Tolerance Of Transgenic Plants Research Articles

Genome-wide identification and expression analysis of the NHX gene family under salt stress in wheat (Triticum aestivum L).

Salt stress affects plant growth and development, resulting in the loss of crop yield across the world, and sodium-proton antiporters (NHXs) are one of the genes known to promote salt tolerance in transgenic plants. In this study, we conducted a comprehensive genome-wide analysis and expression profile of NHX genes in wheat under salinity stress. We identified 30 TaNHX genes in wheat based on the Na+/H+ exchanger domain, with all genes containing an amiloride motif except one, a known for inhibiting Na+ ions in plants. Phylogenetic analysis classified these genes into three classes with subfamilies: 12 were localized in vacuoles, while 18 were in the endoplasmic reticulum and plasma membrane. Promoter analysis revealed stress-related cis-acting elements, indicating their potential role in abiotic stress tolerance. The non-synonymous (Ka)/synonymous (Ks) ratios highlighted that the majority of TaNHX genes experienced robust purifying selection throughout their evolutionary history. Transcriptomis data analysis and qRT-PCR demonstrated distinct expression patterns for TaNHX genes across various tissues when subjected to salt stress. Additionally, we predicted 20 different miRNA candidates targeting the identified TaNHX genes. Protein-protein interaction prediction revealed NHX6's involvement in the SOS1 pathway, while NHX1 gene exhibit proton antiporter activity. Molecular dynamics (MD) simulations were also conducted to examine the interactions of TaNHX1, TaNHX2, and TaNHX3. These results represent a significant advancement in our understanding of the molecular mechanisms governing Na+ transporters. This may also offer promising avenues for future studies aimed at unraveling the intricate details of their biological roles and applications.

The transcription factor SbHY5 mediates light to promote aluminum tolerance by activating SbMATE and SbSTOP1s expression

Aluminum (Al) toxicity is a major factor limiting crop yields in acid soils. Sweet sorghum (Sorghum bicolor L.) is a high-efficient energy crop widely grown in tropical and subtropical regions of the world, where acid soil is common and Al toxicity is widespread. Here, we characterized a transcription factor SbHY5 in sweet sorghum, which mediated light to promote plant Al stress adaptation. The expression of SbHY5 was induced by Al stress and increasing light intensity. The overexpression of SbHY5 improved Al tolerance in transgenic plants, which was associated with increased citrate secretion and reduced Al content in roots. Meanwhile, SbHY5 was found to localize to the nucleus and displayed transcriptional activity. SbHY5 directly activated the expression of SbMATE, indicating that a HY5-MATE-dependent citrate secretion pathway is involved in Al tolerance in plants. SbSTOP1 was reported as a key transcription factor, regulating several Al tolerance genes. Here, inspiringly, we found that SbHY5 directly promoted the transcription of SbSTOP1, implying the existence of HY5-STOP1-Al tolerance genes-mediated regulatory pathways. Besides, SbHY5 positively regulated its own transcription. Our findings revealed a novel regulatory network in which a light signaling factor, SbHY5, confers Al tolerance in plants by modulating the expression of Al stress response genes.

The CCCH zinc finger protein PeC3H74 of Moso bamboo (Phyllostachys edulis) positively regulates drought and salinity tolerances in transgenic plants

Moso bamboo (Phyllostachys edulis) is one of the most important fast growing non-timber forest products in the world, drought and salinity are the main environmental factors limiting its growth and geographical distribution. CCCH zinc finger protein plays a key role in plant response to environmental stresses, but its functional role in Moso bamboo is poorly understood. Here, we found that Moso bamboo PeC3H74 strongly responds to drought and salt stress. PeC3H74 is a transcriptional activator, and it binds to the CT-rich motif. Overexpression of PeC3H74 enhanced ROS scavenging ability in transgenic Arabidopsis thaliana and Oryza sativa plants and improved the tolerance of transgenic plants to drought stress. Transcriptome analysis showed overexpression of PeC3H74 leading to activation of the ABA signaling and stress-related genes in transgenic plants. Overexpression of PeC3H74 reduced the transgenic plant's sensitivity to exogenous ABA. Meanwhile, yeast-one-hybrid and dual-luciferase assays showed that AtDSR1, an abiotic stress regulator of A. thaliana, may act as a downstream target gene of PeC3H74. More importantly, analysis of phenotypic and physiological indicators in transgenic plants showed that PeC3H74 also regulated the tolerance of transgenic A. thaliana and O. sativa plants to salt stress. Overall, our findings provide a key candidate gene for Moso bamboo molecular breeding for abiotic resistance.

