Expression Of Ethylene Biosynthetic Genes Research Articles

Expression of ethylene biosynthetic genes during flower senescence and in response to ethephon and silver nitrate treatments in Osmanthus fragrans.

Sweet osmanthus (Osmanthus fragrans) is an ornamental evergreen tree species in China, whose flowers are sensitive to ethylene. The synthesis of ethylene is controlled by key enzymes and restriction enzymes, 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO), which are encoded by multigene families. However, the key synthase responsible for ethylene regulation in O. fragrans is still unknown. This study aims to screen the key ethylene synthase genes of sweet osmanthus flowers in response to ethylene regulation. In this study, we used the ACO and ACS sequences of Arabidopsis thaliana to search for homologous genes in the O. fragrans petal transcriptome database. These genes were also analyzed bioinformatically. Finally, the expression levels of O. fragrans were compared before and after senescence, as well as after ethephon and silver nitrate treatments. The results showed that there are five ACO genes and one ACS gene in O. fragrans transcriptome database, and the phylogenetic tree revealed that the proteins encoded by these genes had high homology to the ACS and ACO proteins in plants. Sequence alignment shows that the OfACO1-5 proteins have the 2OG-Fe(II) oxygenase domain, while OfACS1 contains seven conserved domains, as well as conserved amino acids in transaminases and glutamate residues related to substrate specificity. Expression analysis revealed that the expression levels of OfACS1 and OfACO1-5 were significantly higher at the early senescence stage compared to the full flowering stage. The transcripts of the OfACS1, OfACO2, and OfACO5 genes were upregulated by treatment with ethephon. However, out of these three genes, only OfACO2 was significantly downregulated by treatment with AgNO3. Our study found that OfACO2 is an important synthase gene in response to ethylene regulation in sweet osmanthus, which would provide valuable data for further investigation into the mechanisms of ethylene-induced senescence in sweet osmanthus flowers.

Differences in ethylene sensitivity, expression of ethylene biosynthetic genes and vase life among carnation varieties

Differences in ethylene sensitivity, expression of ethylene biosynthetic genes and vase life among carnation varieties

Suppression of the target of rapamycin kinase accelerates tomato fruit ripening through reprogramming the transcription profile and promoting ethylene biosynthesis.

Tomato fruit ripening is a unique process of nutritional and energy metabolism. Target of rapamycin (TOR), a conserved serine/threonine protein kinase in eukaryotes, controls cell growth and metabolism by integrating nutrient, energy, and hormone signals. However, it remains unclear whether TOR participates in the modulation of tomato fruit ripening. Here, we showed that the manipulation of SlTOR by chemical or genetic methods greatly alters the process of tomato fruit maturation. Expression pattern analysis revealed that the transcripts of SlTOR declined as fruit ripening progressed. Moreover, suppression of SlTOR by TOR inhibitor AZD8055 or knock down of its transcripts by inducible RNA interference, accelerated fruit ripening, and led to overall effects on fruit maturity, including changes in colour and metabolism, fruit softening, and expression of ripening-related genes. Genome-wide transcription analysis indicated that silencing SlTOR reprogrammed the transcript profile associated with ripening, including cell wall and phytohormone pathways, elevated the expression of ethylene biosynthetic genes, and further promoted ethylene production. In contrast, the ethylene action inhibitor 1-MCP efficiently blocked fruit maturation, even following SlTOR inhibition. These results suggest that accelerated fruit ripening caused by SlTOR inhibition depends on ethylene, and that SlTOR may function as a regulator in ethylene metabolism.

A Glucuronic Acid-Producing Endophyte Pseudomonas sp. MCS15 Reduces Cadmium Uptake in Rice by Inhibition of Ethylene Biosynthesis.

