Ethylene Biosynthesis Genes Research Articles

The miR156/SPL12 module orchestrates fruit colour change through directly regulating ethylene production pathway in blueberry.

Colour change is an important event during fruit ripening in blueberry. It is well known that miR156/SPLs act as regulatory modules mediating anthocyanin biosynthesis and ethylene plays critical roles during colour change, but the intrinsic connections between the two pathways remain poorly understood. Previously, we demonstrated that blueberry VcMIR156a/VcSPL12 affects the accumulation of anthocyanins and chlorophylls in tomato and Arabidopsis. In this study, we first showed that VcMIR156a overexpression in blueberry led to enhanced anthocyanin biosynthesis, decreased chlorophyll accumulation, and, intriguingly, concomitant elevation in the expression of ethylene biosynthesis genes and the level of the ethylene precursor ACC. Conversely, VcSPL12 enhanced chlorophyll accumulation and suppressed anthocyanin biosynthesis and ACC synthesis in fruits. Moreover, the treatment with ethylene substitutes and inhibitors attenuated the effects of VcMIR156a and VcSPL12 on pigment accumulation. Protein-DNA interaction assays indicated that VcSPL12 could specifically bind to the promoters and inhibit the activities of the ethylene biosynthetic genes VcACS1 and VcACO6. Collectively, our results show that VcMIR156a/VcSPL12 alters ethylene production through targeting VcACS1 and VcACO6, therefore governing fruit colour change. Additionally, VcSPL12 may directly interact with the promoter region of the chlorophyll biosynthetic gene VcDVR, thereby activating its expression. These findings established an intrinsic connection between the miR156/SPL regulatory module and ethylene pathway.

MaMADS1-MaNAC083 transcriptional regulatory cascade regulates ethylene biosynthesis during banana fruit ripening.

The hormone ethylene is crucial in the regulation of ripening in climacteric fruit, such as bananas. The transcriptional regulation of ethylene biosynthesis throughout banana fruit ripening has received much study, but the cascaded transcriptional machinery of upstream transcriptional regulators implicated in the ethylene biosynthesis pathway is still poorly understood. Here we report that ethylene biosynthesis genes, including MaACS1, MaACO1, MaACO4, MaACO5, and MaACO8, were upregulated in ripening bananas. NAC (NAM, ATAF, CUC) transcription factor, MaNAC083, a ripening and ethylene-inhibited gene, was discovered as a potential binding protein to the MaACS1 promoter by yeast one-hybrid screening. Further in vitro and in vivo experiments indicated that MaNAC083 bound directly to promoters of the five ethylene biosynthesis genes, thereby transcriptionally repressing their expression, which was further verified by transient overexpression experiments, where ethylene production was inhibited through MaNAC083-modulated transcriptional repression of ethylene biosynthesis genes in banana fruits. Strikingly, MaMADS1, a ripening-induced MADS (MCM1, AGAMOUS, DEFICIENS, SRF4) transcription factor, was found to directly repress the expression of MaNAC083, inhibiting trans-repression of MaNAC083 to ethylene biosynthesis genes, thereby attenuating MaNAC083-repressed ethylene production in bananas. These findings collectively illustrated the mechanistic basis of a MaMADS1-MaNAC083-MaACS1/MaACOs regulatory cascade controlling ethylene biosynthesis during banana fruit ripening. These findings increase our knowledge of the transcriptional regulatory mechanisms of ethylene biosynthesis at the transcriptional level and are expected to help develop molecular approaches to control ripening and improve fruit storability.

Loss of ACO4 in petunia improves abiotic stress tolerance by reducing the deleterious effects of stress-induced ethylene

