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Plasma membrane intrinsic proteins (PIPs), one sub-family of aquaporins (AQPs), are responsible for plant abiotic stress responses. However, little information is currently available about the stress responsiveness of the PIP promoter in vegetable pea. In the present study, one novel promoter of PsPIP2-4 which shared high similarity to the PIP2-type AQPs from other plants, was isolated. Quantitative real-time PCR (qRT-PCR) assays suggested that PsPIP2-4 was predominantly expressed in leaves and abundantly induced by abiotic stress treatments (polyethylene glycol (PEG) 6000, NaCl, and methyl jasmonate (MeJA)). Further, the promoter activity of PsPIP2-4 was verified in transgenic tobacco plants. Beta-glucuronidase (GUS) staining driven by the PsPIP2-4 promoter confirmed that it was mainly detected in the leaves of transgenic seedlings, especially in the guard cells. Exposure of transgenic seedlings to various environmental stimuli proved that the promoter activity of PsPIP2-4 was abundantly strengthened by osmotic, salt, and MeJA stresses. This research provides one stress-inducible promoter enabling targeted gene expression under abiotic stresses and demonstrates its usefulness in the genetic improvement of plant stress resistance. [ABSTRACT FROM AUTHOR]

Climate change and human activities impose severe biotic and abiotic stresses that threaten plant survival. To cope with these extreme conditions, plants have evolved various resistance mechanisms. In recent years, genetic approaches have been increasingly employed to enhance abiotic stress tolerance and develop stress-resistant plants. In this study, we introduced three transporter-related genes from extreme haloarchaea, which thrive in saturated NaCl environments, into the Nicotiana tabacum genome to improve its salt and drought tolerance mechanisms. The U2845, U3508, and CL165 genes were subcloned into the pIPKb004 vector, which includes the cauliflower mosaic virus 35S (CaMV35S) promoter and a hygromycin phosphotransferase II (hptlI) selection marker, using Gateway cloning. The recombinant vector was introduced into the N. tabacum genome via Agrobacterium-mediated transformation. Wild-type and transgenic plants were cultivated under 150 mM NaCl and 200 mM mannitol, along with a no-stress control. The effects of these genes were evaluated through morphological examination, germination rate analysis, proline and malondialdehyde accumulation measurements, and antioxidant enzyme activity assays. The transgenic lines demonstrated enhanced stress tolerance, as evidenced by improved germination rates, increased proline accumulation, and elevated antioxidant enzyme activities. These findings underscore the potential of haloarchaeal genes as valuable genetic resources for developing stress-tolerant plants. [ABSTRACT FROM AUTHOR]

Abstract Background The GT64 subfamily, belonging to the glycosyltransferase family, plays a critical function in plant adaptation to stress conditions and the modulation of plant growth, development, and organogenesis processes. However, a comprehensive identification and systematic analysis of GT64 in cotton are still lacking. Results This study used bioinformatics techniques to conduct a detailed investigation on the GT64 gene family members of eight cotton species for the first time. A total of 39 GT64 genes were detected, which could be classified into five subfamilies according to the phylogenetic tree. Among them, six genes were found in upland cotton. Furthermore, investigated the precise chromosomal positions of these genes and visually represented their gene structure details. Moreover, forecasted cis-regulatory elements in GhGT64s and ascertained the duplication type of the GT64 in the eight cotton species. Evaluation of the Ka/Ks ratio for similar gene pairs among the eight cotton species provided insights into the selective pressures acting on these homologous genes. Additionally, analyzed the expression profiles of the GT64 gene family. Overexpressing GhGT64_4 in tobacco improved its disease resistance. Subsequently, VIGS experiments conducted in cotton demonstrated reduced disease resistance upon silencing of the GhGT64_4, may indicate its involvement in affecting lignin and jasmonic acid biosynthesis pathways, thus impacting cotton resistance. Weighted Gene Co-expression Network Analysis (WGCNA) revealed an early immune response against Verticillium dahliae in G. barbadense compared to G. hirsutum. Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) analysis indicated that some GT64 genes might play a role under various biotic and abiotic stress conditions. Conclusions These discoveries enhance our knowledge of GT64 family members and lay the groundwork for future investigations into the disease resistance mechanisms of this gene in cotton.

