The Production of Ethylene by Whole Apple Fruits and by Tissue 1

It is shown that the production of ethylene by whole apples,discs of peel prepared from them, and extracts prepared fromthe discs and supplied with linolenic acid, increases simultaneouslyduring the development of the respiration climacteric in apples.As the climacteric peak is reached, the ability of the extractsto produce ethylene declines and this is associated with a rapidloss of ethylene-producing activity on short term (up to 24h) ageing of the peel discs from which the extracts were obtained. It is suggested that the gmn.ll ethylene production by extracts(with linolenic acid) from pre-climacteric fruits, which arenot themselves evolving sufficient ethylene for its detectionin the ambient atmosphere, may be explained on the grounds thateven before the climacterio the fruit is producing small amountsof ethylene within the tissue; the tissue, as the climactericphase is approached, becomes more sensitive to ethylene andautocatalysed production of the gas then stimulates the fruitinto the respiration climacteric.     

Metabolic Changes in Excised Fruit Tissue. IV. Changes Occurring in Discs of Apple Peel During the Development of the Respiration Climacteric

It is shown that a sequential development of a series of enzyme systems occurs in the peel of the apple as the respiration climacteric develops in the whole fruit. The sequence of development of these systems, i.e. acetate incorporation into lipid, production of ethylene, incorporation of amino acid into protein and, finally, the decarboxylation of added malate (malate effect) is the same as that shown earlier for the short term (24 hr) aging of peel discs from pre-climacteric apples. As these systems appear in the initial discs from fruit passing through the climacteric they gradually cease to increase during the 24 hour aging period. Uptake studies show that none of the changes in these systems can be due solely to changes in the permeability of the tissue over the climacteric period. On the basis of these results it is tentatively suggested that the aging of discs from pre-climacteric tissue might provide a model system for a detailed study of the physiological and biochemical changes occurring during the climacteric of apple fruits.     

Comparative effects of acetylene and ethylene gas on initiation of banana ripening

Bananas were exposed to acetylene or ethylene at 0·01, 0·1 and 1 ml/litre, under high humidity, for 24 h at 18 °C. They were then transferred to an atmosphere of air alone for a further 4 days and during this period the respiration rate of three fruit from each treatment was measured. Ripeness was then assessed by colour score and soluble solids content. All levels of ethylene initiated ripening. Treatment with ethylene induced a climacteric rise in respiration, an increase in the soluble solids content of the pulp and degreening of the peel. All levels of acetylene, except 0·01 ml/litre, induced a climacteric rise in respiration. Fruit treated with acetylene at 1 ml/litre had a similar colour score and soluble solids content to those ripened by exposure to ethylene. Fruits treated with acetylene at 0·1 ml/litre had a lower soluble solids content and their peel remained green. Treatment with acetylene at 0·01 ml/litre failed to initiate ripening. Sensory evaluation of fruit ripened by acetylene at 1 ml/litre indicated that the acetylene treated fruit ripened slightly more slowly. When compared at the same stage of ripeness fruits from the two treatments were equally palatable.     

Effects of Oxygen Concentration and Ethephon upon the Respiration and Ripening of Banana Fruits

A respiratory climacteric and accumulation of soluble solidsof pulp were induced in banana fruits by (2-chlorethyl) phosphonicacid (ethephon) at oxygen concentrations of 3–21% (v/v).However, induction of peel colour change by ethephon was retardedor prevented at oxygen concentrations of 10% (v/v) or less.Thus pulp tissue ripened whilst the peel remained green. Respirationrate, soluble solids content of pulp, and peel colour were notaffected by ethephon at oxygen concentrations of 1% (v/v) orless. The kinetics of respiratory response to ethephon are consistentwith a model in which ethylene (derived from ethephon) is acoupling-activator of respiration. In this model ethylene decreasesthe affinity of an enzyme system for its substrate, but acceleratesthe release of product.     

Altered sensitivity to ethylene in ‘Tardivo’, a late‐ripening mutant of Clementine mandarin

