The Production of Marker-Free Genetically Engineered Broccoli with Sense and Antisense ACC synthase 1 and ACC oxidases 1 and 2 to Extend Shelf-Life

The production of transgenic broccoli (Brassica oleracea) with increased shelf-life using an Agrobacterium rhizogenes-mediated co-transformation protocol is reported. An Agrobacterium rhizogenes Ri vector, pRi1855:GFP was constructed to allow expression of the green fluorescent protein to identify insertion of Ri T_L-DNA into plant cells. The Brassica oleracea ACC synthase 1 and ACC oxidase 1 and 2 cDNAs in sense and antisense orientations were co-transformed into GDDH33, a doubled haploid calabrese-broccoli cultivar. Transformation efficiency was 3.26%, producing 150 transgenic root lines, of which 18 were regenerated into mature plants. The floral buds from T₀ broccoli heads were assayed for post-harvest production of ethylene and chlorophyll levels. Buds from T₀ lines transformed with ACC oxidase 1 and 2 constructs produced significantly less post-harvest ethylene at 20 °C than the untransformed plants and chlorophyll loss was significantly reduced over a 96 h post-harvest period. The T₀ plants transformed with sense and antisense ACC synthase 1 had a significantly reduced 24 h post-harvest ethylene peak and delayed chlorophyll loss. A positive correlation between post-harvest bud ethylene production and chlorophyll loss was described by a regression. This demonstrates that the shelf-life of a very perishable vegetable may be increased up to 2 days at 20 °C by reducing post-harvest ethylene production.     

Post-harvest alterations in polyamines and ethylene in two diverse rose species

Changes in the concentrations of endogenous polyamines and ethylene were determined in two diverse species of rose viz. Rosa damascena and Rosa bourboniana during post-harvest periods. At full bloom, the concentrations of free putrescine was significantly higher than rest of the polyamines, i.e. spermine and spermidine in both the species. The concentrations of all the polyamines decreased during subsequent periods upto 48 h after full bloom. Similar situation was also observed in conjugated fraction but in bound fraction, during full bloom, the concentration of spermine was higher than rest of the polyamines. In both the species, ethylene showed higher levels during full bloom with maximum in R. damascena, which increased, during post-harvest periods. The possible significance of polyamines, ethylene and their interactions is discussed during post-harvest periods in flowers.     

Fruit abscission and ethylene production of four blackberry cultivars (Rubus spp.)

In all cultivars the force required to harvest fruit declined during ripening. The fruit retention strength (FRS) of ripe fruit varied between cultivars with ‘Ashton Cross’ and ‘Chehalem’ being easier to remove than ‘Bedford Giant’ or ‘Oregon Thornless’. The cultivars ‘Ashton Cross’ and ‘Oregon Thornless’ showed no increase in ethylene production during fruit ripening whereas ‘Bedford Giant’ and ‘Chehalem’ had increased rates of ethylene production (EPR) in the ripe fruit. Exogenous ethylene accelerated abscission, ethylene production and pigment changes in ‘Ashton Cross’ fruit at all stages of development. 1-amino-cyclopropane-1-carboxylic acid (ACC) supplied to fruit at all stages of development was converted to ethylene at levels in excess of those found naturally. The differences between cultivars are discussed with reference to the role of ethylene in both machine harvesting and post-harvest storage of blackberry fruit.     

成串采收对番茄果实采后乙烯合成及贮藏品质的影响

为研究成串采收对番茄果实采后乙烯合成及贮藏品质的影响,对广西田阳县两个栽培区两种嫁接砧木的串收番茄的采后生理指标进行测定,探讨了该采收方式对番茄果实采后保鲜的作用机制。结果表明:整个贮藏期,不同栽培区不同嫁接砧木的番茄成串采收的果实,乙烯生成量明显低于对照的常规单果采收。其中,砧木1号Ⅰ区的串收番茄的乙烯生成量,采后5 d即下降至最低点(0.35 nL·g~(-1)·h~(-1)),显著低于其对照(1.36 nL·g~(-1)·h~(-1)),其他栽培区和砧木组合的串收番茄,在采后15 d乙烯生成量达到最低值。串收番茄的类胡萝卜素、番茄红素和抗坏血酸等果实内天然抗氧化物质的含量,在贮藏前期快速升高,且峰值显著高于对照。此外,成串采收处理还一定程度抑制了果实后熟阶段可溶性糖的积累和可滴定酸的分解。因此,番茄成串采收处理,可能通过抑制果实采后乙烯的合成,同时提高类胡萝卜素、番茄红素和抗坏血酸的水平,并且推迟糖和酸等营养物质的后熟变化进程,实现其延长果实货架期,提高商品品质的作用。     