Wheat WRKY transcription factor TaWRKY24 confers drought and salt tolerance in transgenic plants

Drought and salt stress are major environmental conditions that severely limit plant growth and productivity. WRKY transcription factors play a vital role in the responses against biotic or abiotic stress. In this study, TaWRKY24, a gene of the IIe WRKY family identified in wheat, was cloned and characterized. TaWRKY24 was mainly expressed in wheat leaf and stem and induced by treatment with PEG6000, salt, H2O2, ABA, MeJA, and ethrel. TaWRKY24 transient expression in onion epidermal cells suggested its nuclear localization and its transcriptional activation capability characteristics. Overexpression of TaWRKY24 in tobacco improved the seed germination rate and root growth of seedlings in transgenic lines when subjected to higher mannitol and NaCl concentrations. Further research showed that transgenic lines had higher proline and soluble sugars and lower levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Moreover, compared to normal and negative control plants, TaWRKY24 silenced wheat seedlings had reduced growth under salt and drought stress. This study shows that wheat TaWRKY24 is crucial to plant stress, providing an excellent candidate gene for wheat resistance breeding.

Genome-wide characterization of tobacco B-BOX gene family identified two close members play contrast roles under cold stress

Cold stress influences the growth and geographical distribution of plants. Tobacco (Nicotiana tabacum L.), a model in plant biology studies and a world-widely cultivated economic plant from tropical America, exhibits extreme sensitivity to low temperatures. The conserved B-BOX (BBX) transcription factors play vital roles in plant photomorphogenesis and flowering regulation, and their functions in abiotic stresses, including cold stress, have been uncovered recently. However, few studies on tobacco BBX genes have been conducted to date. In this study, 49 non-redundant BBX genes have been identified in the reference tobacco genome. Gene structure, phylogenetic relationship, prediction of cis-regulatory elements in promoters, tissue-specific expression patterns, and response to cold stress were conducted on these BBX gene members. The expression of NtBBX9 and NtBBX11, two close members in the amplified group II, was up-regulated under cold treatments. Overexpression of NtBBX11 embedded the cold tolerance of transgenic plants, while the NtBBX9 overexpressors were susceptible to cold. The NtBBX9 functions as a negative regulator of tobacco response to cold stress, which was confirmed by the phenotype of NtBBX9 RNA-silenced lines. Furthermore, we demonstrated that NtBBX9 and NtBBX11 regulated the cold stress response by alternating the expression of critical cold-regulated genes such as CBFs, LEA14, and LTI65 and reactive oxygen species (ROS) production. The combination of NtBBX9 and NtBBX11 could facilitate tobacco’s rapid response to and recovery from transient cold stress and regulate plant cold-responsive growth in tropical regions.

Inventory of cadmium-transporter genes in the root of mangrove plant Avicennia marina under cadmium stress

Mangrove Avicennia marina has the importantly potential for cadmium (Cd) pollution remediation in coastal wetlands. Unfortunately, the molecular mechanisms and transporter members for Cd uptake by the roots of A. marina are not well documented. In this study, photosynthetic and phenotypic analysis indicated that A. marina is particularly tolerant to Cd. The content and flux analysis indicated that Cd is mainly retained in the roots, with greater Cd influx in fine roots than that in coarse roots, and higher Cd influx in the root meristem zone as well. Using transcriptomic analysis, a total of 5238 differentially expressed genes were identified between the Cd treatment and control group. Moreover, we found that 54 genes were responsible for inorganic ion transport. Among these genes, AmHMA2, AmIRT1, and AmPCR2 were localized in the plasma membrane and AmZIP1 was localized in both plasma membrane and cytoplasm. All above gene encoding transporters showed significant Cd transport activities using function assay in yeast cells. In addition, the overexpression of AmZIP1 or AmPCR2 in Arabidopsis improved the Cd tolerance of transgenic plants. This is particularly significant as it provides insight into the molecular mechanism for Cd uptake by the roots of mangrove plants and a theoretical basis for coastal wetland phytoremediation.

