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Ethylene production is negatively associated with storage life in sand pear (Pyrus pyrifolia Nakai), particularly at the time of fruit harvest. 1-Aminocyclopropane-1-carboxylic acid synthase (ACS) is the rate-limiting enzyme in ethylene biosynthesis and is considered to be important for fruit storage life. However, the candidate ACS genes and their roles in sand pear remain unclear. The present study identified 13 ACS genes from the sand pear genome. Phylogenetic analysis categorized these ACS genes into four subgroups (type I, type II, type III and putative AAT), and indicated a close relationship between sand pear and Chinese white pear (P. bretschneideri). According to the RNA-seq data and qRT-PCR analysis, PpyACS1, PpyACS2, PpyACS3, PpyACS8, PpyACS9, PpyACS12 and PpyACS13 were differently expressed in climacteric and non-climacteric-type pear fruits, ‘Ninomiyahakuri’ and ‘Eli No.2’, respectively, during fruit ripening. In addition, the expressions of PpyACS2, PpyACS8, PpyACS12 and PpyACS13 were found to be associated with system 1 of ethylene production, while PpyACS1, PpyACS3, and PpyACS9 were found to be associated with system 2, indicating that these ACS genes have different roles in ethylene biosynthesis during fruit development. Overall, our study provides fundamental knowledge on the characteristics of the ACS gene family in sand pear, in addition to their possible roles in fruit ripening. [ABSTRACT FROM AUTHOR]

Drought imposes stress on plants and associated soil microbes, inducing coordinated adaptive responses, which can involve plant–soil signalling via phytohormones. However, we know little about how microbial communities respond to phytohormones, or how these responses are shaped by chronic (long-term) drought. Here, we added three phytohormones (abscisic acid, 1-aminocyclopropane-1-carboxylic acid, and jasmonic acid) to soils from long-term (25-year), field-based climate treatments to test the hypothesis that chronic drought alters soil microbial community responses to plant stress signalling. Phytohormone addition increased soil respiration, but this effect was stronger in irrigated than in droughted soils and increased soil respiration at low phytohormone concentrations could not be explained by their use as substrate. Thus, we show that drought adaptation within soil microbial communities modifies their responses to phytohormone inputs. Furthermore, distinct phytohormone-induced shifts in microbial functional groups in droughted vs. irrigated soils might suggest that drought-adapted soil microorganisms perceive phytohormones as stress-signals, allowing them to anticipate impending drought. Emma Sayer et al. use a 25-year field experiment to investigate how microbial community responses to phytohormones are affected by drought. Phytohormone-induced shifts in microbial functional groups suggest that drought adaptation within soil microbial communities mediates responses to plant stress signalling. [ABSTRACT FROM AUTHOR]

Summary: Ethylene plays a critical role in many diverse processes in plant development. Recent studies have demonstrated that overexpression of the maize ARGOS8 gene reduces the plant's response to ethylene by decreasing ethylene signaling and enhances grain yield in transgenic maize plants. The objective of this study was to determine the effects of ethylene on the development of nodal roots, which are primarily responsible for root‐lodging resistance in maize. Exogenous application of the ethylene precursor 1‐aminocyclopropane‐1‐carboxylic acid (ACC) was found to promote the emergence of nodal roots. Transcriptome analysis of nodal tissues revealed that the expression of genes involved in metabolic processes and cell wall biogenesis was upregulated in response to ACC treatment, supporting the notion that ethylene is a positive regulator for the outgrowth of young root primordia. In BSV::ARGOS8 transgenic plants with reduced ethylene sensitivity due to constitutive overexpression of ARGOS8, nodal root emergence was delayed and the promotional effect of ACC on nodal root emergence decreased. Field tests showed that the BSV::ARGOS8 plants had higher root lodging relative to non‐transgenic controls. When ARGOS8 expression was controlled by the developmentally regulated promoter FTM1, which conferred ARGOS8 overexpression in adult plants but not in the nodal roots and nodes in juvenile plants, the FTM1::ARGOS8 plants had no significant difference in root lodging compared with the wild type but produced a higher grain yield. These results suggest that ethylene has a role in promoting nodal root emergence and that a delay in nodal root development has a negative effect on root‐lodging resistance in maize. Significance Statement: Root‐lodging resistance is a critically important trait for high‐yield maize hybrids and has been selected in maize breeding programs over the past 100 years. In maize, the stem‐borne nodal roots are formed in the normal developmental process and play a major role in root‐lodging resistance. Using the ethylene precursor 1‐aminocyclopropane‐1‐carboxylic acid (ACC) and ARGOS8 transgenic plants which have reduced ethylene sensitivity, we found that ACC promotes nodal root emergence and that ARGOS8 transgenic plants have delayed nodal root development and increased root lodging. These results suggest a positive role for ethylene in root‐lodging resistance. These findings have applications in maize breeding for improved agronomic traits. [ABSTRACT FROM AUTHOR]

Tulip (Tulipa gesneriana L.) holds spectacular and colorful bright floral buds that are widely adorned like no other flower, across the globe. In cut flowers, persistent and prolonged display life is the matter of concern which decides quality index for florists and consumers. Endophytic bacteria, taken up by the plants when colonized, can act as sink for 1-aminocyclopropane- 1-carboxylate (ACC), hence delay senescence. These bacteria, when used as suspension solution for cut flowers, increase the vase life manifolds which signify their application to commercial floriculture. Keeping in view, the exalted status and market value of cut tulips, an experiment was conducted to study the response of four bacterial strains viz. Burkholderia phytofirmans (PsJN), Caulobacter sp. (FA-13), Enterobacter sp. (MN-17) and Bacillus sp. (MN-54) along with control treatment (nontreated stems) in concentrated and diluted forms to estimate the postharvest longevity. 15 mL of suspension solution of bacterial culture was sprayed on each observational unit under controlled conditions in laboratory. In this experiment, (PsJN) in concentrated form, performed best and resulted in the longest vase life (11.2 d), delayed leaf yellowing (8.8 d), more water uptake (70.33 mL), less stem elongation (5.14 cm), more days to open flower bud (2.47 d), maximum flower diameter (50.79 mm) and. Whereas (MN-17) in concentrated form performed best in fresh and dry mass ratio and reduced stem bending. In conclusion, use of plant growth promoting bacteria (PGPB) is a useful tool in escalating vase life of cut tulips with improved floral attributes. [ABSTRACT FROM AUTHOR]

This study examines the association of auxin with ethylene and nitric oxide (NO) in regulating the magnesium (Mg) deficiency-induced root hair development in Arabidopsis thaliana. With Mg deficiency, both ethylene and NO promoted the elevation of root auxin levels in roots by inducing the expression of AUXIN-RESISTANT1 (AUX1), PINFORMED 1 (PIN1) and PIN2 transporters. In turn, auxin stimulated ethylene and NO production by activating the activities of 1-aminocyclopropane-1-carboxylate (ACC) oxidase (ACO), ACC synthase (ACS), nitrate reductase (NR) and NO synthase-like (NOS-L). These processes constituted an NO/ethylene-auxin feedback loop. Interestingly, however, the roles of ethylene and NO in regulating Mg deficiencyinduced root hair development required the action of auxin, but not vice versa. In summary, these results suggest that Mg deficiency induces a positive interaction between the accumulation of auxin and ethylene/NO in roots, with auxin acting downstream of ethylene and NO signals to regulate Mg deficiency-induced root hair morphogenesis. [ABSTRACT FROM AUTHOR]

Camelina sativa treated with plant growth-promoting bacteria (PGPB) producing 1-aminocyclopropane-1-carboxylate deaminase (acdS) or transgenic lines expressing acdS exhibit increased salinity tolerance. AcdS reduces the level of stress ethylene to below the point where it is inhibitory to plant growth. The study determined that several mechanisms appear to be responsible for the increased salinity tolerance and that the effect of acdS on gene expression patterns in C. sativa roots during salt stress is a function of how it is delivered. Growth in soil treated with the PGPB (Pseudomonas migulae 8R6) mostly affected ethylene- and abscisic acid-dependent signaling in a positive way, while expression of acdS in transgenic lines under the control of the broadly active CaMV 35S promoter or the root-specific rolD promoter affected auxin, jasmonic acid and brassinosteroid signaling and/biosynthesis. The expression of genes involved in minor carbohydrate metabolism were also up-regulated, mainly in roots of lines expressing acdS. Expression of acdS also affected the expression of genes involved in modulating the level of reactive oxygen species (ROS) to prevent cellular damage, while permitting ROS-dependent signal transduction. Though the root is not a photosynthetic tissue, acdS had a positive effect on the expression of genes involved in photosynthesis. [ABSTRACT FROM AUTHOR]

