Increased Ethylene Evolution Research Articles

Atypical Climacteric and Functional Ethylene Metabolism and Signaling During Fruit Ripening in Blueberry (Vaccinium sp.).

Climacteric fruits display an increase in respiration and ethylene production during the onset of ripening, while such changes are minimal in non-climacteric fruits. Ethylene is a primary regulator of ripening in climacteric fruits. The ripening behavior and role of ethylene in blueberry (Vaccinium sp.) ripening is controversial. This work aimed to clarify the fruit ripening behavior and the associated role of ethylene in blueberry. Southern highbush (Vaccinium corymbosum hybrids) and rabbiteye (Vaccinium ashei) blueberry displayed an increase in the rate of respiration and ethylene evolution, both reaching a maxima around the Pink and Ripe stages of fruit development, consistent with climacteric fruit ripening behavior. Increase in ethylene evolution was associated with increases in transcript abundance of its biosynthesis genes, AMINOCYCLOPROPANE CARBOXYLATE (ACC) SYNTHASE1 (ACS1) and ACC OXIDASE2 (ACO2), implicating them in developmental ethylene production during ripening. Blueberry fruit did not display autocatalytic system 2 ethylene during ripening as ACS transcript abundance and ACC concentration were not enhanced upon treatment with an ethylene-releasing compound (ethephon). However, ACO transcript abundance was enhanced in response to ethephon, suggesting that ACO was not rate-limiting. Transcript abundance of multiple genes associated with ethylene signal transduction was upregulated concomitant with developmental increase in ethylene evolution, and in response to exogenous ethylene. As these changes require ethylene signal transduction, fruit ripening in blueberry appears to involve functional ethylene signaling. Together, these data indicate that blueberry fruit display atypical climacteric ripening, characterized by a respiratory climacteric, developmentally regulated but non-autocatalytic increase in ethylene evolution, and functional ethylene signaling.

Interactive Effects of Intraspecific Competition and Drought on Stomatal Conductance and Hormone Concentrations in Different Tomato Genotypes

Plant physiological responses to various stresses are characterized by interaction and coupling, while the intrinsic mechanism remains unclear. The effects of intraspecific competition on plant growth, stomatal opening, and hormone concentrations were investigated with three tomato genotypes (WT-wild type, Ailsa Craig; FL-a abscisic acid (ABA) deficient mutant, flacca; NR-a partially ethylene-insensitive genotype) under two water regimes (full irrigation, irrigation amount = daily transpiration; deficit irrigation, 60% of irrigation amount in full irrigation) in this study. Three kinds of competitions were designed, i.e., root and canopy competition, non-root competition, and non-canopy competition, respectively. Intraspecific competition reduced plant leaf area and stomatal conductance (gs) of wild-type tomato, accompanied by ABA accumulation and ethylene evolution. Intraspecific competition-induced decrease in gs was absent in FL and NR, indicating ABA and ethylene involved in plant response to intraspecific competition. As soil water becomes dry, the competition decreased gs by elevating ABA and ethylene accumulations. Under severe drought, the competition-induced decline in gs was covered by the severe drought-induced decrease in gs, as hydraulic signals most probably dominate. The absence of canopy competition insignificantly influenced plant stomatal opening of well-watered tomato, as canopy separation minimized the plant neighbor sensing by ethylene and other signals. Whereas under water deficit condition, the absence of canopy competition significantly reduced ABA accumulation in roots and then stomatal conductance, indicating the belowground neighbor detection signals maybe enhanced by soil drought. The absence of root competition increased ethylene evolution, confirming the importance of ethylene in neighbor detection and plant response to environmental stress.

