ABSTRACT
Raspberry ketone accounts for the characteristic aroma of the raspberry fruit. In order to explore the genes involved in raspberry ketone synthesis, the transcriptome in fruit tissues of two red raspberry varieties "Polka" and "Orange legend", were sequenced and 24213 single genes were obtained. As the red raspberry fruit ripening, genes involved in flavonoid and anthocyanin synthesis were up-regulated, while those associated with lignin synthesis were down-regulated. A gene (RinPKS4) highly related to raspberry ketone synthesis was identified by transcriptome analysis, and RinPKS4 gene was over-expressed in raspberry in order to further understand the function of RinPKS4 gene in raspberry ketone synthesis. The results showed that the gene expression level of RinPKS4 in the leaf tissues of a transgenic lines increased by about 4-fold and the content of raspberry ketone increased by 42.64% compared with the wide type. This study lays a theoretical foundation for further study on the synthesis and regulation of raspberry ketone in red raspberry.
Subject(s)
Butanones , Gene Expression Profiling , Gene Expression Regulation, Plant , Plant Proteins , Rubus , Rubus/genetics , Rubus/metabolism , Rubus/chemistry , Butanones/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Fruit/genetics , Fruit/metabolism , Transcriptome , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Genes, PlantABSTRACT
Ethylene response factors have been shown to be involved in the effects of plant developmental processes and to regulate stress tolerance. The aim of this study was to recognize the regulatory mechanisms of ethylene response factors on tobacco plant height. In this study, a gene-edited mutant (ERF10-KO) and wild type (WT) were utilized as experimental materials. Transcriptome and metabolome analyses were used to investigate the regulatory mechanism of NtERF10 gene editing on plant height in tobacco. Here, through the analysis of differentially expressed genes (DEGs), 2051 genes were upregulated and 1965 genes were downregulated. We characterized the different ERF10-KO and WT plant heights and identified key genes for photosynthesis, the plant hormone signal transduction pathway and the terpene biosynthesis pathway. NtERF10 was found to affect the growth and development of tobacco by regulating the expression levels of the PSAA, PSBA, GLY17 and GGP3 genes. Amino acid metabolism was analyzed by combining analyses of differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs). In addition, we found that members of the bHLH, NAC, MYB, and WRKY transcription factor families have vital roles in regulating plant height. This study not only provides important insights into the positive regulation of the ethylene response factor NtERF10 on plant height during plant growth and development but also provides new research ideas for tobacco molecular breeding.
Subject(s)
Gene Expression Regulation, Plant , Nicotiana , Plant Proteins , Transcription Factors , Nicotiana/genetics , Nicotiana/growth & development , Nicotiana/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Plant Growth Regulators/metabolism , Plant Growth Regulators/genetics , Ethylenes/metabolism , Plants, Genetically Modified/growth & development , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , TranscriptomeABSTRACT
Selenium (Se) is an essential trace element for humans. Low concentrations of Se can promote plant growth and development. Enhancing grain yield and crop Se content is significant, as major food crops generally have low Se content. Studies have shown that Se biofortification can significantly increase Se content in plant tissues. In this study, the genetic transformation of wheat was conducted to evaluate the agronomic traits of non-transgenic control and transgenic wheat before and after Se application. Se content, speciation, and transfer coefficients in wheat grains were detected. Molecular docking simulations and transcriptome data were utilized to explore the effects of selenium-binding protein-A TaSBP-A on wheat growth and grain Se accumulation and transport. The results showed that TaSBP-A gene overexpression significantly increased plant height (by 18.50%), number of spikelets (by 11.74%), and number of grains in a spike (by 35.66%) in wheat. Under normal growth conditions, Se content in transgenic wheat grains did not change significantly, but after applying sodium selenite, Se content in transgenic wheat grains significantly increased. Analysis of Se speciation revealed that organic forms of selenomethionine (SeMet) and selenocysteine (SeCys) predominated in both W48 and transgenic wheat grains. Moreover, TaSBP-A significantly increased the transfer coefficients of Se from solution to roots and from flag leaves to grains. Additionally, it was found that with the increase in TaSBP-A gene overexpression levels in transgenic wheat, the transfer coefficient of Se from flag leaves to grains also increased.