Heterosis is widely applied in agriculture; however, the underlying molecular mechanisms for superior performance are not well understood. Ethylene biosynthesis and signaling genes are shown to be down-regulated in Arabidopsis interspecific hybrids. Ethylene is a plant hormone that promotes fruit ripening and maturation but inhibits hypocotyl elongation. Here we report that application of exogenous ethylene could eliminate biomass vigor in Arabidopsis thaliana F1 hybrids, suggesting a negative role of ethylene in heterosis. Ethylene biosynthesis is mediated by the rate-limiting enzyme, 1-aminocyclopropane-1-carboxylate synthase (ACS). Downregulation of ACS genes led to the decrease of ethylene production, which was associated with the high-vigor F1 hybrids, but not with the low-vigor ones. At the mechanistic level, expression of ACS genes was down-regulated diurnally and indirectly by Circadian Clock Associated 1 (CCA1) during the day and directly by Phyotochrome-Interacting Factor 5 (PIF5) at night. Consistent with the negative role of ethylene in plant growth, biomass vigor was higher in the acs mutants than in wild-type plants, while increasing endogenous ethylene production in the hybridizing parents reduced growth vigor in the hybrids. Thus, integrating circadian rhythms and light signaling into ethylene production is another regulatory module of complex biological networks, leading to biomass heterosis in plants. [ABSTRACT FROM AUTHOR]

Endophytes are bacterial or fungal organisms associated with plants in an obligate or facultative manner. In order to maintain a stable symbiosis, many of the endophytes produce compounds that promote plant growth and help them adapt better to the environment. This study was conducted to explore the potential of jute bacterial endophytes for their growth promotion ability in direct and indirect ways. A total of 27 different bacterial species were identified from different varieties of a jute plant ( Corchorus olitorius ) and different parts of the plant (leaf, root, seed, and seedling) based on 16S rRNA gene sequence. Two of the isolates showed ACC deaminase activity with Staphylococcus pasteuri strain MBL_B3 and Ralstonia solanacearum strain MBL_B6 producing 18.1 and 8.08 μM mg −1  h −1 α-ketobutyrate respectively while eighteen had the ACC deaminase gene ( acdS ). Fourteen were positive for siderophore activity while Kocuria sp. strain MBL_B19 (133.36 μg/ml) and Bacillus sp. strain MBL_B17 (124.72 μg/ml) showed high IAA production ability. Seven bacterial strains were able to fix nitrogen with only one testing positive for nifH gene. Five isolates exhibited phosphorus utilization ability with Bacillus sp. strain MBL_B17 producing 218.47 μg P/ml. Three bacteria were able to inhibit the growth of a phytopathogen, Macrophomina phaseolina and among them Bacillus subtilis strain MBL_B4 was found to be the most effective, having 82% and 53% of relative inhibition ratio (RIR) and percent growth inhibition (PGI) values respectively. Nine bacteria were tested for their in vivo growth promotion ability and most of these isolates increased seed germination potential and vigour index significantly. Bacillus subtilis strain MBL_B13 showed 26.8% more vigour index than the control in which no bacterial inoculum was used. All inoculants were found to increase the dry weight of jute seedlings in comparison to the control plants and the most increase in fresh weight was found for Staphylococcus saprophyticus strain MBL_B9. Staphylococcus pasteuri strain MBL_B3 exhibited diverse in vitro growth promotion activity and significant growth promoting effect in in vivo pot experiments. These bacterial strains with plant growth enhancing abilities have the potential to be used as bioinoculants. [ABSTRACT FROM AUTHOR]

Lack of regulated expression and tissue specificity are the major drawbacks of plant and virus-derived constitutive promoters. A precise tissue or site-specific expression, facilitate regulated expression of proteins at the targeted time and site. Publically available microarray data on whitefly and aphid infested Arabidopsis thaliana L. was used to identify whitefly and aphid-inducible genes. The qRT-PCR further validated the inducible behaviour of these genes under artificial infestation. Promoter sequences of genes were retrieved from the Arabidopsis Information Resources database with their corresponding 5′UTR and cloned from the A. thaliana genome. Promoter reporter transcriptional fusions were developed with the beta-glucuronidase (GUS) gusA gene in a binary expression vector to validate the inducible behaviour of these promoters in eight independent transgenic Nicotiana tabaccum lines. Histochemical analysis of the reporter gene in T2 transgenic tobacco lines confirmed promoter driven expression at the sites of aphid and whitefly infestation. The qRT-PCR and GUS expression analysis of transgenic lines revealed that abscisic acid largely influenced the expression of both aphid and whitefly inducible promoters. Further, whitefly-specific promoter respond to salicylic acid and jasmonic acid (JA), whereas aphid-specific promoters to JA and 1-aminocyclopropane carboxylic acid. The response of promoters to phytohormones correlated to the presence of corresponding conserved cis-regulatory elements. [ABSTRACT FROM AUTHOR]

Ethylene and its precursor 1-aminocyclopropane-1-carboxylate (ACC) actively participate in plant developmental, defense and symbiotic programs. In this sense, ethylene and ACC play a central role in the regulation of bacterial colonization (rhizospheric, endophytic, and phyllospheric) by the modulation of plant immune responses and symbiotic programs, as well as by modulating several developmental processes, such as root elongation. Plant-associated bacterial communities impact plant growth and development, both negatively (pathogens) and positively (plantgrowth promoting and symbiotic bacteria). Some members of the plant-associated bacterial community possess the ability to modulate plant ACC and ethylene levels and, subsequently, modify plant defense responses, symbiotic programs and overall plant development. In this work, we review and discuss the role of ethylene and ACC in several aspects of plant-bacterial interactions. Understanding the impact of ethylene and ACC in both the plant host and its associated bacterial community is key to the development of new strategies aimed at increased plant growth and protection. [ABSTRACT FROM AUTHOR]

Drought is one of the major constraints limiting agricultural production worldwide and is expected to increase in the future. Limited water availability causes significant effects to plant growth and physiology. Plants have evolved different traits to mitigate the stress imposed by drought. The presence of plant growth-promoting rhizobacteria (PGPR) could play an important role in improving plant performances and productivity under drought. These beneficial microorganisms colonize the rhizosphere of plants and increase drought tolerance by lowering ethylene formation. In the present study, we demonstrate the potential to improve the growth of velvet bean under water deficit conditions of two different strains of PGPR with ACCd (1-Aminocyclopropane-1-Carboxylate deaminase) activity isolated from rainfed farming system. We compared uninoculated and inoculated plants with PGPR to assess: a) photosynthetic performance and biomass; b) ACC content and ethylene emission from leaves and roots; c) leaf isoprene emission. Our results provided evidence that under drought conditions inoculation with PGPR containing the ACCd enzyme could improve plant growth compared to untreated plants. Ethylene emission from roots and leaves of inoculated velvet bean plants was significantly lower than uninoculated plants. Moreover, isoprene emission increased with drought stress progression and was higher in inoculated plants compared to uninoculated counterparts. These findings clearly illustrate that selected PGPR strains isolated from rainfed areas could be highly effective in promoting plant growth under drought conditions by decreasing ACC and ethylene levels in plants. [ABSTRACT FROM AUTHOR]

Ethylene influences many aspects of plant growth and development. The biosynthesis of ethylene is highly regulated by a variety of internal and external cues. A key target of this regulation is 1-aminocyclopropane-1-carboxylic acid ( ACC) synthases ( ACS), generally the rate-limiting step in ethylene biosynthesis, which is regulated both transcriptionally and post-transcriptionally. Prior studies have demonstrated that cytokinin and brassinosteroid ( BR) act as regulatory inputs to elevate ethylene biosynthesis by increasing the stability of ACS proteins. Here, we demonstrate that several additional phytohormones also regulate ACS protein turnover. Abscisic acid, auxin, gibberellic acid, methyl jasmonic acid, and salicylic acid differentially regulate the stability of ACS proteins, with distinct effects on various isoforms. In addition, we demonstrate that heterodimerization influences the stability of ACS proteins. Heterodimerization between ACS isoforms from distinct subclades results in increased stability of the shorter-lived partner. Together, our study provides a comprehensive understanding of the roles of various phytohormones on ACS protein stability, which brings new insights into crosstalk between ethylene and other phytohormones, and a novel regulatory mechanism that controls ACS protein stability through a heterodimerization of ACS isoforms. [ABSTRACT FROM AUTHOR]