Overexpression of OsPIN9 Impairs Chilling Tolerance via Disturbing ROS Homeostasis in Rice.

The auxin efflux transporter PIN-FORMED (PIN) family is one of the major protein families that facilitates polar auxin transport in plants. Here, we report that overexpression of OsPIN9 leads to altered plant architecture and chilling tolerance in rice. The expression profile analysis indicated that OsPIN9 was gradually suppressed by chilling stress. The shoot height and adventitious root number of OsPIN9-overexpressing (OE) plants were significantly reduced at the seedling stage. The roots of OE plants were more tolerant to N-1-naphthylphthalamic acid (NPA) treatment than WT plants, indicating the disturbance of auxin homeostasis in OE lines. The chilling tolerance assay showed that the survival rate of OE plants was markedly lower than that of wild-type (WT) plants. Consistently, more dead cells, increased electrolyte leakage, and increased malondialdehyde (MDA) content were observed in OE plants compared to those in WT plants under chilling conditions. Notably, OE plants accumulated more hydrogen peroxide (H2O2) and less superoxide anion radicals (O2-) than WT plants under chilling conditions. In contrast, catalase (CAT) and superoxide dismutase (SOD) activities in OE lines decreased significantly compared to those in WT plants at the early chilling stage, implying that the impaired chilling tolerance of transgenic plants is probably attributed to the sharp induction of H2O2 and the delayed induction of antioxidant enzyme activities at this stage. In addition, several OsRboh genes, which play a crucial role in ROS production under abiotic stress, showed an obvious increase after chilling stress in OE plants compared to that in WT plants, which probably at least in part contributes to the production of ROS under chilling stress in OE plants. Together, our results reveal that OsPIN9 plays a vital role in regulating plant architecture and, more importantly, is involved in regulating rice chilling tolerance by influencing auxin and ROS homeostasis.

The wheat VQ motif-containing protein TaVQ4-D positively regulates drought tolerance in transgenic plants.

VQ motif-containing proteins play important roles in plant abiotic and biotic stresses. In this study, we cloned the VQ-protein gene TaVQ4-D that is induced by drought stress. Arabidopsis and wheat plants overexpressing TaVQ4-D showed increased tolerance to drought stress. In contrast, wheat lines in which TaVQ4-D expression had been silenced showed decreased drought tolerance. Under drought stress conditions, the contents of superoxide dismutase and proline increased and the content of malondialdehyde decreased in transgenic wheat plants overexpressing TaVQ4-D compared with WT. At the same time, the expression of ROS scavenging-related genes and stress-related genes was up-regulated. However, plants of TaVQ4-D-silenced wheat lines showed decreased activities of antioxidant enzymes and reduced expression of some stress-related and antioxidant-related genes. In addition, the TaVQ4-D protein physically interacts with two mitogen-activated protein kinases (MPK3 and MPK6) and plays a role in plant drought stress as the phosphorylated substrates of MPK3 and MPK6. In summary, the results of our study suggest that TaVQ4-D can positively regulate drought stress tolerance in wheat.

Overexpression of the Purple Perilla (Perilla frutescens (L.)) FAD3a Gene Enhances Salt Tolerance in Soybean.