Dynamic regulation of phytohormone levels is pivotal for plant adaptation to harmful conditions. It is increasingly evidenced that endophytic bacteria can regulate plant hormone levels to help their hosts counteract adverse effects imposed by abiotic and biotic stresses, but the mechanisms underlying the endophyte-induced stress resistance of plants remain largely elusive. In this study, a glucuronic acid-producing endophyte Pseudomonas sp. MCS15 alleviated cadmium (Cd) toxicity in rice plants. Inoculation with MCS15 significantly inhibited the expression of ethylene biosynthetic genes including OsACO3, OsACO4, OsACO5, OsACS2, and OsACS5 and thus reduced the content of ethylene in rice roots. In addition, the expression of iron uptake-related genes including OsIRT1, OsIRT2, OsNAS1, OsNAS2 and OsYSL15 was significantly downregulated in the MCS15-inoculated roots under Cd stress. Similarly, glucuronic acid treatment also remarkably inhibited root uptake of Cd and reduced the production of ethylene. However, treatment with 1-aminocyclopropyl carboxylic acid (ACC), a precursor of ethylene, almost abolished the MCS15 or glucuronic acid-induced inhibition of Cd accumulation in rice plants. Conversely, treatment with aminoethoxyvinyl glycine (AVG), an inhibitor of ethylene biosynthesis, markedly reduced the Cd accumulation in plants. Taken together, our results revealed that the endophytic bacteria MCS15-secreted glucuronic acid inhibited the biosynthesis of ethylene and thus weakened iron uptake-related systems in rice roots, which contributed to preventing the Cd accumulation.

The LcKNAT1-LcEIL2/3 Regulatory Module Is Involved in Fruitlet Abscission in Litchi.

Large and premature organ abscission may limit the industrial development of fruit crops by causing serious economic losses. It is well accepted that ethylene (ET) is a strong inducer of organ abscission in plants. However, the mechanisms underlying the control of organ abscission by ET are largely unknown. We previously revealed that LcKNAT1, a KNOTTED-LIKE FROM ARABIDOPSIS THALIANA1 (KNAT1)-like protein, acted as a negative regulator in control of fruitlet abscission through suppressing the expression of ET biosynthetic genes in litchi. In this study, we further reported that LcKNAT1 could also directly repress the transcription of LcEIL2 and LcEIL3, two ETHYLENE INSENSITIVE 3-like (EIL) homologs in litchi, which functioned as positive regulators in ET-activated fruitlet abscission by directly promoting the expression of genes responsible for ET biosynthesis and cell wall degradation. The expression level of LcKNAT1 was downregulated, while LcEIL2/3 was upregulated at the abscission zone (AZ) accompanying the fruitlet abscission in litchi. The results of electrophoretic mobility shift assays (EMSAs) and transient expression showed that LcKNAT1 could directly bind to the promoters of LcEIL2 and LcEIL3 and repress their expression. Furthermore, the genetic cross demonstrated that the β-glucuronidase (GUS) expression driven by the promoters of LcEIL2 or LcEIL3 at the floral AZ was obviously suppressed by LcKNAT1 under stable transformation in Arabidopsis. Taken together, our findings suggest that the LcKNAT1-LcEIL2/3 regulatory module is likely involved in the fruitlet abscission in litchi, and we propose that LcKNAT1 could suppress both ET biosynthesis and signaling to regulate litchi fruit abscission.

Transcriptional and post-transcriptional regulation of ethylene biosynthesis by exogenous acetylsalicylic acid in kiwifruit.

Levels of ethylene, implicated in the induction of fruit ripening in a diverse array of plants, are influenced by genetic and environmental factors, such as other plant hormones. Among these, salicylic acid (SA) and its derivative, acetylsalicylic acid (ASA), have been demonstrated to inhibit ethylene biosynthesis in fruit, yet the underlying regulatory mechanisms remain elusive. Here, we showed that treatment with exogenous ASA dramatically reduced ethylene production, as well as activities of ACC synthase (ACS) and ACC oxidase (ACO), in kiwifruit tissues. Comparative transcriptome analysis indicated the differential expression of ethylene biosynthetic genes (AdACS1/2 and AdACO5). A screen of transcription factors indicated that AdERF105L and AdWRKY29 were ASA-responsive regulators of AdACS1/2 and AdACO5, respectively. In addition to these genes, AdACS3 and AdACO3 were abundantly expressed in both ASA-treated and control tissues. AdACS3 protein was phosphorylated and stabilized by AdMPK16, a mitogen-activated protein kinase, while AdACO3 activity was enhanced by AdAP, an aspartic peptidase. Exogenous ASA downregulated AdMPK16 and AdAP, thereby influencing ethylene biosynthesis at a post-transcriptional level. These findings led us to propose a multidimensional system for inhibition of ethylene biosynthesis by ASA, inducing differential expression of some ethylene biosynthesis genes, as well as differential effects on protein activity on other targets.