Background: Ethylene plays an important role in the regulation of floral organ development in soybean, and 1-aminocyclopropane-1-carboxylate synthase (ACS) is a rate-limiting enzyme for ethylene biosynthesis. However, whether ACS also regulates floral organ differentiation in soybean remains unknown. To address this, we constructed an RNAi vector to inhibit ACS expression in cotyledonary nodes. Linear DNA cassettes of RNAi-ACS obtained by PCR were used to transform soybean cotyledonary nodes. Results: In total, 131 of 139 transiently transformed plants acquired herbicide resistance and displayed GUS activities in the new buds. In comparison to untransformed seedling controls, a greater number of flower buds were differentiated at the cotyledonary node; GM-ACS1 mRNA expression levels and ethylene emission in the transformed buds were reduced. Conclusion: These results indicate that the cotyledonary node transient transformation system may be suitable for stable transformation and that the inhibition of ACS expression may be an effective strategy for promoting floral organ differentiation in soybean. [ABSTRACT FROM AUTHOR]

Camelina sativa (camelina) is an oilseed crop touted for use on marginal lands; however, it is no more tolerant of soil salinity than traditional crops, such as canola. Plant growth-promoting bacteria (PGPB) that produce 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) facilitate plant growth in the presence of abiotic stresses by reducing stress ethylene. Rhizospheric and endophytic PGPB and the corresponding acdS- mutants of the latter were examined for their ability to enhance tolerance to salt in camelina. Stimulation of growth and tolerance to salt was correlated with ACC deaminase production. Inoculation of soil with wild-type PGPB led to increased shoot length in the absence of salt, and increased seed production by approximately 30-50% under moderately saline conditions. The effect of ACC deaminase was further examined in transgenic camelina expressing a bacterial gene encoding ACC deaminase (acdS) under the regulation of the CaMV 35S promoter or the root-specific rolD promoter . Lines expressing acdS, in particular those using the rolD promoter, showed less decline in root length and weight, increased seed production, better seed quality and higher levels of seed oil production under salt stress. This study clearly demonstrates the potential benefit of using either PGPB that produce ACC deaminase or transgenic plants expressing the acdS gene under the control of a root-specific promoter to facilitate plant growth, seed production and seed quality on land that is not normally suitable for the majority of crops due to high salt content. [ABSTRACT FROM AUTHOR]

The expression and accumulation of members of the 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) and ACC oxidase (ACO) gene families was examined in white clover roots grown in either Pi (phosphate) sufficient or Pi-deprived defined media. The accumulation of one ACO isoform, TR-ACO1, was positively influenced after only 1 h of exposure to low Pi, and this was maintained over a 7-day time-course. Up-regulation of TR-ACS1, TR-ACS2 and TR-ACS3 transcript abundance was also observed within 1 h of exposure to low Pi in different tissue regions of the roots, followed by a second increase in abundance of TR-ACS2 after 5-7 days of exposure. An increase in transcript abundance of TR-ACO1 and TR-ACO3, but not TR-ACO2, was observed after 1 h of exposure to low Pi, with a second increase in TR-ACO1 transcripts occurring after 2-5 days. These initial increases of the TR-ACS and TR-ACO transcript abundance occurred before the induction of Trifolium repens PHOSPHATE TRANSPORTER 1 (TR-PT1), and the addition of sodium phosphite did not up-regulate TR-ACS1 expression over 24 h. In situ hybridization revealed some overlap of TR-ACO mRNA accumulation, with TR-ACO1 and TR-ACO2 in the root tip regions, and TR-ACO1 and TR-ACO3 mRNA predominantly in the lateral root primordia. TR-ACO1p-driven GFP expression showed that activation of the TR-ACO1 promoter was initiated within 24 h of exposure to low Pi (as determined by GFP protein accumulation). These results suggest that the regulation of ethylene biosynthesis in white clover roots is biphasic in response to low Pi supply. [ABSTRACT FROM AUTHOR]

Sex determination in plants gives rise to unisexual flowers that facilitate outcrossing and enhance genetic diversity. In cucumber and melon, ethylene promotes carpel development and arrests stamen development. Five sex-determination genes have been identified, including four encoding 1-aminocyclopropane-1-carboxylate (ACC) synthase that catalyzes the rate-limiting step in ethylene biosynthesis, and a transcription factor gene CmWIP1 that corresponds to the Mendelian locus gynoecious in melon and is a negative regulator of femaleness. ACC oxidase (ACO) converts ACC into ethylene; however, it remains elusive which ACO gene in the cucumber genome is critical for sex determination and how CmWIP1 represses development of female flowers. In this study, we discovered that mutation in an ACO gene, CsACO2 , confers androecy in cucumber that bears only male flowers. The mutation disrupts the enzymatic activity of CsACO2, resulting in 50% less ethylene emission from shoot tips. CsACO2 was expressed in the carpel primordia and its expression overlapped with that of CsACS11 in female flowers at key stages for sex determination, presumably providing sufficient ethylene required for proper CsACS2 expression. CmACO3 , the ortholog of CsACO2 , showed a similar expression pattern in the carpel region, suggesting a conserved function of CsACO2/CmACO3. We demonstrated that CsWIP1, the ortholog of CmWIP1, could directly bind the promoter of CsACO2 and repress its expression. Taken together, we propose a presumably conserved regulatory module consisting of WIP1 transcription factor and ACO controls unisexual flower development in cucumber and melon. [ABSTRACT FROM AUTHOR]

Soybean is stress-sensitive crop that exhibits markedly reduced growth under flooding and drought conditions. Three S-adenosylmethionine synthetases (SAMs) proteins were identified as flooding and drought responsive proteins in soybean using a proteomic technique. To better understand the role of these SAMs proteins in soybean under flooding and drought stresses, temporal, organ, and stress specificities were examined at mRNA and enzyme activity levels. The activity of SAMs decreased in response to the flooding, however, it was not significantly changed by NaCl, cold, gibberellic acid, and calcium in soybean roots. The activity of SAMs was induced in roots and hypocotyls under drought. The mRNA expression of the S- adenosylmethionine synthetase ( SAMs) family was down-regulated in root tips and roots under the flooding and the drought, and SAMs 1 and SAMs 2 were down-regulated in roots under both stresses. A gene 1-aminocyclopropane-1-carboxylate synthase was up-regulated in root tips, roots, and hypocotyls under drought, however, it was not changed in root tips and roots under the flooding. In addition, 1-aminocyclopropane-1-carboxylate oxidase was induced in root tips under flooding and drought. These results suggest that SAMs was involved in the response to the flooding and drought and it might affect ethylene biosynthesis in soybean. [ABSTRACT FROM AUTHOR]

Background: Oligogalacturonic acids (OGs) are oligomers of alpha-1,4-linked galacturonosyl residues that are released from cell walls by the hydrolysis of polygalacturonic acids upon fruit ripening and under abiotic/biotic stress. OGs may induce ethylene production and fruit ripening, however, the mechanism(s) behind these processes is unknown. Results: Tomato cultivar 'Ailsa Craig' (AC) and mutant Neverripe, ripening inhibitor, non-ripening, and colorless non-ripening fruits were treated with OGs at different stages. Only AC fruits at mature green stage 1 showed an advanced ripening phenomenon, although transient ethylene production was detected in all of the tomato fruits. Ethylene synthesis genes LeACS2 and LeACO1 were rapidly up-regulated, and the phosphorylated LeACS2 protein was detected after OGs treatment. Protein kinase/phosphatase inhibitors significantly affected the ripening process induced by the OGs. As a potential receptor of OGs, LeWAKL2 was also up-regulated in their presence. Conclusions: We demonstrated that OGs promoted tomato fruit ripening by inducing ethylene synthesis through the regulation of LeACS2 at transcriptional and post-translational levels. [ABSTRACT FROM AUTHOR]

This study was conducted during 2011 and 2012 to evaluate the effect of 1-aminocyclopropane-1-carboxylate (ACC) deaminase-containing, N2-fixing and P-solubilizing bacteria on the yield and morpho-physiological parameters of strawberry. A total of 8 applications at the trial set, with four water regimes were randomly distributed into the pots. The diminishing water supply caused a gradual decrease in the plant growth, chlorophyll content and berry yield, accompanied by increasing activities of drought stress markers such as total phenolics content (TPC), trolox equivalent antioxidant capacity (TEAC), malondialdehyde (MDA) content, hydrogen peroxide (H2O2), glutathione reductase (GR), glutathione S-transferase (GST), catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and ascorbate peroxidase (APX) in the leaves of strawberry. The multi-trait bacteria also increased plant growth and yield as well as TPC, TEAC, antioxidant enzymes (GR, GST, CAT, POD, SOD and APX) activity, phytohormone (GA, SA and IAA) and the contents of N, P, K, Ca, Fe, Mn, Zn and Cu, but decreased MDA and H2O2 contents which may contribute in part to activation of physiological and biochemical processes involved in the alleviation of the effect of drought stress. [ABSTRACT FROM AUTHOR]