Role of Ethylene in Avocado Fruit Development and Ripening: II. ETHYLENE PRODUCTION AND RESPIRATION BY HARVESTED FRUITS

Avocado (Persea americana Mill.) fruits were harvested at successivedevelopment stages during a period of 10 months. Ethylene productionand respiration were determined during the post-harvest period. Detached immature fruits were found to have a preclimactericincrease in ethylene production and respiration without anysigns of ripening. In fruits larger than 20 g a second phaseof climacteric ethylene production and respiration, associatedwith ripening, ensued. The preclimacteric ethylene was produced mainly by the seedcoat. It is suggested that the high ethylene production potentialof the seed coat may serve as a means for inducing abscissionin young fruits.     

Non-invasive photoacoustic spectroscopic determination of relative endogenous nitric oxide and ethylene content stoichiometry during the ripening of strawberries Fragaria anannasa (Duch.) and avocados Persea americana (Mill.)

Employing non-invasive photoacoustic spectrometry, emissions of nitric oxide (NO) and ethylene in post-harvest strawberries and avocados were monitored. A clear-cut stoichiometric relationship was found between the two gases: unripe fruit manifesting high NO and low ethylene levels-the converse in ripe fruit. Findings are discussed in the light of putative control of ethylene-promoted fruit senescence by endogenous NO.     

Glucose Inhibits ACC Oxidase Activity and Ethylene Biosynthesis in Ripening Tomato Fruit

Experiments were carried out to evaluate the effect of glucose on ripening and ethylene biosynthesis in tomato fruit (Lycopersicon esculentum Mill.). Fruit at the light-red stage were vacuum infiltrated with glucose solutions post-harvest and changes in 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, ACC, ACC oxidase, and ethylene production monitored over time. ACC oxidase activity was also measured in pericarp discs from the same fruits that were treated either with glucose, fructose, mannose, or galactose. While control fruit displayed a typical peak of ethylene production, fruit treated with glucose did not. Glucose appeared to exert its effect on ethylene biosynthesis by suppressing ACC oxidase activity. Fructose, mannose, and galactose did not inhibit ACC oxidase activity in tomato pericarp discs. Glucose treatment inhibited ripening-associated colour development in whole fruit. The extent of inhibition of colour development was dependent upon the concentration of glucose. These results indicate that glucose may play an important role in ethylene-associated regulation of fruit ripening.     

Ethylene-Induced Increase in Glutamine Synthetase Activity and mRNA Levels in Hevea brasiliensis Latex Cells

Ethylene, used as a stimulant of latex production in Hevea brasiliensis, significantly activates the regenerating metabolism within the laticiferous cells. In this context, attention was focused on glutamine synthetase (GS; EC 6.3.1.2), a key enzyme in nitrogen metabolism. A specific and significant activation of the cytosolic glutamine synthetase (GScyt) in the laticiferous cells after ethylene treatment parallels the increase of latex yield. A marked accumulation of the corresponding mRNA was found, but in contrast, a slight and variable increase of the polypeptide level is at the limit of detection by western blotting. The GS response to ethylene might be mediated by ammonia that increases in latex cytosol following ethylene treatment. The physiological significance for such a regulation by ethylene of the GScyt is discussed in terms of the nitrogen requirement for protein synthesis associated with latex regeneration.     

Ethylene Production by Attached Leaves or Intact Shoots of Tobacco Cultivars Differing in Their Speed of Yellowing during Curing

Using an open air flow system, differences in the yellowing rate of leaves during curing were assessed in relation to ethylene production by shoots of intact seedlings or attached mature leaves of 60 day old tobacco (Nicotiana tabacum L.) plants. The rate of ethylene evolution from the leaves of the fast yellowing cultivars was significantly higher than in the slow yellowing ones. The same differences were obtained with shoots of intact seedlings. The findings suggest that it is possible to use ethylene production by seedlings as a selection criterion in screening for genotypic differences in the rate of yellowing. The ability of carbon dioxide (1%) to enhance ethylene production by attached leaves was significant in a slow, but not in a fast yellowing cultivar. However, similar amounts of ethylene were produced on administration of 1-aminocyclopropane-1-carboxylic acid to a slow and a fast yellowing cultivar. Exposure of attached leaves to exogenous ethylene (0.1 microliter per liter) accelerated the loss of chlorophyll and protein. This treatment was effective only for slightly yellow leaves and not for fully expanded green ones. The significance and possible use of ethylene in the flue-curing process are discussed.     