The increasingly serious trend of soil salinization inhibits the normal growth and development of soybeans, leading to reduced yields and a serious threat to global crop production. Microsomal ω-3 fatty acid desaturase encoded by the FAD3 gene is a plant enzyme that plays a significant role in α-linolenic acid synthesis via regulating the membrane fluidity to better accommodate various abiotic stresses. In this study, PfFAD3a was isolated from perilla and overexpressed in soybeans driven by CaMV P35S, and the salt tolerance of transgenic plants was then evaluated. The results showed that overexpression of PfFAD3a increased the expression of PfFAD3a in both the leaves and seeds of transgenic soybean plants, and α-linolenic acid content also significantly increased; hence, it was shown to significantly enhance the salt tolerance of transgenic plants. Physiological and biochemical analysis showed that overexpression of PfFAD3a increased the relative chlorophyll content and PSII maximum photochemical efficiency of transgenic soybean plants under salt stress; meanwhile, a decreased accumulation of MDA, H2O2, and O2•-, increased the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbic acid peroxidase (APX), as well as the production of proline and soluble sugar. In summary, the overexpression of PfFAD3a may enhance the salt tolerance in transgenic soybean plants through enhanced membrane fluidity and through the antioxidant capacity induced by C18:3.

Two Sweet Sorghum (Sorghum bicolor L.) WRKY Transcription Factors Promote Aluminum Tolerance via the Reduction in Callose Deposition.

Aluminum (Al) toxicity is a primary limiting factor for crop production in acidic soils. The WRKY transcription factors play important roles in regulating plant growth and stress resistance. In this study, we identified and characterized two WRKY transcription factors, SbWRKY22 and SbWRKY65, in sweet sorghum (Sorghum bicolor L.). Al induced the transcription of SbWRKY22 and SbWRKY65 in the root apices of sweet sorghum. These two WRKY proteins were localized in the nucleus and exhibited transcriptional activity. SbWRKY22 showed the significant transcriptional regulation of SbMATE, SbGlu1, SbSTAR1, SbSTAR2a, and SbSTAR2b, which are major known Al tolerance genes in sorghum. Interestingly, SbWRKY65 had almost no effect on the aforementioned genes, but it significantly regulated the transcription of SbWRKY22. Therefore, it is speculated that SbWRKY65 might indirectly regulate Al-tolerance genes mediated by SbWRKY22. The heterologous expression of SbWRKY22 and SbWRKY65 greatly improved the Al tolerance of transgenic plants. The enhanced Al tolerance phenotype of transgenic plants is associated with reduced callose deposition in their roots. These findings suggest the existence of SbWRKY22- and SbWRKY65-mediated Al tolerance regulation pathways in sweet sorghum. This study extends our understanding of the complex regulatory mechanisms of WRKY transcription factors in response to Al toxicity.

The ERF-VII transcription factor AvERF75 positively regulates the waterlogging tolerance by interacting with AvLOB41 in kiwifruit (Actinidia valvata)

The ethylene-responsive VII (ERF-VII) transcription factors are crucial in regulating plant waterlogging tolerance. However, the positive role of ERF-VII genes in kiwifruit waterlogging tolerance is rarely reported. In this study, we cloned the AvERF75 gene from the waterlogging-tolerant genotype KR5 (Actinidia valvata) and explored its roles in response to waterlogging stress. Subcellular localization showed AvERF75 protein was localized in cytoplasm and nucleus, and transactivation assays indicated it exhibited transactivation activity. Overexpression of AvERF75 significantly improved the waterlogging tolerance of transgenic plants (both tobacco and kiwifruit) by scavenging ROS. furthermore, we identified AvLOB41 as an interacting partner of AvERF75 protein by Y2H screening and BiFC assay. Taken together, this study confirms the positive role of AvERF75 in regulating waterlogging tolerance and reveals a new regulatory network in response to waterlogging stress.

Dehydrin AmDH1 improves drought tolerance in transgenic maize plants

Dehydrin is a large protein group to have protective functions to plant cells under osmotic stress. Overexpression of dehydrin encoded genes isolated from mangrove plants had improved drought tolerance of transgenic plants. Therefore, in this research, a coding sequence with the length of 573bp that encodes Dehydrin1 (DH1) protein was isolated from a mangrove plant namely Mam bien (Avicennia marina) which naturally grows in Xuan Thuy national park, Vietnam. The coding sequence was denoted as AmDH1 and cloned into the plant transformation vector pCAM 35S-AmDH1. The transgenic vector was transformed into A. tumefiaciens cells and the vector was transferred into the K7 maize line to generate transgenic maize seedlings. The AmDH1 transgenic maize T2 seedlings have shown an improved drought tolerance than the original K7 maize line.