The study evaluates the effect of rhizobacteria having 1-aminocyclopropane-1-carboxylate deaminase (ACCd) on the development of wheat seedlings. This enzyme has been proposed to play a key role in microbe-plant association. Three fluorescent pseudomonads containing this deaminase were selected from 70 strains of pseudomonads isolated from rhizosphere of wheat ( Triticum aestivum L.) and rape ( Brassica napus L.). These bacteria, varied significantly in the ability to both biosynthesize auxins and hydrolyze ACC. Among them, Pseudomonas brassicacearum subsp. brassicacearum strain RZ310 presented the highest activities of ACC deaminase during 96 h of growth in liquid Dworkin and Foster (DF) salt medium. Additionally, this rape rhizosphere strain did not produce indoles. Two other isolates, Pseudomonas sp. PO283 and Pseudomonas sp. PO366, secreted auxins only in the presence of their precursor. Phylogenetic analysis of the 16S rRNA gene and four other protein-encoding genes indicated that these wheat rhizosphere isolates belonged to the fluorescent Pseudomonas group. Moreover, the effects of these strains on wheat seedling growth under in vitro conditions were markedly dependent on both their cell suspensions used to grain inoculation and nutrient conditions. Strains tested had beneficial influence on wheat seedlings mainly at low cell densities. In addition, access to nutrients markedly changed bacteria action on cereal growth. Their presence generally favored the positive effects of pseudomonads on length and the estimated biomasses of wheat coleoptiles. Despite these general rules, impacts of each isolate on the growth parameters of cereal seedlings were unique. [ABSTRACT FROM AUTHOR]

Glutathione (GSH) plays a fundamental role in plant defense-signaling network. Recently, we have established the involvement of GSH with ethylene (ET) to combat environmental stress. However, the mechanism of GSH-ET interplay still remains unexplored. Here, we demonstrate that GSH induces ET biosynthesis by modulating the transcriptional and posttranscriptional regulations of its key enzymes, 1-aminocyclopropane-1-carboxylate synthase (ACS) and 1-aminocyclopropane-1-carboxylate oxidase (ACO). Transgenic Arabidopsis (Arabidopsis thaliana) plants with enhanced GSH content (AtECS) exhibited remarkable up-regulation of ACS2, ACS6, and ACO1 at transcript as well as protein levels, while they were down-regulated in the GSH-depleted phytoalexin deficient2-1 (pad2-1) mutant. We further observed that GSH induced ACS2 and ACS6 transcription in a WRKY33-dependent manner, while ACO1 transcription remained unaffected. On the other hand, the messenger RNA stability for ACO1 was found to be increased by GSH, which explains our above observations. In addition, we also identified the ACO1 protein to be a subject for S-glutathionylation, which is consistent with our in silico data. However, S-glutathionylation of ACS2 and ACS6 proteins was not detected. Further, the AtECS plants exhibited resistance to necrotrophic infection and salt stress, while the pad2-1 mutant was sensitive. Exogenously applied GSH could improve stress tolerance in wild-type plants but not in the ET-signaling mutant ethylene insensitive2-1, indicating that GSH-mediated resistance to these stresses occurs via an ET-mediated pathway. Together, our investigation reveals a dual-level regulation of ET biosynthesis by GSH during stress. [ABSTRACT FROM AUTHOR]

Petroleum is potentially toxic to living organisms, and there are worldwide efforts to develop methods for bioremediation of petroleum-contaminated soils. Phytoremediation is an effective method to reduce the concentration of petroleum in soils, and plant growth-promoting rhizobacteria (PGPR) play an important role in the phytoremediation. Two PGPR, Pseudomonas aeruginosa SLC-2 and Serratia marcescens BC-3, were isolated from the rhizophere of Echinochloa grown in petroleum-contaminated soil. These isolates showed capacities for 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, indole acetic acid production, siderophore synthesis, and the degradation of petroleum. The ACC deaminase activity of SLC-2 and BC-3 was 2.52 ± 0.03 μmol α-KA (mg Pr·h) and 38.52 ± 0.37 μmol α-KA (mg Pr·h), respectively. On the other hand, when the concentration of l-Trp increased, the IAA synthesis of BC-3 also increased, while the synthesis of SLC-2 did not change significantly. The ability of synthesized siderophore of SLC-2 was much higher than that of BC-3. The petroleum degradations of SLC-2 and BC-3 increased 4.78 and 7.36 %, respectively. The pot experiment of oat was performed to evaluate the plant growth-promoting abilities of SLC-2 and BC-3. Compared with non-inoculated controls, the height and fresh weight of stems increased (23.64 and 42.57) % and (16.98 and 28.3) %, respectively, whereas the length and fresh weight of roots also increased (10.34 and 20.84) and (24.13 and 43.11) %, respectively, when inoculated with SLC-2 and BC-3. The results indicated that P. aeruginosa SLC-2 and S. marcescens BC-3 can serve as promising microbes for increasing plant growth in petroleum-contaminated soil to improve the phytoremediation efficiency. [ABSTRACT FROM AUTHOR]

Drought is a major abiotic stress that affects the development and growth of most plants, and limits crop yield worldwide. Although the response of plants to drought has been well documented, much less is known about how plants respond to the water recovery process, namely rehydration. Here, we describe the spatio-temporal response of plant reproductive organs to rehydration using rose flowers as an experimental system. We found that rehydration triggered rapid and transient ethylene production in the gynoecia. This ethylene burst serves as a signal to ensure water recovery in flowers, and promotes flower opening by influencing the expression of a set of rehydration-responsive genes. An in-gel kinase assay suggested that the rehydration-induced ethylene burst resulted from transient accumulation of RhACS1/2 proteins in gynoecia. Meanwhile, RhMPK6, a rose homolog of Arabidopsis thaliana MPK6, is rapidly activated by rehydration within 0.5 h. Furthermore, RhMPK6 was able to phosphorylate RhACS1 but not RhACS2 in vitro. Application of the kinase inhibitor K252a suppressed RhACS1 accumulation and rehydration-induced ethylene production in gynoecia, and the protein phosphatase inhibitor okadaic acid had the opposite effect, confirming that accumulation of RhACS1 was phosphorylation-dependent. Finally, silencing of RhMPK6 significantly reduced ethylene production in gynoecia when flowers were subjected to rehydration. Taken together, our results suggest that temporal- and spatial-specific activation of an RhMPK6-RhACS1 cascade is responsible for rehydration-induced ethylene production in gynoecia, and that the resulting ethylene-mediated signaling pathway is a key factor in flower rehydration. [ABSTRACT FROM AUTHOR]

Our research shows that there is isoform specificity and redundancy among 6 out of 13 14-3-3 members in root growth under control and abiotic stress conditions.Arabidopsis 14-3-3 proteins are a family of conserved proteins that interact with numerous partner proteins in a phospho-specific manner, and can affect the target proteins in a number of ways; e.g. modification of enzymatic activity. We isolated T-DNA insertion lines in six 14-3-3 genes within the non-epsilon group that phylogenetically group in three closely related gene pairs. In total, 6 single, 3 double, 12 triple, and 3 quadruple mutants were generated. The mutants were phenotyped for primary root growth on control plates: single and double mutants were indistinguishable from WT, whereas six triples and all quadruples showed a shorter primary root. In addition, length of the first epidermal cell with a visible root hair bulge (LEH) was used to determine primary root elongation on medium containing mannitol and 1-aminocyclopropane-1-carboxylic acid (ACC). This analysis showed clear differences depending on the stress and 14-3-3 gene combinations. Next to the phenotypic growth analyses, a 14-3-3 pull-down assay on roots treated with and without mannitol showed that mannitol stress strongly affects the 14-3-3 interactome. In conclusion, we show gene specificity and functional redundancy among 14-3-3 proteins in primary root elongation under control and under abiotic stress conditions and changes in the 14-3-3 interactome during the onset of stress adaptation. [ABSTRACT FROM AUTHOR]