To investigate the role of ethylene (ET) in abiotic stress tolerance in petunia cv. ‘Mirage Rose’, petunia plants in which the ET biosynthesis gene 1-aminocyclopropane-1-carboxylic acid oxidase 4 (ACO4) was knocked out (phaco4 mutants) and wild-type (WT) plants were exposed to heat and drought conditions. Loss of function of ACO4 significantly delayed leaf senescence and chlorosis under heat and drought stress by maintaining the SPAD values and the relative water content, indicating a greater stress tolerance of phaco4 mutants than that of WT plants. This tolerance was related to the lower ET and reactive oxygen species levels in the mutants than in WT plants. Furthermore, the stress-induced expression of genes related to ET signal transduction, antioxidant and proline activities, heat response, and biosynthesis of abscisic acid was higher in the mutants than in WT plants, indicating a greater stress tolerance in the former than in the latter. These results demonstrate the deleterious effects of stress-induced ET on plant growth and provide a better physiological and molecular understanding of the role of stress ET in the abiotic stress response of petunia. Because the loss of function of ACO4 in petunia improved stress tolerance, we suggest that ACO4 plays a vital role in stress-induced leaf senescence and acts as a negative regulator of abiotic stress tolerance.

Transcriptomic and Metabolomic Analyses of Seedlings of Two Grape Cultivars with Distinct Tolerance Responses to Flooding and Post-Flooding Stress Conditions

Grapes, an important and widespread fruit crop providing multiple products, face increasing flooding risks due to intense and frequent extreme rainfall. It is thus imperative to fully understand the flood-tolerance mechanisms of grapevines. Here, RNA-seq and LC-MS/MS technologies were used to analyze the transcriptome and metabolome changes in the roots of SO4 (tolerant to flooding) and Kyoho (sensitive to flooding) grapes under flooding and post-flooding conditions. The results showed that the abundance of many metabolites in the phenylpropanoids and polyketides, organic acids and their derivatives, and organic oxygen compounds superclasses changed in different patterns between the Kyoho and SO4 grapes under flooding and post-flooding conditions. Jasmonic acid and the ascorbic acid–glutathione cycle played a pivotal role in coping with both hypoxia stress and reoxygenation stress incurred during flooding and post-flooding treatments in the SO4 cultivar. Under flooding stress, the regulatory mechanistic shift from aerobic respiration to anaerobic fermentation under hypoxia is partly missing in the Kyoho cultivar. In the post-flooding stage, many genes related to ethylene, gibberellins, cytokinins, and brassinosteroids biosynthesis and brassinosteroids-responsive genes were significantly downregulated in the Kyoho cultivar, adversely affecting growth recovery; however, their expression was not reduced in the SO4 cultivar. These findings enhance our understanding of the flooding-tolerance mechanisms in grapes.

Identifying genes regulated during natural, on-plant senescence in broccoli (Brassica oleracea) in contrast to postharvest senescence

The nutritional value of broccoli (Brassica oleracea L. var. Italica), its health benefits, as well as its economic value in world production make broccoli an ideal target for improvement given its current postharvest problems related to rapid senescence. Broccoli florets are harvested when their flowers are still developing and photosynthetically active. Harvest causes disruption of supply of water, hormones, nutrients, and energy, leads to imbalance of source-to-sink transition, and triggers stress-induced senescence. There is a clear gap in knowledge about the mechanisms of developmentally regulated, natural (on-plant) senescence and stress-induced postharvest senescence in broccoli. In this study, we aimed at elucidating the molecular mechanisms of natural (on-plant) senescence in broccoli in contrast to postharvest senescence. The genetic progression of natural senescence in broccoli florets was profiled in this study through the RNA-sequencing approach. Candidate genes were identified at each developmental stage of broccoli floret for the involvement of senescence by developmental signals. We further conducted comparative transcriptome analysis on those genes under developmentally regulated senescence and stress-induced postharvest senescence. We found that many genes behave differently in the two conditions, such as sugar transport family, senescence-associated genes (SAGs), ethylene biosynthesis and signaling genes, and flower senescence genes. A comparison of two SWEET family genes SWEET 3 and SWEET 9 in natural and postharvest senescence demonstrated opposite roles in source-sink transition in broccoli. Moreover, we found previously unreported transcription factors including stomatal patterning genes MUTE, SCREAM2, and FAMA and brassinosteroid signaling gene BR ENHANCED EXPRESSION 1 (BEE) that were regulated during natural senescence. In this research, we identified genes involved in natural senescence and further compared with postharvest senescence of commercial maturity broccoli to understand the dynamic senescence networks regulated by both developmental cues and stress signals.

An insertion of transposon in DcNAP inverted its function in the ethylene pathway to delay petal senescence in carnation (Dianthus caryophyllus L.).

