ACC Synthase Activity Research Articles

The Potential of Essential Oils from Active Packaging to Reduce Ethylene Biosynthesis in Plant Products. Part 2: Fruits (Blueberries and Blackberries).

Plant essential oils (EOs) have an important ability to inhibit ethylene biosynthesis. Nevertheless, the effects of EOs on the key components of ethylene biosynthesis (l-aminocyclopropane-1-carboxylic (ACC) oxidase activity, ACC synthase activity, and ACC content) have not yet been thoroughly studied. Accordingly, this study focused on the effects of emitted EOs from active packaging (EO doses from 100 to 1000 mg m-2) on the key components of ethylene biosynthesis of blueberries and blackberries under several storage temperatures. Anise EO and lemon EO active packaging induced the greatest inhibitory effects (60-76%) on the ethylene production of blueberries and blackberries, respectively, even at high storage temperatures (22 °C). In terms of EO doses, active packaging with 1000 mg m-2 of anise EO or lemon EO led to the highest reduction of ethylene production, respectively. At 22 °C, the investigated EO active packing reduced the activities of ACC synthase and ACC oxidase up to 50%. In order to minimise ethylene biosynthesis in blueberries and blackberries when they are stored even under improper temperature scenarios at high temperatures, this EO active packaging is a natural and efficient technological solution.

The Potential of Essential Oils from Active Packaging to Reduce Ethylene Biosynthesis in Plant Products. Part 1: Vegetables (Broccoli and Tomato).

Essential oils (EOs) extracted from plants have a high potential to reduce ethylene biosynthesis, although their effects have not been deeply studied yet on the key components of the ethylene biosynthesis pathway: l-aminocyclopropane-1-carboxylic (ACC) oxidase activity, ACC synthase activity, and ACC content. Hence, the present study aimed to elucidate the effects of released EOs from active packaging (with different EO doses ranging from 100 to 1000 mg m-2) on the ethylene biosynthesis key components of broccoli and tomato under different storage temperature scenarios. The largest ethylene inhibitory effects on broccoli and tomatoes were demonstrated by grapefruit EO and thyme essential EO (up to 63%), respectively, which were more pronounced at higher temperatures. Regarding EO doses, active packaging with a thyme EO dose of 1000 mg m-2 resulted in the strongest reduction (33-38%) of ethylene production in tomatoes. For broccoli, identical results were shown with a lower grapefruit EO dose of 500 mg m-2. The studied EO-active packaging decreased ACC synthase and ACC oxidase activities by 40-50% at 22 °C. Therefore, this EO-active packaging is a natural and effective technology to reduce ethylene biosynthesis in broccoli and tomatoes when they are stored, even in unsuitable scenarios at high temperatures.

Domain Swapping between AtACS7 and PpACL1 Results in Chimeric ACS-like Proteins with ACS or Cβ-S Lyase Single Enzymatic Activity.

The gaseous hormone ethylene plays a pivotal role in plant growth and development. In seed plants, the key rate-limiting enzyme that controls ethylene biosynthesis is ACC synthase (ACS). ACS has, for a long time, been believed to be a single-activity enzyme until we recently discovered that it also possesses Cβ-S lyase (CSL) activity. This discovery raises fundamental questions regarding the biological significance of the dual enzymatic activities of ACS. To address these issues, it is highly necessary to obtain ACS mutants with either ACS or CSL single activity. Here, domain swapping between Arabidopsis AtACS7 and moss CSL PpACL1 were performed. Enzymatic activity assays of the constructed chimeras revealed that, R10, which was produced by replacing AtACS7 box 6 with that of PpACL1, lost ACS but retained CSL activity, whereas R12 generated by box 4 substitution lost CSL and only had ACS activity. The activities of both chimeric proteins were compared with previously obtained single-activity mutants including R6, AtACS7Q98A, and AtACS7D245N. All the results provided new insights into the key residues required for ACS and CSL activities of AtACS7 and laid an important foundation for further in-depth study of the biological functions of its dual enzymatic activities.