Unusual ethylene-related behavior in senescing flowers of the carnation Sandrosa

The time course of ethylene production by senescing carnation ( Dianthus caryophyllus L. cv. Sandrosa) flowers was studied. These flowers are unusual in that they do not exhibit an autocatalytic increase in ethylene production nor do they develop petal in-rolling. Exposure of the flowers to exogenous ethylene resulted in a rise in their ethylene-forming enzyme (EFE) activity and ethylene production, and at the same time a marked decline in their fresh weight. Natural senescence was also accompanied by a rise in EFE activity, but with no concomitant rise in 1-amino cyclopropane carboxylic acid synthase activity nor in ethylene production. A shift in responsiveness to ethylene was observed, with young flowers more responsive to exogenous ethylene than older flowers. The results are discussed in terms of a proposed mechanism allowing for the decline in competence of this cultivar to respond to ethylene during senescence.     

Properties of the proteinaceous inhibitor of ethylene synthesis: Action on ethylene production and indoleacetylaspartate formation

Several properties of the proteinaceous inhibitor of ethylenebiosynthesis are described. The inhibitor reversibly inhibitedethylene production by auxin-treated hypocotyl segments of etiolatedmungbean seedlings. It also inhibited endogenous ethylene formation.Fluorescence microscopy of tissue treated with the inhibitorlabeled with a fluorescent dye, FITC, revealed that the actionof the inhibitor only on epidermal cells was enough to achieveinhibition. The site of auxin-induced ethylene production wasassumed to be the epidermis. The inhibitor inhibits neitherrespiration of hypocotyl segments nor auxin-induced elongationgrowth of wheat coleoptiles. Relative levels of free IAA intissue were not altered by the inhibitor, but IA-Asp formationwas reversively suppressed. A possible interrelation between ethylene production and IA-Aspformation is discussed (Received March 30, 1973; )     

Commercial techniques for preserving date palm (Phoenix dactylifera) fruit quality and safety: A review

The popularity of date palm (Phoenix dactylifera) fruit is increasing, therefore the demand for high-quality date palm fruit with less or no chemical treatment is the topic of interest for date producers and consumers. The quality of date palm fruit is much dependent on its postharvest handling and processing. For preventing the degradation and maintenance of the high quality of dates during the storage an appropriate harvest and post-harvest processes are required. The process should control the biotic and abiotic factors like insects, fungus, temperature, as well as handling and processing of dates. Therefore, in this work, we reviewed the literature related to the protection of date fruits during their post-harvest life. The commercially viable advance and updated techniques that can be used to avoid storage losses and problems while keeping fruit quality (nutritional, color, flavor, and texture) and microbial safety under optimal conditions are discussed.     

Endogenous and exogenous ethylene induces needle abscission and cellulase activity in post-harvest balsam fir (Abies balsamea L.)

Post-harvest needle loss is a major problem for balsam fir and other Christmas tree species. Recent evidence has implicated ethylene as a signal responsible for post-harvest needle abscission, but enzymological changes remain unknown. The objective of this study was to identify and quantify cellulase activity associated with endogenous and exogenous ethylene-induced abscission. An experiment was designed with three treatments (control, endogenous ethylene, or exogenous ethylene) with five replicates. Key response variables include needle retention duration, xylem pressure potential, ethylene evolution rate, and cellulase activity. Two complimentary methods were used to assess cellulase activity: a cellulose plate digestion and zymography. The results confirm ethylene as a signal for post-harvest abscission and identify ethylene-induced cellulase. Ethylene evolution was typically between 15 and 16 μL g⁻¹ h⁻¹, but there was no difference among the three treatments. However, exogenous ethylene significantly decreased needle retention by 60% and resulted in a sixfold decrease in xylem pressure potential. In addition, cellulase activity increased by 8- and 12-fold in endogenous and exogenous ethylene-induced abscission, respectively, compared to the control. Identification of ethylene-induced cellulase activity has increased our understanding of the post-harvest needle abscission process and confirms ethylene’s role as a signal molecule.     

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.     