Petal senescence is the final stage of flower development. Transcriptional regulation plays key roles in this process. However, whether and how post-transcriptional regulation involved is still largely unknown. Here, we identified an ethylene-induced NAC family transcription factor DcNAP in carnation (Dianthus caryophyllus L.). One allele, DcNAP-dTdic1, has an insertion of a dTdic1 transposon in its second exon. The dTdic1 transposon disrupts the structure of DcNAP and causes alternative splicing, which transcribes multiple domain-deleted variants (DcNAP2 and others). Conversely, the wild type allele DcNAP transcribes DcNAP1 encoding an intact NAC domain. Silencing DcNAP1 delays and overexpressing DcNAP1 accelerates petal senescence in carnation, while silencing and overexpressing DcNAP2 have the opposite effects, respectively. Further, DcNAP2 could interact with DcNAP1 and interfere the binding and activation activity of DcNAP1 to the promoters of its downstream target ethylene biosynthesis genes DcACS1 and DcACO1. Lastly, ethylene signalling core transcriptional factor DcEIL3-1 can activate the expression of DcNAP1 and DcNAP2 in the same way by binding their promoters. In summary, we discovered a novel mechanism by which DcNAP regulates carnation petal senescence at the post-transcriptional level. It may also provide a useful strategy to manipulate the NAC domains of NAC transcription factors for crop genetic improvement.

Fruit ripening under heat stress: The intriguing role of ethylene-mediated signaling

Crop production is significantly influenced by climate, and even minor climate changes can have a substantial impact on crop yields. Rising temperature due to climate change can lead to heat stress (HS) in plants, which not only hinders plant growth and development but also result in significant losses in crop yields. To cope with the different stresses including HS, plants have evolved a variety of adaptive mechanisms. In response to these stresses, phytohormones play a crucial role by generating endogenous signals that regulate the plant’s defensive response. Among these, Ethylene (ET), a key phytohormone, stands out as a major regulator of stress responses in plants and regulates many plant traits, which are critical for crop productivity and nutritional quality. ET is also known as a ripening hormone for decades in climacteric fruit and many studies are available deciphering the function of different ET biosynthesis and signaling components in the ripening process. Recent studies suggest that HS significantly affects fruit quality traits and perturbs fruit ripening by altering the regulation of many ethylene biosynthesis and signaling genes resulting in substantial loss of fruit yield, quality, and postharvest stability. Despite the significant progress in this field in recent years the interplay between ET, ripening, and HS is elusive. In this review, we summarized the recent advances and current understanding of ET in regulating the ripening process under HS and explored their crosstalk at physiological and molecular levels to shed light on intricate relationships

Transcriptome sequencing reveals the senescence mechanism of ethylene-insensitive cut Rhododendron flowers

Cut rhododendron flowers have high ornamental value and are loved by consumers. It is important to clarify the senescence mechanism of cut rhododendron flowers to prolong their vase life. Transcriptome sequencing was used to investigate flower senescence in four stages of Rhododendron ‘Hong Shanhu.’ Using RNA sequencing, we generated approximately 100.17 Gb clean data and 173,502 annotated unigenes. Ethylene biosynthesis genes, ACOs , had different expression trends, and most ethylene signaling genes had low expression levels. The ethylene release rate and the activities of ACS and ACO were on the rise during cut flower development. The effects of 200 mg/L ethephon and 0.1 mM/L spermidine on vase life and fresh weight were limited. Most of the LEA and Dehydrin were upregulated with cut flower development and the fresh weight was decreased during fully flowering stage. Based on the different expression trends, differentially expressed genes were classified into 10 clusters. Cluster 2 showed significant upregulation between Z4 and Z3, suggesting that the genes in this cluster were upregulated, probably due to the senescence of cut flowers. The overall trend of Cluster 7 was positively correlated with the opening process of cut flowers. Cluster 9 had a peak at Z3, which indicated that the unigenes started the senescence process. bZIP53, RAP2.4, UIF1, HAT5, ILR3, DEAR3, SCL8, TZF10, MYB70, ERF5, ERF9, and CBF1 had higher expression levels than the other transcription factors in these clusters, indicating that they are probably the key transcription factors involved in petal senescence. Our results indicated that ‘Hong Shanhu’ cut flowers were ethylene-insensitive, and their senescence was probably related to water stress. These results contribute to understanding the mechanism of senescence in cut rhododendron flowers.