Subject(s)
Gene Expression Regulation, Plant , Plant Proteins , Plants, Genetically Modified , Selenium-Binding Proteins , Selenium , Sodium Selenite , Triticum , Triticum/genetics , Triticum/metabolism , Triticum/growth & development , Selenium-Binding Proteins/metabolism , Selenium-Binding Proteins/genetics , Selenium/metabolism , Plants, Genetically Modified/growth & development , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Sodium Selenite/metabolism , Edible Grain/metabolism , Edible Grain/genetics , Edible Grain/growth & development , Molecular Docking Simulation , Seeds/growth & development , Seeds/metabolism , Seeds/genetics , Seeds/drug effectsABSTRACT
Cadmium (Cd) accumulates in rice and then moves up the food chain, causing serious health problems for humans. Glutathione S-transferase (GST) binds exogenous hazardous compounds to glutathione (GSH), which performs a variety of roles in plant responses to Cd stress. Here, Cd stimulated the transcripts of a novel OsGST gene, and the OsGST protein, which was localized in the nucleus and cytoplasm, was also induced by Cd. In OsGST deletion mutant lines generated by CRISPR/Cas9, more Cd was accumulated, and Cd hypersensitive phenotypes were observed, while transgenic lines overexpressing OsGST exhibited enhanced Cd tolerance and less Cd accumulation. Further analysis indicated that the osgst mutants exhibited considerably greater reactive oxygen species (ROS) and higher GSH level, and the antioxidant activity associated genes' expression were down-regulated, imply that OsGST controlled rice Cd accumulation and resistance through preserving the equilibrium of the GSH and redox in rice.
Subject(s)
Cadmium , Glutathione , Oryza , Plants, Genetically Modified , Oryza/genetics , Oryza/metabolism , Cadmium/metabolism , Cadmium/toxicity , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Glutathione/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Regulation, Plant , Reactive Oxygen Species/metabolism , Glutaredoxins/genetics , Glutaredoxins/metabolismABSTRACT
Multiprotein bridging factor 1 (MBF1) is a very important transcription factor (TF) in plants, whose members influence numerous defense responses. Our study found that MBF1c in Cucurbitaceae was highly conserved. CsMBF1c expression was induced by temperature, salt stress, and abscisic acid (ABA) in cucumber. Overexpressed CsMBF1c enhanced the heat resistance of a cucumber, and the Csmbf1c mutant showed decreased resistance to high temperatures (HTs). CsMBF1c played an important role in stabilizing the photosynthetic system of cucumber under HT, and its expression was significantly associated with heat-related TFs and genes related to protein processing in the endoplasmic reticulum (ER). Protein interaction showed that CsMBF1c interacted with dehydration-responsive element binding protein 2 (CsDREB2) and nuclear factor Y A1 (CsNFYA1). Overexpression of CsNFYA1 in Arabidopsis improved the heat resistance. Transcriptional activation of CsNFYA1 was elevated by CsMBF1c. Therefore, CsMBF1c plays an important regulatory role in cucumber's resistance to high temperatures.
Subject(s)
Cucumis sativus , Gene Expression Regulation, Plant , Plant Proteins , Thermotolerance , Transcription Factors , Cucumis sativus/genetics , Cucumis sativus/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Thermotolerance/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Hot Temperature , Arabidopsis/genetics , Arabidopsis/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolismABSTRACT
Scopoletin and chlorogenic acid (CGA) are important polyphenols that regulate plant growth, development, and stress resistance. The ERF transcription factor WAX INDUCER1 (WIN1) promotes the biosynthesis of cutin, suberine, and wax. However, its full roles in regulating the accumulation of plant secondary metabolites still remain to be further clarified. In this study, NtWIN1 gene encoding a SHINE-type AP2/ERF transcription factor of the Va subgroup was identified from N. tabacum. NtWIN1 showed high expression levels in tobacco stems, sepals, and pistils. Overexpression (OE) and knock-out of NtWIN1 showed that it promoted the accumulation of total polyphenols and altered their composition. Compare to that of WT plants, the CGA contents significantly increased by 25%-50% in the leaves, flowers, and capsules of OE lines, while the scopoletin contents in the OE plants significantly decreased by 30%-67%. In contrast, the CGA contents in ntwin1 lines reduced by 23%-26%, and the scopoletin contents in ntwin1 increased by 38%-75% compare to that of WT plants. Chromatin immunoprecipitation and Dual-Luc transcription activation assays showed that NtWIN1 could bind to the promoters of NtF6'H1 and NtCCoAMT, thereby modulating their expression. The scopoletin content in ntwin1/ntf6'h1 double mutant was significantly lower than that in ntwin1 and WT plants, but showed no significant differences with that in ntf6'h1 mutant, further indicating that the inhibition of NtWIN1 on scopoletin accumulation depends on the activity of NtF6'H1. Our study illustrates the new roles of NtWIN1, and provides a possible target for regulating the synthesis of polyphenols in tobacco.