UV RESISTANCE LOCUS8 mediates ultraviolet-B-induced stomatal closure in an ethylene-dependent manner

Although the UV RESISTANCE LOCUS 8 (UVR8)-CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1)-ELONGATED HYPOCOTYL5 (HY5) signaling pathway, ethylene, hydrogen peroxide (H2O2), and nitric oxide (NO) all participate in ultraviolet-B (UV-B)-triggered stomatal closing, their interrelationship is not clear. Here, we found that UV-B-induced the expression of ethylene biosynthetic genes, production of ethylene, H2O2, and NO, and stomata closing were impaired in uvr8, cop1, and hy5 mutants. UV-B-induced NO production and stomata closing were also defective in mutants for ETHYLENE RESPONSE 1 (ETR1), ETHYLENE INSENSITIVE 2 (EIN2), and EIN3, but UV-B-triggered H2O2 generation was only inhibited in etr1. In either the absence or presence of UV-B, ethylene triggered H2O2 production but not NO generation and stomatal closure in cop1 and hy5, and stomata closing in cop1 and hy5 was induced by NO but not H2O2. Moreover, NO production and stomatal closure were constitutively caused by over-expression of COP1 or HY5 in ein2 and ein3, but not by over-expression of EIN2 or EIN3 in cop1 and hy5. Our data indicate that the UVR8-COP1-HY5 signaling module mediates UV-B-induced ethylene production, ethylene is then perceived by ETR1 to induce H2O2 synthesis. H2O2 induces NO generation and subsequent stomata closing via an EIN2, EIN3, COP1, and HY5-dependent pathway(s).

Auxin-activated MdARF5 induces the expression of ethylene biosynthetic genes to initiate apple fruit ripening.

Summary The gaseous plant hormone ethylene induces the ripening of climacteric fruit, including apple (Malus domestica). Another phytohormone, auxin, is known to promote ethylene production in many horticultural crops, but the regulatory mechanism remains unclear.Here, we found that auxin application induces ethylene production in apple fruit before the stage of commercial harvest, when they are not otherwise capable of ripening naturally.The expression of MdARF5, a member of the auxin response factor transcription factor (TF) family involved in the auxin signaling pathway, was enhanced by treatment with the synthetic auxin naphthaleneacetic acid (NAA). Further studies revealed that MdARF5 binds to the promoter of MdERF2, encoding a TF in the ethylene signaling pathway, as well as the promoters of two 1‐aminocyclopropane‐1‐carboxylic acid synthase (ACS) genes (MdACS3a and MdACS1) and an ACC oxidase (ACO) gene, MdACO1, all of which encode key steps in ethylene biosynthesis, thereby inducing their expression. We also observed that auxin‐induced ethylene production was dependent on the methylation of the MdACS3a promoter.Our findings reveal that auxin induces ethylene biosynthesis in apple fruit through activation of MdARF5 expression.

Blue light postpones senescence of carnation flowers through regulation of ethylene and abscisic acid pathway-related genes

Endogenous signals in response to exogenous factors determine the senescence of flowers. Interactions among phytohormones especially abscisic acid (ABA) and ethylene are the major determinant of the senescence. In the present study, complex expression patterns of the genes related to ABA and ethylene as endogenous signals were investigated on cut carnations (Dianthus caryophyllus L.) that were exposed to different light spectra. Expression of ethylene biosynthetic (DcACS and DcACO), and signaling (DcETR and DcEin2) genes and also genes involved in ABA biosynthesis (DcZEP1 and DcNCED1), transport (DcABCG25 and DcABCG40) and catabolism (DcCYP707A1) were evaluated in petals of carnations exposed to three light spectra [white, blue and red]. Lowest relative membrane permeability (RMP) was detected in flowers that exposed to Blue light (BLFs), as a consequence, the longest vase life was found in BLFs. The Red and White lights markedly accelerated flower senescence and increased expression of DcACS and DcACO on day 6 and 10 of vase life assessment respectively; while Blue light inhibited the expression of ethylene biosynthetic genes. Expression of the genes involved in the production and transport of ABA and in signal transduction of ethylene was elevated during vase life of flowers irrespective of exposure to different light spectra. In conclusion, Blue light can be an effective environmental factor to extend the vase life of carnation flowers by delaying the petal senescence through down-regulation of ethylene biosynthetic genes and up-regulation of ABA biosynthetic genes.