Overexpression of a Fragaria vesca 1R-MYB Transcription Factor Gene (FvMYB114) Increases Salt and Cold Tolerance in Arabidopsis thaliana

The MYB (v-MYB avian myeloblastosis viral oncogene homolog) transcription factor (TF) family has numerous members with complex and diverse functions, which play an indispensable role in regulating the response of plants to stress. In this study, a new 1R-MYB TF gene was obtained from Fragaria vesca (a diploid strawberry) by cloning technology and given a new name, FvMYB114. According to the subcellular localization results, FvMYB114 protein was a nuclear localization protein. Overexpression of FvMYB114 greatly enhanced the adaptability and tolerance of Arabidopsis thaliana to salt and low temperature. Under salt and cold stress, the transgenic plants had greater proline and chlorophyll contents and higher activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) than the wild-type (WT) and unloaded-line (UL) A. thaliana. However, malondialdehyde (MDA) was higher in the WT and UL lines. These results suggested that FvMYB114 may be involved in regulating the response of A. thaliana to salt stress and cold stress. FvMYB114 can also promote the expression of genes, such as the genes AtSOS1/3, AtNHX1 and AtLEA3 related to salt stress and the genes AtCCA1, AtCOR4 and AtCBF1/3 related to cold stress, further improving the tolerance of transgenic plants to salt and cold stress.

Overexpression of the Wheat TaPsb28 Gene Enhances Drought Tolerance in Transgenic Arabidopsis

Psb28 is a soluble protein in the photosystem II (PSII) complex, but its role in the drought stress response of wheat remains unclear. Here, we functionally characterized the TaPsb28 gene, which positively regulates drought tolerance in wheat. When the full-length 546-bp TaPsb28 cDNA was transferred into Arabidopsis thaliana, it was located in the guard cell chloroplast around the stroma. Overexpression of TaPsb28 conferred drought tolerance, as exhibited by the increases in the survival rate. Transgenic plants maintained lower MDA content and higher chlorophyll content by inducing chlorophyll synthase (ChlG) gene transcription. The content of abscisic acid (ABA) and zeatin increased significantly in wild-type (WT) plants under drought stress, and the transcriptional expression levels of RD22, dihydroflavonol 4-reductase (DFR) and anthocyanin reductase (ANR) genes were induced, thus enhancing the contents of endogenous cyanidin, delphinidin, and proanthocyanidins. However, in transgenic plants, although anthocyanins were further aggregated, the ABA increase was inhibited, zeatin was restored to the control level under drought stress, and stomatal closure was promoted. These findings indicate ABA and zeatin have opposite synergistic effects in the process of drought tolerance caused by TaPsb28 because only after the effect of zeatin is alleviated can ABA better play its role in promoting anthocyanin accumulation and stomatal closure, thus enhancing the drought tolerance of transgenic plants. The results suggest that overexpression of TaPsb28 exerts a positive role in the drought response by influencing the functional metabolism of endogenous hormones. The understanding acquired through the research laid a foundation for further in-depth investigation of the function of TaPsb28 in drought resistance in wheat, especially its relationship with anthocyanidin accumulation.

Overexpression of RtSYP121 confers cadmium colerance by promoting vesicle trafficking, maintaining ion homeostasis, and alleviating photosynthetic inhibition in Arabidopsis