Subject(s)
Chlorogenic Acid , Gene Expression Regulation, Plant , Nicotiana , Plant Proteins , Scopoletin , Nicotiana/genetics , Nicotiana/metabolism , Scopoletin/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Chlorogenic Acid/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Methionine Adenosyltransferase/metabolism , Methionine Adenosyltransferase/genetics , Plants, Genetically Modified/metabolism , Plants, Genetically Modified/genetics , Genes, PlantABSTRACT
In plants, abiotic stressors are frequently encountered during growth and development. To counteract these challenges, zinc finger proteins play a critical role as transcriptional regulators. The EgrZFP6 gene, which codes for a zinc finger protein of the C2H2 type, was shown to be considerably elevated in the leaves of Eucalyptus grandis seedlings in the current study when they were subjected to a variety of abiotic stimuli, including heat, salinity, cold, and drought. Analysis conducted later showed that in EgrZFP6 transgenic Arabidopsis thaliana, EgrZFP6 was essential for causing hyponastic leaves and controlling the stress response. Furthermore, the transgenic plants showed elevated levels of reactive oxygen species (ROS), such as superoxide and hydrogen peroxide (H2O2). Additionally, in EgrZFP6-overexpressing plants, transcriptome sequencing analysis demonstrated a considerable downregulation of many genes involved in photosynthesis, decreasing electron transport efficiency and perhaps promoting the buildup of ROS. Auxin levels were higher and auxin signal transduction was compromised in the transgenic plants. Stress-related genes were also upregulated in Arabidopsis as a result of EgrZFP6 overexpression. It is hypothesized that EgrZFP6 can downregulate photosynthesis, which would cause the production of ROS in chloroplasts. As a result, this protein may alter plant stress responses and leaf morphology via a retrograde mechanism driven by ROS. These results highlight the significance of zinc finger proteins in this sophisticated process and advance our understanding of the complex link between gene regulation, ROS signaling, and plant stress responses.
Subject(s)
Arabidopsis , Eucalyptus , Gene Expression Regulation, Plant , Photosynthesis , Plant Proteins , Plants, Genetically Modified , Reactive Oxygen Species , Photosynthesis/genetics , Reactive Oxygen Species/metabolism , Eucalyptus/genetics , Eucalyptus/metabolism , Plants, Genetically Modified/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Down-Regulation/genetics , Plant Leaves/metabolism , Plant Leaves/genetics , Stress, Physiological , Hydrogen Peroxide/metabolism , CYS2-HIS2 Zinc FingersABSTRACT
Minimizing cadmium (Cd) contamination in rice grains is crucial for ensuring food security and promoting sustainable agriculture. Utilizing genetic modification to generate rice varieties with low Cd accumulation is a promising strategy due to its cost-effectiveness and operational simplicity. Our study demonstrated that the CRISPR-Cas9-mediated quadruple mutation of the multicopper oxidase genes OsLPR1/3/4/5 in the japonica rice cultivar Tongjing 981 had little effect on yields. However, a notable increase was observed in the cell wall functional groups that bind with Cd. As a result, the quadruple mutation of OsLPR1/3/4/5 enhanced Cd sequestration within the cell wall while reducing Cd concentrations in both xylem and phloem sap, thereby inhibiting Cd transport from roots to shoots. Consequently, Cd concentrations in brown rice and husk in oslpr1/3/4/5 quadruple mutants (qm) decreased by 52% and 55%, respectively, compared to the wild-type. These findings illustrate that the quadruple mutation of OsLPR1/3/4/5 is an effective method for minimizing Cd contamination in rice grains without compromising yields. Therefore, the quadruple mutation of OsLPR1/3/4/5 via biotechnological pathways may represent a valuable strategy for the generation of new rice varieties with low Cd accumulation.
