Ozone-induced inhibition of kiwifruit ripening is amplified by 1-methylcyclopropene and reversed by exogenous ethylene

Background

Understanding the mechanisms involved in climacteric fruit ripening is key to improve fruit harvest quality and postharvest performance. Kiwifruit (Actinidia deliciosa cv. ‘Hayward’) ripening involves a series of metabolic changes regulated by ethylene. Although 1-methylcyclopropene (1-MCP, inhibitor of ethylene action) or ozone (O₃) exposure suppresses ethylene-related kiwifruit ripening, how these molecules interact during ripening is unknown.

Results

Harvested ‘Hayward’ kiwifruits were treated with 1-MCP and exposed to ethylene-free cold storage (0?°C, RH 95%) with ambient atmosphere (control) or atmosphere enriched with O₃ (0.3?μL?L^??1) for up to 6?months. Their subsequent ripening performance at 20?°C (90% RH) was characterized. Treatment with either 1-MCP or O₃ inhibited endogenous ethylene biosynthesis and delayed fruit ripening at 20?°C. 1-MCP and O₃ in combination severely inhibited kiwifruit ripening, significantly extending fruit storage potential. To characterize ethylene sensitivity of kiwifruit following 1-MCP and O₃ treatments, fruit were exposed to exogenous ethylene (100?μL?L^??1, 24?h) upon transfer to 20?°C following 4 and 6?months of cold storage. Exogenous ethylene treatment restored ethylene biosynthesis in fruit previously exposed in an O₃-enriched atmosphere. Comparative proteomics analysis showed separate kiwifruit ripening responses, unraveled common 1-MCP- and O₃-dependent metabolic pathways and identified specific proteins associated with these different ripening behaviors. Protein components that were differentially expressed following exogenous ethylene exposure after 1-MCP or O₃ treatment were identified and their protein-protein interaction networks were determined. The expression of several kiwifruit ripening related genes, such as 1-aminocyclopropane-1-carboxylic acid oxidase (ACO1), ethylene receptor (ETR1), lipoxygenase (LOX1), geranylgeranyl diphosphate synthase (GGP1), and expansin (EXP2), was strongly affected by O₃, 1-MCP, their combination, and exogenously applied ethylene.

Conclusions

Our findings suggest that the combination of 1-MCP and O₃ functions as a robust repressive modulator of kiwifruit ripening and provide new insight into the metabolic events underlying ethylene-induced and ethylene-independent ripening outcomes.

     

Effect of 1-MCP and low-temperature storage on postharvest conservation of camu–camu

Camu–camu, a native fruit from the Amazon region, is a rich source of bioactive compounds. However, its intense metabolic activity and high-water content limit the fruit’s postharvest storage and marketing. The aim of this study, conducted in two parts, was to evaluate the effects of 1-MCP and storage temperature on the physiology and postharvest preservation of camu–camu fruit. In part 1 of the study, fruit harvested at maturity stage 3 were divided into groups: control, 1-methylcyclopropene (1-MCP; 900 nL L^?1; 12 h) and ethylene (1000 µL L^?1; 24 h) and were stored at 22?±?1 °C and 85?±?5% RH for 9 days. In part 2, fruit harvested at maturity stage 3 were stored at 5, 10, 15, 20, or 25?±?1 °C and 85?±?5% RH for 9 days. During storage, fruit were evaluated daily for decay, mass loss, respiratory activity, and ethylene production, and every 3 days they were evaluated for peel color, pulp firmness, soluble solids content, total titratable acidity, ascorbic acid, total chlorophyll, and total anthocyanins. Fruit treated with 1-MCP exhibited delayed ripening due to lower metabolic activity, as evidenced by delay to softening, reduced mass loss and no decay. Storage at 5 °C prevented ethylene production, mass loss, color changes, and maintained pulp firmness, while did not affect soluble solids content. The results indicated that storage of camu–camu fruit at 5 °C or at 25 °C following application of 900 nL L^?1 1-MCP were effective strategies to delay ripening and maintain fruit quality up to 9 days.     