Cadmium (Cd) is a toxic heavy metal in soil that seriously threatens crop production, food security, and human health. Syntaxins, a prototype family of Soluble N-ethyl-maleimide-associated protein receptors (SNAREs) involved in vesicle trafficking, are implicated in resistance to abiotic stresses, including Cd stress, but the molecular mechanisms underlying the involvement of syntaxins in Cd tolerance in plants are unclear. In this study, we isolated and functionally characterized the syntaxin gene RtSYP121 from Reaumuria trigyna to evaluate its potential for phytoremediation. RtSYP121 resides in the plasma membrane. The transcriptional level of RtSYP121 was strongly increased by salt, drought, and Cd stress. Overexpression of RtSYP121 significantly enhanced the Cd tolerance of transgenic Arabidopsis. The Cd tolerance of transgenic plants mainly depended on elevated vesicle trafficking, which increased the content of K+ and Ca2+ and thus decreased the accumulation of Cd2+ by regulating the delivery or activity of ion transporters, channels, and pumps. Moreover, overexpression of RtSYP121 in Arabidopsis ameliorated Cd stress–induced phytotoxic effects, including growth inhibition, ROS burst, photosynthetic impairment, and cell death. Therefore, we suggest that RtSYP121 plays multiple roles in the plant response to Cd stress by promoting vesicle trafficking, maintaining ion homeostasis, and alleviating photosynthetic inhibition.

Constitutive expression of AtSINA2 from Arabidopsis improves grain yield, seed oil and drought tolerance in transgenic soybean

The SEVEN IN Absentia (SINA), a typical member of the RING E3 ligase family, plays a crucial role in plant growth, development and response to abiotic stress. However, its biological functions in oil crops are still unknown. Previously, we reported that overexpression of AtSINA2 in Arabidopsis positively regulated the drought tolerance of transgenic plants. In this work, we demonstrate that ectopic expression of AtSINA2 in soybean improved the shoot growth, grain yield, drought tolerance and seed oil content in transgenic plants. Compared to wild type, transgenic soybean produced greater shoot biomass and grain yield, and showed improved seed oil and drought tolerance. Physiological analyses exhibited that the increased drought tolerance of transgenic plants was accompanied with a higher chlorophyll content, and a lower malondialdehyde accumulation and water loss during drought stress. Further transcriptomic analyses revealed that the expressions of genes related to plant growth, flowering and stress response were up- or down-regulated in transgenic soybean under both normal and drought stress conditions. Our findings imply that AtSINA2 improved both agricultural production and drought tolerance, and it can be used as a candidate gene for the genetic engineering of new soybean cultivars with improved grain yield and drought resistance.

Integration of chromatin accessibility and gene expression reveals new regulators of cold hardening to enhance freezing tolerance in Prunus mume.

Low temperature is one of the most important abiotic factors limiting the growth, development and geographical distribution of plants. Prunus mume is an attractive woody ornamental plant that blooms in early spring in Beijing. However, the molecular mechanisms underlying cold hardening to enhance freezing tolerance in Prunus genus remains elusive. This study examined the dynamic physiological responses induced by cold hardening, and identified freezing-tolerance genes by RNA-seq and ATAC-seq analyses. Cold hardening elevated the content of soluble substances and enhanced freezing resistance in P. mume. Transcriptome analysis indicated that the candidate differentially expressed genes (DEGs) were those enriched in Ca2+ signalling, mitogen-activated protein kinase (MAPK) cascade, abscisic acid signalling, and inducer of CBF expression 1 (ICE)-C-repeat binding factor (CBF) signalling pathways. The openness of gene chromatin positively correlated with the expression level of these genes. Thirteen motifs were identified in the open chromatin regions in the treatment group subjected to freezing after cold hardening. The chromatin opening of transcription start site at the proximal -177 region of cold-shock protein CS120-like (PmCSL) was markedly increased, while the expression level of PmCSL was significantly up-regulated. Overexpression of PmCSL in Arabidopsis significantly improved the freezing tolerance of transgenic plants. These findings provide new insights into the regulatory mechanism of freezing tolerance to improve breeding of cold-hardy P. mume plants.

Functional Characterization of Sugar Beet M14 Antioxidant Enzymes in Plant Salt Stress Tolerance.