Subject(s)
Cadmium , Mutation , Oryza , Plant Proteins , Oryza/genetics , Oryza/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Soil Pollutants/metabolism , Edible Grain , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , CRISPR-Cas Systems , Oxidoreductases/genetics , Oxidoreductases/metabolism , Food Contamination/analysisABSTRACT
Soybean (Glycine max [Linn.] Merr.) is an important oilseed crop. Although transcription factors (TFs) can coordinate the expression of mRNA and lncRNA, their coordination in the soybean oil synthesis pathway remains unclear. This study examined the interaction between the TF GmDof11 and lncRNA13082 and found that overexpression of GmDof11 led to an increase in the number of Arabidopsis seeds, thousand seed weight, crude protein, hydrolysis amino acid, and soluble sugar. Additionally, it reduced the triglyceride and starch contents and affected the proportion of fatty acids, increasing the contents of palmitic acid, stearic acid, and linolenic acid. The yeast two-hybrid experiments revealed that GmDof11 interacts with GmBCCP1, GmLEC1b, and GmFAB2 proteins. In the RT-qPCR analysis of transgenic soybean roots, it was found that GmDof11 can activate the production of lncRNA13082 and work in conjunction with lncRNA13082 to oversee oil synthesis and nutrient storage. Our research provides robust theoretical evidence for a comprehensive resolution of TF-lncRNA regulation in the soybean oil synthesis network.
Subject(s)
Arabidopsis , Gene Expression Regulation, Plant , Glycine max , Plant Proteins , Plants, Genetically Modified , RNA, Long Noncoding , Transcription Factors , Glycine max/genetics , Glycine max/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Seeds/genetics , Seeds/metabolism , Seeds/chemistry , Soybean Oil/metabolism , Soybean Oil/genetics , Fatty Acids/metabolism , Fatty Acids/biosynthesisABSTRACT
LBD (LATERAL ORGAN BOUNDARIES DOMAIN) transcription factors are key regulators of plant growth and development. In this study, we functionally characterized the PagLBD4 gene in Populus (Populus alba × Populus glandulosa). Overexpression of PagLBD4 (PagLBD4OE) significantly repressed secondary xylem differentiation and secondary cell wall (SCW) deposition, while CRISPR/Cas9-mediated PagLBD4 knockout (PagLBD4KO) significantly increased secondary xylem differentiation and SCW deposition. Consistent with the functional analysis, gene expression analysis revealed that SCW biosynthesis pathways were significantly down-regulated in PagLBD4OE plants but up-regulated in PagLBD4KO plants. We also performed DNA affinity purification followed by sequencing (DAP-seq) to identify genes bound by PagLBD4. Integration of RNA sequencing (RNA-seq) and DAP-seq data identified 263 putative direct target genes (DTGs) of PagLBD4, including important regulatory genes for SCW biosynthesis, such as PagMYB103 and PagIRX12. Together, our results demonstrated that PagLBD4 is a repressor of secondary xylem differentiation and SCW biosynthesis in Populus, which possibly lead to the dramatic growth repression in PagLBD4OE plants.
Subject(s)
Cell Differentiation , Cell Wall , Gene Expression Regulation, Plant , Plant Proteins , Populus , Transcription Factors , Xylem , Populus/genetics , Populus/metabolism , Cell Wall/metabolism , Cell Wall/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Cell Differentiation/genetics , Xylem/metabolism , Xylem/genetics , Plants, Genetically Modified/metabolismABSTRACT
Spatholobus suberectus Dunn (Leguminosae) has been used for medicinal purposes for a long period. Flavonoids are the major bioactive components of S. suberectus. However, there is still limited knowledge of the exact method via which transcription factors (TFs) regulate flavonoid biosynthesis. The full-length transcriptome of S. suberectus was analyzed using SMRT sequencing; 61,548 transcripts were identified, including 12,311 new gene loci, 53,336 novel transcripts, 44,636 simple sequence repeats, 36,414 complete coding sequences, 871 long non-coding RNAs and 6781 TFs. The SsMYB158 TF, which is associated with flavonoid biosynthesis, belongs to the R2R3-MYB class and is localized subcellularly to the nucleus. The overexpression of SsMYB158 in Nicotiana benthamiana and the transient overexpression of SsMYB158 in S. suberectus resulted in a substantial enhancement in both flavonoids and catechin levels. In addition, there was a remarkable upregulation in the expression of essential enzyme-coding genes associated with the flavonoid biosynthesis pathways. Our study revealed SsMYB158 as a critical regulator of flavonoid biosynthesis in S. suberectus and laying the foundation for its molecular breeding.