Background: The GT64 subfamily, belonging to the glycosyltransferase family, plays a critical function in plant adaptation to stress conditions and the modulation of plant growth, development, and organogenesis processes. However, a comprehensive identification and systematic analysis of GT64 in cotton are still lacking. Results: This study used bioinformatics techniques to conduct a detailed investigation on the GT64 gene family members of eight cotton species for the first time. A total of 39 GT64 genes were detected, which could be classified into five subfamilies according to the phylogenetic tree. Among them, six genes were found in upland cotton. Furthermore, investigated the precise chromosomal positions of these genes and visually represented their gene structure details. Moreover, forecasted cis-regulatory elements in GhGT64s and ascertained the duplication type of the GT64 in the eight cotton species. Evaluation of the Ka/Ks ratio for similar gene pairs among the eight cotton species provided insights into the selective pressures acting on these homologous genes. Additionally, analyzed the expression profiles of the GT64 gene family. Overexpressing GhGT64_4 in tobacco improved its disease resistance. Subsequently, VIGS experiments conducted in cotton demonstrated reduced disease resistance upon silencing of the GhGT64_4, may indicate its involvement in affecting lignin and jasmonic acid biosynthesis pathways, thus impacting cotton resistance. Weighted Gene Co-expression Network Analysis (WGCNA) revealed an early immune response against Verticillium dahliae in G. barbadense compared to G. hirsutum. Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) analysis indicated that some GT64 genes might play a role under various biotic and abiotic stress conditions. Conclusions: These discoveries enhance our knowledge of GT64 family members and lay the groundwork for future investigations into the disease resistance mechanisms of this gene in cotton. [ABSTRACT FROM AUTHOR]

To elucidate the function of the cold-resistance regulatory gene FmNAC1 from Fraxinus mandshurica Rupr., this study identified the role that overexpression of the FmNAC1 gene plays in tobacco growth and cold-stress regulation. The cloned FmNAC1 gene from F. mandshurica is 891 bp in length and encodes 296 amino acids. Our subcellular localization analysis confirmed that FmNAC1 is primarily located in the nucleus and functions as a transcription factor. FmNAC1 is responsive to cold and NaCl stress, as well as to the induction of IAA, GA, and ABA hormone signals. To further elucidate its function in cold resistance, four transgenic tobacco lines expressing FmNAC1 (FmNAC1-OE) were generated through tissue culture after the Agrobacterium-mediated transformation of wild-type (WT) Nicotiana tabacum L. These FmNAC1-OE plants exhibited accelerated growth after transplantation. When exposed to low-temperature conditions at −5 °C for 24 h, the rates of wilting and yellowing of the FmNAC1-OE plants were significantly lower than those of the WT tobacco plants. Additionally, the membrane integrity, osmotic regulation, and reactive oxygen species (ROS)-scavenging abilities of the FmNAC1-OE tobacco lines were better than those of the WT plants, indicating the potential of the FmNAC1 gene to improve plant cold resistance. The gene expression results further revealed that the FmNAC1 transcription factor exhibits regulatory interactions with growth-related genes such as IAA and AUX1; cold-resistance-related genes such as ICE, DREB, and CBF1; and genes involved in the clearance of reactive oxygen species (ROS), such as CAT and SOD. All of this evidence shows that the FmNAC1 transcription factor from F. mandshurica plays a key role in contributing to the enhancement of growth, cold resistance, and ROS clearance in transgenic tobacco plants. [ABSTRACT FROM AUTHOR]

bZIP transcription factors play important roles in regulating plant development and stress responses. Although bZIPs have been identified in many plant species, there is little information on the bZIPs in Chrysanthemum. In this study, bZIP TFs were identified from the leaf transcriptome of C. mongolicum, a plant naturally tolerant to drought. A total of 28 full-length bZIP family members were identified from the leaf transcriptome of C. mongolicum and were divided into five subfamilies based on their phylogenetic relationships with the bZIPs from Arabidopsis. Ten conserved motifs were detected among the bZIP proteins of C. mongolicum. Subcellular localization assays revealed that most of the CmbZIPs were predicted to be localized in the nucleus. A novel bZIP gene, designated as CmbZIP9, was cloned based on a sequence of the data of the C. mongolicum transcriptome and was overexpressed in tobacco. The results indicated that the overexpression of CmbZIP9 reduced the malondialdehyde (MDA) content and increased the peroxidase (POD) and superoxide dismutase (SOD) activities as well as the expression levels of stress-related genes under drought stress, thus enhancing the drought tolerance of transgenic tobacco lines. These results provide a theoretical basis for further exploring the functions of the bZIP family genes and lay a foundation for stress resistance improvement in chrysanthemums in the future. [ABSTRACT FROM AUTHOR]