Proanthocyanidin delays litchi peel browning by inhibiting ethylene biosynthesis, respiratory metabolism, and phenol oxidase activities

Proanthocyanidin (PA) promotes antioxidant capacity and enhances resistance against oxidative stress-induced senescence in fruits after harvest. However, the effects of PA on the postharvest physiology of litchi fruit remain unclear. This study aimed to determine the effect of PA on ethylene and respiration physiology, and on browning enzymes of ‘feizixiao’ litchi stored at 25 °C for 8 d. Proanthocyanidin treatment delayed peel browning and color change, maintained higher peel lightness, and reduced ethylene production and respiration rates in litchi fruit. Subsequently, the key genes and/or enzymes involved in the ethylene biosynthesis pathway and respiration metabolism were determined. The PA-treated fruit exhibited lower levels of 1-aminocyclopropanecarboxylic acid (ACC) content, ACC synthase (ACS) and ACC oxidase (ACO) activity, and an inhibition of LcACS7/9 and LcACO1/2 genes on the peel and pulp compared to that of the control. In addition, the application of PA reduced respiration rate by downregulating the expression of LcAtpB, LcAOX1, LcUCP1, LcAAC1, and LcSnRK2. Furthermore, the phenol oxidase including peroxidase (POD), polyphenol oxidase (PPO), and laccase (LAC), were inhibited during litchi peel browning and senescence. Therefore, the process of PA-delayed browning of litchi may occur by delaying ethylene production and respiratory rate and inhibiting phenol oxidase activities in litchi fruit.

Effects of Flooding and Endogenous Hormone on the Formation of Knee Roots in Taxodium ascendens.

Taxodium ascendens is a typical tree species with high flood tolerance, and it can generate knee roots in the wetlands. This study investigated the number and size of knee roots and the soil flooding conditions. Furthermore, we also measured physiology, biochemical responses, and the anatomical structure of knee roots and underground roots at different developmental stages. This study aimed to understand the adaptation mechanism of T. ascendens to flooding stress and the formation mechanism of the knee roots. The results showed that the formation of knee roots was significantly affected by the soil water table (P < 0.05). The middle water table was more conducive to the formation of knee roots. In the middle water table, the 1-aminocyclopropane-1-carboxylic acid (ACC) content and ACC synthase activity were significantly lower in the knee roots than in the underground roots. The knee roots at the young-aged stage showed the highest ACC oxidase activity among the development stages of the knee roots. The ethylene release rate was significantly higher in the knee roots than in the underground roots (P < 0.05). Indole-3-acetic acid (IAA) content first increased, then decreased with knee root development. The periderm cells at the apex of the knee roots were dead and had many intercellular spaces, which was beneficial for the growth of T. ascendens. In conclusion, the middle water table induced the ethylene and IAA production, which promoted the formation of knee roots, which improved roots ventilation and flooding tolerance of T. ascendens. The results obtained can provide information about mechanisms of knee roots formation and provide scientific evidence for the afforestation and management under wetland conditions.

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.

Activation of 1-Aminocyclopropane-1-Carboxylic Acid Synthases Sets Stomatal Density and Clustered Ratio on Leaf Epidermis of Arabidopsis in Response to Drought.