Effects of 1-MCP on the vase life and ethylene response of cut flowers

Pretreatment for 6 h with low concentrations of 1-MCP (1-Methylcyclopropene, formerly designated as SIS-X), a cyclic ethylene analog, inhibits the normal wilting response of cut carnations exposed continuously to 0.4 μl·l^?1 ethylene. The response to 1-MCP was a function of treatment concentration and time. Treatment with 1-MCP was as effective in inhibiting ethylene effects as treatment with the anionic silver thiosulfate complex (STS), the standard commercial treatment. Other ethylene-sensitive cut flowers responded similarly to carnations. In the presence of 1 μl·l^?1 ethylene, the vase life of 1-MCP-treated flowers was up to 4 times that of the controls.     

Comparison of cyclopropene, 1-methylcyclopropene, and 3,3-dimethylcyclopropene as ethylene antagonists in plants

A comparison has been made of cyclopropene (CP), 1-methylcyclopropene (1-MCP), and 3,3-dimethyl-cyclopropene (3,3-DMCP) in their ability to protect plants against ethylene. In bananas, both CP and 1-MCP are effective around 0.5 nL L^–1, and 3,3-DMCP was effective at 1 L L^–1. Bananas treated with CP and 1-MCP again become sensitive to ethylene at 12 days and those treated with 3,3-DMCP at 7 days. Mature green tomatoes are protected by 5–7 nL L^–1 of 1-MPC for 8 days at 25°C and tomatoes treated with 3,3-DMCP at 5–10 L L^–1 are protected for 5 days. Carnation flowers are protected with CP or 1-MCP after exposure to 0.5 nL L^–1 for 24 hours and by 1 L L^–1 of 3,3-DMCP. The display life of Campanula flowers is increased from 3.3 to 5.4 days by 10 L L^–1 of 3,3-DMCP and to 9 days by 20 nL L^–1 of 1-MCP. Ethylene inhibition of pea seedlings is reduced by treatment with 1-MCP at 10 L L^–1 of ethylene but as ethylene is increased to 3000 L L^–1 growth inhibition increases. 3,3-DMCP treatment causes very little reduction of the ethylene effect even at very low concentrations.     

Effect of cyanobacterial extracellular products on high-frequency in vitro induction and elongation of Gossypium hirsutum L organs through shoot apex explants

The challenging task of bringing high efficiency transformed plants attracts lot of attention in recent times. In search for this, there have been many attempts made using, different techniques like tissue culture and plant breeding methods. Here we report a suitable alternative facile route, where cyanobacterial extracellular products are utilized as growth regulators and its performance validated on Gossypium hirsutum L. MS medium is tested with cyanobacterial extracellular products of Nostoc ellipsosporum, Dolichospermum flos-aquae and Oscillatoria acuminata .Our best results show that the addition of O. acuminata extracellular product with plant growth hormones gives the excellent induction and elongation in cotton. In addition to this, the multiple shoot was obtained on MS medium fortified with 1.0 mg L^?1 BA with 8% O. acuminata and 1.5 mg L^?1 TDZ with 12% O. acuminata. High frequency of shoot elongation supplemented with MS medium, iP 2.5 mg L^?1 and 16% O. acuminata and root production MS medium fortified with 12% O. acuminata best responsible for regeneration in cotton plants. The rooted plants were hardened and transferred to soil with 90% survival rate.     