Abscisic Acid Antagonizes Ethylene Production through the ABI4-Mediated Transcriptional Repression of ACS4 and ACS8 in Arabidopsis

Increasing evidence has revealed that abscisic acid (ABA) negatively modulates ethylene biosynthesis, although the underlying mechanism remains unclear. To identify the factors involved, we conducted a screen for ABA-insensitive mutants with altered ethylene production in Arabidopsis. A dominant allele of ABI4, abi4-152, which produces a putative protein with a 16-amino-acid truncation at the C-terminus of ABI4, reduces ethylene production. By contrast, two recessive knockout alleles of ABI4, abi4-102 and abi4-103, result in increased ethylene evolution, indicating that ABI4 negatively regulates ethylene production. Further analyses showed that expression of the ethylene biosynthesis genes ACS4, ACS8, and ACO2 was significantly decreased in abi4-152 but increased in the knockout mutants, with partial dependence on ABA. Chromatin immunoprecipitation–quantitative PCR assays showed that ABI4 directly binds the promoters of these ethylene biosynthesis genes and that ABA enhances this interaction. A fusion protein containing the truncated ABI4-152 peptide accumulated to higher levels than its full-length counterpart in transgenic plants, suggesting that ABI4 is destabilized by its C terminus. Therefore, our results demonstrate that ABA negatively regulates ethylene production through ABI4-mediated transcriptional repression of the ethylene biosynthesis genes ACS4 and ACS8 in Arabidopsis.

Genome-wide digital transcript analysis of putative fruitlet abscission related genes regulated by ethephon in litchi.

The high level of physiological fruitlet abscission in litchi (Litchi chinensis Sonn.) causes severe yield loss. Cell separation occurs at the fruit abscission zone (FAZ) and can be triggered by ethylene. However, a deep knowledge of the molecular events occurring in the FAZ is still unknown. Here, genome-wide digital transcript abundance (DTA) analysis of putative fruit abscission related genes regulated by ethephon in litchi were studied. More than 81 million high quality reads from seven ethephon treated and untreated control libraries were obtained by high-throughput sequencing. Through DTA profile analysis in combination with Gene Ontology and KEGG pathway enrichment analyses, a total of 2730 statistically significant candidate genes were involved in the ethephon-promoted litchi fruitlet abscission. Of these, there were 1867 early-responsive genes whose expressions were up- or down-regulated from 0 to 1 d after treatment. The most affected genes included those related to ethylene biosynthesis and signaling, auxin transport and signaling, transcription factors (TFs), protein ubiquitination, ROS response, calcium signal transduction, and cell wall modification. These genes could be clustered into four groups and 13 subgroups according to their similar expression patterns. qRT-PCR displayed the expression pattern of 41 selected candidate genes, which proved the accuracy of our DTA data. Ethephon treatment significantly increased fruit abscission and ethylene production of fruitlet. The possible molecular events to control the ethephon-promoted litchi fruitlet abscission were prompted out. The increased ethylene evolution in fruitlet would suppress the synthesis and polar transport of auxin and trigger abscission signaling. To the best of our knowledge, it is the first time to monitor the gene expression profile occurring in the FAZ-enriched pedicel during litchi fruit abscission induced by ethephon on the genome-wide level. This study will contribute to a better understanding for the molecular regulatory mechanism of fruit abscission in litchi.

Effects of elevated CO2 and water stress on physiological responses of Perilla frutescens var. japonica HARA

The aim of the present study was to investigate the interactive effects of expected environmental constraints, specifically elevated CO2 and drought conditions, on the physiological responses of Perilla frutescens var. japonica Arum. Perilla frutescens var. japonica ‘Arum’ was exposed to 700 μmol mol−1 of CO2 under both well-watered and water-stressed conditions. Photosynthetic rate was higher under elevated CO2 conditions. Stomatal resistance increased while transpiration rates declined, which suggests that water-use efficiency rose under elevated CO2 conditions. Under water-stressed conditions, elevated CO2 concentrations induced much higher stomatal resistance than ambient CO2 levels. This result implies that elevated CO2 concentrations might increase plant sensitivity to water stress, thereby providing plants with increased protection against drought. Furthermore, elevated CO2 concentrations alleviated drought-induced photosynthetic decline in the early stages of drought, although this effect was not sustainable. Water stress also elevated ascorbate peroxidase (APX) activity but had little effect on glutathione reductase activity. Water stress-induced APX activity was much lower under elevated CO2 conditions compared to ambient CO2 levels. Therefore, elevated CO2 concentrations might increase plant resistance to water stress. Moreover, there is a possibility that increased ethylene evolution under elevated CO2 conditions could improve the capacity of plants to scavenge reactive oxygen species by enhancing APX activity.