Abstract Plant cellulose synthase is one of the important glycosyltransferases, which catalyzes the synthesis of the paracrystalline of H-bonded-β-(1,4)-glucose chains. This study isolated the cellulose synthase 3 (Ces3) sequence from sugarcane (Saccharum officinarum L.) leaves. SoCes3 (GenBank accession No. MG324347) has a full-length cDNA sequence of 3625 bp. It contains an open reading frame (3225 bp), encoding 1074 amino acids with a molecular weight of about 120.89 kDa and isoelectric point of 6.26. SoCes3 protein showed high activity with other plant cellulose synthases. The recombinant protein contains plant cellulose synthase (Ces) protein conservative domains. In subcellular localization experiments, the fusion protein of SoCes3 with green fluorescent protein (GFP) was specifically localized in the cell membrane. The gene expression of SoCes3 was found in the leaf, leaf sheath, and internodes of the sugarcane stem. The highest expression level was found in the internode, especially with the highest expression level in the 5th internode and lowest in the leaves, and the gene expression level of SoCes3 was upregulated by PP333 and not in gibberellic acid-treated plants. It was conducted in tobacco plants to understand the biotechnological potential of SoCes3. The contents of cellulose and lignin were increased in SoCes3-overexpressing tobacco. Transcriptomic analysis showed that the transgenic tobacco induced different genes associated with different biological regulatory processes. Differentially expressed genes (DEGs) mediated plant hormone signal transduction, starch and sucrose metabolism signaling pathways were widely induced and mostly upregulated. The transcription levels in SoCes3-overexpressing transgenic lines were higher than wild-type. Graphical Abstract

[ Objective ] Monogalactose diglyceride synthase ( MGD ) is a key enzyme in the synthesis of mo- nogalactose diglyceride ( MGDG ) and plays an important role in plant response to low phosphorus toler- ance. We aim to understand the role of the OsMGD2 and OsMGD3 genes of rice in response to phosphorus ( P ) deficiency. [ Methods ] A pot experiment was conducted to analyze physiological responses and lipid composition changes of the wild-type ( SR1 ) and transgenic tobacco under normal and low P condition. [ Results ] There was no difference in P content between the transgenic and wild-type tobacco under normal and P deficiency conditions. However , the biomass , chlorophyll content , and photosynthetic electronic transfer of the transgenic tobacco were higher than wild-type. The phospholipid ( PL ) content , digalactose diglyceride ( DGDG ) content , DGDG / MGDG ratio , and GL / PL ratio of the transgenic tobacco were higher than those of the wild-type tobacco under low phosphorus deficiency , and the lipid content and ratio of the tobacco expressing OsMGD3 were higher than the tobacco expressing OsMGD2 . [ Conclusion ] Regulation of OsMGD2 / 3 gene expression in rice can improve lipid remodeling ability and maintain higher photosyn- thetic and growth capacity under phosphorus deficiency , increasing plant tolerance to low phosphorus stress.

Background: Heat stress is a detrimental abiotic stress that limits the development of many plant species and is linked to a variety of cellular and physiological problems. Heat stress affects membrane fluidity, which leads to negative effects on cell permeability and ion transport. Research reveals that heat stress causes severe damage to cells and leads to rapid accumulation of reactive oxygen species (ROS), which could cause programmed cell death. Methods and results: This current study aimed to validate the role of Triticum aestivum Salt Stress Root Protein (TaSSRP) in plants' tolerance to heat stress by modulating its expression in tobacco plants. The Relative Water Content (RWC), total chlorophyll content, and Membrane Stability Index (MSI) of the seven distinct transgenic lines (T0 − 2, T0 − 3, T0 − 6, T0 − 8, T0 − 9, T0 − 11, and T0 − 13), increased in response to heat stress. Despite the fact that the same tendency was detected in wild-type (WT) plants, changes in physio-biochemical parameters were greater in transgenic lines than in WT plants. The expression analysis revealed that the transgene TaSSRP expressed from 1.00 to 1.809 folds in different lines in the transgenic tobacco plants. The gene TaSSRP offered resistance to heat stress in Nicotiana tabacum, according to the results of the study. Conclusion: These findings could help to improve our knowledge and understanding of the mechanism underlying thermotolerance in wheat, and the novel identified gene TaSSRP could be used in generating wheat varieties with enhanced tolerance to heat stress. [ABSTRACT FROM AUTHOR]