Subject(s)
Fabaceae , Flavonoids , Gene Expression Regulation, Plant , Plant Proteins , Transcription Factors , Transcriptome , Flavonoids/biosynthesis , Flavonoids/metabolism , Flavonoids/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Transcriptome/genetics , Fabaceae/genetics , Fabaceae/metabolism , Nicotiana/genetics , Nicotiana/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Genes, PlantABSTRACT
Nicotine constitutes approximately 90% of the total alkaloid content in leaves within the Nicotiana species, rendering it the most prevalent alkaloid. While the majority of genes responsible for nicotine biosynthesis express in root tissue, the influence of light on this process through shoot-to-root mobile ELONGATED HYPOCOTYL 5 (HY5) has been recognized. CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1), a key regulator of light-associated responses, known for its role in modulating HY5 accumulation, remains largely unexplored in its relationship to light-dependent nicotine accumulation. Here, we identified NtCOP1, a COP1 homolog in Nicotiana tabacum, and demonstrated its ability to complement the cop1-4 mutant in Arabidopsis thaliana at molecular, morphological, and biochemical levels. Through the development of NtCOP1 overexpression (NtCOP1OX) plants, we observed a significant reduction in nicotine and flavonol content, inversely correlated with the down-regulation of nicotine and phenylpropanoid pathway. Conversely, CRISPR/Cas9-based knockout mutant plants (NtCOP1CR) exhibited an increase in nicotine levels. Further investigations, including yeast-two hybrid assays, grafting experiments, and Western blot analyses, revealed that NtCOP1 modulates nicotine biosynthesis by targeting NtHY5, thereby impeding its transport from shoot-to-root. We conclude that the interplay between HY5 and COP1 functions antagonistically in the light-dependent regulation of nicotine biosynthesis in tobacco.
Subject(s)
Gene Expression Regulation, Plant , Nicotiana , Nicotine , Nicotiana/metabolism , Nicotiana/genetics , Nicotine/biosynthesis , Nicotine/metabolism , Plant Proteins/metabolism , Plant Proteins/genetics , Arabidopsis/metabolism , Arabidopsis/genetics , Ubiquitin-Protein Ligases/metabolism , Ubiquitin-Protein Ligases/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Basic-Leucine Zipper Transcription Factors/genetics , Plants, Genetically Modified/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/geneticsABSTRACT
Potassium (K+) plays a role in enzyme activation, membrane transport, and osmotic regulation processes. An increase in potassium content can significantly improve the elasticity and combustibility of tobacco and reduce the content of harmful substances. Here, we report that the expression analysis of Nt GF14e, a 14-3-3 gene, increased markedly after low-potassium treatment (LK). Then, chlorophyll content, POD activity and potassium content, were significantly increased in overexpression of Nt GF14e transgenic tobacco lines compared with those in the wild type plants. The net K+ efflux rates were severely lower in the transgenic plants than in the wild type under LK stress. Furthermore, transcriptome analysis identified 5708 upregulated genes and 2787 downregulated genes between Nt GF14e overexpressing transgenic tobacco plants. The expression levels of some potassium-related genes were increased, such as CBL-interacting protein kinase 2 (CIPK2), Nt CIPK23, Nt CIPK25, H+-ATPase isoform 2 a (AHA2a), Nt AHA4a, Stelar K+ outward rectifier 1(SKOR1), and high affinity K+ transporter 5 (HAK5). The result of yeast two-hybrid and luciferase complementation imaging experiments suggested Nt GF14e could interact with CIPK2. Overall, these findings indicate that NtGF14e plays a vital roles in improving tobacco LK tolerance and enhancing potassium nutrition signaling pathways in tobacco plants.