Ethylene mediates salicylic-acid-induced stomatal closure by controlling reactive oxygen species and nitric oxide production in Arabidopsis

Both salicylic acid (SA) and ethylene induce stomatal closure and positively regulate stomatal immunity, but their interactions in guard cell signaling are unclear. Here, we observed that SA induced the expression of ethylene biosynthetic genes; the production of ethylene, reactive oxygen species (ROS) and nitric oxide (NO); and stomatal closure in Arabidopsis thaliana. However, SA-induced stomatal closure was inhibited by an ethylene biosynthetic inhibitor and mutations in ethylene biosynthetic genes, ethylene-signaling genes [RESPONSE TO ANTAGONIST 1 (RAN1), ETHYLENE RESPONSE 1 (ETR1), ETHYLENE INSENSITIVE 2 (EIN2), EIN3 and ARABIDOPSIS RESPONSE REGULATOR 2 (ARR2)], NADPH oxidase genes [ATRBOHD and ATRBOHF], and nitrate reductase genes (NIA1 and NIA2). Furthermore, SA-triggered ROS production in guard cells was impaired in ran1, etr1, AtrbohD and AtrbohF, but not in ein2, ein3 or arr2. SA-triggered NO production was impaired in all ethylene-signaling mutants tested and in nia1 and nia2. The stomata of mutants for CONSTITUTIVE TRIPLE RESPONSE1 (CTR1) showed constitutive ROS and NO production and closure. These results indicate that ethylene mediates SA-induced stomatal closure by activating ATRBOHD/F-mediated ROS synthesis in an RAN1-, ETR1- and CTR1-dependent manner. This in turn induces NIA1/2-mediated NO production and subsequent stomatal closure via the ETR1, EIN2, EIN3 and ARR2-dependent pathway(s).

Silicon alleviates long-term copper toxicity and influences gene expression in Nicotiana tabacum

Silicon (Si) is beneficial for plant growth and aids in stress tolerance. In this study, the effects of Si on long-term copper (Cu) toxicity in the low Si accumulator Nicotiana tabacum were evaluated. Silicon supplementation alleviated growth inhibition in roots and shoots of N. tabacum exposed to Cu toxicity. Alleviation of Cu toxicity correlated with increased Si accumulation in roots and leaves, suggesting N. tabacum contains a stress-regulated mechanism for Si transport. Root Cu concentration decreased in Si-supplemented plants exposed to Cu toxicity. Interestingly, Copper Transporter 1 (COPT1) expression decreased in roots of Si-supplemented plants exposed to Cu toxicity, which may contribute to Cu uptake reduction. Decreases in ethylene (ET) biosynthetic gene expression were previously implicated in Si-mediated stress alleviation. In the present study, Si-mediated alleviation of Cu toxicity corresponded with increased ET biosynthetic gene expression.

Mulberry EIL3 confers salt and drought tolerances and modulates ethylene biosynthetic gene expression.

Ethylene regulates plant abiotic stress responses and tolerances, and ethylene-insensitive3 (EIN3)/EIN3-like (EIL) proteins are the key components of ethylene signal transduction. Although the functions of EIN3/EIL proteins in response to abiotic stresses have been investigated in model plants, little is known in non-model plants, including mulberry (Morus L.), which is an economically important perennial woody plant. We functionally characterized a gene encoding an EIN3-like protein from mulberry, designated as MnEIL3. A quantitative real-time PCR analysis demonstrated that the expression of MnEIL3 could be induced in roots and shoot by salt and drought stresses. Arabidopsis overexpressing MnEIL3 exhibited an enhanced tolerance to salt and drought stresses. MnEIL3 overexpression in Arabidopsis significantly upregulated the transcript abundances of ethylene biosynthetic genes. Furthermore, MnEIL3 enhanced the activities of the MnACO1 and MnACS1 promoters, which respond to salt and drought stresses. Thus, MnEIL3 may play important roles in tolerance to abiotic stresses and the expression of ethylene biosynthetic genes.