Plant cellulose synthase is one of the important glycosyltransferases, which catalyzes the synthesis of the paracrystalline of H-bonded-β-(1,4)-glucose chains. This study isolated the cellulose synthase 3 (Ces3) sequence from sugarcane (Saccharum officinarum L.) leaves. SoCes3 (GenBank accession No. MG324347) has a full-length cDNA sequence of 3625 bp. It contains an open reading frame (3225 bp), encoding 1074 amino acids with a molecular weight of about 120.89 kDa and isoelectric point of 6.26. SoCes3 protein showed high activity with other plant cellulose synthases. The recombinant protein contains plant cellulose synthase (Ces) protein conservative domains. In subcellular localization experiments, the fusion protein of SoCes3 with green fluorescent protein (GFP) was specifically localized in the cell membrane. The gene expression of SoCes3 was found in the leaf, leaf sheath, and internodes of the sugarcane stem. The highest expression level was found in the internode, especially with the highest expression level in the 5th internode and lowest in the leaves, and the gene expression level of SoCes3 was upregulated by PP333 and not in gibberellic acid-treated plants. It was conducted in tobacco plants to understand the biotechnological potential of SoCes3. The contents of cellulose and lignin were increased in SoCes3-overexpressing tobacco. Transcriptomic analysis showed that the transgenic tobacco induced different genes associated with different biological regulatory processes. Differentially expressed genes (DEGs) mediated plant hormone signal transduction, starch and sucrose metabolism signaling pathways were widely induced and mostly upregulated. The transcription levels in SoCes3-overexpressing transgenic lines were higher than wild-type. [ABSTRACT FROM AUTHOR]

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Chromatin remodelers are essential for regulating plant growth, development, and responses to environmental stresses. HIT4 (HEAT-INTOLERANT 4) is a novel stress-induced chromatin remodeling factor that has been less studied in abiotic stress and stress resistance, particularly in cotton. In this study, we conducted a comprehensive analysis of the members of the HIT4 gene family in Gossypium hirsutum using bioinformatics methods, including phylogenetic relationships, gene organization, transcription profiles, phylogenetic connections, selection pressure, and stress response. A total of 18 HIT4 genes were identified in four cotton species, with six HIT4 gene members in upland cotton. Based on the evolutionary relationships shown in the phylogenetic tree, the 18 HIT4 protein sequences were classified into four distinct subgroups. Furthermore, we conducted chromosome mapping to determine the genomic locations of these genes and visually represented the structural characteristics of HIT4 in G. hirsutum. In addition, we predicted the regulatory elements in HIT4 in G. hirsutum and conducted an analysis of repetitive sequences and gene collinearity among HIT4 in four cotton species. Moreover, we calculated the Ka/Ks ratio for homologous genes to assess the selection pressure acting on HIT4. Using RNA-seq, we explored the expression patterns of HIT4 genes in G. hirsutum and Gossypium barbadense. Through weighted gene co-expression network analysis (WGCNA), we found that GHHIT4_4 belonged to the MEblue module, which was mainly enriched in pathways such as DNA replication, phagosome, pentose and glucuronate interconversions, steroid biosynthesis, and starch and sucrose metabolism. This module may regulate the mechanism of upland cotton resistance to Verticillium wilt through DNA replication, phagosome, and various metabolic pathways. In addition, we performed heterologous overexpression of GH_D11G0591 (GHHIT4_4) in tobacco, and the results showed a significant reduction in disease index compared to the wild type, with higher expression levels of disease resistance genes in the transgenic tobacco. After conducting a VIGS (virus-induced gene silencing) experiment in cotton, the results indicated that silencing GHHIT4_4 had a significant impact, the resistance to Verticillium wilt weakened, and the internode length of the plants significantly decreased by 30.7% while the number of true leaves increased by 41.5%. qRT-PCR analysis indicated that GHHIT4_4 mainly enhanced cotton resistance to Verticillium wilt by indirectly regulating the PAL, 4CL, and CHI genes. The subcellular localization results revealed that GHHIT4_4 was predominantly distributed in the mitochondria and nucleus. This study offers preliminary evidence for the involvement of the GHHIT4_4 in cotton resistance to Verticillium wilt and lays the foundation for further research on the disease resistance mechanism of this gene in cotton. [ABSTRACT FROM AUTHOR]