Subject(s)
14-3-3 Proteins , Gene Expression Regulation, Plant , Nicotiana , Plant Proteins , Plants, Genetically Modified , Potassium , Nicotiana/genetics , Nicotiana/metabolism , 14-3-3 Proteins/metabolism , 14-3-3 Proteins/genetics , Potassium/metabolism , Plant Proteins/metabolism , Plant Proteins/genetics , Plants, Genetically Modified/metabolism , Stress, Physiological/geneticsABSTRACT
BACKGROUND: γ-Hexachlorocyclohexane (γ-HCH), an organochlorine insecticide of anthropogenic origin, is a persistent organic pollutant (POP) that causes environmental pollution concerns worldwide. Although many γ-HCH-degrading bacterial strains are available, inoculating them directly into γ-HCH-contaminated soil is ineffective because of the low survival rate of the exogenous bacteria. Another strategy for the bioremediation of γ-HCH involves the use of transgenic plants expressing bacterial enzyme for γ-HCH degradation through phytoremediation. RESULTS: We generated transgenic Arabidopsis thaliana expressing γ-HCH dehydrochlroninase LinA from bacterium Sphingobium japonicum strain UT26. Among the transgenic Arabidopsis T2 lines, we obtained one line (A5) that expressed and accumulated LinA well. The A5-derived T3 plants showed higher tolerance to γ-HCH than the non-transformant control plants, indicating that γ-HCH is toxic for Arabidopsis thaliana and that this effect is relieved by LinA expression. The crude extract of the A5 plants showed γ-HCH degradation activity, and metabolites of γ-HCH produced by the LinA reaction were detected in the assay solution, indicating that the A5 plants accumulated the active LinA protein. In some A5 lines, the whole plant absorbed and degraded more than 99% of γ-HCH (10 ppm) in the liquid medium within 36 h. CONCLUSION: The transgenic Arabidopsis expressing active LinA absorbed and degraded γ-HCH in the liquid medium, indicating the high potential of LinA-expressing transgenic plants for the phytoremediation of environmental γ-HCH. This study marks a crucial step toward the practical use of transgenic plants for the phytoremediation of POPs.
Subject(s)
Arabidopsis , Biodegradation, Environmental , Hexachlorocyclohexane , Plants, Genetically Modified , Sphingomonadaceae , Arabidopsis/genetics , Arabidopsis/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Hexachlorocyclohexane/metabolism , Sphingomonadaceae/genetics , Sphingomonadaceae/metabolism , Sphingomonadaceae/enzymology , Soil Pollutants/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Lyases/genetics , Lyases/metabolismABSTRACT
Pteris vittata is the first-reported arsenic (As) hyperaccumulator, which has been applied to phytoremediation of As-contaminated soil. PvACR3, a key arsenite (AsIII) antiporter, plays an important role in As hyperaccumulation in P. vittata. However, its functions in plants are not fully understood. In this study, the PvACR3 gene was heterologously expressed in tobacco, driven by its native promoter (ProPvACR3). After growing at 5 µM AsIII or 10 µM AsV in hydroponics for 1-5 days, PvACR3-expression enhanced the As levels in leaves by 66.4-113 and 51.8-101%, without impacting the As contents in the roots or stems. When cultivated in As-contaminated soil, PvACR3-expressed transgenic plants accumulated 47.9-85.5% greater As in the leaves than wild-type plants. In addition, PvACR3-expression increased the As resistance in transgenic tobacco, showing that enhanced leaf As levels are not detrimental to its overall As tolerance. PvACR3 was mainly expressed in tobacco leaf veins and was likely to unload AsIII from the vein xylem vessels to the mesophyll cells, thus elevating the leaf As levels. This work demonstrates that heterologously expressing PvACR3 under its native promoter specifically enhances leaf As accumulation in tobacco, which helps to reveal the As-hyperaccumulation mechanism in P. vittata and to enhance the As accumulation in plant leaves for phytoremediation.