Trihelix transcription factor SlGT31 regulates fruit ripening mediated by ethylene in tomato.

Trihelix proteins are plant-specific transcription factors that play crucial roles in plant development and stress responses. However, the involvement of trihelix proteins in fruit ripening and transcriptional regulatory mechanisms remains largely unclear. In this study, we cloned a trihelix gene SlGT31, whose relative expression was significantly induced by the application of exogenous ethylene but repressed by 1-methylcyclopropene (1-MCP). Suppression of SlGT31 resulted in delayed fruit ripening, decreased accumulation of total carotenoids and ethylene content, and inhibition of relative expression of genes related to ethylene and fruit ripening. Conversely, the opposite results were observed in SlGT31-overexpression lines. Yeast one-hybrid and dual-luciferase assays suggested that SlGT31 could bind to the promoters of two key ethylene biosynthesis genes ACO1 and ACS4. These results indicate that SlGT31 may act as a positive modulator during fruit ripening.

A novel NAC transcription factor, PpNAP6, is involved in peach ripening by activating ethylene synthesis

Peach (Prunus persica) is a typical climacteric fruit, having an obvious peak of ethylene release during ripening. Transcriptional regulators play key roles in fruit ripening. NAP (NAC-like, activated by AP3/P1) protein is a subfamily of the NAC (NAM, ATAF1/2 and CUC2) transcription factor (TF) family. Here, we discovered the function of peach NAP subfamily NAC TF, PpNAP6, homologous to the tomato ripening-related TF NOR. In the pulp of ‘Zaofengwang’ peach, PpNAP6 is highly expressed, and its transcript level decreased gradually throughout the whole development period, however, it is up-regulated at onset of peach ripening. Transient transformation experiments showed that PpNAP6 over-expression in peach fruit increased ethylene release and accelerated fruit ripening, while PpNAP6 down-expression inhibited ethylene biosynthesis. Dual-luciferase reporter assay (DLR) and electrophoretic mobility shift assay (EMSA) proved that PpNAP6 can directly bind to the promoters of key ethylene biosynthesis genes PpACS1 and PpACO1, and activate their transcription. Moreover, PpNAP6 was heterologously overexpressed in tomato nor mutant, and the transgenic lines showed the overexpression of PpNAP6 could partially recover its ripening-inhibited phenotype. Interestingly, PpNAP6 activates ethylene biosynthesis during peach ripening, whereas its transcription was inhibited treated with ethephon and enhanced with 1-MCP treatment. In summary, our work revealed that PpNAP6 regulates peach fruit ripening through ethylene biosynthesis, and may be involved in the fine regulation of ethylene via a feedback mechanism.

Ethanol fermentation- and ethylene physiology-related gene expression profiles in Red Delicious apples stored under variable hypoxic conditions and protocols

Dynamic Controlled Atmosphere (DCA) is beneficial in maintaining specific quality parameters but, due to the extreme oxygen levels applied, can cause adverse effects on the fruit by inducing excessive anaerobic metabolism and the production of off-flavors. The metabolic adaptation and responses of apples (Malus domestica Borkh.) cv. Red Delicious to static or dynamic oxygen concentrations (0.3 and 0.8%, with sequential shifts) during cold storage for 7 months were studied by monitoring quality parameters and the expression of genes involved in sugar, fermentative metabolism, and ethylene physiology. Ethanol content reached the highest levels (around 400 mg/kg FW) under 0.3% oxygen concentration and fruit firmness appeared to be reduced in samples accumulating the highest levels of ethanol. Oxygen switch was effective in reducing the ethanol concentrations with timing-dependent variable effects. The expression of fermentative (alcohol dehydrogenase, lactate dehydrogenase, pyruvate decarboxylase) and sugar metabolism (β-amylase; phosphofructokinase; sucrose synthase) genes resulted to be differently affected by the hypoxic conditions imposed, in particular during the early stages of storage. Sucrose synthase expression appeared to be highly sensitive to changes in low oxygen concentration. Ethylene biosynthesis (ACC synthase and oxidase) genes showed marked differences in their expression in relation to the static and dynamic protocols and the hypoxic conditions, as well as six Ethylene Responsive Factors (ERF) genes, some of them possibly involved in the oxygen sensing mechanism operating in fruit tissues.