A patatin-like protein synergistically regulated by jasmonate and ethylene signaling pathways plays a negative role in Nicotiana attenuata resistance to Alternaria alternata

Although patatin was initially identified as a major storage protein in potato tubers, patatin-like proteins (PLPs) have been recently reported to be widely present in many plant species and shown to be involved in plant-pathogen interactions. However, it is not clear whether PLPs are involved in Nicotiana attenuata resistance against the necrotrophic fungal pathogen, Alternaria alternata. In this study we identified a NaPLP gene, whose expression was highly elicited by A. alternata inoculation. Silencing NaPLP enhanced N. attenuata resistance to A. alternata, which was associated with higher induction levels of JA and ethylene biosynthetic genes, NaACS1, NaACO1 and NaLOX3. The induction of NaPLP expression by the fungus was abolished in JA-deficient plants and significantly reduced in ethylene-insensitive plants. In addition, NaPLP transcripts were highly induced by exogenous treatment with either methyl jasmonate (MeJA) or ethephon. Co-treatment with MeJA and ethephon led to a much higher induction level of NaPLP transcripts, and this synergistic induction was largely dependent on endogenous JA and ethylene signaling pathways. Thus, we conclude that the NaPLP gene is elicited by A. alternata via JA and ethylene signaling pathways in a synergistic way; however, unlike other JA- and ethylene-induced defense genes, NaPLP negatively affects plant resistance to the fungus likely by suppressing JA and ethylene biosynthetic gene expression.

The regulatory module MdPUB29‐MdbHLH3 connects ethylene biosynthesis with fruit quality in apple

The plant hormone ethylene is critical for climacteric fruit ripening, while glucose and anthocyanins determine the fruit quality of climacteric fruits such as apple. Understanding the exact molecular mechanism for this process is important for elucidating the interconnection of ethylene and fruit quality. Overexpression of apple MdbHLH3 gene, an anthocyanin-related basic helix-loop-helix transcription factor (bHLH TF) gene, promotes ethylene production, and transgenic apple plantlets and trees exhibit ethylene-related root developmental abnormalities, premature leaf senescence, and fruit ripening. Biochemical analyses demonstrate that MdbHLH3 binds to the promoters of three genes that are involved in ethylene biosynthesis, including MdACO1, MdACS1, and MdACS5A, activating their transcriptional expression, thereby promoting ethylene biosynthesis. High glucose-inhibited U-box-type E3 ubiquitin ligase MdPUB29, the ortholog of Arabidopsis AtPUB29 in apple, influences the expression of ethylene biosynthetic genes and ethylene production by direct ubiquitination of the MdbHLH3 protein. Our findings provide new insights into the ubiquitination of MdbHLH3 by glucose-inhibited ubiquitin E3 ligase MdPUB29 in the regulation of ethylene biosynthesis as well as indicate that the regulatory module MdPUB29-MdbHLH3 connects ethylene biosynthesis with fruit quality in apple.

Comparative Physiological, Biochemical, and Genetic Responses to Prolonged Waterlogging Stress in Okra and Maize Given Exogenous Ethylene Priming.