Subject(s)
Arsenic , Nicotiana , Plant Leaves , Plants, Genetically Modified , Nicotiana/metabolism , Nicotiana/genetics , Arsenic/metabolism , Plant Leaves/metabolism , Plants, Genetically Modified/metabolism , Promoter Regions, Genetic , Biodegradation, Environmental , Soil Pollutants/metabolismABSTRACT
The study of the mechanisms by which melatonin protects against cadmium (Cd) toxicity in plants is still in its infancy, particularly at the molecular level. In this study, the gene encoding a novel serotonin N-acetyltransferase 3 (SNAT3) in rice, a pivotal enzyme in the melatonin biosynthetic pathway, was cloned. Rice (Oryza sativa) OsSNAT3 is the first identified plant ortholog of archaeon Thermoplasma volcanium SNAT. The purified recombinant OsSNAT3 catalyzed the conversion of serotonin and 5-methoxytryptamine to N-acetylserotonin and melatonin, respectively. The suppression of OsSNAT3 by RNAi led to a decline in endogenous melatonin levels followed by a reduction in Cd tolerance in transgenic RNAi rice lines. In addition, the expression levels of genes encoding the endoplasmic reticulum (ER) chaperones BiP3, BiP4, and BiP5 were much lower in RNAi lines than in the wild type. In transgenic rice plants overexpressing OsSNAT3 (SNAT3-OE), however, melatonin levels were higher than in wild-type plants. SNAT3-OE plants also tolerated Cd stress, as indicated by seedling growth, malondialdehyde, and chlorophyll levels. BiP4 expression was much higher in the SNAT3-OE lines than in the wild type. These results indicate that melatonin engineering could help crops withstand Cd stress, resulting in high yields in Cd-contaminated fields.
Subject(s)
Arylalkylamine N-Acetyltransferase , Cadmium , Gene Expression Regulation, Plant , Melatonin , Oryza , Plant Proteins , Plants, Genetically Modified , Stress, Physiological , Oryza/genetics , Oryza/metabolism , Oryza/drug effects , Melatonin/metabolism , Melatonin/pharmacology , Cadmium/metabolism , Cadmium/toxicity , Plant Proteins/metabolism , Plant Proteins/genetics , Plants, Genetically Modified/metabolism , Arylalkylamine N-Acetyltransferase/metabolism , Arylalkylamine N-Acetyltransferase/genetics , Molecular Chaperones/metabolism , Molecular Chaperones/genetics , Serotonin/metabolismABSTRACT
Plant synthetic biology has significant theoretical advantages in exploration and production of plant natural products. However, its contribution to the field of biosynthesis is currently limited due to the lack of efficient chassis systems and related enabling technologies. Synthetic biologists often avoid tobacco as a chassis system because of its long operation cycle, difficulties in genetic and metabolic modification, complex metabolism and purification background, nicotine toxicity, and challenges in accurately controlling for agricultural production. Nevertheless, the tobacco suspension cell chassis system offers a viable solution to these challenges. The objective of this research was to develop a tobacco suspension cell chassis with high scientific and industrial potential. This chassis should exhibit rapid growth, high biomass, excellent dispersion, high transformation efficiency, and minimal nicotine content. Nicotiana benthamiana, which has high applicability in molecular technology, was used to induce suspension cells. The induced suspension cells, named NBS-1, exhibited rapid growth, excellent dispersion, and high biomass, reaching a maximum biomass of 476.39 g/L (fresh weight), which was significantly higher than that of BY-2. The transformation efficiency of the widely utilized pEAQ-HT transient expression system in NBS-1 reached 81%, which was substantially elevated compared to BY-2. The metabolic characteristics and bias of BY-2 and NBS-1 were analyzed using transcriptome data. It was found that the gene expression of pathways related to biosynthesis of flavonoids and their derivatives in NBS-1 was significantly higher, while the pathways related to alkaloid biosynthesis were significantly lower compared to BY-2. These findings were further validated by the total content of flavonoid and alkaloid. In summary, our research demonstrates NBS-1 possesses minimal nicotine content and provides valuable guidance for selecting appropriate chassis for specific products. In conclusion, this study developed NBS-1, a tobacco suspension cell chassis with excellent growth and transformation, high flavonoid content and minimal nicotine content, which has important guiding significance for the development of tobacco suspension cell chassis.