Extracellular self-DNA induced a PTI-related local defence against Rhizopus rot in postharvest peach fruit

The plant immune response against various pathogens relies on a convoluted recognition system based on perceiving exogenous or endogenous stimuli, such as pathogen- or damage-associated molecular patterns (PAMPs or DAMPs). Extracellular self-DNA (sDNA) is an established DAMP source, but the plant intracellular signaling mechanism after sDNA perception remains largely unclear. Here, we observed that peach fruit was more resistant to the fungus Rhizopus stolonifer after sDNA perception. We identified a complete MAPK cascade (MAPKKK1, MAPKK2 and MAPK1) that functions downstream of the FLS2-BAK1 receptor complex following the perception of sDNA-derived DAMPs by means of transcriptome sequencing. The results of yeast two-hybrid (Y2H) and glutathione S-transferase (GST) pull-down assays confirmed the protein interaction of FLS2 and BAK1; meanwhile, the interaction of MAPKKK1 and MAPKK2 and its downstream interaction of MAPKK2 and MAPK1 were both exhibited following the another Y2H results. Accordingly, MAPK1 interacted with 1-aminocyclopropane-1-carboxylate oxidase (ACO1), which was directly associated with increased ethylene production and upregulation of the transcription profile of ethylene synthesis-related genes (PpSAMDC, PpACS1 and PpACO1), ethylene receptors (ETRs and ERS1), ethylene-responsive transcription factors (ERFs) and three ethylene-inducible marker genes, PLANT DEFENSIN1.2 (PDF1.2), CHITINASE (CHI) and HOOKLESS1 (HLS1). Accordingly, the overexpression of MAPK1 enhanced the disease resistance of transgenic tomato against the fungus R. stolonifer and increased ethylene biosynthesis and ethylene signaling-related gene transcription. Taken together, our data provide insight into the putative mechanism underlying the recognition of sDNA components by the FLS2-BAK1 complex and its downstream events in the transduction of ethylene signaling by the MAPK cascade to mediate local resistance in postharvest peach fruit.

Senescence in dahlia flowers is regulated by a complex interplay between flower age and floret position.

Mechanisms regulating flower senescence are not fully understood in any species and are particularly complex in composite flowers. Dahlia (Dahlia pinnata Cav.) florets develop sequentially, hence each composite flower head includes florets of different developmental stages as the whole flower head ages. Moreover, the wide range of available cultivars enables assessment of intraspecific variation. Transcriptomes were compared amongst inner (younger) and outer (older) florets of two flower head ages to assess the effect of floret vs. flower head ageing. More gene expression, including ethylene and cytokinin pathway expression changed between inner and outer florets of older flower heads than between inner florets of younger and older flower heads. Additionally, based on Arabidopsis network analysis, different patterns of co-expressed ethylene response genes were elicited. This suggests that changes occur in young inner florets as the whole flower head ages that are different to ageing florets within a flower head. In some species floral senescence is orchestrated by the plant growth regulator ethylene. However, there is both inter and intra-species variation in its importance. There is a lack of conclusive data regarding ethylene sensitivity in dahlia. Speed of senescence progression, effects of ethylene signalling perturbation, and patterns of ethylene biosynthesis gene expression differed across three dahlia cultivars ('Sylvia', 'Karma Prospero' and 'Onesta') suggesting differences in the role of ethylene in their floral senescence, while effects of exogenous cytokinin were less cultivar-specific.

Morphophysiological and transcriptome analysis reveal that reprogramming of metabolism, phytohormones and root development pathways governs the potassium (K+) deficiency response in two contrasting chickpea cultivars.