The adjustment of stomatal density and clustered ratio on the epidermis is the important strategy for plants to respond to drought, because the stoma-based water loss is directly related to plant growth and survival under drought conditions. But the relevant adjustment mechanism still needs to be explored. 1-Aminocyclopropane-1-carboxylate (ACC) is disclosed to promote stomatal development, while in vivo ACC levels depend on activation of ACC synthase (ACS) family members. Based on the findings of ACS expression involving in drought response and several ACS activity inhibitors reducing stomatal density and cluster in drought response, here we examined how ACS activation is involved in the establishment of stomatal density and cluster on the epidermis under drought conditions. Preliminary data indicated that activation of ACS2 and/or ACS6 (ACS2/6) increased stomatal density and clustered ratio on the Arabidopsis leaf epidermis by accumulating ACC under moderate drought, and raised the survival risk of seedlings under escalated drought. Further exploration indicated that, in Arabidopsis seedlings stressed by drought, the transcription factor SPEECHLESS (SPCH), the initiator of stomatal development, activates ACS2/6 expression and ACC production; and that ACC accumulation induces Ca2+ deficiency in stomatal lineage; this deficiency inactivates a subtilisin-like protease STOMATAL DENSITY AND DISTRIBUTION 1 (SDD1) by stabilizing the inhibition of the transcription factor GT-2 Like 1 (GTL1) on SDD1 expression, resulting in an increases of stomatal density and cluster ratio on the leaf epidermis. This work provides a novel evidence that ACS2/6 activation plays a key role in the establishment of stomatal density and cluster on the leaf epidermis of Arabidopsis in response to drought.

Dual activities of ACC synthase: Novel clues regarding the molecular evolution of ACS genes.

Ethylene plays profound roles in plant development. The rate-limiting enzyme of ethylene biosynthesis is 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS), which is generally believed to be a single-activity enzyme evolving from aspartate aminotransferases. Here, we demonstrate that, in addition to catalyzing the conversion of S-adenosyl-methionine to the ethylene precursor ACC, genuine ACSs widely have Cβ-S lyase activity. Two N-terminal motifs, including a glutamine residue, are essential for conferring ACS activity to ACS-like proteins. Motif and activity analyses of ACS-like proteins from plants at different evolutionary stages suggest that the ACC-dependent pathway is uniquely developed in seed plants. A putative catalytic mechanism for the dual activities of ACSs is proposed on the basis of the crystal structure and biochemical data. These findings not only expand our current understanding of ACS functions but also provide novel insights into the evolutionary origin of ACS genes.

Phomopsis liquidambaris reduces ethylene biosynthesis in rice under salt stress via inhibiting the activity of 1-aminocyclopropane-1-carboxylate deaminase.

The endophytic fungus Phomopsis liquidambaris is characterized as a plant growth-promoting agent under salt stress, but its mechanism is unknown. Herein, 1-aminocyclopropane-1-carboxylatedeaminase (ACCD) from the strain was confirmed that it had the ability of utilizing1-aminocyclopropane-1-carboxylate as the sole nitrogen source. The full-length ACCD gene was 1152bp, which encodes a mature protein of 384 amino acids with a molecular mass of 41.53kDa. The ACCD activity was 3.9-fold in 3mmol L-1 ACC by qRT-PCR under salt stress comparing with no salt tress. Ethylene production was increased to 34.55-70.60% and reduced the growth of rice by 23-69.73% under salt stress. Inoculation of P. liquidambaris increased root-shoot length, fresh and dry weight, and overall growth of stressed rice seedlings. ACC accumulation, ACC synthase and ACC oxidase activities increased in salt-treated rice seedlings, while they were significantly reduced when P. liquidambaris was inoculated into rice by qRT-PCR. It therefore can be concluded that P. liquidambaris can be used as a plant growth promoting fungus against salt stress and other biotic or abiotic stresses.

Delaying Broccoli Floret Yellowing by Phytosulfokine α Application During Cold Storage.