Subject(s)
Nicotiana , Nicotiana/metabolism , Nicotiana/genetics , Synthetic Biology , Plants, Genetically Modified/metabolism , Metabolic Engineering/methods , Cell Culture Techniques/methods , Nicotine/metabolism , Nicotine/biosynthesis , BiomassABSTRACT
Aluminum (Al) toxicity is one of the environmental stress factors that affects crop growth, development, and productivity. MYB transcription factors play crucial roles in responding to biotic or abiotic stresses. However, the roles of MYB transcription factors in Al tolerance have not been clearly elucidated. Here, we found that GmMYB183, a gene encoding a R2R3 MYB transcription factor, is involved in Al tolerance. Subcellular localization studies revealed that GmMYB183 protein is located in the nucleus, cytoplasm and cell membrane. Overexpression of GmMYB183 in Arabidopsis and soybean hairy roots enhanced plant tolerance towards Al stress compared to the wild type, with higher citrate secretion and less Al accumulation. Furthermore, we showed that GmMYB183 binds the GmMATE75 gene promoter encoding for a plasma-membrane-localized citrate transporter. Through a dual-luciferase reporter system and yeast one hybrid, the GmMYB183 protein was shown to directly activate the transcription of GmMATE75. Furthermore, the expression of GmMATE75 may depend on phosphorylation of Ser36 residues in GmMYB183 and two MYB sites in P3 segment of the GmMATE75 promoter. In conclusion, GmMYB183 conferred Al tolerance by promoting the secretion of citrate, which provides a scientific basis for further elucidating the mechanism of plant Al resistance.
Subject(s)
Aluminum , Arabidopsis , Gene Expression Regulation, Plant , Glycine max , Plant Proteins , Promoter Regions, Genetic , Transcription Factors , Aluminum/toxicity , Aluminum/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis/drug effects , Glycine max/genetics , Glycine max/metabolism , Glycine max/drug effects , Transcription Factors/metabolism , Transcription Factors/genetics , Gene Expression Regulation, Plant/drug effects , Plant Proteins/genetics , Plant Proteins/metabolism , Promoter Regions, Genetic/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Stress, Physiological/genetics , Stress, Physiological/drug effects , Plant Roots/metabolism , Plant Roots/genetics , Plant Roots/drug effects , Carrier ProteinsABSTRACT
Atlantic salmon were fed either a diet reflecting current commercial feeds with added oil supplied by a blend of fish oil and rapeseed oil (COM), or a diet formulated with oil from transgenic Camelina sativa containing 20% EPA + DHA (TCO). Salmon were grown from smolt to market size (>3 kg) in sea pens under semi-commercial conditions. There were no differences in growth, feed efficiency or survival between fish fed the TCO or COM diets at the end of the trial. Levels of EPA + DHA in flesh of salmon fed TCO were significantly higher than in fish fed COM. A 140 g fillet from TCO-fed salmon delivered 2.3 g of EPA + DHA, 67% of the weekly requirement level recommended by many health agencies, and 1.5-fold more than the 1.5 g of EPA + DHA for COM-fed fish. Oil from transgenic Camelina supported growth and improved the nutritional quality of farmed salmon in terms of increased "omega-3" supply for human consumers.
Subject(s)
Animal Feed , Brassicaceae , Docosahexaenoic Acids , Eicosapentaenoic Acid , Plant Oils , Plants, Genetically Modified , Salmo salar , Animals , Salmo salar/metabolism , Salmo salar/growth & development , Docosahexaenoic Acids/analysis , Docosahexaenoic Acids/metabolism , Animal Feed/analysis , Eicosapentaenoic Acid/analysis , Eicosapentaenoic Acid/metabolism , Brassicaceae/chemistry , Brassicaceae/metabolism , Brassicaceae/growth & development , Plant Oils/metabolism , Plants, Genetically Modified/chemistry , Plants, Genetically Modified/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/growth & development , Fish Oils/metabolism , Seawater/chemistry , AquacultureABSTRACT
Phytoremediation is an environmental safety strategy that might serve as a viable preventative approach to reduce soil contamination in a cost-effective manner. Using plants to remediate pollution from the environment is referred to as phytoremediation. In the past few decades, plants have undergone genetic manipulation to overcome inherent limitations by using genetically modified plants. This review illustrates the eco-friendly process of cleaning the environment using transgenic strategies combined with omics technologies. Herbicides tolerance and phytoremediation abilities have been established in genetically modified plants. Transgenic plants have eliminated the pesticides atrazine and metolachlor from the soil. To expand the application of genetically engineered plants for phytoremediation process, it is essential to test strategies in the field and have contingency planning. Omics techniques were used for understanding various genetic, hormonal, and metabolic pathways responsible for phytoremediation in soil. Transcriptomics and metabolomics provide useful information as resources to understand the mechanisms behind phytoremediation. This review aims to highlight the integration of transgenic strategies and omics technologies to enhance phytoremediation efficiency, emphasizing the need for field testing and comprehensive planning for successful implementation.