Use of 1-Aminocyclopropane Carboxylic Acid and Metamitron for Delayed Thinning of Apple Fruit

The normal window for application of thinning chemicals in apple extends from bloom until 3 weeks after bloom, when the fruit reach a mean diameter of ≈16 mm. After this time fruit are generally insensitive to standard chemical thinning sprays. The potential for the photosystem II (PSII) inhibitor metamitron and the ethylene precursor 1-aminocyclopropane carboxylic acid (ACC) to thin apple fruit after the traditional thinning window was investigated in field experiments over three years. A standard rescue thinning spray of carbaryl plus ethephon plus naphthaleneacetic acid (NAA) reduced fruit set of Gale ‘Gala’ if applied when the mean fruit diameter was 18, 20, and 27 mm in 2010, 2011, and 2012, respectively. The thinning activity of 400 mg·L−1 ACC was equivalent to the standard rescue thinning spray in 2010, whereas 350 mg·L−1 metamitron reduced fruit set more effectively than either the standard or ACC in 2010. Application of 400 mg·L−1 ACC plus 350 mg·L−1 metamitron when the mean fruit diameter was 18 mm reduced fruit set to almost no crop in 2010. The combination of metamitron plus ACC exhibited thinning activity after application at 25 and 33 mm mean fruit diameter in 2011 and 2012, respectively. Increased ethylene evolution was found in detached ‘GoldRush’ fruit 24 h after applications of ACC from 11 mm to 27 mm mean fruit diameter, but not when ACC was applied at 31 mm mean fruit diameter. Ethylene evolution was much higher after application of ACC at the 11 mm or 17 mm mean fruit diameter stage compared with application when fruit diameter was 23 mm or 27 mm. The thinning activity of ACC was related to the period of maximum ethylene response. The effects of delayed applications of ACC and metamitron on fruit set tended to be greater when these two chemicals were combined, suggesting that the creation of a carbohydrate stress and the capacity to convert ACC to ethylene are both required to trigger abscission of apple fruit larger than 18 mm in diameter.

Apple Fruitlet Ethylene Evolution and MdACO1, MdACS5A, and MdACS5B Expression after Application of Naphthaleneacetic Acid, 6-Benzyladenine, Ethephon, or Shading

Abscission of apple (Malus ×domestica Borkh.) fruitlets is associated with increased ethylene evolution, although the role of ethylene is not clear. Fruitlet ethylene evolution and expression of ethylene-related MdACO1, MdACS5A, and MdACS5B genes were followed in the abscission zone (AZ) and fruit cortex (FC) of king and lateral fruitlets of ‘Golden Delicious’/M.9 after spraying with 15 mg·L−1 naphthaleneacetic acid (NAA), 150 mg·L−1 6-benzyladenine (BA), 500 mg·L−1 ethephon, and after 3 days 96% shading, all applied at a mean 10 mm fruitlet diameter. Lateral fruitlets (LF) were more susceptible to abscission after treatment with chemical thinners or shading compared with king fruitlets (KF) in which only ethephon-treated KF abscised later. On KF, only ethephon increased fruitlet ethylene evolution, MdACO1 expression in FC and AZ 8 days after treatment (DAT), and abscission of KF 10 DAT, but the other treatments did not. Although ethephon increased MdACO1 expression in FC of KF 8 DAT, the expression of MdACS5A and MdACS5B remained unchanged. On LF, all treatments at 8 DAT increased ethylene evolution and MdACO1 expression in FC and enhanced abscission 10 to 22 DAT. Expression of MdACS5A and MdACS5B in FC of LF 8 DAT was expressed at a lower level compared with MdACO1. In the AZ of both KF and LF, only ethephon increased expression of MdACO1, MdACS5A, and MdACS5B.