Soil microorganisms with phytase- and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activities are widely studied as plant growth-promoting rhizobacteria (PGPR). Here, we explored the bacterial community structure and occurrence of putative PGPR in plants grown in agro-ecosystems and undisturbed ecosystems from northern, central, and southern Chile. Total rhizobacterial community structure was evaluated by denaturing gradient gel electrophoresis, and dominant bands present in diverse ecosystems were sequenced. Significant differences in total bacterial communities were shown with some bacterial orders ( Enterobacteriales, Actinomycetales, and Rhizobiales) being highly similar to both ecosystems. Twenty-nine putative PGPR, showing phytate- and ACC-degrading activities and production of auxin, were selected from across the sites. Based on 16S rRNA gene sequencing, the putative PGPR were characterized as Enterobacteriales ( Enterobacter, Serratia, Pantoea, Rahnella, Leclercia), Pseudomonas, and Bacillus, consistent with previously reported PGPR and endophytic bacteria. Beta-propeller phytase genes with similarity to Bacillus were also identified. PGPR from agro-ecosystems appeared to show higher auxin production compared to those from undisturbed ecosystems. This study demonstrates that putative PGPR are widely distributed across Chilean soils. Further understanding of their contribution to the growth and adaptation of plant hosts to local soil conditions may provide opportunity for development of new PGPR in Chilean agriculture. [ABSTRACT FROM AUTHOR]

Whole-genome duplications ( WGDs) are widespread in plants, and three lineage-specific WGDs occurred in the banana ( Musa acuminata) genome. Here, we analysed the impact of WGDs on the evolution of banana gene families involved in ethylene biosynthesis and signalling, a key pathway for banana fruit ripening., Banana ethylene pathway genes were identified using comparative genomics approaches and their duplication modes and expression profiles were analysed., Seven out of 10 banana ethylene gene families evolved through WGD and four of them (1-aminocyclopropane-1-carboxylate synthase ( ACS), ethylene-insensitive 3-like ( EIL), ethylene-insensitive 3-binding F-box ( EBF) and ethylene response factor ( ERF)) were preferentially retained. Banana orthologues of At EIN3 and At EIL1, two major genes for ethylene signalling in Arabidopsis, were particularly expanded. This expansion was paralleled by that of EBF genes which are responsible for control of EIL protein levels. Gene expression profiles in banana fruits suggested functional redundancy for several Ma EBF and Ma EIL genes derived from WGD and subfunctionalization for some of them., We propose that EIL and EBF genes were co-retained after WGD in banana to maintain balanced control of EIL protein levels and thus avoid detrimental effects of constitutive ethylene signalling. In the course of evolution, subfunctionalization was favoured to promote finer control of ethylene signalling. [ABSTRACT FROM AUTHOR]

Ethylene is an essential plant hormone also known as a stress hormone because its synthesis is accelerated by induction of a variety of biotic and abiotic stress. The plant growth promoting bacteria containing the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase enhances plant growth by decreasing plant ethylene levels under stress conditions. The expression of ACC deaminase ( acdS) gene in transgenic plants is an alternative approach to overcome the ethylene-induced stress. Several transgenic plants have been engineered to express both bacterial/plant acdS genes which then lowers the stress-induced ethylene levels, thus efficiently combating the deleterious effects of environmental stresses. This review summarizes the current knowledge of various transgenic plants overexpressing microbial and plant acdS genes and their potential under diverse biotic and abiotic stresses. Transcription regulation mechanism of acdS gene from different bacteria, with special emphasis to nitrogen fixing bacteria is also discussed in this review. [ABSTRACT FROM AUTHOR]

The changes in abscissic acid (ABA) and jasmonic acid (JA) contents during fruit development of dragon fruit (Hylocereus undaatus) were determined. Associated changes in respiration, 1-aminocyclopropane-1-carboxylic acid (ACC) contents, and sugar levels were also measured. ACC content and respiration rate were high at early fruit development stages and gradually decreased as the fruit matured and ripened. In the pulp, ABA content increased before the increase in glucose and fructose contents. JA contents in the peel and pulp were high at the immature stage and decreased toward maturation. The results indicate that dragon fruit is non-climacteric and that JA may have a role in cell division in the developing fruit while ABA may induce fruit maturation. [ABSTRACT FROM AUTHOR]

Background Climacteric fruit exhibit high ethylene and respiration levels during ripening but these levels are limited in non-climacteric fruit. Even though capsicum is in the same family as the wellcharacterised climacteric tomato (Solanaceae), it is non-climacteric and does not ripen normally in response to ethylene or if harvested when mature green. However, ripening progresses normally in capsicum fruit when they are harvested during or after what is called the 'Breaker stage'. Whether ethylene, and components of the ethylene pathway such as 1- aminocyclopropane 1-carboxylate (ACC) oxidase (ACO), ACC synthase (ACS) and the ethylene receptor (ETR), contribute to non-climacteric ripening in capsicum has not been studied in detail. To elucidate the behaviour of ethylene pathway components in capsicum during ripening, further analysis is therefore needed. The effects of ethylene or inhibitors of ethylene perception, such as 1-methylcyclopropene, on capsicum fruit ripening and the ethylene pathway components may also shed some light on the role of ethylene in nonclimacteric ripening. Results The expression of several isoforms of ACO, ACS and ETR were limited during capsicum ripening except one ACO isoform (CaACO4). ACS activity and ACC content were also low in capsicum despite the increase in ACO activity during the onset of ripening. Ethylene did not stimulate capsicum ripening but 1-methylcyclopropene treatment delayed the ripening of Breaker-harvested fruit. Some of the ACO, ACS and ETR isoforms were also differentially expressed upon treatment with ethylene or 1-methylcyclopropene. Conclusions ACS activity may be the rate limiting step in the ethylene pathway of capsicum which restricts ACC content. The differential expression of several ethylene pathway components during ripening and upon ethylene or 1-methylclopropene treatment suggests that the ethylene pathway may be regulated differently in non-climacteric capsicum compared to the climacteric tomato. Ethylene independent pathways may also exist in non-climacteric ripening as evidenced by the up-regulation of CaACO4 during ripening onset despite being negatively regulated by ethylene exposure. However, some level of ethylene perception may still be needed to induce ripening especially during the Breaker stage. A model of capsicum ripening is also presented to illustrate the probable role of ethylene in this non-climacteric fruit. [ABSTRACT FROM AUTHOR]

Prunus is an economically important genus with a wide range of physiological and biological variability. Using the peach genome as a reference, sequencing reads from four almond accessions and one sweet cherry cultivar were used for comparative analysis of these three Prunus species. Reference mapping enabled the identification of many biological relevant polymorphisms within the individuals. Examining the depth of the polymorphisms and the overall scaffold coverage, we identified many potentially interesting regions including hundreds of small scaffolds with no coverage from any individual. Non-sense mutations account for about 70 000 of the 13 million identified single nucleotide polymorphisms ( SNPs). Blast2 GO analyses on these non-sense SNPs revealed several interesting results. First, non-sense SNPs were not evenly distributed across all gene ontology terms. Specifically, in comparison with peach, sweet cherry is found to have non-sense SNPs in two 1-aminocyclopropane-1-carboxylate synthase ( ACS) genes and two 1-aminocyclopropane-1-carboxylate oxidase ( ACO) genes. These polymorphisms may be at the root of the nonclimacteric ripening of sweet cherry. A set of candidate genes associated with bitterness in almond were identified by comparing sweet and bitter almond sequences. To the best of our knowledge, this is the first report in plants of non-sense SNP abundance in a genus being linked to specific GO terms. [ABSTRACT FROM AUTHOR]

The plant hormone ethylene is known to affect various developmental processes including dormancy and growth. Yet, little information is available about the role of ethylene during paradormancy release in underground adventitious buds of leafy spurge. In this study, we examined changes in ethylene evolution and the ethylene biosynthetic enzyme ACC oxidase following paradormancy release (growth induction). Our results did not show an obvious increase in ethylene during bud growth. However, when buds were incubated with 1 mM ACC, ethylene levels were higher in growing than non-growing buds, suggesting that the levels of ACC oxidase increased in growing buds. Real-time qPCR indicated that the transcript of a Euphorbia esula ACC oxidase ( Ee-ACO) increased up to threefold following growth induction. In addition, a 2.5- to 4-fold increase in ACO activity was observed 4 days after decapitation, and the Ee-ACO accounted for 40 % of the total ACO activity. Immunoblot analyses identified a 36-kD Ee-ACO protein that increased in expression during bud growth. This protein was highly expressed in leaves, moderately expressed in crown buds, stems and meristems, and weakly expressed in roots and flowers. Immunolocalization of Ee-ACO on growing bud sections revealed strong labeling of the nucleus and cytoplasm in cells at the shoot apical meristem and leaf primordia. An exception to this pattern occurred in cells undergoing mitosis, where labeling of Ee-ACO was negligible. Taken together, our results indicated an increase in the levels of Ee-ACO during paradormancy release of leafy spurge that was not correlated with an increase in ethylene synthesis. [ABSTRACT FROM AUTHOR]