‘Tardivo’ mandarin is a mutant of ‘Comune’ Clementine with a delay in peel degreening and coloration, allowing late harvesting. In this work, we have explored if the late‐harvesting phenotype of ‘Tardivo’ mandarin is related to altered perception and sensitivity to ethylene. The peel degreening rate was examined after a single ethephon treatment or during a continuous ethylene application in fruits at two maturation stages. In general, ethylene‐induced peel degreening was considerably delayed and reduced in fruits of ‘Tardivo’, as well as the concomitant reduction of chlorophyll (Chl) and chloroplastic carotenoids, and the accumulation of chromoplastic carotenoids. Analysis of the expression of genes involved in Chl degradation, carotenoids, ABA, phenylpropanoids and ethylene biosynthesis revealed an impairment in the stimulation of most genes by ethylene in the peel of ‘Tardivo’ fruits with respect to ‘Comune’, especially after 5 days of ethylene application. Moreover, ethylene‐induced expression of two ethylene receptor genes, ETR1 and ETR2, was also reduced in mutant fruits. Expression levels of two ethylene‐responsive factors, ERF1 and ERF2, which were repressed by ethylene, were also impaired to a different extent, in fruits of both genotypes. Collectively, results suggested an altered sensitivity of the peel of ‘Tardivo’ to ethylene‐induced physiological and molecular responses, including fruit degreening and coloration processes, which may be time‐dependent since an early moderated reduction in the responses was followed by the latter inability to sustain ethylene action. These results support the involvement of ethylene in the regulation of at least some aspects of peel maturation in the non‐climacteric citrus fruit.     

香蕉果实XET cDNA的克隆及其在成熟软化的果实果皮和果肉中的表达分析

木葡聚糖内糖基转移酶(Xyloglucan endotransglycosylase,XET)通过分解细胞壁半纤维素多糖的主要成分--木葡聚糖而参与果实软化.为了阐明香蕉(Musa acuminata.Colla cv.GrandNain)果实成熟过程中的软化与细胞壁代谢酶XET基因表达模式的关系,采用RT-PCR和RACE-PCR方法,首次从成熟香蕉果实果肉中分离了编码XT基因的全长cDNA(MA-XET1,全长1 095 bp).序列分析表明,MA-XET1的5'端和3'端的非翻译区分别为66 bp和1 89bp,该片段含有一个完整的开放读码框,编码280个氨基酸,推导的MA-XET1蛋白质中存在XET蛋白的催化活性部位DEIDFEFL.Southern杂交表明,MA-XET1在香蕉基因组中由多拷贝基因编码.Northern分析显示,跃变前期的果肉中,不能检测MA-XET1基因的表达,跃变期的果实果肉中MA-XET1表达增加,跃变后期该基因表达略有减弱;在跃变前期的果实果皮中,MA-XET1的积累较低,跃变期的果实果皮中积累大幅增加,而后迅速下降.Propylene(丙烯,乙烯的类似物)处理降低香蕉果实果皮和果肉的硬度,而且propylene促进MA-XET1在果皮和果肉中的积累.这些结果表明,MA-XET1参与香蕉果实成熟过程中的果皮和果肉软化,并且,MA-XET1的表达在转录水平上受乙烯调控.     

Influence of Plant Hormones on Ethylene Production in Apple, Tomato, and Avocado Slices during Maturation and Senescence

Ethylene production by tissue slices from preclimacteric, climacteric, and postclimacteric apples was significantly reduced by isopentenyl adenosine (IPA), and by mixtures of IPA and indoleacetic acid, and of IPA, indoleacetic acid, and gibberellic acid after 4 hours of incubation. Ethylene production by apple (Pyrus malus L.) slices in abscisic acid was increased in preclimacteric tissues, decreased in climacteric peak tissues, and little affected in postclimacteric tissues. Indoleacetic acid suppressed ethylene production in tissues from preclimacteric apples but stimulated ethylene production in late climacteric rise, climacteric, and postclimacteric tissue slices. Gibberellic acid had less influence in suppressing ethylene production in preclimacteric peak tissue, and little influenced the production in late climacteric rise, climacteric peak, and postclimacteric tissues. IPA also suppressed ethylene production in pre- and postclimacteric tissue of tomatoes (Lycopersicon esculentum) and avocados (Persea gratissima). If ethylene production in tissue slices of ripening fruits is an index of aging, then IPA would appear to retard aging in ripening fruit, just as other cytokinins appear to retard aging in senescent leaf tissue.     

Localization of the Ethylene-synthesizing System in Apple Tissue

Apple (Malus sp.) slices gradually lost the ability to synthesize ethylene when incubated with a mixture of enzymes that digest cell walls. The released protoplasts did not produce ethylene. The release of protoplasts was faster from climacteric fruit slices than from preclimacteric tissue. In protoplast suspension culture, as new cell wall was deposited (as judged by the intensity of fluorescence of regenerating protoplasts stained with Calcofluor White and the incorporation of labeled myo-inositol into their ethanol-insoluble residue), ethylene synthesis was gradually regained. Restored ethylene synthesis reached a maximum after 80 hours in protoplasts from preclimacteric fruit and in 120 hours in those from climacteric tissue. Addition of methionine (1 mm) to the culture medium was essential for appreciable synthesis of ethylene; and this synthesis was inhibited by the aminoethoxy analogue of rhizobitoxine and by propyl gallate, inhibitors of ethylene synthesis in higher plants. We suggest that the ethylene-synthesizing enzyme system is highly structured in the apple cell and is localized in a cell wall-cell membrane complex.     

