findings revealed in this study imply that the competition of ethylene and the analogues for the ethylene site of binding is of a non-competitive nature. The analogues effectively inhibited ethylene action only if applied before the plant material was exposed to ethylene, or in the case of fruits shortly after harvest. Simultaneous application of the analogues and ethylene reduced the inhibitory effect of the analogues. Application of the analogues after exposure to ethylene or after fruit ripening had nullified the inhibitory effect of the analogues. Ripening of fruits, treated with the analogues, was inhibited for a finite period of time after which the fruits ripened normally. This resumption of ripening ability is attributed to presence of free binding sites on the ethylene receptor at the point of recovery from the inhibition. As the analogues are volatile, non-corrosive, non-toxic, odorless compounds and effective at minute concentrations, they can be considered promising candidates for practical use.     

Apple 1-Aminocyclopropane-1-Carboxylic Acid Synthase Genes, MdACS1 and MdACS3a, are Expressed in Different Systems of Ethylene Biosynthesis

1-Aminocyclopropane-1-carboxylic acid synthase (ACS) is one of the key regulatory enzymes involved in the synthesis of ethylene. Climacteric fruit ripening is accompanied by increased ethylene production, in which ethylene biosynthesis is changed from system 1 to system 2. In apple, at least four members of the ACS gene family have been identified, two of which, MdACS1 and MdACS3a, have been studied extensively due to their specific expression in fruit tissue. However, the regulatory role of MdACS1 and MdACS3a in the ethylene biosynthesis system is unknown. Here we addressed this issue by investigating ACS expression in ripening apple fruits. Expression analysis in ‘Golden Delicious’ and ‘Red Fuji’ fruits, in combination with treatments of 1-MCP (1-methylcyclopropene, an ethylene inhibitor) and Ethephon (an ethylene releaser) has demonstrated that MdACS3a and MdACS1operate in system 1 and system 2 ethylene biosynthesis, respectively.     

1-甲基环丙烯采后处理对樱桃番茄果实成熟过程的影响

研究了不同浓度(0、0.035、0.07和0.11μL/L)的乙烯受体竞争性抑制剂1-甲基环丙烯(1-MCP)采后处理对绿熟期樱桃番茄的乙烯合成、果实软化、果实色素(叶绿素、茄红素、β-胡萝卜素)含量消长的影响.0.07 μL/L及其以上浓度的1-MCP降低了前期乙烯合成,同时推迟了乙烯释放高峰,但0.035 μL/L浓度的1-MCP处理并不能抑制内源乙烯合成.1-MCP显著延迟了果实软化和叶绿素降解,但并不影响这两个过程的启动.茄红素合成的启动和积累均受到了1-MCP抑制,而1-MCP并不推迟β-胡萝卜素合成的启动,只抑制其积累.这些结果提示了乙烯调节成熟生理过程的不同机制.对于绿熟期的樱桃番茄,0.07～0.11μL/L的1-MCP是实用的有效处理浓度.1-MCP有效浓度可能用于了解果实的乙烯受体水平和乙烯敏感性.     

Analysis of physiological and biochemical changes in kiwifruit (Actinidia deliciosa cv. Allison) after the postharvest treatment with 1-Methylcyclopropene

Kiwifruits have limited shelf life under ambient conditions. However, it is necessary to extend its life so as to make it available for longer time in the market and to make it commercial fruits in India. Hence, the present study was designed to observe the effect of different concentrations of 1-Methylcyclopropene (1-MCP) on physiological and biochemical parameters which have great influence on post harvest life and quality of kiwifruits. Kiwifruits cv. Allison was treated with different concentrations of 1-Methylcyclopropene (0.5 μL/L, 1 μL/L, 2 μL/L) and un-treated fruits served as control. 1-MCP treatment was given for 24 h at 20°C. After treatments, the fruits were transferred to ambient storage, and observations on different physiological and biochemical parameters were recorded at 3 days interval. Our results indicated that all concentrations of 1-MCP delayed ripening of kiwifruits but 2 μL/L concentrations was the most effective in doing so. Fruits treated with 1-MCP at 2 μL/L started ripening after 12th day of storage whereas untreated fruits started ripening even on 6th day. Polygalactouronase (PG) and lipoxygenase (LOX) enzyme activities were lesser in 1-MCP treated fruits than control. 1-MCP treated fruits respired less and evolved lesser ethylene.     