The 1-aminocyclopropane-1-carboxylate oxidase (ACO) gene in the final step of ethylene biosynthesis was isolated from ethylene-sensitive Vanda Miss Joaquim flowers. This consists of 1,242 base pairs (bp) encoding for 326 amino acid residues. To investigate the specific divergence in orchid ACO sequences, the deduced Vanda ACO was aligned with five other orchid ACOs. The results reveal that the ACO sequences within Doritaenopsis, Phalaenopsis and Vanda show highly conserved and almost 95% identical homology, while the ACOs isolated from Cymbidium, Dendrobium and Cattleya are 87-88% identical to Vanda ACO. In addition, the 2-oxoglutarate- Fe(II)_oxygenase (Oxy) domain of orchid ACOs consists of a higher degree of amino acid conservation than that of the non-haem dioxygenase (DIOX_N) domain. The overall homology regions of Vanda ACO are commonly folded into 12 α-helices and 12 β-sheets similar to the three dimensional template-structure of Petunia ACO. This Vanda ACO cloned gene is highly expressed in flower tissue compared with root and leaf tissues. In particular, there is an abundance of ACO transcript accumulation in the column followed by the lip and the perianth of Vanda Miss Joaquim flowers at the fullyopen stage. [ABSTRACT FROM AUTHOR]

Pollination induced an increase in ethylene production concomitantly with ovary growth, epinasty, drooping, changes in venation and fading of the colour of the flowers of Dendrobium 'Pompadour'. Removal of stigmatic fluid prior to pollination significantly reduced ovary growth, while ethylene production, epinasty, drooping, venation and colour of pollinated flowers were not significantly affected. Removal of stigma fluid had no effect on ovary growth of non-pollinated flowers treated with 1-aminocyclopropane-1-carboxylic acid (ACC ), but significantly reduced the ovary growth of non-pollinated flowers induced by the application of napthaleneacetic acid (NAA). The effect of stigmatic fluid on ovary growth of pollinated flowers is discussed. [ABSTRACT FROM AUTHOR]

The fruit of melting-flesh peach (Prunus persica L. Batsch) cultivars produce high levels of ethylene caused by high expression of PpACS1 (an isogene of 1-aminocyclopropane-1-carboxylic acid synthase), resulting in rapid fruit softening at the late-ripening stage. In contrast, the fruit of stony hard peach cultivars do not soften and produce little ethylene due to low expression of PpACS1. To elucidate the mechanism for suppressing PpACS1 expression in stony hard peaches, a microarray analysis was performed. Several genes that displayed similar expression patterns as PpACS1 were identified and shown to be indole-3-acetic acid (IAA)-inducible genes (Aux/IAA, SAUR). That is, expression of IAA-inducible genes increased at the late-ripening stage in melting flesh peaches; however, these transcripts were low in mature fruit of stony hard peaches. The IAA concentration increased suddenly just before harvest time in melting flesh peaches exactly coinciding with system 2 ethylene production. In contrast, the IAA concentration did not increase in stony hard peaches. Application of 1-naphthalene acetic acid, a synthetic auxin, to stony hard peaches induced a high level of PpACS1 expression, a large amount of ethylene production and softening. Application of an anti-auxin, α-(phenylethyl-2-one)-IAA, to melting flesh peaches reduced levels of PpACS1 expression and ethylene production. These observations indicate that suppression of PpACS1 expression at the late-ripening stage of stony hard peach may result from a low level of IAA and that a high concentration of IAA is required to generate a large amount of system 2 ethylene in peaches. [ABSTRACT FROM AUTHOR]

Flowering plants utilize different floral structures to develop flesh tissue in fruits. Here we show that suppression of the homeologous SEPALLATA1/2-like genes MADS8 and MADS9 in the fleshy fruit apple ( Malus x domestica) leads to sepaloid petals and greatly reduced fruit flesh. Immunolabelling of cell-wall epitopes and differential staining showed that the developing hypanthium (from which the apple flesh develops) of MADS8/9-suppressed apple flowers lacks a tissue layer, and the remaining flesh tissue of fully developed apples has considerably smaller cells. From these observations, it is proposed that MADS8 and MADS9 control the development of discrete zones within the hypanthium tissue, and therefore fruit flesh, and also act as foundations for development of different floral organs. At fruit maturity, the MADS8/9-suppressed apples do not ripen in terms of both developmentally controlled ripening characters, such as starch degradation, and ethylene-modulated ripening traits. Transient assays suggest that, like the RIN gene in tomato, the MADS9 gene acts as a transcriptional activator of the ethylene biosynthesis enzyme, 1-aminocyclopropane-1-carboxylate (ACC) synthase 1. The existence of a single class of genes that regulate both flesh formation and ripening provides an evolutionary tool for controlling two critical aspects of fleshy fruit development. [ABSTRACT FROM AUTHOR]

Ethylene and light act through specific signal transduction mechanisms to coordinate the development of higher plants. Application of 1-aminocyclopropane-1-carboxylic acid (ACC, an ethylene precursor) suppresses the hypocotyl elongation of Arabidopsis seedlings in dark, but stimulates it in light. However, the mechanisms of opposite effects of ethylene on hypocotyl elongation in light and dark remain unclear. In the present study, we investigated the key factors involved in the opposite effects of ethylene on hypocotyl elongation in Arabidopsis seedlings. The effects of ACC on hypocotyl elongation of IAA-insensitive mutants including tir1- 1, axr1- 3, and axr1- 12 seedlings were reduced in light but not in dark. The DR5 promoter, a synthetic auxin-response promoter, was used to quantify the level of IAA responses. There was a marked increase in DR5-GFP signals in response to ACC treatment in hypocotyls of DR5- GFP seedlings in light, but not in dark. CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) is an important downstream component of light signaling. ETHYLENE-INSENSITIVE3 (EIN3, an ethylene-stabilized transcription factor) directly regulates ETHYLENE-RESPONSE-FACTOR1 (ERF1). The cop1- 4 mutant treated with ACC and cop1- 4/EIN3ox plants developed long hypocotyls in darkness. Expression of ERF1 in the cop1- 4 mutant was induced by ACC treatment in dark, but the expression of ERF1 in the wild type was not affected. Taken together, ethylene-promoting hypocotyl via IAA is mediated by light, and COP1 has a significant impact on the transcription of some genes downstream of EIN3. Thus, COP1 plays a crucial role in the opposite effects of ethylene on hypocotyl elongation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Sorghum is an energy crop with high biomass production potential and low input requirement. To identify markers linked to grain and biomass production traits, 43 SSR markers were mapped for association with tiller number and kernel weight using the sorghum mini core of 242 landraces. While kernel weight was evaluated in two environments, tiller number was evaluated in four environments. The number of SSR alleles was positively correlated with polymorphism information content for the markers. Association mapping found one marker (4-162) linked to kernel weight and two (40-1896 and 81-108) to tiller numbers. 4-162 and 40-1896 co-localized with previously mapped quantitative trait loci. Localized association mapping around 81-108 identified an amino-cyclopropane-carboxylate (ACC) oxidase gene as a candidate for tiller number. ACC oxidase is an ethylene forming enzyme and increased ethylene level has been shown to increase the number of tillers in the grass family. The results provide the groundwork to identify genes regulating kernel weight and tiller number in sorghum in the future. [ABSTRACT FROM AUTHOR]