Potassium (K+) is an essential macronutrient for plant growth and development. K+ deficiency hampers important plant processes, such as enzyme activation, protein synthesis, photosynthesis and stomata movement. Molecular mechanism of K+ deficiency tolerance has been partly understood in model plants Arabidopsis, but its knowledge in legume crop chickpea is missing. Here, morphophysiological analysis revealed that among five high yielding desi chickpea cultivars, PUSA362 shows stunted plant growth, reduced primary root growth and low K+ content under K+ deficiency. In contrast, PUSA372 had negligible effect on these parameters suggesting that PUSA362 is K+ deficiency sensitive and PUSA372 is a K+ deficiency tolerant chickpea cultivar. RNA-seq based transcriptome analysis under K+ deficiency revealed a total of 820 differential expressed genes (DEG's) in PUSA362 and 682 DEGs in PUSA372. These DEGs belongs to different functional categories, such as plant metabolism, signal transduction components, transcription factors, ion/nutrient transporters, phytohormone biosynthesis and signalling, and root growth and development. RNA-seq expression of randomly selected 16 DEGs was validated by RT-qPCR. Out of 16 genes, 13 showed expression pattern similar to RNA-seq expression, that verified the RNA-seq expression data. Total 258 and 159 genes were exclusively up-regulated, and 386 and 347 genes were down-regulated, respectively in PUSA362 and PUSA372. 14 DEGs showed contrasting expression pattern as they were up-regulated in PUSA362 and down-regulated in PUSA372. These include somatic embryogenesis receptor-like kinase 1, thaumatin-like protein, ferric reduction oxidase 2 and transcription factor bHLH93. Nine genes which were down-regulated in PUSA362 found to be up-regulated in PUSA372, including glutathione S-transferase like, putative calmodulin-like 19, high affinity nitrate transporter 2.4 and ERF17-like protein. Some important carbohydrate metabolism related genes, like fructose-1,6-bisphosphatase and sucrose synthase, and root growth related Expansin gene were exclusively down-regulated, while an ethylene biosynthesis gene 1-aminocyclopropane-1-carboxylate oxidase 1 (ACO1) was up-regulated in PUSA362. Interplay of these and several other genes related to hormones (auxin, cytokinin, GA etc.), signal transduction components (like CBLs and CIPKs), ion transporters and transcription factors might underlie the contrasting response of two chickpea cultivars to K+ deficiency. In future, some of these key genes will be utilized in genetic engineering and breeding programs for developing chickpea cultivars with improved K+ use efficiency (KUE) and K+ deficiency tolerance traits.

DcWRKY33 promotes petal senescence in carnation (Dianthus caryophyllus L.) by activating genes involved in the biosynthesis of ethylene and abscisic acid and accumulation of reactive oxygen species.

Carnation (Dianthus caryophyllus L.) is one of the most famous and ethylene-sensitive cut flowers worldwide, but how ethylene interacts with other plant hormones and factors to regulate petal senescence in carnation is largely unknown. Here we found that a gene encoding WRKY family transcription factor, DcWRKY33, was significantly upregulated upon ethylene treatment. Silencing and overexpression of DcWRKY33 could delay and accelerate the senescence of carnation petals, respectively. Abscisic acid (ABA) and H2 O2 treatments could also accelerate the senescence of carnation petals by inducing the expression of DcWRKY33. Further, DcWRKY33 can bind directly to the promoters of ethylene biosynthesis genes (DcACS1 and DcACO1), ABA biosynthesis genes (DcNCED2 and DcNCED5), and the reactive oxygen species (ROS) generation gene DcRBOHB to activate their expression. Lastly, relationships are existed between ethylene, ABA and ROS. This study elucidated that DcWRKY33 promotes petal senescence by activating genes involved in the biosynthesis of ethylene and ABA and accumulation of ROS in carnation, supporting the development of new strategies to prolong the vase life of cut carnation.

Wounding rapidly alters transcription factor expression, hormonal signaling, and phenolic compound metabolism in harvested sugarbeet roots.