Inhibition of the ethylene response by 1-MCP in tomato suggests that polyamines are not involved in delaying ripening, but may moderate the rate of ripening or over-ripening

Ethylene initiates the ripening and senescence of climacteric fruit, whereas polyamines have been considered as senescence inhibitors. Ethylene and polyamine biosynthetic pathways share S-adenosylmethionine as a common intermediate. The effects of 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, on ethylene and polyamine metabolism and associated gene expression was investigated during ripening of the model climacteric fruit, tomato (Solanum lycopersicum L.), to determine whether its effect could be via polyamines as well as through a direct effect on ethylene. 1-MCP delayed ripening for 8 d compared with control fruit, similarly delaying ethylene production and the expression of 1-aminocyclopropane-1-carboxylic acid (ACC)-synthase and some ethylene receptor genes, but not that of ACC oxidase. The expression of ethylene receptor genes returned as ripening was reinitiated. Free putrescine contents remained low while ripening was inhibited by 1-MCP, but increased when the fruit started to ripen; bound putrescine contents were lower. The activity of the putrescine biosynthetic enzyme, arginine decarboxylase, was higher in 1-MCP-treated fruit. Activity of S-adenosylmethionine-decarboxylase peaked at the same time as putrescine levels in control and treated fruit. Gene expression for arginine decarboxylase peaked early in non-treated fruit and coincident with the delayed peak in putrescine in treated fruit. A coincident peak in the gene expression for arginase, S-adenosylmethionine-decarboxylase, and spermidine and spermine synthases was also seen in treated fruit. No effect of treatment on ornithine decarboxylase activity was detected. Polyamines are thus not directly associated with a delay in tomato fruit ripening, but may prolong the fully-ripe stage before the fruit tissues undergo senescence.     

1-MCP对室温贮藏下不同成熟度猕猴桃的生理效应

以适期采收(成熟度Ⅰ)和晚采收(成熟度Ⅱ)的'海沃德'猕猴桃果实为材料,研究0.5 μL·L~(-1) 1-甲基环丙烯(1-MCP)处理对室温贮藏(20℃)条件下两种果实的生理生化指标的影响.结果显示:1-甲基环丙烯(1-MCP)处理能有效延缓贮藏期间两种成熟度果实的硬度、可滴定酸含量、维生素C含量的下降,显著降低二者的乙烯释放速率和呼吸强度及其峰值,提高果实的POD、CAT和SOD活性.与成熟度Ⅰ相比,成熟度Ⅱ果实的硬度下降较快,且丙二醛含量和呼吸速率却明显较高,呼吸速率和乙烯释放量高峰时间相对提前.研究表明,在相同的贮藏条件下,晚采收的'海沃德'猕猴桃果实成熟衰老进程明显快于适期采收果实,贮藏效果较差;1-MCP能够明显延长猕猴桃果实的贮藏时间,表现出显著的保鲜效果.     

ABA和乙烯在草莓采后成熟衰老中的作用

随着草莓果实采后成熟衰老,ABA和乙烯生成迅速增长,乙烯累积与果实的变质腐烂程度呈正相关。ABA处理能增高纤维素酶活性和呼吸,而GA有抑制作用。ABA能促进乙烯、ACC生成,对MACC则无影响。GA_3抑制乙烯、ACC生成,促进MACC积累。CO_2对草莓有良好保鲜效果,并有效地抑制ABA和乙烯生成,低温下效果更为显著。     

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.     

1-Methylcyclopropene prevents cotton physiological and molecular responses to ethylene

The phytohormone ethylene can cause adverse effects in plants, including inhibition of shoot elongation and abscission of leaves, flowers and fruits. 1-Methylcyclopropene (1-MCP) is a competitive inhibitor of ethylene binding with the ethylene receptors and prevents ethylene responses. To determine the effectiveness of 1-MCP as an ethylene prophylactic for cotton, an assay system was developed that integrated analyses of leaf declination, shoot elongation and ethylene-responsive gene expression (GhACS6, GhACO5, GhEIN4, GhLTL1). Using a flow-through controlled growth system, the optimal parameters of ethylene treatment in eliciting responses and of 1-MCP treatment in preventing these responses, were determined. Shoot elongation and gene expression changes in response to ethylene correlated well, but gene expression changes were found to offer a more robust estimate of ethylene responsiveness. The effect of ethylene treatment on the expression of the marker genes was determined to be optimal with 3?h of 10???L?L^?1 fumigation. 250?nL?L^?1 of gaseous 1-MCP provided for 1?h immediately before ethylene fumigation was highly efficacious at preventing ethylene responses, but the duration of protection did not exceed 48?h. A liquid formulation of 1-MCP designed for field use was found to be equally effective as the gas. The results, and the system described, may prove useful in increasing 1-MCP efficacy as a tool to manage the negative effects of stress ethylene in cotton crops.     