Ameliorative effect of brassinosteroid and ethylene on germination of cucumber seeds in the presence of sodium chloride

Brassinosteroids are a class of plant polyhydroxysteroids with a diverse of functions in plant growth and development, while ethylene is a gaseous hormone involved in regulation of numerous physiological processes. To evaluate the roles of BR and ethylene in seed germination under conditions of salt stress, effects of 24-Epibrassinolide (EBR) and 1-aminocyclopropane-1-carboxylic acid (ACC) on seed germination of cucumber (Cucumis sativus) seeds in the presence of 250 mM NaCl were investigated. Seed germination was significantly inhibited by the presence of NaCl in the incubation medium, and the inhibitory effect was significantly alleviated by addition of EBR and ACC to the incubation medium containing NaCl. There was an increase in ethylene evolution during seed germination and this increase was suppressed by salt stress. The reduction in ethylene evolution from imbibed seeds by salt stress was attenuated by EBR. Salt stress inhibited ACC oxidase (ACO) activity and EBR reversed the salt stress-induced decrease in ACO activity. Salt stress reduced expression of gene encoding ACO (CsACO2), and EBR reversed the salt stress-induced down-regulation of CsACO2. The alleviative effect of EBR on seed germination in the presence of NaCl was diminished by antagonist of ethylene synthesis, aminoethoxyvinylglycine. These results indicate that both ethylene and BR are likely to be associated with suppression of seed germination under salt stress and that the mitigating effect of BR on salt stress-induced inhibition of seed germination may occur through its interaction with ethylene synthesis.

Involvement of ethylene in growth induction of stationary tobacco pith tissue in vitro

The effects of ethylene on growth initiation of tobacco pith tissue in vitro were investigated. Pith explants were incubated on a double inorganic modified White’s media containing 0.2 mg l−1 kinetin with and without indole-3-acetic acid (IAA) and the ethylene synthesis inhibitor aminooxyacetic acid (AOA). The burst of wound ethylene had no effect on growth initiation, was not affected by the AOA, and decreased to its minimum level during the initial 24 h in culture. Tissue growth was initiated after 72 h and continued on IAA-containing media only. A marked increase in ethylene evolution occurred only in tissues subjected to an IAA-containing medium prior to growth initiation. AOA inhibited this ethylene synthesis and the following growth of the tissues. The initial water uptake by the pith explants occurring even in the absence of IAA was also inhibited by AOA. The metabolic indicators for growth initiation such as enhanced respiration, increased activity of nitrate reductase, and initiation of cathodic isoperoxidases were all inhibited by AOA. It was concluded that the primer function of IAA in growth initiation is via inducing the biosynthesis of a marked ethylene signal, which in the absence of which active growth will not occur. The inhibiting effect of AOA is continuous and a transfer of the pith explants to fresh IAA-containing media did not result in a new ethylene burst nor tissue growth induction. The morphological changes in the tissues and cells during the initial stages of their development on the different media are demonstrated.

1-Aminocyclopropane-1-carboxylic acid (ACC) concentration and ACC synthase expression in soybean roots, root tips, and soybean cyst nematode (Heterodera glycines)-infected roots

Colonization of plant roots by root knot and cyst nematodes requires a functional ethylene response pathway. However, ethylene plays many roles in root development and whether its role in nematode colonization is direct or indirect, for example lateral root initiation or root hair growth, is not known. The temporal requirement for ethylene and localized synthesis of ethylene during the life span of soybean cyst nematode (SCN) on soybean roots was further investigated. Although a significant increase in ethylene evolution was not detected from SCN-colonized roots, the concentration of the immediate precursor to ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC), was higher in SCN-colonized root pieces and root tips than in other parts of the root. Moreover, expression analysis of 17 ACC synthase (ACS) genes indicated that a select set of ACS genes is expressed in SCN-colonized root pieces that is clearly different from the set of genes expressed in non-colonized roots or root tips. Semi-quantitative real-time PCR indicated that ACS transcript accumulation correlates with the high concentration of ACC in root tips. In addition, an ACS-like sequence was found in the public SCN nucleotide database. Acquisition of a full-length sequence for this mRNA (accession GQ389647) and alignment with transcripts for other well-characterized ACS proteins indicated that the nematode sequence is missing a key element required for ACS activity and therefore probably is not a functional ACS. Moreover, no significant amount of ACC was found in any growth stage of SCN that was tested.