Salt stress can cause cellular dehydration, which induces oxidative stress by increasing the production of reactive oxygen species (ROS) in plants. They may play signaling roles and cause structural damages to the cells. To overcome the negative impacts, the plant ROS scavenging system plays a vital role in maintaining the cellular redox homeostasis. The special sugar beet apomictic monosomic additional M14 line (BvM14) showed strong salt stress tolerance. Comparative proteomics revealed that six antioxidant enzymes (glycolate oxidase (GOX), peroxiredoxin (PrxR), thioredoxin (Trx), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), and dehydroascorbate reductase3 (DHAR3)) in BvM14 were responsive to salt stress. In this work, the full-length cDNAs of genes encoding these enzymes in the redox system were cloned from the BvM14. Ectopic expression of the six genes reduced the oxidative damage of transgenic plants by regulating the contents of hydrogen peroxide (H2O2), malondialdehyde (MDA), ascorbic acid (AsA), and glutathione (GSH), and thus enhanced the tolerance of transgenic plants to salt stress. This work has charecterized the roles that the antioxidant enzymes play in the BvM14 response to salt stress and provided useful genetic resources for engineering and marker-based breeding of crops that are sensitive to salt stress.

EARLY FLOWERING3 Gene Confers Earlier Flowering and Enhancement of Salt Tolerance in Woody Plant Osmanthus fragrans

Osmanthus fragrans Lour. is popular in landscaping and gardening in Asia. In recent years, growing attention has been given to evergreen tree flowering and adaptation. EARLY FLOWERING3 (ELF3) plays an essential role in plant flowering regulation and abiotic stress tolerance. However, there is very little known about how the ELF3 gene affects flowering time and salt tolerance in O. fragrans. To elucidate the potential role of the flowering-related gene ELF3 in responding to salt tolerance, a significantly upregulated gene OfELF3 was obtained by RNA sequencing (RNA-seq) after salt treatment in O. fragrans. Our results showed that OfELF3 is a nuclear protein, which did not have a transcriptional activation ability. OfELF3 accumulation was determined in different tissues and the differentiation process of floral buds by qRT–PCR, and the gene was also significantly induced by salt stress treatment. In addition, overexpression of OfELF3 accelerated the flowering time of transgenic Arabidopsis lines, and an increase in the expression of flowering integrators such as AtFT, AtSOC1, and AtAP1 was investigated. Moreover, OfELF3 overexpression significantly improved the salt tolerance of transgenic plants, seed germination and root length of transgenic plants and was superior to those of the wild type (WT) under NaCl treatment at 4 days post-germination and the 5-day-old seedling stage, respectively. Similarly, phenotype and physiological indexes (REL, MDA and soluble protein) of 3-week-old transgenic plants were superior to the WT plants as well. Together, our results suggest that OfELF3 is not only a positive regulator in the regulation of flowering but is also involved in the salt tolerance response in O. fragrans.

Overexpression of acdS in Petunia hybrida Improved Flower Longevity and Cadmium-Stress Tolerance by Reducing Ethylene Production in Floral and Vegetative Tissues.

The role of acdS, which encodes the 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzyme, in extending flower longevity and improving tolerance to cadmium (Cd) stress was assessed using transgenic Petunia hybrida cv. ‘Mirage Rose’ overexpressing acdS and wild-type (WT) plants. The overexpression of acdS reduced ethylene production in floral tissue via suppression of ethylene-related genes and improved flower longevity, approximately 2 to 4 days longer than WT flowers. Under Cd stress, acdS significantly reduced Cd-induced ethylene production in vegetable tissues of transgenic plants through suppression of ethylene-related genes. This resulted in a lower accumulation of ethylene-induced reactive oxygen species (ROS) in the transgenic plants than in WT plants. In addition, expression of the genes involved in the activities of antioxidant and proline synthesis as well as the metal chelation process was also higher in the former than in the latter. Moreover, Cd accumulation was significantly higher in WT plants than in the transgenic plants. These results are linked to the greater tolerance of transgenic plants to Cd stress than the WT plants, which was determined based on plant growth and physiological performance. These results highlight the potential applicability of using acdS to extend flower longevity of ornamental bedding plants and also reveal the mechanism by which acdS improves Cd-stress tolerance. We suggest that acdS overexpression in plants can extend flower longevity and also help reduce the negative impact of Cd-induced ethylene on plant growth when the plants are unavoidably cultivated in Cd-contaminated soil.