Waterlogging is an environmental challenge affecting crops worldwide. Ethylene induces the expression of genes linked to important agronomic traits under waterlogged conditions. The ability of okra (Abelmoschus esculentus L. Moench.) and maize (Zea mays L.) given exogenous ethylene priming to tolerate prolonged waterlogged conditions was investigated in this study. The investigation was carried out as field experiments using 3 week-old plants grouped into four treatments; control, waterlogged plants, ethylene priming of plants before waterlogging, and ethylene priming of plants after waterlogging. Different growth parameters were recorded. Soil chemical and bacterial analyses were performed. The activity and gene expression of antioxidant enzymes were studied. The ethylene biosynthetic genes expression analysis and root anatomy of surviving okra plants were also carried out. Results revealed that okra and maize plants showed increase in their height under waterlogged conditions. Ethylene priming and waterlogged conditions induced early production of adventitious roots in okra and maize. Maize survival lasted between 5 and 9 weeks under waterlogging without reaching the flowering stage. However, okra survived up to 15 weeks under waterlogging producing flower buds and fruits in all treatments. Variable changes were also recorded for total soluble phenolics of soil. Cross sections of waterlogged okra roots showed the formation of a dark peripheral layer and numerous large aerenchyma cells which may have assisted in trapping oxygen required for survival. The activity and gene expression levels of antioxidant enzymes were studied and showed higher increases in the root and leaf tissues of okra and maize subjected to both waterlogging and ethylene priming, as compared to control or waterlogged condition. Quantitative RT-PCR analysis also showed that the ethylene biosynthetic gene expression levels in all okra and maize tissues were up-regulated and showed much higher levels under ethylene-treated waterlogged conditions than those expressed under control or waterlogged conditions at all time points. These results indicate that okra and maize tissues respond to the conditions of waterlogging and exogenous ethylene priming by inducing their ethylene biosynthetic genes expression in order to enhance ethylene production and tolerate the prolonged waterlogging stress. In conclusion, this study revealed that exogenously generated ethylene gas as a priming treatment before or after waterlogging could enhance waterlogging tolerance in maize and okra crops.

Silencing of histone deacetylase SlHDT3 delays fruit ripening and suppresses carotenoid accumulation in tomato

The acetylation levels of histones on lysine residues are regulated by histone acetyltransferases and histone deacetylases, which play an important but understudied role in the control of gene expression in plants. There is an increasing research focus on histone deacetylation in crops, but to date, there is little information regarding tomato. With the aim of characterizing the tomato HD2 family of histone deacetylases, an RNA interference (RNAi) expression vector of SlHDT3 was constructed and transformed into tomato plants. The time of fruit ripening was delayed and the shelf life of the fruit was prolonged in SlHDT3 RNAi lines. The accumulation of carotenoid was decreased by altering of the carotenoid pathway flux. Ethylene content was also reduced and expression of ethylene biosynthetic genes (ACS2, ACS4 and ACO1, ACO3) and ripening-associated genes (RIN, E4, E8, PG, Pti4 and LOXB) was significantly down-regulated in SlHDT3 RNAi lines. The expression of genes involved in fruit cell wall metabolism (HEX, MAN, TBG4, XTH5 and XYL) was inhibited compared with wild type. These results indicate that SlHDT3 functions as a positive regulator of fruit ripening by affecting ethylene synthesis and carotenoid accumulation and that SlHDT3 lies upstream of SlMADS-RIN in the fruit ripening regulatory network.

A histone deacetylase gene, SlHDA3, acts as a negative regulator of fruit ripening and carotenoid accumulation.

SlHDA3 functions as an inhibitor and regulates tomato fruit ripening and carotenoid accumulation. Post-translational modifications, including histones acetylation, play a pivotal role in the changes of chromatin structure dynamic modulation and gene activity. The regulation of histone acetylation is achieved by the action of histone acetyltransferases and deacetylases, which play crucial roles in the regulation of transcription activation. There is an increasing research focus on histone deacetylation in crops, but the role of histone deacetylase genes (HDACs) in tomato has not been elucidated. With the aim of characterizing the tomato RPD3/HDA1 family histone deacetylase genes, SlHDA3 was isolated and its RNA interference (RNAi) lines was obtained. The fruit of SlHDA3 RNAi lines exhibited accelerated ripening process along with short shelf life characteristics. The accumulation of carotenoid was increased due to the alteration of the carotenoid pathway flux. Climacteric ethylene production also stimulated along with significantly up-regulated expression of ethylene biosynthetic genes (ACS2, ACS4, ACO1 and ACO3) and fruit ripening-associated genes (RIN, E4, E8, PG, Pti4, LOXB, Cnr and TAGL1) in SlHDA3 RNAi lines. Besides, fruit cell wall metabolism-associated genes (HEX, MAN, TBG4, XTH5 and XYL) were enhanced in transgenic lines. Relative to wild type (WT) plants, SlHDA3 RNAi seedlings displayed shorter hypocotyls and more sensitivity to ACC (1-aminocyclopropane-1-carboxylate). These results indicated that SlHDA3 is involved in the regulation of fruit ripening by affecting ethylene biosynthesis and carotenoid accumulation.