The application of ethephon (an ethylene releaser) increases growth, photosynthesis and nitrogen accumulation in mustard (Brassica juncea L.) under high nitrogen levels

Ethephon (2-chloroethyl phosphonic acid), an ethylene-releasing compound, influences growth and photosynthesis of mustard (Brassica juncea L. Czern & Coss.). We show the effect of nitrogen availability on ethylene evolution and how this affects growth, photosynthesis and nitrogen accumulation. Ethylene evolution in the control with low N (100 mg N kg(-1) soil) was two-times higher than with high N (200 mg N kg(-1) soil). The application of 100-400 microl x l(-1) ethephon post-flowering, i.e. 60 days after sowing, on plants receiving low or high N further increased ethylene evolution. Leaf area, relative growth rate (RGR), photosynthesis, leaf nitrate reductase (NR) activity and leaf N reached a maximum with application of 200 microl x l(-1) ethephon and high N. The results suggest that the application of ethephon influences growth, photosynthesis and N accumulation, depending on the amount of nitrogen in the soil.

Benzylaminopurine Application on Two Different Apple Cultivars (Malus domestica) Displays New and Unexpected Fruitlet Abscission Features

It has been previously shown that abscission of apple fruitlets is preceded by an increase in ethylene evolution and in the amount of transcripts for 1-aminocyclopropane-1-carboxylate oxidase (ACO), an enzyme catalysing the final step in ethylene biosynthesis. These events are concomitant with shedding induction and chemical thinning. There are several thinners but their mode of action and efficacy is poorly understood. One of them is benzylaminopurine (BA), a cytokinin believed to act by enhancing vegetative activity and stressing the competition between shoots and fruitlets, thus leading to fruitlet shedding. Nevertheless, the specific mechanism of action of BA and the variable effect depending on apple cultivar (easy or difficult to thin) are poorly understood. Abscission, the amount of MdACO1 transcripts and other parameters were followed in immature apple fruits during the period of physiological drop. The cultivars studied were 'Golden Delicious' and the 'spur' type 'Red Delicious'. BA was used as a thinning agent and was sprayed 14 d after petal fall (DAPF). Fruitlets were divided into central (C) and lateral (L) fruitlet populations. Fruitlet size was significantly different between C and L fruitlets but it did not differ much between the populations within the same cultivar. C fruitlets were characterized by basal ethylene evolution while L fruitlets displayed an increase in hormone biosynthesis during abscission induction. Cluster composition evaluated by the L/C ratio differed in the two varieties, being almost unchanged throughout abscission induction in 'Golden Delicious' and progressively decreasing in 'Red Delicious'. Shoot growth activity evaluated at the end of the season indicated a possible connection with both the ongoing abscission and BA application. MDACO1 transcripts were mainly detected in L fruitlets and the accumulation was related to total abscission in 'Golden Delicious', while in 'Red Delicious' expression was observed in both C and L fruitlets. BA probably exerts its thinning effect through vegetative growth. In the 'spur' type 'Red Delicious' the chemical is ineffective, probably due to a limited action on shoot growth due to genetic characteristics. The amount of MdACO1 transcripts in seeds is a good indicator of abscission.

Stem and leaf growth of alpine sun and prairie shade ecotypes of Stellaria longipes under different photoperiods: role of ethylene