Physiological responses,tolerance efficiency,and phytoextraction potential of Hylotelephium spectabile (Boreau) H. Ohba under Cd stress in hydroponic condition

Abstract

A sand hydroponic experiment with different concentrations of 0, 5, 10, 20, 40?mg L^?1 Cd was used to study the growth and physiological response of Hylotelephium spectabile (Boreau) H. Ohba. and its phytoextraction potential for Cd. The results showed that total plant biomass under 5?mg L^?1 Cd treatment was slightly affected. The content of malondialdehyde (MDA) in leaf exposed to Cd was higher, and the POD and CAT activity exhibited a positive response to the low level of Cd addition (5?mg·L^?1). The photosynthesis pigments were slightly inhibited, and the ultrastructure of chloroplast remained intact after treatment with 10?mg L^?1 Cd. The maximum leaf Cd content (603?mg·kg^?1) was found in 5?mg L^?1 Cd treatment, then decreased with the Cd level increased. The maximum Cd content in the shoots far exceeds the threshold level (100?mg kg^?1) for a Cd-hyperaccumulator plant with the value of translocation factor (TF_shoot/root) for Cd reaching up to 5.62. In conclusion, H. spectabile showed normal growth and physiological response and high shoot Cd accumulation under 5?mg L^?1 Cd stress, which made it to be a good candidate for phytoextraction of low-level Cd polluted environment.     

Effect of 1-methylcyclopropene on postharvest physiological changes of ‘Zaohong’ plum

Plum is a highly perishable fruit and postharvest fruit softening limits its shelf life. The aim of this work was to study the specific effects of 1-methylcyclopropene (1-MCP) treatment on physiological changes in ‘Zaohong’ plums. Plums were treated with 500 nL L⁻¹ 1-MCP at 20°C for 18 h followed by 20°C storage. The results showed that 1-MCP treatment significantly reduced endogenous ethylene production and the activities of ethylene biosynthetic enzymes’ (1-aminocyclopropane-1-carboxylic acid synthase, ACS and 1-aminocyclopropane-1-carboxylic acid oxidase, ACO) in plum fruit during storage when compared with untreated fruit. Although 1-MCP treatment inhibited ethylene production and 1-aminocyclopropane-1-carboxylic acid (ACC) accumulation, it did not inhibit the accumulation of N-malonyl-ACC (MACC). Higher firmness was also found in 1-MCP-treated plums than in controls. During storage, superoxide anion (O₂^−·) and hydrogen peroxide (H₂O₂) levels decreased in 1-MCP-treated fruit. 1-MCP treatment also regulated superoxide dismutase (SOD) and catalase (CAT) activities during storage. Xylanase activity was upregulated while activities of polygalacturonase (PG), pectin methyl esterase (PME) and cellulase enzymes in the fruit were downregulated by 1-MCP treatment. In conclusion, 1-MCP might be a potent compound for extending both storage period and shelf life of ‘Zaohong’ plums by suppressing ethylene biosynthesis, regulating cell wall degradation enzymes and reducing fruit softening.     

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.     

Effect of orchard and postharvest application of daminozide on ethylene synthesis by apple fruit

The effects of daminozide (butanedioic acid-2,2-dimethylhydrazide) on ethylene synthesis by apple fruits were investigated. The objective was to determine the effects of postharvest applications as compared to the standard application of diaminozide in the orchard. Immersion in a solution containing 4.25 g L^?1 active ingredient for 5 min delayed the rise in ethylene production in individual “Cox” apples at 15°C by about 2 days, whereas orchard application of 0.85 g L^?1 caused delays of about 3 days. Both modes of application depressed the maximal rate of ethylene production attained by ripe apples by about 30%. Daminozide did not affect the stimulation of respiration by ethylene treatment of “Gloster” apples, but it delayed the increase in ethylene synthesis. Daminozide applied immediately after harvest delayed the rise in ethylene synthesis in “Golden Delicious” held at 15°C, but it was less effective when applied 48 h after harvest or when apples were held at 5°C. Exposure to 1–2 μl L^?1 ethylene for 48 h was less effective in promoting the rise in ethylene in daminozidetreated “Cox” and “Gloster” apples than in untreated fruit. High (100–1000 μl L^?1) concentrations of ethylene more or less overcame the daminozide effect. Apples absorbed about 40% of surface-applied [¹⁴C]daminozide in 48 h, but more than 90% of the radioactivity in the fruit was recovered from the peel and outer 1 cm of the cortex. Daminozide was partly converted to carbon dioxide and other metabolites.     