NADPH-dependent thioredoxin reductase (NTR), which is a flavin protein family of disulfide oxidoreductases present in all living cells, reduces thioredoxin (Trx). The roles of NTR B under stress conditions have not been well characterized. In this study, a tomato NTR B (SlNTRB) gene was cloned and functionally characterized. The mRNA transcript level of SlNTRB was induced significantly after nitrate treatment. The seed germination rate of SlNTRB overexpressed transgenic tobacco was higher than wild type (WT) under nitrate stress. The growth of SlNTRB overexpressed plants was better than WT and the reactive oxygen species (ROS) accumulation were lower than WT under nitrate stress. The NTR activity, superoxide dismutase (SOD) activity, the GSH/GSSG ratio, and the expression of Trx were significantly higher than WT after nitrate stress treatment. The SlNTRB overexpressed transgenic tobacco seeds showed higher tolerance to H2O2 and methyl viologen (MV) treatments than WT. The above results indicated that the SlNTRB gene improved the plant nitrate stress tolerance by alleviating the oxidative damage.Key message: Overexpression of tomato SlNTRB gene in tobacco enhances the excess nitrate stress tolerance with less oxidative damage. [ABSTRACT FROM AUTHOR]

Spodoptera frugiperda is a notorious insect-pest that causes massive yield losses to several agriculturally important food crops every year. S. frugiperda decimates food crops like tomato, potato, cotton, sugarcane, brinjal globally. Various pest resistance genes have been deployed to improve crop resistance towards insect pests, mainly through the transgenic expression of Cry toxins. In this study, we showed that a chimeric Bt gene cry1AcF developed by swapping the domains of cry1Ac and cry1F genes, was effective against S. frugiperda. The T-DNA region harbouring cry1AcF gene under the transcriptional control of CaMV35S promoter was introduced into tobacco (Nicotiana tabacum L. cv. Petit Havana) using Agrobacterium-mediated plant transformation. The transgenic tobacco plants were initially screened on kanamycin and characterized by PCR and Southern hybridization to confirm the transgene integration. Cry1AcF expression was detected with strip test and western blot experiments. Insect bioassay was performed with detached leaves to evaluate the extent of leaf damage and larval mortality in order to select transgenic events with superior performance. Further, insect bioassay conducted using selected transgenic lines, resulted up to 82% larval mortality, indicating high efficacy against S. frugiperda. Therefore, in this study, the transgenic tobacco lines expressing cry1AcF gene showed resistance against S. frugiperda insect. Further, the cry1AcF gene could be effectively utilized to combat resistance against S. frugiperda in other susceptible crops. [ABSTRACT FROM AUTHOR]

[Objective] The Larix kaempferi flavanone 3-hydroxylase 2 (LkF3H2) gene in Larix kaempferi plays a crucial role in flavonoid biosynthesis. This study is aimed to investigate the function of the LkF3H2 in L. kaempferi and plant flavonoid biometabolism. [Method] The LkF3H2 gene of L. kaempferi was cloned based on preliminary transcriptome data, and bioinformatics analysis and tissue expression analysis were performed. The gene was then stably transformed into tobacco to examine the relationship between gene expression and flavonoid content using tissue expression analysis and flavonoid content determination. [Result] The cDNA sequence length of LkF3H2 gene is 1 074 bp, and its protein encodes 358 amino acids with a molecular formula of C1785H2807N481O541S18 and a molecular weight of 40.24 kD, indicating that it is an unstable hydrophilic protein lacking a signal peptide. Moreover, phylogenetic analysis revealed that LkF3H2 is closely related to the F3H genes of Pinus taeda, Pinus radiata, Picea abies, and Picea glauca. In addition, the LkF3H2 gene had the highest expression in the leaves of one-month-old L. kaempferi seedlings and the stems of one-year L. kaempferi seedlings, as determined by tissue expression analysis. Moreover, it had the highest expression in the transgenic tobacco leaves; notably, transgenic tobacco leaves had the highest flavonoid content, followed by stems and roots, indicating a positive relationship between flavonoid content and LkF3H2 gene expression in transgenic tobacco. [Conclusion] The LkF3H2 gene of L. kaempfer belongs to the 2-oxoglutarate-dependent dioxygenase family and is pivotal in flavonoid biosynthesis. [ABSTRACT FROM AUTHOR]