E3 ubiquitin ligases select specific proteins for ubiquitin conjugation, and the modified proteins are commonly degraded through the 26S proteasome. XBAT32 is a RING-type E3 ligase involved in maintaining appropriate levels of ethylene. Previous work has suggested that XBAT32 modulates ethylene production by ubiquitinating two ethylene biosynthesis enzymes, ACS4 (type-II isoform) and ACS7 (type-III isoform). In Arabidopsis, conserved sequences within the C-terminal tail of type-I and -II 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) isoforms influence ubiquitin-dependent proteolysis. ACS7, the sole Arabidopsis type-III ACS, contains a truncated C-terminal tail that lacks all known regulatory sequences, which suggests that this isoform may not be subject to ubiquitin-mediated proteasomal degradation. Here we demonstrate in planta that ACS7 is turned over in a 26S proteasome-dependent manner and that degradation of ACS7 requires the E3 ligase XBAT32. Furthermore, the ethylene-related phenotypes that result from overexpression of ACS7 in wild-type plants are greatly exaggerated in xbat32-1, suggesting that XBAT32 is required to attenuate the effect of overexpression of ACS7. This observation is consistent with a role for XBAT32 in the ubiquitin-mediated degradation of ACS7. The dark-grown phenotype of xbat32-1 seedlings overexpressing ACS7 can be effectively rescued by aminoethoxyvinylglycine, an inhibitor of ACS activity. The degradation rate of ACS4 is also significantly slower in the absence of XBAT32, further implicating XBAT32 in the ubiquitin-mediated degradation of ACS4. Altogether, these results demonstrate that XBAT32 targets ethylene biosynthetic enzymes for proteasomal degradation to maintain appropriate levels of hormone production. [ABSTRACT FROM AUTHOR]

Maintenance of high bacterial population in the rhizosphere improves the efficiency of these organisms. This high bacterial population can be maintained by the application of enriched compost which supports their growth and activities. Thus integrated use of Rhizobium, plant growth promoting rhizobacteria (PGPR) containing 1-aminocyclopropane-1-carboxylate deaminase (ACC-deaminase) and P-enriched compost (PEC) could be highly effective for promoting growth, nodulation, and yield of lentil (Lens culinaris Medik.). A field study was conducted to evaluate the potential of Rhizobium, PGPR containing ACC-deaminase and PEC for promoting growth of lentil. For this study, the soil type was sandy clay loam soil having pH 7.6; EC (electrical conductivity) 2.8 dS m-1; organic matter (OM) 0.59%; total N 0.032%; available P 7.9 mg kg-1, and extractable K 129 mg kg-1. Treatments were replicated thrice, using randomized complete block (RCB) design. Results showed that the integrated use of R. leguminosarum with Pseudomonas spp. containing ACC-deaminase along with PEC was highly effective and caused up to 73.5, 73.9, 74.4, 67.5, 73.3, 65.8, 40.5, and 52.5% increase in fresh biomass, grain yield, straw yield, pods plant-1, nodule plant-1, nodule dry weight plant-1, 1000-grain weight, and N content in grain of lentil, respectively, as compared to respective control. It is concluded that integrated use of R. leguminosarum with Pseudomonas spp. having trait ACC-deaminase plus PEC would be an effective approach for better nodulation which consequently improved yield of lentil under natural conditions. [ABSTRACT FROM AUTHOR]

1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) produced by rhizobacteria could be remove the ethylene precursor and stimulate plant growth. Aim of the work was investigation on effect of rhizosphere bacteria Pseudomonas mendocina containing plasmid carrying gene encoding ACC deaminase on resistance of tomato plant to salinity. Amplification of acds gene in selected Pseudomonas was performed; the gene was cloned in Escherichia coli and was cloned subsequently in P. mendocina. Enzyme activity was determined in cloned Escherichia coli and cloned P. mendocina for confirmation of gene expression. Effect of bacterial ACC deaminase on resistance of tomato plants to NaCl was studied in Pot and Greenhouse. In pot experiment, tomato plant treated by cloned P. mendocina was compared with plants treated by P. mendocina (without plasmid) and control group. Salinity were established by adding 172 and 207 mM of NaCl to irrigated water. Greenhouse experiments were conducted in similar groups of bacteria in 207 mM of NaCl. Results obtained from pot experiment revealed that plants treated by cloned P. mendocina in 172 mM of NaCl was showed increasing content of growth than ones treated by P. mendocina and control as 11%, 18.4% growth for the shoot, 16.6%, 3.7% for roots and 9.6%, 27.5% for wet weight after five weeks, respectively. In 207 mM of NaCl, the results were as 14.9 %, 9.7% for shoot, 94.3%, 15.7% for roots and 96.4%, 50.6% for wet weight, respectively. In greenhouse experiment, results in same parameter in 207 mM of NaCl were revealed as 63.7%, 7 times for shoot, 2.8, 14 times for roots and 66.1%, 154 times for wet weight, respectively. We concluded that recombinant P. mendocina producing ACC deaminase by reduction of ethylene content of tomato plant in high salt concentrations could result in improvement of plant resistance to salinity. [ABSTRACT FROM AUTHOR]

1-Aminocyclopropane-1-carboxylate (ACC) synthase was rapidly induced in mesocarp tissue of Cucurbita maxima after wounding in the cut surface layer in 1 mm thickness (ca. 9 cells) (first layer) in both the enzyme activity and the levels of transcript. This led to a rapid accumulation of ACC and hence ethylene production. In the inside tissue (1–2 mm) (second layer), no significant induction of ACC synthase was observed, which resulted in a low level of ACC, although ethylene was evolved at a much lower rate than the first one. In contrast to ACC synthase, ACC oxidase was induced markedly in both the first and second layers and the development of its activity and the levels of mRNA remained high until later stages. It was considered that wound ethylene was closely associated with the development of ACC oxidase, since 2,5-norbornadiene (NBD), an inhibitor of ethylene action, substantially suppressed it. Phenylalanine ammonia-lyase (PAL) greatly increased in activity after wounding similarly to that of ACC synthase, in which increase in PAL activity occurred predominantly in the first layer. Induction of peroxidase activity after wounding had a close correlation in profile with that of ACC oxidase in that marked increases in the activity were observed in both the first and second layers and were strongly suppressed by NBD application. Four peroxidase isozymes were found by PAGE, among which a fraction was newly detected after wounding. [ABSTRACT FROM AUTHOR]

Two cDNA clones, pOS-ACO2 and pOS-ACO3, encoding 1-aminocyclopropane-1-carboxylate (ACC) oxidase were isolated from rice seedling cDNA library. pOS-ACO3 is a 1,299 bp full-length clone encoding 321 amino acids (Mr=35.9 kDa), while pOS-ACO2 is 1,072 bp long and is a partial cDNA clone encoding 314 amino acids. These two deduced amino acid sequences share 70% identity, and display a high degree of sequence identity (72-92%) with previously isolated pOS-ACOl of deepwater rice. The chromosomal location studies show that OS-ACO2 is positioned on the long arm of chromosome 9, while OSACO3 on the long arm of chromosome 2 of rice genome. A marked increase in the level of OS-ACO2 transcript was observed in IAA-treated etiolated rice seedlings, whereas the OS-ACO3 mRNA was greatly accumulated by ethylene treatment. Results of ethylene inhibitor studies indicated that auxin promotion of the OS-ACO2 transcription was not mediated through the action of auxin-induced ethylene. Thus, it appears that there are two groups of ACC oxidase transcripts in rice plants, either auxin-induced or ethylene-induced. The auxin-induced OS-ACO2 expression was partially inhibited by ethylene, while ethylene induction of OS-ACO3 transcription was completely blocked by auxin. These results indicate that the expression of ACC oxidase genes is regulated by complex hormonal networks in a gene specific manner in rice seedlings. Okadaic acid, a potent inhibitor of protein phosphatase, effectively suppressed the IAA induction of OS-ACO2 expression, suggesting that protein dephosphorylation plays a role in the induction of ACC oxidase by auxin. A scheme of the multiple regulatory pathways for the expression of ACC oxidase gene family by auxin, ethylene and protein phosphatase is presented. [ABSTRACT FROM AUTHOR]

We investigated the expression pattern of three 1-aminocyclopropane-1-carboxylate (ACC) synthase genes, CS-ACS1, CS-ACS2 and CS-ACS3 in cucumber (Cucumis sativus L.) fruit under CO2 stress. CO2 stress-induced ethylene production paralleled the accumulation of only CS ACS1 transcripts which disappeared upon withdrawal of CO2 Cycloheximide inhibited the CO2 stress-induced ethylene production but superinduced the accumulation of CS-ACS1 transcript. At higher concentrations, cycloheximide also induced the accumulation of CS-ACS2 and CS-ACS3 transcripts. In the presence of CO2 and cycloheximide, the accumulation of CS-ACS2 transcript occurred within 1 h, disappeared after 3 h and increased greatly upon withdrawal of CO2 Inhibitors of protein kinase and types 1 and 2A protein phosphatases which inhibited and stimulated, respectively, CO2 stress-induced ethylene production had little effect on the expression of these genes. The results presented here identify CS-ACS1 as the main ACC synthase gene responsible for the increased ethylene biosynthesis in cucumber fruit under CO2 stress and suggest that this gene is a primary response gene and its expression is under negative control since it is expressed by treatment with cycloheximide. The results further suggest that the regulation of CO2 stress-induced ethylene biosynthesis by reversible protein phosphorylation does not result from enhanced ACC synthase transcription. [ABSTRACT FROM AUTHOR]