GhXB38D represses cotton fibre elongation through ubiquitination of ethylene biosynthesis enzymes GhACS4 and GhACO1

Ethylene plays an essential role in the development of cotton fibres. Ethylene biosynthesis in plants is elaborately regulated by the activities of key enzymes, 1-aminocyclopropane-1-carboxylate oxidase (ACO) and 1-aminocyclopropane-1-carboxylate synthase (ACS); however, the potential mechanism of post-translational modification of ACO and ACS to control ethylene synthesis in cotton fibres remains unclear. Here, we identify an E3 ubiquitin ligase, GhXB38D, that regulates ethylene biosynthesis during fibre elongation in cotton. GhXB38D gene is highly expressed in cotton fibres during the rapid elongation stage. Suppressing GhXB38D expression in cotton significantly enhanced fibre elongation and length, accompanied by the up-regulation of genes associated with ethylene signalling and fibre elongation. We demonstrated that GhXB38D interacts with the ethylene biosynthesis enzymes GhACS4 and GhACO1 in elongating fibres and specifically mediates their ubiquitination and degradation. The inhibition of GhXB38D gene expression increased the stability of GhACS4 and GhACO1 proteins in cotton fibres and ovules, resulting in an elevated concentration of ethylene. Our findings highlight the role of GhXB38D as a regulator of ethylene synthesis by ubiquitinating ACS4 and ACO1 proteins and modulating their stability. GhXB38D acts as a negative regulator of fibre elongation and serves as a potential target for enhancing cotton fibre yield and quality through gene editing strategy.     

Quercetin and silver nitrate modulate organogenesis in <Emphasis Type="Italic">Carissa carandas</Emphasis> (L.)

An in vitro organogenesis protocol for Carissa carandas L. was developed using an auxin transport inhibitor (quercetin) and silver nitrate (AgNO₃), an inhibitor of ethylene action, in association with cytokinins in the culture medium. This protocol produced the maximum number of shoots from aseptic seedling-derived shoot apex explants of C. carandas. The highest rate of shoot multiplication was recorded on MS medium containing 2.0 mg L^?1 6-benzylaminopurine; 0.5 mg L^?1 kinetin, and 0.75 mg L^?1 quercetin at after 4 wk of culture. Similar results were obtained when MS medium fortified with 2.0 mg L^?1 BAP, 0.5 mg L^?1 kinetin, and 1.5 mg L^?1 AgNO₃ was used. However, successful rooting was achieved on quarter strength MS medium with 0.5 mg L^?1 indole-3-acetic acid. In this study, an inhibitor of auxin transport and ethylene action maximized shoot multiplication in medium fortified with cytokinins. The established rapid micropropagation method could be used to conserve elite genotypes of C. carandas.     

Effect of 1-methylcyclopropene and methylenecyclopropane on ethylene binding and ethylene action on cut carnations

1-Methylcyclopropene (1-MCP), formerly designated as Sis-X, has been shown to be an effective inhibitor of ethylene responses in carnation flowers in either the light or the dark. The binding appears to be to the receptor and to be permanent. A 6 h treatment at 2.5 nl l^–1 is sufficient to protect against ethylene, and 0.5 nl l^–1 is sufficient if exposure is for 24 h. As carnation flowers age, a little higher concentration appears to be needed. Most of the natural increase in ethylene production during senescence is prevented by treatment with 1-MCP. A closely related compound, methylenecyclopropane shows ethylene activity. A tritium labelled 1-MCP (60 mCi mmol^–1) has been prepared. A higher specific activity is needed for more critical studies.     