Plant ecotypes of Stellaria longipes Goldie from competitive, shade-adapted prairie habitat and less competitive, nonshaded alpine habitat were subjected to shortened or extended photoperiod conditions. Increasing daylength was positively correlated to increased stem elongation in both ecotypes. Leaf length and area for shade (prairie) plants was significantly altered with increased photoperiods, whereas sun (alpine) plants exhibited minimal leaf expansion in response to increased photoperiod. Increased ethylene evolution in the alpine genotype during rapid stem elongation and extended photoperiods suggests that ethylene plays a growth regulatory role in this sun-adapted genotype. The prairie genotype evolved less ethylene during these same periods, indicating either a diminished requirement for elevated ethylene to effect elongation and leaf expansion responses or possibly increased ethylene sensitivity because of interactions with other hormones, such as gibberellin or auxin. The sun genotype consistently produced more ethylene than the shade genotype under all photoperiod treatments. We conclude that photoperiod alters stem elongation and leaf expansion responses; similar trends were observed for extended photoperiods as were observed for shaded conditions, specifically low light intensity (photosynthetically active radiation) conditions. Further, ethylene levels altered during these responses, especially in sun-adapted alpine plants, which suggests that ethylene is involved in these growth responses.

Water relations, activities of antioxidants, ethylene evolution and membrane integrity of pigeonpea roots as affected by soil moisture

The plants of pigeonpea (Cajanus cajan L.) cv. H77-216 were subjected to moderate [soil moisture content (SMC) = 7.3 ± 0.5 %] and severe (SMC = 4.3 ± 0.5 %) drought by withholding the irrigation at vegetative stage (45 d after sowing). The control plants were maintained at SMC of 11.0 ± 0.5 %. Half of the stressed plants were re-irrigated and their recovery was studied after 2 d. Leaf water potential, osmotic potential, and relative water content of leaf and root decreased significantly while a sharp rise in proline and total soluble sugars contents were noticed. Drought induced a significant increase in 1-aminocyclopropane 1-carboxylic acid (ACC) content and ACC oxidase activity which caused a considerable increase in ethylene evolution. Malondialdehyde content and relative stress injury were increased under drought whereas reverse was true for ascorbic acid content. The membrane integrity of roots decreased during stress and recovered on rehydration. The specific activity of total superoxide dismutase, ascorbate peroxidase, glutathione reductase, and glutathione transferase decreased to 37 - 78 %, 17 - 62 %, 29 - 36 % and 57 - 79 % at moderate and severe drought, respectively. The increase in activity of catalase and peroxidase could not overcome the accumulation of H2O2 content in the roots.

Regulatory Role of Cystathionine-γ-Synthase and de novo Synthesis of Methionine in Ethylene Production during tomato Fruit Ripening

The essential amino acid methionine is a substrate for the synthesis of S-adenosyl-methionine (SAM), that donates its methyl group to numerous methylation reactions, and from which polyamines and ethylene are generated. To study the regulatory role of methionine synthesis in tomato fruit ripening, which requires a sharp increase in ethylene production, we cloned a cDNA encoding cystathionine gamma-synthase (CGS) from tomato and analysed its mRNA and protein levels during tomato fruit ripening. CGS mRNA and protein levels peaked at the "turning" stage and declined as the fruit ripened. Notably, the tomato CGS mRNA level in both leaves and fruit was negatively affected by methionine feeding, a regulation that Arabidopsis, but not potato CGS mRNA is subject to. A positive correlation was found between elevated ethylene production and increased CGS mRNA levels during the ethylene burst of the climacteric ripening of tomato fruit. In addition, wounding of pericarp from tomato fruit at the mature green stage stimulated both ethylene production and CGS mRNA level. Application of exogenous methionine to pericarp of mature green fruit increased ethylene evolution, suggesting that soluble methionine may be a rate limiting metabolite for ethylene synthesis. Moreover, treatment of mature green tomato fruit with the ethylene-releasing reagent Ethephon caused an induction of CGS mRNA level, indicating that CGS gene expression is regulated by ethylene. Taken together, these results imply that in addition to recycling of the methionine moieties via the Yang pathway, operating during synthesis of ethylene, de novo synthesis of methionine may be required when high rates of ethylene production are induced.