During postharvest life, broccoli suffers from floret yellowing confining its economic and nutritional value. The objective of the present study was to explore the mechanisms employed by phytosulfokine α (PSKα) at 150 nM for delaying floret yellowing in broccoli during storage at 4°C for 28 days. Our results showed that the higher endogenous accumulation of hydrogen sulfide (H2S) resulting from the higher gene expression and activities of l-cysteine desulfhydrase (LCD) and d-cysteine desulfhydrase (DCD) in broccoli floret treated with 150 nM PSKα may serve as an endogenous signaling molecule for delaying senescence. Moreover, the suppressed ethylene biosynthesis in broccoli floret treated with 150 nM PSKα might be ascribed to lower gene expression and activities of ACC synthase (ACS) and ACC oxidase (ACO). Furthermore, lower gene expression and activities of Mg2+ dechelatase (MDC), pheophytinase (PPH), and pheophorbide a oxygenase (PaO) might be the reasons for the higher accumulation of chlorophyll in broccoli floret treated with 150 nM PSKα. Based on our findings, exogenous PSKα application could be employed as signaling bioactive hormone for retarding floret yellowing of broccoli during storage at 4°C for 28 days.

Reduced ethylene synthesis of mangoes under high CO2 atmosphere storage

. High CO2 atmospheres have been reported to be accountable for slower ripening processes of many fruit species. In modified or controlled atmosphere storage of mangoes (Mangifera indica L.) delayed ripening is attributed to the effects of CO2 on ethylene biosynthesis, which is reduced under CO2 concentrations beyond 10%. In the present work the objective was to determine if those elevated CO2 atmospheres on ethylene synthesis could be attributed to the action of CO2 upon ACC oxidase. Mature green or tree ripe `Tommy Atkins` mangoes were, in four experiments, held in a flow through system of either 10 or 25% CO2 mixed to 5% O2 or only air for 14 or 21 days at 5, 8 or 12°C. Mangoes in the 25% CO2 atmosphere did not produce detectable levels of ethylene, whereas under 10% CO2 the production rates were significantly suppressed at 5 or 8°C. However, 1-aminocyclopropane-1-carboxylic acid (ACC) concentrations in mango mesocarp tissue at retrieval from storage were similar to the air controls and ACC synthase activity was not completely inhibited. The direct effects of CO2 concentrations on ACC oxidase activity is to be considered the most important factor in inhibiting ethylene biosynthesis of mangoes under 25% CO2 atmospheres.

Effect of modified and controlled atmosphere storage on enzyme activity and senescence of Dendrobium orchids

This research investigated the effect of different atmosphere storage conditions on 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase enzyme activity; and senescence of cut Dendrobium orchid flowers. The atmosphere storage conditions under study were normal atmosphere, modified atmosphere packaging, and controlled atmosphere. Under the normal atmosphere, carbon dioxide (CO2) and oxygen (O2) concentrations were 0.03 and 21 %, respectively. For the modified atmosphere packaging, cut orchid flowers were wrapped in polypropylene film prior to filling with 5 % CO2 and 2 % O2, while under the controlled atmosphere, CO2 and O2 concentrations were maintained at 5 and 2 %, respectively. The storage temperature and relative humidity were 13 °C and 95 %, respectively. The ACC synthase and ACC oxidase activity and ethylene-induced electrolyte leakage were determined and results compared. The controlled atmosphere substantially lowered ACC synthase and ACC oxidase activity and was effective in delaying senescence of cut orchid flowers, as indicated by the longest storage life of 28.33 days, followed by the modified atmosphere packaging (18.15 days) and normal atmosphere (11.67 days). The longer storage life enables suppliers of orchid flowers to efficiently manage the demand and supply and also provides exporters with new opportunities to expand into distant overseas markets. The novelty of this research lies in the use of different storage environments to investigate the senescence mechanisms at tissue level of Dendrobium orchid flowers in response to ACC synthase and ACC oxidase enzyme activity.