Identification of somaclonal variants in proliferating shoot cultures of Senecio cruentus cv. Tokyo Daruma

A protocol for in vitro propagation of cineraria (Senecio cruentus) was developed. The highest frequency of shoot proliferation was obtained from nodal explants cultured on Murashige and Skoog (MS) medium supplemented with 2.0?mg L^?1 6-benzyladenine (BA) and 0.5?mg L^?1 ??-naphthalene acetic acid (NAA), with a mean number of 14 shoots per explant. A high concentration of BA (4.0?mg L^?1) and repeated subcultures resulted in hyperhydric shoots. Decreasing the BA concentration to 1.0?mg L^?1 in the culture medium eliminated hyperhydricity. The concentration of ammonium nitrate (NH₄NO₃) and temperature had marked effects on somaclonal variation. Variation was observed when the cultures were maintained at 15?°C but not at 25?°C. Variants with blue-colored leaves and stems were identified; whereas, normal plants maintained their green-colored leaves and stems. The highest frequency of variation (67.5?%), with a mean number of 3.0 variant shoots per explants, was obtained on shoot proliferation medium (MS?+?2.0?mg L^?1 BA and 0.5?mg L^?1 NAA) devoid of NH₄NO₃. The best rooting (100?%), with the highest number of roots per shoot (10.8) and the greatest root length (6.8?cm) was obtained on medium supplemented with 0.1?mg L^?1 NAA. In vitro-grown plantlets were successfully acclimatized in a greenhouse, and transferred to the field.     

Phytoremediative potential of salt-tolerant grass species for cadmium and lead under contaminated nutrient solution

Abstract

Phytoremediation of heavy metal contaminated soils represents a promising technique and salt-tolerant hyperaccumulators for multiple metals are the need of time. Therefore, phytoremediation potential of four salt-tolerant grass species [Dhab (Desmostachya bipinnata), Kallar (Leptochloa fusca), Para (Brachiaria mutica) and Sporobolus (Sporobolus arabicus Boiss)] was evaluated for cadmium (Cd) and lead (Pb) in a hydroponic study. The plants were harvested after a growth period of 3 months in a nutrient solution containing different levels of Cd (0, 5, and 25?mg?L^?1) and Pb (0, 25, and 125?mg L^?1). Results indicated that Dhab grass showed the highest root and shoot dry matter yield followed by Para, Kallar and Sporobolus grass irrespective of metal or its level under which they were grown. All the grass species showed considerable Cd-accumulating potential with an accumulation of >150?mg kg^?1of shoot dry matter at a higher level of Cd-contamination (25?mg?L^?1). While in case of shoot Pb-accumulation only Para grass performed well and accumulated Pb >1000?mg kg^?1 of shoot dry matter at the higher level of Pb-contamination (125?mg?L^?1). Moreover, Para and Dhab grasses performed better for shoot Cd-uptake, while only Para grass showed promising shoot Pb uptake potential. In conclusion, these grass species could be penitentially used for phytoremediation of salt-affected Cd and Pb contaminated soils.     

Physiological Effects of 1-Methylcyclopropene on Well-Watered and Water-Stressed Cotton Plants

The current study investigated the effect of 1-methylcyclopropene (1-MCP), an ethylene inhibiting compound, in alleviating the detrimental effect of drought on cotton plants. The experiment was conducted in a growth chamber in 2006 and 2007. Treatments consisted of (T1) an untreated control well-watered, (T2) 1-MCP at 10 g ai/ha well-watered, (T3) an untreated control water-stressed, and (T4) 1-MCP at 10 g ai/ha water-stressed. Water-stress treatment consisted of withholding water from the pots until stomatal closure. The water-stress regime and the 1-MCP treatments were imposed at the pinhead-square stage, approximately 4 weeks after planting. Water-stressed plants treated with 1-MCP had a higher stomatal resistance, less negative water potential, higher activity of antioxidant enzymes, and better maintenance of membrane integrity. The greatest effects on stomatal resistance were observed at 5 days after treatment initiation, in which water-stressed 1-MCP-treated plants exhibited stomatal resistance of 0.079 m² s mmol⁻¹, whereas water-stressed untreated plants exhibited only 0.047 m² s mmol⁻¹. There was no significant effect of 1-MCP on water-use efficiency, transpiration, and dry matter production. These results indicated that application of 1-MCP to water-stressed cotton may have the potential to lower levels of stress in treated plants.     