Three full length 1-aminocyclopropane-l-carboxylate (ACC) oxidase cDNA clones (pNG-ACOl, 1,254 bp; pNG-ACO2,1,198 bp; and pNG-ACO3,1,053 bp) were isolated from the TMV-treated leaf cDNA library of Nicotiana glutinosa plant. They share a high degree of sequence identity (78–81%) throughout the coding regions but are divergent within the 3′-untranslated regions. The gene-specific probes were prepared using these regions to investigate the differential expression of the ACC oxidase gene family in various organs and in response to a multitude of biotic and abiotic stresses in N. glutinosa plants. All three genes were transcriptionally active displaying unique patterns of expression. Both the pNG-ACOl and pNG-ACO3 transcripts highly accumulated during the senescence of leaves, while the pNG-ACO2 mRNA was constitutively present. In addition, the NG-ACO1 and NG-ACO3 transcripts were predominantly found in roots whereas the NG-ACO2 mRNA was mainly in stems. Upon TMV infection, both NG-ACO1 and NG-ACO3 were markedly induced, but in mock treatment which has an effect of mild wounding, only the NG-ACO3 gene was induced. Furthermore, salicylic acid and CuSO4 treatments of leaves increased the level of NG-ACO1 and NG-ACO3 transcripts, while they did not affect the NG-ACO2 gene expression. Results showed that both the NG-ACO1 and NG-ACO3 genes were highly inducible by ethylene and methyl jasmonate treatments, with NG-ACO3 being more responsive. By contrast, NG-ACO2 did not respond to these growth regulators. Thus, it appears that there are two groups of ACC oxidase transcripts expressed in leaf tissue of N. glutinosa, either stress-induced or constitutive. The possible molecular mechanism of differential regulation of ACC oxidase gene expression and its physiological significance are discussed. [ABSTRACT FROM AUTHOR]

In cucumber (Cucumis sativus L.) plants, it has been reported that a high correlation existed between the evolution of ethylene from the apices and the development of female flowers. We isolated three distinct cDNA encoding 1-aminocyclopropane-1-carboxylate (ACC) synthase from cucumber. Among of these, only CS-ACS2 mRNA was detected at the apices of gynoecious cucumber in which female flowers were developing. The expression of the CS-ACS2 gene was examined in the apices of three cucumber cultivars (Rensei, Ougonmegami 2-gou and Shimoshirazu) by RNA gel blot analysis. In these cultivars, both the timing and the levels of expression of the CS-ACS2 transcript were correlated with the development of female flowers at the nodes. Furthermore, the timing of the induction of expression of the CS-ACS2 gene at the apex corresponded to that of the action of ethylene in induction of the first female flower at the apex of gynoecious cucumber plants. These results suggest that the development of female flowers might be regulated by the level of CS-ACS2 mRNA at the apex. [ABSTRACT FROM AUTHOR]

We have examined whether or not a positive feedback regulation of gene expression for l-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase also operates in ripening tomato (Lycopersicon esculentum) fruit during the burst of ethylene production. Two cDNA fragments for ACC synthase and one for ACC oxidase were cloned with high homology to already known genes involved in ethylene biosynthesis in ripening tomato fruit. Accumulation of mRNAs which hybridize to these cDNA probes were induced in mature green fruit within two days by treatment with propylene. In the fruit ripened from the turning stage, red color development, ethylene production, ACC content, and activities of ACC synthase and ACC oxidase increased as maturity progressed. The abundance of two ACC synthase and one ACC oxidase mRNAs in the fruit increased from the turning to pink stage and were followed by a slight decline towards the red stage. These increases in mRNAs abundance with ripening were prevented to a large extent by treatment with the ethylene action inhibitor, 1-methylcyclopropene (MCP). This was most pronounced in the fruit treated with MCP at the turning stage, in which the accumulation of ACC synthase and ACC oxidase transcripts was almost completely eliminated in the first two d, precisely the same stage at which the control fruit had the greatest level of each mRNA accumulation. The inhibition of transcript accumulation recovered to the control level within two to four d. MCP also decreased ethylene biosynthetic activity, although this decrease did not reflect the reduction in the mRNAs accumulation. These results suggest that a strong positive feedback regulation is involved in ethylene biosynthesis at the gene transcriptional level in tomato fruit, even at the stage with a burst of ethylene production. [ABSTRACT FROM AUTHOR]

We have isolated four cDNA clones of ACC synthase from etiolated mungbean seedlings treated with auxin. PVR-ACS2, pVR-ACS3 and pVR-ACS6 contained the same sequences as the previously reported DNA fragments, pMAC2, pMAC3 (Botella et al. 1992b) and pMBAl (Kim et al. 1992), respectively. pVR-ACSl was identical with pAIM-1 (Botella et al. 1992a).VR-ACS6 was specifically induced in response to the auxin signal. The IAA-induction of VR-ACS6 was very rapid (within 30 min) and insensitive to cycloheximide treatment at concentrations up to 100 μM. Significant accumulation of VR-ACS6 mRNA was detected at 1 μM.IAA.The IAA-induced expression of VR-ACS6 was suppressed by ABA and ethylene, but enhanced by BA. These characteristics of VR-ACS6 expression were well correlated with the physiological data of auxin-induced ethylene production in mungbean hypocotyls.VR-ACS1 was strongly induced by cycloheximide, but was found to be not auxin-specific. Inhibitors of either ethylene biosynthesis (AOA) or action (NBD) increased the basal level of VR-ACS1 mRNA. [ABSTRACT FROM AUTHOR]

Partial submergence greatly stimulates internodal growth in deepwater rice (Oryza sativa L.). Previous work has shown that the effect of submergence is, at least in part, mediated by ethylene, which accumulates in the air spaces of submerged internodes. To investigate the expression of the genes encoding ethylene biosynthetic enzymes during accelerated growth of deepwater rice, we cloned a 1-aminocyclopropane- 1-carboxylate (ACC) oxidase cDNA (OSACO1) from internodes of submerged plants and measured the activity of the enzyme in tissue extracts with an improved assay. We found an increase in ACC oxidase mRNA levels and enzyme activity after 4 to 24 h of submergence. Thus, it is likely that ethylene biosynthesis in internodes of deepwater rice is controlled, at least in part, at the level of ACC oxidase. [ABSTRACT FROM AUTHOR]

Background: Fig fruit are highly perishable at the tree-ripe (TR) stage. Commercial-ripe (CR) fruit, which are harvested before the TR stage for their postharvest transportability and shelf-life advantage, are inferior to TR fruit in size, color and sugar content. The succulent urn-shaped receptacle, serving as the protective structure and edible part of the fruit, determines fruit quality. Quantitative iTRAQ and RNA-Seq were performed to reveal the differential proteomic and transcriptomic traits of the receptacle at the two harvest stages. Results: We identified 1226 proteins, of which 84 differentially abundant proteins (DAPs) were recruited by criteria of abundance fold-change (FC) ≥1.3 and p < 0.05 in the TR/CR receptacle proteomic analysis. In addition, 2087 differentially expressed genes (DEGs) were screened by ≥2-fold expression change: 1274 were upregulated and 813 were downregulated in the TR vs. CR transcriptomic analysis. Ficin was the most abundant soluble protein in the fig receptacle. Sucrose synthase, sucrose-phosphate synthase and hexokinase were all actively upregulated at both the protein and transcriptional levels. Endoglucanase, expansin, beta-galactosidase, pectin esterase and aquaporins were upregulated from the CR to TR stage at the protein level. In hormonal synthesis and signaling pathways, high protein and transcriptional levels of aminocyclopropane-1-carboxylate oxidase were identified, together with a few diversely expressed ethylene-response factors, indicating the potential leading role of ethylene in the ripening process of fig receptacle, which has been recently reported as a non-climacteric tissue. Conclusions: We present the first delineation of intra- and inter-omic changes in the expression of specific proteins and genes of TR vs. CR fig receptacle, providing valuable candidates for further study of fruit-quality formation control and fig cultivar innovation to accommodate market demand. [ABSTRACT FROM AUTHOR]