Injuries sustained by sugarbeet (Beta vulgaris L.) roots during harvest and postharvest operations seriously reduce the yield of white sugar produced from stored roots. Although wound healing is critically important to reduce losses, knowledge of these processes is limited for this crop as well as for roots in other species. To better understand the metabolic signals and changes that occur in wounded roots, dynamic changes in gene expression were determined by RNA sequencing and the activity of products from key genes identified in this analysis were determined in the 0.25 to 24h following injury. Nearly five thousand differentially expressed genes that contribute to a wide range of cellular and molecular functions were identified in wounded roots. Highly upregulated genes included transcription factor genes, as well as genes involved in ethylene and jasmonic acid (JA) biosynthesis and signaling and phenolic compound biosynthesis and polymerization. Enzyme activities for key genes in ethylene and phenolic compound biosynthesis and polymerization also increased due to wounding. Results indicate that wounding causes a major reallocation of metabolism in sugarbeet taproots. Although both ethylene and JA are likely involved in triggering wound responses, the greater and more sustained upregulation of ethylene biosynthesis and signaling genes relative to those of JA, suggest a preeminence of ethylene signaling in wounded sugarbeet roots. Changes in gene expression and enzymes involved in phenolic compound metabolism additionally indicate that barriers synthesized to seal off wounds, such as suberin or lignin, are initiated within the first 24h after injury.

The mechanism of gibberellins treatment suppressing kiwifruit postharvest ripening processes by transcriptome analysis

Gibberellins (GAs) control numerous processes in plants, including fruit ripening. However, the role of GAs in kiwifruit ripening is not well characterized. Here, we noticed that GA3 treatment in postharvest kiwifruit delayed the ripening process and many physiological traits were detected such as firmness, starch/total soluble solid content, cell-wall components (pectin, cellulose and hemicellulose), ethylene and ester production, and ascorbic acid (vitamin C) concentration. Transcriptome analysis supplied a molecular basis for investigating the inhibiting mechanism of GA3 and metabolic variations during kiwifruit ripening. A total of 3116 differently expressed genes (DEGs) were identified, including 51 genes involved in plant hormone signal transduction pathways and 36 structural genes related to these physiological traits. 16 major DEGs were analyzed by real-time PCR during the whole storage period. The expression of most of them was significantly suppressed by GA3 treatment, such as two ethylene biosynthesis genes (ACOs ), two β-amylase (BAMs), five cell-wall modification genes, one GA2ox, and one GA receptor GID1, whereas the expression of Actinidia29942 (JAZ) was enhanced by GA3 treatment. Correlation analysis between multiple physiological indicators and candidate genes showed that GA3 could regulate kiwifruit postharvest ripening effectively and comprehensively. Together, these GA-responsive genes may play important roles in controlling kiwifruit ripening, and GA treatment could be a promising approach for the preservation of postharvest kiwifruit.

Variation in longevity of cut and in planta flowers of potted carnation varieties affected by their relationship with ethylene and water

Carnation (<italic>Dianthus caryophyllus</italic> L.) is a typical ethylene-sensitive cut flower, but a few differences in ethylene sensitivity have been reported for different potted carnation species. In this study, we investigated the relationship between vase life, ethylene sensitivity, ethylene biosynthesis gene expression, and flower volatile substance content of 17 different potted carnation varieties. It was found that under the same post-harvest environmental conditions, the vase life of different varieties ranged from 6.2−14.2 d. Among the 17 varieties, 'Cherry' and 'Grace Bay' were highly sensitive to ethylene, and qRT-PCR analysis showed that their ethylene biosynthesis genes <italic>DcACS1</italic> and <italic>DcACO1</italic> expression increased the most, while 'Pink and Purple', 'Bondi Beach', and 'Grane Beach' showed the opposite pattern. In addition, the lower ethylene release was important in leading to longer vase life of potted carnations, and that ethylene release from shorter vase life varieties was 2.5−4.5 times greater than that of longer vase life varieties. Varieties that are more sensitive to ethylene are more likely to have a shorter vase life due to early disruption of water relations, which is mainly the result of reduced stem hydraulic conductivity and transpiration water loss. Analysis of volatile substances showed that ethylene had no significant effect on the release of volatile substances from potted carnations. Therefore, a better understanding of petal senescence in potted carnations will help us to improve measures to extend flower longevity according to the ethylene sensitivity of different varieties.