Cadmium uptake kinetics and plants factors of shoot Cd concentration

Background and aims

Accumulation of Cd in the shoots of plants grown on Cd contaminated soils shows considerable variation. A previous preliminary experiment established that one major reason for this variation was the rate of Cd influx into the roots (mol Cd cm^?2 root s^?1). However, this experiment did not distinguish between solubilization of soil Cd on the one hand and difference in Cd uptake kinetics on the other. The main objectives of the present study were thus to characterize Cd uptake kinetics of plants continuously exposed to Cd concentrations similar to those encountered in soils. Furthermore we determined the factors responsible for differences in shoot Cd concentration such as net Cd influx, root area-shoot dry weight ratio, shoot growth rate and proportion of Cd translocated to the shoot.

Materials and methods

Maize, sunflower, flax and spinach were grown in nutrient solution with five constant Cd concentrations varying from 0 to 1.0 μmol?L^?1. Root and shoot parameters as well as Cd uptake were determined at two harvest dates and from these data Cd net influx and shoot growth rates were calculated.

Results and conclusions

Cadmium uptake kinetics, i.e. the net Cd influx vs. Cd solution concentration followed a straight line. Its slope is the root absorbing power, α, $ \left( {\alpha ={{{\mathrm{Cd}\;\mathrm{net}\;\mathrm{influx}}} \left/ {{\mathrm{Cd}\;\mathrm{solution}\;\mathrm{concentration}}} \right.}} \right) $ . The α values of spinach and flax were about double that of maize and sunflower (5?×?10^?6?cm?s^?1 vs. 2.5?×?10^?6?cm?s^?1). Spinach and flax had a 3–5 times higher shoot Cd concentration than maize and sunflower. The difference in shoot Cd concentration was partly due to the higher Cd influx but also to a higher translocation of Cd from root to shoot and also to a slower shoot growth rate.     

Effect of 1-methylcyclopropene on ethylene-induced abscission in citrus

Pre-treatment of citrus leaves and leaf explants ( Citrus sinensis [L.] Osbeck cv. Shamouti), with 1-methylcyclopropene (1-MCP), induced endogenous ethylene production when leaves were further incubated in air. The induction of ethylene production was 1-MCP concentration-dependent. Abscission was concomitantly delayed. In leaves pre-treated with 1-MCP followed by exposure to ethylene, abscission was significantly delayed in comparison with those without 1-MCP pre-treatment. When leaf explants were co-treated for 24 h with ethylene and 1-MCP, abscission was delayed quite efficiently. The Lineweaver-Burke plot yielded a half-maximal value of 0.234 μl l⁻¹ for the effect of ethylene on abscission. 1-MCP⁻¹ competed kinetically with ethylene with a K_i value of approximately 1.4−5.5 nl l⁻¹ 1-MCP. Under these experimental conditions there was some competition between 1-MCP and ethylene. However, ethylene was not able to completely counteract the inhibitory effect of 1-MCP. Pre-treatment with 1-MCP, followed by exogenous ethylene treatment, suppressed the induction of endo- β -glucanase (EG) activity at the laminar abscission zone. The ethylene-dependent accumulation of the hydrolyse gene was demonstrated by blocking the accumulation of CsCel a1 mRNA by 1-MCP. Six hours of exposure of leaves to 1-MCP at various times during a total of 24 h ethylene treatment efficiently reversed ethylene induction of CsCel a1 gene at mRNA level up to 18 h. The results demonstrate that the induction of abscission by ethylene is controlled at mRNA level at the abscission zone.