Perception of the plant hormone ethylene: known-knowns and known-unknowns

The gaseous phytohormone ethylene is implicated in virtually all phases of plant growth and development and thus has a major impact on crop production. This agronomic impact makes understanding ethylene signaling the Philosopher’s Stone of the plant biotechnology world in applications including post-harvest transport of foodstuffs, consistency of foodstuff maturity pre-harvest, decorative flower freshness and longevity, and biomass production for biofuel applications. Ethylene is biosynthesized by plants in response to environmental factors and plant life-cycle events, and triggers a signaling cascade that modulates over 1000 genes. The key components in the perception of ethylene are a family of copper dependent receptors, the bioinorganic chemistry of which has been largely ignored by the chemical community. Since identification of these receptors two decades ago, there has been tremendous growth in knowledge in the biological community on the signal transduction pathways and mechanisms of ethylene signaling. In this review, we highlight these advances and key chemical voids in knowledge that are overdue for exploration, and which are required to ultimately regulate and control ethylene signaling.     

Senescence in cut carnation flowers: Temporal and physiological relationships among water status, ethylene, abscisic acid and membrane permeability

Changes in water status, membrane permeability, ethylene production and levels of abscisic acid (ABA) were measured during senescence of cut carnation flowers ( Dianthus caryophyllus L. cv. White Sim) in order to clarify the temporal sequence of physiological events during this post-harvest period. Ethylene production and ABA content of the petal tissue rose essentially in parallel during natural senescence and after treatment of young flowers with exogenous ethylene, indicating that their syntheses are not widely separated in time. However, solute leakage, reflecting membrane deterioration, was apparent well before the natural rise in ethylene and ABA had begun. In addition, there were marked changes in water status of the tissue, including losses in water potential (ψ_w), and turgor (ψ_p), that preceded the rise in ABA and ethylene. As senescence progressed, ψ_w continued to decline, but ψ_p returned to normal levels. These temporal relationships were less well resolved when senescence of young flowers was induced by treatment with ethylene, presumably because the time-scale had been shortened. Thus changes in membrane permeability and an associated water stress in petal tissue appear to be earlier symptoms of flower senescence than the rises in ABA or ethylene. These observations support the contention that the climacteric-like rise in ethylene production is not the initial or primary event of senescence and that the rise in ABA titre may simply be a response to changes in water status.     

Submergence-Induced Ethylene Synthesis,Entrapment, and Growth in Two Plant Species with Contrasting Flooding Resistances

Submergence-induced ethylene synthesis and entrapment were studied in two contrasting Rumex species, one flood-resistant (Rumex palustris) and the other flood-sensitive (Rumex acetosa). The application of a photoacoustic method to determine internal ethylene concentrations in submerged plants is discussed. A comparison with an older technique (vacuum extraction) is described. For the first time ethylene production before, during, and after submergence and the endogenous concentration during submergence were continuously measured on a single intact plant without physical perturbation. Both Rumex species were characterized by enhanced ethylene concentrations in the shoot after 24 h of submergence. This was not related to enhanced synthesis but to continued production and physical entrapment. In R. palustris, high endogenous ethylene levels correlated with enhanced petiole and lamina elongation. No dramatic change in leaf growth rate was observed in submerged R. acetosa shoots. After desubmergence both species showed an increase in ethylene production, the response being more pronounced in R. palustris. This increase was linked to the enhanced postsubmergence growth rate of leaves of R. palustris. Due to the very rapid escape of ethylene out of desubmerged plants to the atmosphere (90% disappeared within 1 min), substantial underestimation of internal ethylene concentrations can be expected using more conventional vacuum extraction techniques.     

Ethylene Synthesis and Floral Senescence following Compatible and Incompatible Pollinations in Petunia inflata

Ethylene production and floral senescence following compatible and incompatible pollinations were studied in a self-incompatible species, Petunia inflata. Both compatible and incompatible pollinations resulted in a burst of ethylene synthesis that peaked 3 hours after pollination. P. inflata pollen was found to carry large amounts of the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC). The amount of pollen-held ACC varied in different genetic backgrounds, and the magnitude of the peak correlated with the amount of ACC borne by the pollen. Aminooxyacetic acid (AOA), an inhibitor of ACC synthesis, had no inhibitory effect on this ethylene response, indicating that pollen-borne ACC was largely responsible for the early synthesis of ethylene. After compatible pollination, a second increase in ethylene synthesis began at 18 hours, and the first sign of senescence appeared at 36 hours. Upon treatment with AOA, the second phase of ethylene production was reduced by 95%, indicating that endogenous ACC synthesis was required for this phase of ethylene synthesis. AOA treatment also delayed senescence to 6 days after anthesis. After incompatible pollination, a second increase in ethylene production did not occur until 3 days, and the first sign of senescence occurred 12 hours later. Unpollinated flowers showed an increase in ethylene production 3 to 4 days after anthesis and displayed signs of senescence 1 day later. The significance of the early and late phases of pollination-induced ethylene synthesis is discussed.