ACC deaminase-producing rhizosphere competent Bacillus spp. mitigate salt stress and promote Zea mays growth by modulating ethylene metabolism

Plant growth-promoting rhizobacteria (PGPR) are known for growth promotion and mitigating environmental stresses. Here, we examined the propitiousness of three indigenous salt-tolerant PGPR, i.e., Bacillus subtilis (NBRI 28B), B. subtilis (NBRI 33N), and B. safensis (NBRI 12M) for plant growth promotion and salt stress amelioration in Zea mays. Results of the in vitro plant growth-promoting attribute revealed NBRI 12M demonstrated the highest values at 1M salt (NaCl) concentration. Furthermore, the greenhouse experiment using three Bacillus strains confirmed plant growth-promoting and salt stress-ameliorating ability, through colonizing successfully and mitigating the adverse effects of ethylene by modulating 1-aminocyclopropane-1-carboxylic acid (ACC) accumulation, ACC-oxidase (ACO), and ACC-synthase (ACS) activities under salt stress. Bacillus sp. inoculation has also induced plant response for defense enzymes, chlorophyll, proline and soluble sugar under salt stress. Among three Bacillus strains, NBRI 12M not only demonstrated higher values for plant growth-promoting (PGP) attributes but also the same was observed in the greenhouse experiment. Thus, the outcomes of this comparative study represent for the first time that salt-tolerant Bacillus strains exhibiting multiple PGP attributes under salt stress along with high rhizosphere competence can alleviate salt stress by reducing the stress ethylene level in the host plant.

The Role of 1-Methylcyclopropene in the regulation of ethylene biosynthesis and ethylene receptor gene expression in Mangifera indica L. (Mango Fruit).

Mango (Mangifera indica L.) is respiratory climacteric fruit that ripens and decomposes quickly following their harvest. 1‐methylcyclopropene (1‐MCP) is known to affect the ripening of fruit, delaying the decay of mango stored under ambient conditions. The objective of this study was to clarify the role of 1‐MCP in the regulation of ethylene biosynthesis and ethylene receptor gene expression in mango. 1‐MCP significantly inhibited the 1‐aminocyclopropane‐1‐carboxylic acid (ACC) content. The activity of ACC oxidase (ACO) increased on days 6, 8, and 10 of storage, whereas delayed ACC synthase (ACS) activity increased after day 4. The two homologous ethylene receptor genes, ETR1 and ERS1 (i.e., MiETR1 and MiERS1), were obtained and deposited in GenBank® (National Center for Biotechnology Information‐National Institutes of Health [NCBI‐NIH]) (KY002681 and KY002682). The MiETR1 coding sequence was 2,220 bp and encoded 739 amino acids (aa). The MiERS1 coding sequence was 1,890 bp and encoded 629 aa, similar to ERS1 in other fruit. The tertiary structures of MiETR1 and MiERS1 were also predicted. MiERS1 lacks a receiver domain and shares a low homology with MiETR1 (44%). The expression of MiETR1 and MiERS1 mRNA was upregulated as the storage duration extended and reached the peak expression on day 6. Treatment with 1‐MCP significantly reduced the expression of MiETR1 on days 4, 6, and 10 and inhibited the expression of MiETR1 on days 2, 4, 6, and 10. These results indicated that MiETR1 and MiERS1 had important functions in ethylene signal transduction. Treatment with 1‐MCP might effectively prevent the biosynthesis of ethylene, as well as ethylene‐induced ripening and senescence. This study presents an innovative method for prolonging the storage life of mango after their harvest through the regulation of MiETR1 and MiERS1 expression.

Hydrogen gas alleviates postharvest senescence of cut rose 'Movie star' by antagonizing ethylene.