Dof (DNA binding with one finger) proteins are plant-specific transcription factors involved in gene regulation during abiotic and biotic stresses. Our previous studies suggested that tobacco Dof proteins, including BBF3, can upregulate the expression of the virus resistance N gene as well as resistance-related genes. In this study, we generated and characterized transgenic tobacco lines carrying the N gene that overexpressed BBF3 cDNA constitutively. In the absence of virus challenge, the BBF3 overexpression caused no developmental defects and no changes in the N transcript level, although the transcript levels of some defense-related genes were affected compared with levels in non-transgenic plants. TMV infection induced a hypersensitive reaction (HR) with larger necrotic lesions in transgenic lines than in non-transgenic plants, but there was no change in total virus accumulation per lesion in a standardized area. Activation of the N gene expression in non-transgenic plants during synchronously-induced HR was followed by downregulation of BBF3 gene expression. Constitutive overexpression of the BBF3 transgene resulted in higher accumulation of the N gene as well as HR marker genes (Hin1 and Hsr203j) and lower accumulation of salicylic acid- and jasmonic acid-signaling marker genes (PR1-a and PR-1b). In particular, the expression of PR-1b was remarkably suppressed in the BBF3-overexpressing transgenic lines. Dof-binding motifs were confirmed in the upstream regions of all target genes examined. Our results suggest that BBF3 has a role in modulating the expression of genes associated with virus elicitor recognition and defense responses resulting in larger areas of HR-associated cell death. [ABSTRACT FROM AUTHOR]

Plant protease inhibitors (PI's) inhibit the activity of gut proteases and thus provide resistance against insect attack. Previously we have published first report on cloning and characterization of a novel Bowman–Birk protease inhibitor gene (RbTI) from ricebean (Vigna umbellata). In this study, the RbTI gene was further characterized and validated as a potential candidate for transferring insect resistance in economically important crops. We have successfully generated transgenic tobacco plants expressing RbTI gene constitutively under CaMV35S promoter using Agrobacterium transformation. Genomic PCR and GUS analysis confirmed the successful integration of RbTI gene into tobacco plant genome. qRT-PCR analysis revealed highest RbTI gene expression in transformed tobacco leaves nearing maturity. Feeding of transformed tobacco leaf tissue showed prominent effect on larval mortality throughout the larval growth stages mainly during first three days of feeding. For functional analysis of RbTI gene, we estimated the inhibitory activity of protein extracts from normal and transformed tobacco plants against gut proteases of Spodoptera litura and H. armigera larval instars. Maximum inhibition of trypsin (82.42% and 73.25%) and chymotrypsin (69.50% and 60.64%) enzymes was recorded at early larval stages of both insects. The results of this study validated the future use of RbTI gene from ricebean legume as a potential candidate for transferring insect resistance in economically important crops. Insight, innovation, integration : Present study was conducted with the aim to utilize the state of art biotechnological techniques for transferring key pest resistant genes from underutilized promising crop ricebean. The tobacco plant has been utilized as modern plant for proof of concept where a protease inhibitor gene from Ricebean has been transferred to tobacco plant which induced larval mortality within first three days of feeding at all larval developmental stages. The biochemical assays on mid-gut total protein extract showed that the transgenic tobacco leaves have inhibiting effect on trypsin and chymotrypsin enzymes of insect which is otherwise required for digestion of food by them. Hence, we provide a novel gene that could be utilized for pest resistance in other crops different developmental stages. [ABSTRACT FROM AUTHOR]

Salinity is a prominent environmental stressor that significantly impacts plant growth and development. Here, we conducted research on the physiological and transcriptomic mechanism of a wild cucumber, Cucumis hystrix Chakr, under NaCl stress. Physiological data showed that contents of malondialdehyde, peroxide (H2O2), proline, soluble sugar, and activities of antioxidant enzymes of superoxide dismutase, peroxidase, ascorbate peroxidase, and glutathione reductase in wild cucumber plants were increased significantly after NaCl treatment. Transcriptomic analysis revealed that 3509 transcripts were differentially expressed in leaves and 5516 transcripts in roots after NaCl treatment. Numerous genes were related to the signal transduction, transcription factor, ion transport, osmotic metabolism, and reactive oxygen species scavenging. Moreover, the thioredoxin H type gene of Cucumis hystrix Chakr (ChTrxh) was isolated and characterized. Our study demonstrated that the transgenic tobacco plants overexpressing ChTrxh exhibited enhanced tolerance to NaCl stress compared to wild-type plants. These findings contribute valuable insights into the functional characteristics of important genes in wild cucumber under NaCl stress. [ABSTRACT FROM AUTHOR]