Superoxide dismutase and catalase activities in apple fruit during ripening and post‐harvest and with special reference to ethylene

Oxidative stress is involved in many biological systems, among which are fruit ripening and senescence. Free radicals play an important role in senescence and ageing processes. Plants have evolved antioxidative strategies in which superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6) are the most efficient antioxidant enzymes, influencing patterns of fruit ripening. Variations in total SOD and CAT activities were determined at regular intervals during ripening and senescence in on‐tree and cold‐stored apple fruits of the cultivars Fuji and Golden Delicious. In all fruits, internal ethylene concentration was also measured. The results suggest that the onset of ripening, signalled by ethylene burst, is closely related to SOD and CAT activities. In on‐tree fruits the climacteric peak in ethylene was associated with the peaks of SOD and CAT activity in both cultivars. Quite different results were obtained in cold‐stored fruits: Ethylene concentration increased in both cultivars during the storage. CAT activity doubled in both cultivars. SOD activity decreased in Golden Delicious and peaked in Fuji.     

Proteomic analysis of differentially accumulated proteins during ripening and in response to 1-MCP in papaya fruit

Papaya (Carica papaya L.) is a climacteric fruit susceptible to postharvest losses due to the ethylene-induced ripening. The inhibitor of ethylene action, 1-methylcyclopropene (1-MCP), has been used worldwide as a safe postharvest non-toxic agent, but the physiological and biochemical modifications induced by 1-MCP are not well understood. Using the 2-DE analysis, we report the changes in the protein profiles after 6 and 18 days of postharvest and the effect of the effect of 1-MCP treatment on fruits. Twenty seven protein spots showing differences in abundance during ripening were successfully identified by nano-LC-ESI/MS/MS. Some spots corresponded to the cell wall degrading enzymes related to fruit ripening; others were involved in oxidative damage protection, protein folding, and cell growth and survival that were induced by 1-MCP. This is the first proteomic report analyzing the effect of 1-MCP in papaya ripening. The present data will help to shed light on papaya fruit ripening process.     

Inhibition of ethylene-induced cellular senescence symptoms by 1-methylcyclopropene, a new inhibitor of ethylene action

Ethylene is known to accelerate flower senescence, but the sequence of events that links its interaction with the tissue and the final senescence symptoms is still obscure. Recently, 1-methylcyclopropene (1-MCP) was found to inhibit ethylene-induced wilting in flowers. This work was carried out in order to investigate the effects of 1-MCP on cellular senescence symptoms in petunia flowers following expossure to ethylene. Cut petunia ( Petunia hybrida ) flowers that were exposed to ethylene for 12 h at concentrations of 1–12 ppm wilted sooner than their untreated counterparts. This effect was abolished by a 6-h pre-treatment with 1-MCP. Immediately following the ethylene treatment, decreases in petal fresh weight and total protein content were measured, along with higher electrolyte leakage, and lower membrane lipid fluidity and protein content. When applied alone, 1-MCP had relatively little impact on these parameters. However, when the flowers were treated with 1-MCP prior to the ethylene treatment, ethylene had no effect. These results indicate that while ethylenes effects on wilting were obvious 3 days after the treatment, cellular parameters were affected already at the end of the treatment. Since 1-MCP repressed these early ethylene effects, it was concluded that it interferes with ethylene action in petunia flowers at a rather early stage, long before apparent wilting.     

Control of ethylene synthesis by expression of a bacterial enzyme in transgenic tomato plants.

Synthesis of the phytohormone ethylene is believed to be essential for many plant developmental processes. The control of ripening in climacteric fruits and vegetables is among the best characterized of these processes. One approach to reduce ethylene synthesis in plants is metabolism of its immediate precursor, 1-aminocyclopropane-1-carboxylic acid (ACC). Soil bacteria containing an enzyme, ACC deaminase, were identified by their ability to grow on ACC as a sole nitrogen source. The gene encoding ACC deaminase was cloned and introduced into tomato plants. Reduction in ethylene synthesis in transgenic plants did not cause any apparent vegetative phenotypic abnormalities. However, fruits from these plants exhibited significant delays in ripening, and the mature fruits remained firm for at least 6 weeks longer than the nontransgenic control fruit. These results indicated that ACC deaminase is useful for examining the role of ethylene in many developmental and stress-related processes in plants as well as for extending the shelf life of fruits and vegetables whose ripening is mediated by ethylene.