H2 prolonged the vase life and improved the vase quality of cut roses through repressing endogenous ethylene production and alleviating ethylene signal transduction during the entire senescing period. Recently, the application of hydrogen gas (H2) was shown to improve postharvest quality and longevity in perishable horticultural products, but the specific regulation mechanism remains obscure. Here, endogenous ethylene production and the expression of genes in ethylene biosynthesis and signalling pathway were investigated to explore the crosstalk between H2 and ethylene during the senescence of cut roses. Our results revealed that addition of exogenous ethylene by ethephon accelerated the senescence of cut roses, in which 100mg L-1 ethephon displayed the most obvious senescent phenotype. While the applied different concentrations (1%, 10%, 50% and 100%) of hydrogen-rich water (HRW) conducted different affects in alleviating the senescence of cut roses, and 1% HRW displayed the best ornamental quality and the longest vase life by reducing ethylene production, supported by the decrease of 1-aminocyclopropene-1-carboxylate (ACC) accumulation, ACC synthase (ACS) and ACC oxidase (ACO) activities, and Rh-ACS3 and Rh-ACO1 expressions in ethylene biosynthesis. In addition, HRW increased the transcripts of ethylene receptor genes Rh-ETR1 at blooming period from day 4 to day 6 and suppressed Rh-ETR3 at senescence phase at day 8 after harvest. Furthermore, the relevant affection of HRW on Rh-ETR1 and Rh-ETR3 expressions still existed when the ethylene production was compromised by adequate addition of exogenous ethylene in HRW-treated cut rose petals, and HRW directly repressed the protein level of Rh-ETR3 in a transient expression assay. Overall, the results suggested that H2 is involved in neutralizing ethylene-mediated postharvest in cut flowers.

Synergistic effect of nitric oxide with hydrogen sulfide on inhibition of ripening and softening of peach fruits during storage

The effects of fumigation with NO, H2S and combination of both on the quality, ethylene biosynthesis and softening of postharvest ‘Dahong’ peach fruit were investigated in current study. Results showed that NO or H2S significantly inhibited the increase of rot index, soluble solid contents, decrease of firmness along with titratable acid contents (P < 0.05). Combinatorial treatment (NO+H2S) further inhibited ripening of peach fruits, moreover it also showed significant decline in ACC content, ACC synthase and oxidase activities compared to individual treatments, was mainly linked with reduced ethylene production. The combined treatment significantly inhibited the increase of water-soluble and CDTA-soluble cell wall fractions, the decrease of Na2CO3-soluble fractions in peach fruit with significantly reduced softening related enzymes activities. These observations suggested the existence of synergistic functions between NO and H2S in inhibiting ethylene biosynthesis and cell wall metabolism in order to maintain superior quality of peach fruits.

Induction of postharvest resistance in melon using pulsed light as abiotic stressor

The aim of this study was use the pulsed light (PL) as tool to booster the postharvest resistance in melons. Cantaloupe melons at mature stage were submitted to the treatments as follows: control, pulsed light (PL; 9 J cm−2), 1-methylcyclopropene (1-MCP; 600 nL L-1) and 1-MCP + PL (600 nL L-1 + 9 J cm−2). Postharvest resistance measurements according survival analysis developed at 4 °C, included: ethylene production, 1-aminocyclopropane-l-carboxylic acid (ACC) oxidase activity, ACC synthase activity, firmness, pH, soluble solids, titratable acidity, SS/AT ratio, soluble sugars, vitamin C, total polyphenols, phenylalanine ammonia lyase (PAL) activity, colour epicarp, free polyamines (putrescine-Put, spermine-Spe and spermidine-Spd) and total polyamines (TPo). Combined effect of the PL+1-MCP promoted an additional of 12 days in the shelf life of melons. PL limited the ethylene biosynthesis leading to an immediate upregulation of polyamines (Put + Spe + Spd), hence, associated with high amounts of TPo (56%). PL treatment promoted an increase of PAL activity (67%) that synthesized high content of phenolic compounds (55%). The results suggest that PL with a fluence of 9 J cm−2 stimulate the postharvest defense mechanisms in melon, due to the ethylene production limitation, which was influenced by the increased polyamines, maintaining the quality attributes.