The respiratory climacteric is present in Charentais (Cucumis melo cv. Reticulatus F1 Alpha) melons ripened on or off the plant

Ripening of climacteric fruit is accompanied by an increasein respiration and autocatalytic ethylene synthesis. In harvestedmelons, there is variation in the magnitude and duration ofthe respiratory climacteric depending on the cultivar. It hasrecently been reported that, while the ripening-associated increasein ethylene production is present, the respiratory climactericis absent in ripening melon fruit attached to the plant, leadingto the suggestion that climacteric respiration is an artifactof harvest. To address the universality of this phenomenon,ripening behaviour in the melon cultivar Charentais (Cucumismelo cv. Reticulatus F1 Alpha), was investigated and the resultsshow that the respiratory climacteric occurs in fruit ripenedboth on and off the plant. Key words: Cucumis melo, ethylene, respiratory climacteric     

Ethylene biosynthesis in fruit tissues

Tracer studies with avocado tissues indicate that methionine is converted to ethylene at stages of the climacteric rise and the climacteric peak, but not at the preclimacteric stage. The results suggest that the control of ethylene biosynthesis is at a step after methionine is synthesized. The endogenous content of methionine was found to be so low that methionine must be actively turned over for ethylene biosynthesis during the stages when the rate of ethylene production is high. Oxygen was found to be essential for this conversion, indicating that at least one of the steps in conversion of methionine to ethylene is oxygen-dependent. The ability of methionine and its keto analogue (α-keto-γ-methylthiobutyric acid) to serve as ethylene precursors by apple tissues was compared. Chemical and kinetic evidence support the view that methionine is a closer precursor of ethylene than its keto analogue.     

Biochemical aspects of the developing and ripening banana

The peel and pulp of the banana fruit and the pseudostem were examined for glutamate-oxaloacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and aldolase activities and protein, phenolics, chlorophyll and starch. The peel-pulp ratio at various stages of fruit development on the plant and in detached fruits showing incipient ripening were used as an index of the physiological age of the fruit. The enzymes exhibited maximum activity at a stage corresponding to the initiation of the climacteric. GPT level at this stage was higher than that of GOT. An initial increase in the protein content was followed by a decline in both peel and pulp, the level reaching a minimum in climacteric fruits. Astringency, measured in terms of total phenolics, decreased with development; in mature fruits, peel contained 4–5 × as much phenolics as pulp. Chlorophyll in mature fruits was 10 × higher than in young fruits and decreased in ripe fruits. The onset of ripening was attended with a pronounced decrease in the starch. The various analyses were carried out also on the pseudostem removed from the plant soon after flower formation.     

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.     

Metabolism of 1-aminocyclopropane-1-carboxylic acid in ripening apple fruits

In preclimacteric apple fruits ( Malus × domestica Borkh. cv. Golden Delicious) ethylene production is controlled by the rates of 1-aminocyclopropane-1-carboxylic acid (ACC) synthesis, and by its metabolism to ethylene by the ethylene-forming enzyme and to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC) by malonyl CoA-ACC transferase. The onset of the climacteric in ethylene production is associated with an increase in the activity of the ethylene-forming enzyme in the pulp and with a rise in the activity of ACC synthase. Malonyl transferase activity is very high in the skin of immature fruit, decreases sharply before the onset of the climacteric, and remains nearly constant thereafter. More than 40% of the ACC synthesized in the skin and around 5% in the flesh, are diverted to MACC at early climacteric. At the climacteric peak there are substantial gradients in ethylene production between different portions of the tissue, the inner cortical tissues producing up to twice as much as the external tissues. This increased production is associated with, and apparently due to, increased content of ACC synthase. Less than 1% of the synthesized ACC is diverted to MACC in the flesh of climacteric apples. In contrast, the skin contains high activity of malonyl transferase, and correspondingly high levels [1000 nmol (g dry weight)⁻¹] of MACC.     

The Effect of the Date of Rindite Treatment on the Concentration of Potato Virus Y in Tubers

The main purpose of this work was to study the host-parasite relationship between attacking fungi and the infected climacteric fruit. The study dealt mainly with physiological changes such as respiration and ethylene evolution pattern occurring in fruit inoculated with different fungi after picking. These physiological changes in infected fruit were found to be basically similar to those of uninfected fruit, but they were substantially accelerated. A direct correlation was obtained between the rate of fungal development in fruit (incubation period and rot development) and the extent of acceleration of respiration and ethylene evolution; in general, a faster development of the fungus was related to greater acceleration of ethylene and respiration rate of the infected fruit. Fungal infection was found to have an effect on fruits similar to that of exogenic ethylene treatment on sound fruit.     