Pawpaw [Asimina triloba (L.) Dunal] Fruit Ripening. I. Ethylene Biosynthesis and Production

Pawpaw fruit ethylene production, 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) and ACC oxidase (ACO) activities, and tissue content of the ethylene precursor ACC and conjugate malonyl-ACC (MACC) were measured during postharvest ripening. Fruit were harvested near the advent of the ripening process and were ripened at room temperature. The fruit displayed increases in ethylene production and respiration rate during ripening with maxima for both 3 days after harvest. Mean ethylene maxima on a fresh weight basis were 4.7 and 7.6 μg·kg-1·h-1 and mean respiratory (CO2 production) maxima on a fresh weight basis were 220 and 239 mg·kg-1·h-1 in 1999 and 2001, respectively. The increase in ethylene evolution coincided with an increase in respiration and a rapid decline in fruit firmness. Internal and external fruit firmness declined in a parallel manner. The ethylene climacteric peak occurred after the greatest decline in fruit firmness, indicating that low levels of ethylene may be sufficient to initiate the ripening process. The ethylene climacteric peak also coincided with the highest activities of both ACS and ACO as well as the maximum tissue ACC content. As ACC content increased, MACC content declined, suggesting a regulation of ethylene production via free ACC levels by malonylation of ACC. Thus, the climacteric development of ethylene production may be regulated by an increase of ACS activity and a decrease in ACC malonyltransferase activity, making more free ACC available for the production of ethylene by increased activity of ACO.

Plant water status, H<sub>2</sub>O<sub>2</sub> scavenging enzymes, ethylene evolution and membrane integrity of Cicer arietinum roots as affected by salinity

The chickpea genotype, CSG-8962 was raised in screenhouse to study salinity induced changes in ethylene evolution, antioxidative defence system and membrane integrity in relation to changes in plant water and mineral content. At vegetative stage (60 d after sowing), the plants were exposed to single saline irrigation (0, 2.5, 5.0 and 10.0 dS m-1). Sampling was done 3 d after saline treatments. The other sets of treated plants were re-irrigated with water and sampled after further 3 d. The Ψw of leaf and Ψs of leaf and roots decreased from -0.47 to -0.61 MPa, -0.67 to -1.23 MPa and from -0.57 to -0.95 MPa, respectively, with increasing salinity. Similarly, RWC of leaf and roots reduced from 87.5 to 72.3 % and 96.7 to 84.35 %, respectively. The decline in Ψs of roots was mainly due to accumulation of proline and total soluble sugar. With salinity, increase in ethylene evolution, 1-aminocyclopropane-1-carboxylic acid (ACC) content and ACC oxidase activity was reported. Similarly, marked increase in H2O2 content (20 - 182 %) and lipid peroxidation (43 - 170 %) was observed. The defense mechanism activated in roots was confirmed by the increased activities of superoxide dismutase (SOD), peroxidase (POX), ascorbate peroxidase (APX), glutathione transferase (GTase), glutathione reductase (GR) and catalase (CAT) but ascorbic acid (AA) content was decreased. About 3-fold increase in Na+/K+ ratio and 2.5 fold increase in Cl- content was observed. Upon desalinization, a partial recovery was observed in most of the parameters studied.

Ethylene Evolution and Ammonium Accumulation by Tomato Plants after Root-Applied Glufosinate-Ammonium Treatment in the Presence of Ethylene Inhibitors

In previous research in hydroponics, ethylene evolution by tomato shoots (Lycopersicon esculentum Mill.) was promoted by the herbicidal action of root-applied glufosinate-ammonium (GLA). Ethylene evolution was preceded by accumulation and was detected before or concurrent with development of toxicity symptoms following GLA application. In the current experiment, aminooxyacetic acid (AOA) or silver thiosulfate (STS) was added in nutrient solutions to elucidate the relationship between accumulation, ethylene evolution, and symptom development. Aminooxyacetic acid is an inhibitor of ethylene synthesis, and STS is an inhibitor of ethylene action. In hydroponics, 4-week-old tomato seedlings were used. The GLA was added at 25 mg/L into Hoagland no. 1 nutrient solution. For the AOA and STS treatments, AOA or STS was added to concentrations of 1 mM or 25 mM, respectively. With GLA alone, foliar symptoms of injury appeared at 4 days after treatment (DAT), and plant death occurred at 10 DAT. Adding STS did not affect this process; however, AOA delayed the appearance of symptoms for 2 days, but plants died at 10 DAT. Neither AOA nor STS significantly affected plant mass relative to growth with GLA alone. With GLA alone, accumulated in shoots and roots at 2 DAT. The AOA inhibited accumulation in shoots until 6 DAT and in roots until 8 DAT. The STS had no effect on shoot accumulation relative to GLA alone but inhibited the accumulation in roots until 6 DAT. Herbicide-induced ethylene evolution occurred at 4 DAT without inhibitors and then fell. The AOA inhibited an increase in ethylene evolution during the period of the experiment, but STS had no inhibitory effect on the rise in ethylene production. This result suggests that AOA can inhibit ethylene evolution after accumulation and delay the development of toxicity symptoms. The STS had no effect on either accumulation or symptom appearance. The inhibitors did not prevent death of the plants exposed to GLA.