1-Methylcyclopropene retards pak choi (Brassica rapasubsp. chinensis) yellowing via BcNAC055-, BcMYB44-, and BcOBF1-mediated regulation of the key chlorophyll degrading geneBcNYC1during storage at 20 °C

AbstractThe objective of this study was to investigate the molecular regulatory mechanism of 1-methylcyclopropene (1-MCP) treatment on leaf yellowing of pak choi during storage at 20 °C. In the present study, compared with the control and 10 μL/L ethylene-treated pak choi, 5.0 μL/L 1-MCP fumigation alleviated the yellowing process of pak choi as proved by the maintenance of higher levels of colour, chlorophyll content, and appearance. 1-MCP treatment decreased chlorophyll breakdown by downregulating the activity of chlorophyll-degrading enzymes (chlorophyll b reductase (CBR) and pheophytinase (PPH)) and the expression of their related genes (BcNYC1 and BcPPH). The application of 1-MCP also inhibited the transcript abundance of ethylene biosynthesis and signal transduction genes (BcACO, BcEIN3, and BcERF) and reduced ethylene production, thus helping to maintain the green colour of pak choi; however, ethylene treatment exerted the opposite function. Meanwhile, three transcription factors (BcNAC055, BcMYB44, and BcOBF1) were successfully cloned. Experiments performed using yeast one-hybrid assay, dual-luciferase reporter system and in vivo bioluminescence imaging assay, confirmed that BcNAC055, BcMYB44, and BcOBF1 directly bound to the BcNYC1 promoter. 1-MCP inhibited the expression levels of BcNAC055, BcMYB44, BcOBF1, and BcNYC1 during storage, while the application of ethylene activated their expression. In conclusion, 1-MCP downregulated the expression of the key chlorophyll degradation gene BcNYC1 by inhibiting the transcript level of its three positive regulators (BcNAC055, BcMYB44, and BcOBF1) and lowered the activity of chlorophyll-degrading enzyme CBR, thereby helping mitigate chlorophyll breakdown to retard leaf yellowing in pak choi. The results of this work extend our understanding of 1-MCP-delayed yellowing in postharvest pak choi.

ACC deaminase-producing endophytic fungal consortia promotes drought stress tolerance in M.oleifera by mitigating ethylene and H2O2.

Drought has become more prevalent due to dramatic climate change worldwide. Consequently, the most compatible fungal communities collaborate to boost plant development and ecophysiological responses under environmental constraints. However, little is known about the specific interactions between non-host plants and endophytic fungal symbionts that produce growth-promoting and stress-alleviating hormones during water deficits. The current research was rationalized and aimed at exploring the influence of the newly isolated, drought-resistant, ACC deaminase enzyme-producing endophytic fungi Trichoderma gamsii (TP), Fusarium proliferatum (TR), and its consortium (TP+TR) from a xerophytic plant Carthamus oxycantha L. on Moringa oleifera L. grown under water deficit induced by PEG-8000 (8% osmoticum solution). The current findings revealed that the co-inoculation promoted a significant enhancement in growth traits such as dry weight (217%), fresh weight (123%), root length (65%), shoot length (53%), carotenoids (87%), and chlorophyll content (76%) in comparison to control plants under water deficit. Total soluble sugars (0.56%), proteins (132%), lipids (43%), flavonoids (52%), phenols (34%), proline (55%), GA3 (86%), IAA (35%), AsA (170%), SA (87%), were also induced, while H2O2 (-45%), ABA (-60%) and ACC level (-77%) was decreased by co-inoculation of TP and TR in M. oleifera plants, compared with the non-inoculated plants under water deficit. The co-inoculum (TP+TR) also induced the antioxidant potential and enzyme activities POX (325%), CAT activity (166%), and AsA (21%), along with a lesser decrease (-2%) in water potential in M. oleifera plants with co-inoculation under water deficit compared with non-inoculated control. The molecular analysis for gene expression unraveled the reduced expression of ethylene biosynthesis and signaling-related genes up to an optimal level, with an induction of antioxidant enzymatic genes by endophytic co-inoculation in M. oleifera plants under water deficit, suggesting their role in drought stress tolerance as an essential regulatory function. The finding may alert scientists to consider the impacts of optimal reduction of ethylene and induction of antioxidant potential on drought stress tolerance in M. oleifera. Hence, the present study supports the use of compatible endophytic fungi to build a bipartite mutualistic symbiosis in M. oleifera non-host plants to mitigate the negative impacts of water scarcity in arid regions throughout the world.