Effect of spermidine and D-arginine treatments on ethylene synthesis in Xinjiang ‘Saimaiti’ apricot

Xinjiang ‘Saimaiti’ apricots in the green ripe stage were used as the experimental materials. The apricots were treated with spermidine (Spd) and d-arginine (D-Arg). Fruit firmness, ethylene production, 1-aminocyclopropane-1-carboxylic-acid (ACC) synthase activity, ACC oxidase activity and gene expression were studied. The results show that Spd treatment could delay the decline of fruit firmness and reduce ACC synthase and ACC oxidase activity through the inhibition of the relative expression of PaACS2 and PaACO, thus further reducing ethylene production and delaying senescence. Inhibition of polyamine synthesis due to d-Arg treatment can increase the activity of ethylene related enzymes and promote ethylene production. The results suggested that Spd treatment inhibits ethylene production. PaACS2 was the key gene involved in regulating the maturation and senescence of ‘Saimaiti’ apricots.

Expression of grape ACS1 in tomato decreases ethylene and alters the balance between auxin and ethylene during shoot and root formation

Ethylene plays an important role in the grape rachis, where its production can be 10 times higher than in the berry. VvACS1 is the only rachis-specific ACC synthase (ACS) gene, and its expression is coincident with ethylene production in the rachis of Vitis vinifera ‘Thompson seedless’. VvACS1 was cloned and ectopically expressed in tomato (Solanum lycopersicum ‘Moneymaker’). Lateral buds were increased in two- or four-week-old 35s∷VvACS1 transgenic tomato plants after transplanting. Compared with wild-type (WT) plants, the transgenic tomato plants showed higher expression of the VvACS1 gene in the flowers, leaves, rachis, and fruits. There was no obvious difference of ACS activity in the fruit of tomato, and only increased ACS activity in the rachis of tomato. Ethylene production was decreased in flowers, leaves, and fruits (seven weeks after full bloom), while the relative expression of endogenous tomato ACS1 and ACS6 genes was not down-regulated by the ectopic expression of VvACS1. These results imply that post-transcriptional or post-translational regulation of ACS may occur, resulting in lower ethylene production in the transgenic tomato plants. Moreover, expression of VvACS1 in tomato resulted in decreased auxin and increased zeatin contents in the lateral buds, as well as reduced or delayed formation of adventitious roots in lateral bud cuttings. RNA-Seq and qRT-PCR analyses of rooted lateral bud cuttings indicated that the relative expression levels of the genes for zeatin O-glucosyltransferase-like, auxin repressed/dormancy-associated protein, and ERF transcription factors were higher in transgenic tomatoes than in WT, suggesting that ethylene may regulate auxin transport and distribution in shoots and that adventitious root formation employs coordination between auxin and ethylene.

Effects of the inoculations using bacteria producing ACC deaminase on ethylene metabolism and growth of wheat grown under different soil water contents

Crop growth and productivity are often impacted by the increased ethylene content induced by adverse environmental conditions such drought. Inoculations with bacteria producing ACC deaminase is considered as a potential biological approach to improve the growth and tolerance of stressed plants by lowering endogenous ethylene level. In this study, germinated wheat seeds were inoculated using three species of the rhizobacteria, which were isolated from the rhizosphere of wheat growing in dryland, and sown in pots. After three weeks, wheat seedlings were exposed to non-limiting water condition, medium drought and severe drought, respectively, for six weeks. The results showed that, irrespective of rhizobacterial inoculations, decreased soil water contents stimulated wheat ethylene metabolism, which was reflected by the significantly increased activity of ACC synthetase and ACC oxidase, besides an increased content of ACC both in the roots and leaves, and an enhanced capacity of leaves to release ethylene, concomitant with a significant decline in shoot and roots biomass. The inoculations of all three rhizobacterial species under each water condition reduced ACC content in wheat leaves, but effects of the inoculations on ACC synthase and ACC oxidase activity in the leaves and roots, ACC content in the roots, the capacity of leaves to release ethylene, and wheat growth varied with water conditions and bacterial species. Hence, both soil water conditions and rhizobacterial inoculations acted on all the processes of ethylene metabolism, with the former being dominant. The inoculations under non-limiting water condition and medium drought promoted shoot and root growth of wheat plants.