Relationships between the longevity, water relations, ethylene sensitivity, and gene expression of cut roses

Cut roses (Rosa hybrida L.) have been classified as ethylene-sensitive, but the nature of the ethylene sensitivity changes in these flowers has not been well characterized. In the present study, the relationships between vase life, ethylene sensitivity, and expression of ethylene biosynthetic genes were determined in 33 cultivars. Despite the same growing season and identical postharvest environments, the vase life of cultivars ranged from 5.5 to 15.5 d. Ethylene primarily accelerated petal wilting, which is the main factor shortening the longevity in long vase life cultivars. qRT-PCR analysis revealed that among five ethylene synthesis genes, RhACO1 showed the most distinct expression patterns in response to ethylene. ACS genes were differentially expressed in response to ethylene, and only RhACS2 and RhACS4 are induced by ethylene and during flower senescence. Correlation analysis revealed that the major factors that terminate vase life vary depending on flower sensitivity to ethylene. The vase life of ethylene-sensitive cultivars was strongly related to RhACO1 transcript level, which was induced by ethylene exposure. In contrast, the vase life of ethylene-insensitive cultivars was mostly reduced due to an early failure of water relations, as a consequence of the decreased hydraulic conductance of stems and water loss via transpiration. The current study revealed that postharvest treatments to improve the vase life should be differently applied based on the ethylene sensitivity of each cultivar.

Transcriptomic and metabolic analyses provide new insights into chilling injury in peach fruit.

Low temperature conditioning (LTC) alleviates peach fruit chilling injury but the underlying molecular basis is poorly understood. Here, changes in transcriptome, ethylene production, flesh softening, internal browning and membrane lipids were compared in fruit maintained in constant 0°C and LTC (pre-storage at 8°C for 5d before storage at 0°C). Low temperature conditioning resulted in a higher rate of ethylene production and a more rapid flesh softening as a result of higher expression of ethylene biosynthetic genes and a series of cell wall hydrolases. Reduced internal browning of fruit was observed in LTC, with lower transcript levels of polyphenol oxidase and peroxidase, but higher lipoxygenase. Low temperature conditioning fruit also showed enhanced fatty acid content, increased desaturation, higher levels of phospholipids and a preferential biosynthesis of glucosylceramide. Genes encoding cell wall hydrolases and lipid metabolism enzymes were coexpressed with differentially expressed ethylene response factors (ERFs) and contained ERF binding elements in their promoters. In conclusion, LTC is a special case of cold acclimation which increases ethylene production and, operating through ERFs, promotes both softening and changes in lipid composition and desaturation, which may modulate membrane stability, reducing browning and contributing to alleviation of peach fruit chilling injury.

Expression analysis by RT-PCR of genes involved in ethylene synthesis and signal transduction in miniature roses

Ethylene is one of many factors that affect the quality, appearance and longevity of miniature roses. In this study RT-PCR is used to compare the expression of ethylene biosynthetic genes and ethylene signal transduction genes at different stages of flower development in the two cultivars ‘Vanilla’ and ‘Lavender’, which show low and high ethylene-sensitivity, respectively, and their F1 offspring. The genes for the ethylene receptors RhETR1, RhETR2 and RhETR3, the genes for the receptor-associated signaling proteins RhCTR1 and RhCTR2, the genes for the transcription factors RhEIN3 and RhEIL, and the genes for the ACC synthases RhACS1 and RhACS2 each had an expression pattern that varied between the tested plants and tissues, but could not be correlated with the ethylene sensitivity of the plants. RhETR1, RhETR2, RhETR3 and RhEIN3 were, e.g., expressed more in ‘Vanilla’ than in ‘Lavender’ in most, but not all, of the investigated tissues, but were in general not expressed more in progeny with low sensitivity than in progeny with high sensitivity. No expression was detected for the 4 other genes that were investigated, i.e. genes for RhETR4, RhACS3, RhACS4 and RhACS5. It is concluded that the precise transcriptional activities of the tested genes do not appear to be crucial in determining the ethylene sensitivity of miniature roses. It therefore appears likely that transcription of other genes involved in ethylene signal transduction, posttranscriptional or posttranslational control, or crosstalk with other signal transduction pathways may be important for the degree of ethylene sensitivity of miniature roses.