Mechanical wounding and abscission in citrus

Fruit detachment force (FDF), ethylene evolution, fruit and leaf drop were determined in Citrus sinensis for periods up to 96 h after mechanical wounding. Injury by removing a thin section of mature fruit flavedo reduced FDF, increased ethylene evolution and promoted abscission. Injuring flavedo 1 cm below the calyx was more effective at reducing FDF than injuring flavedo at the equator or the blossom‐end of mature fruit. Injuring the calyx or peduncle of mature fruit, or injuring three leaves closest to the mature fruit did not reduce FDF. Immature fruitlets either did not abscise or underwent low rates of abscission in response to mechanical wounding, depending on age. Inhibiting ethylene binding in wounded mature fruit with 1‐methylcyclopropene (1‐MCP) increased ethylene evolution compared with wounded fruit alone, but the reduction in FDF was similar. When an ethylene biosynthesis inhibitor (aminoethoxyvinylglycine, AVG) was used, reduction in FDF of wounded mature fruit exposed to AVG was similar to that of wounded fruit alone but ethylene production was markedly reduced. Wounding mature leaf blades in the presence or absence of 1‐MCP resulted in elevated but equal ethylene evolution up to 48 h after wounding, however, no leaf drop occurred. Thereafter, ethylene evolution was higher in 1‐MCP‐treated wounded leaves. Removing up to 77% of the total mature leaf area did not cause leaf drop, nor did wounding tissue across the laminar or petiolar abscission zones. Leaflets of 5 mm length reached nearly 100% abscission after mechanical wounding, whereas wounding leaves 20 mm length resulted in 15% abscission. The data suggest that mechanical wounding of flavedo results in mature fruit abscission, and ethylene binding may not be mandatory to initiate abscission in citrus fruit. The differential response of fruit and leaves at different ages to wounding may be related to potential contribution to carbohydrate accumulation, and production and sensitivity of tissues to an abscission signal(s).

Ripening of mountain papaya (Vasconcellea pubescens) and ethylene dependence of some ripening events

The first noticeable event during the ripening of mountain papaya ( Vasconcellea pubescens ) is the rapid degreening of the skin, followed by climacteric ethylene and flesh softening. When ethylene perception is blocked by 1-methylcyclopropene (1-MCP, 0.3 μl l −1 for 16 h at 20 °C), the increase in ethylene evolution is avoided, and softening and color development are partially delayed. This indicates that ethylene perception is an absolute requirement for ethylene production during ripening and that softening and color development are only partially dependent on ethylene. Our data also suggest that the changes in pH, titratable acidity, and soluble solids are not dependent on ethylene. Mountain papaya fruit displayed the typical climacteric rise in ethylene production, reaching maximum values of 1.8–3.2 μmol kg −1 h −1 , and so confirmed the climacteric nature of the fruit. The activity of ACC oxidase paralleled changes in ethylene production. Nevertheless, when 1-MCP treated fruit recover ethylene sensitivity, a big increase in ACC oxidase activity is observed followed by a further increase, but at a reduced level, of ethylene; the reduced ACC content suggests that in these fruit the availability of ACC is limiting ethylene biosynthesis. Finally, our results indicate that treatment of mountain papaya with 1-MCP delays the onset of the climacteric phase, and therefore can be used commercially to extend effectively the short shelf life of this fruit.