Reversible inhibition of ethylene action and interruption of petal senescence in carnation flowers by norbornadiene

The inhibitory effects of the cyclic olefin 2,5-norbornadiene (NBD) on ethylene action were tested in carnation (Dianthus caryophyllus L. cv White Sim) flowers. Treatment of flowers at anthesis with ethylene in the presence of 500 microliters per liter NBD increased the concentration of ethylene required to elicit a response (petal senescence), indicating that NBD behaves as a competitive inhibitor of ethylene action. Transfer of flowers producing autocatalytic ethylene and exhibiting evidence of senescence (petal in-rolling) to an atmosphere of NBD resulted in a rapid reduction in ethylene production, petal 1-aminocyclopropane-1-carboxylic acid synthase activity, 1-aminocyclopropane-1-carboxylic acid content, and ethylene forming enzyme activity. Removal of NBD resulted in recovery of ethylene biosynthesis. These results support the autocatalytic regulation of ethylene production during the climacteric stage of petal senescence and suggest that continued perception of ethylene is required for maintenance of ethylene biosynthesis. The inhibition of ethylene action by NBD after the flowers had reached the climacteric peak was associated with interruption of petal senescence as evidenced by reversal of senescence symptoms. This result is in contrast to the widely held belief that the rate of petal senescence is fixed and irreversible once petals enter into the ethylene climacteric.     

Interrelationship of Gene Expression, Polysome Prevalence, and Respiration during Ripening of Ethylene and/or Cyanide-Treated Avocado Fruit

Upon initiation of ripening in avocado fruit (Persea americana Mill. cv Hass) with 10 microliters/liter ethylene, polysome prevalence and associated poly(A)⁺ mRNA increase approximately 3-fold early in the respiratory climacteric and drop off to preclimacteric levels at the peak of the respiratory climacteric. The increase in poly(A)⁺ mRNA on polysomes early in the respiratory climacteric constitutes a generic increase in constitutive mRNAs. New gene expression associated with ripening is minimal but evident after 10 hours of ethylene treatment and continues to increase relative to constitutive gene expression throughout the climacteric. The respiratory climacteric can be temporally separated into two phases. The first phase is associated with a general increase in protein synthesis, whereas the second phase reflects new gene expression and accumulation of corresponding proteins which may be responsible for softening and other ripening characteristics. A major new message on polysomes that arises concomitantly with the respiratory climacteric codes for an in vitro translation product of 53 kilodaltons which is immunoprecipitated by antiserum against avocado fruit cellulase.

Cyanide at 500 microliters/liter fails to affect the change in polysome prevalance or new gene expression associated with the ethylene-evoked climacteric in avocado fruit. Treatment of fruit with 500 microliters/liter cyanide alone initiates a respiratory increase within 4 hours, ethylene biosynthesis within 18 hours, and new gene expression akin to that educed by ethylene within 20 hours of exposure to cyanide.

     

Effects of ethylene on potato tuber respiration

Treatment of potato tubers (Solanum tuberosum L.) with ethylene gas causes a rapid rise in their respiration rate, reaching 5 to 10 times the rate of untreated tubers over 30 hours of treatment and then falling slowly. The response shows a lag of 8 hours, and more than 24 hours of exposure is required for maximum effect; the temperature optimum is near 25 C. In sensitivity to low concentrations and dependence on temperature, the phenomenon is similar to the effect of ethylene on the respiration of climacteric and nonclimacteric fruits. Treated potato tubers returned to air recover their sensitivity to ethylene more slowly than do nonclimacteric fruits (e.g., mature green oranges). It is proposed that the respiratory rise characteristic of ripening in climacteric fruits and of the wound response in plant tissues is induced by a rise in endogenous tissue ethylene.     

香蕉果实成熟软化时果皮和果肉中α-L-阿拉伯呋喃糖苷酶活性的变化

阿拉伯糖是果实软化过程中变化最明显的细胞壁糖残基之一,α-L-阿拉伯呋喃糖苷酶是导致细胞壁多糖中阿拉伯糖残基降解的主要糖苷酶。为阐明该酶在香蕉果实成熟软化中的作用,实验对香蕉贮藏过程中果皮和果肉中该酶活性以及果实硬度、呼吸强度和乙烯释放量的变化进行了研究。结果表明:α-L-阿拉伯呋喃糖苷酶在果实初期的变化很小,到果实硬度开始急剧下降时达到最大,增加量达10倍以上,且果肉中的酶活性大于果皮中;乙烯吸收剂处理延缓了香蕉果实呼吸和乙烯高峰的出现时间,降低了果实硬度、果皮和果肉中α-L-阿拉伯呋喃糖苷酶活性变化的速度和幅度。以上结果表明α-L-阿拉伯呋喃糖苷酶起诱导香蕉果实成熟的作用,在果实的软化中起着十分重要的作用,且其活性受乙烯的调节。