[Objective] This study explored the biological function of nuclear factor GmNF-YA 13 gene in soybean responding to abiotic stress to provide basis for the application of GmNF-YA 13 in resistance breeding. [Method] The seeds of soybean varieties Beidou 9 and tobacco NC89 were selected for drought, salt, cold and ABA treatments after the first terately compound leaf unfolded. The relative expression of GmNF-YAlS, was detected by real-time fluorescence quantitative RT-PCR (qRT-PCR). GmNF-YA 1 3 was cloned and analyzed by bioinformatics. GmNF-YA 13 was transformed into tobacco, and the expression levels of stress-related genes (NtABA2, NtOsmotin, NtAPX2, NtSOD, NtCATl and NtCaMKS.) in transgenic tobacco were detected by qRT-PCR. [Result] Drought, salt, cold and ABA treatments could induce the expression of GmNF-YA 13 at different degrees, with especially significant effects in drought and ABA treatments. Gm NF-YA 13 was located on chromosome 13 of soybean genome with 915 bp open reading frame and it encoded a protein containing 304 amino acids with a molecular weight of 75.14 ku and an isoelectric point of 5.10. GmNF-YA13 protein contained a conserved DNA binding domain. The protein phylogenetic analysis showed that GmNF-YA13 was closely related to GmNF-YA7 and Arabidopsis AtNF-YA1. The promoter cis-elements prediction analysis showed that the promoter region of GmNF-YA13 contained three ABA responsive elements (ABRE), one anaerobic induction element (ARE), one defense and stress responsive element (TC-nch repeats) and one MYB binding site involved in drought-inducibility (MBS). The relative expression of GmNF-YA 13 was the highest in leaves. The GmNF-YA 13 plant expression vector was constructed and transformed into tobacco to obtain three transgenic tobacco plants. The relative expression levels of NtABA2, NtOsmotin and NtAPX2 in transgenic tobacco were significantly higher than those in wild-type tobacco, while those of NtSOD, NtCAT1 and NtCaMK3 were not significantly different. [Conclusion] GmNF-YA 13 was closely related to abiotic stress such as drought, salt and cold in soybean. GmNF-YA 13 may enhance the stress resistance of transgenic tobacco by promoting the expression of stress-related genes. [ABSTRACT FROM AUTHOR]

bZIP transcription factors play important roles in regulating plant development and stress responses. Although bZIPs have been identified in many plant species, there is little information on the bZIPs in Chrysanthemum. In this study, bZIP TFs were identified from the leaf transcriptome of C. mongolicum, a plant naturally tolerant to drought. A total of 28 full-length bZIP family members were identified from the leaf transcriptome of C. mongolicum and were divided into five subfamilies based on their phylogenetic relationships with the bZIPs from Arabidopsis. Ten conserved motifs were detected among the bZIP proteins of C. mongolicum. Subcellular localization assays revealed that most of the CmbZIPs were predicted to be localized in the nucleus. A novel bZIP gene, designated as CmbZIP9, was cloned based on a sequence of the data of the C. mongolicum transcriptome and was overexpressed in tobacco. The results indicated that the overexpression of CmbZIP9 reduced the malondialdehyde (MDA) content and increased the peroxidase (POD) and superoxide dismutase (SOD) activities as well as the expression levels of stress-related genes under drought stress, thus enhancing the drought tolerance of transgenic tobacco lines. These results provide a theoretical basis for further exploring the functions of the bZIP family genes and lay a foundation for stress resistance improvement in chrysanthemums in the future.

Flavonoid-3′,5′-hydroxylase (F3′5′H) is the key enzyme for the biosynthesis of delphinidin-based anthocyanins, which are generally required for purple or blue flowers. Previously, we isolated a full-length cDNA of PgF3′5′H from Platycodon grandiflorus, which shared the highest homology with Campanula medium F3′5′H. In this study, PgF3′5′H was subcloned into a plant over-expression vector and transformed into tobacco via Agrobacterium tumefaciens to investigate its catalytic function. Positive transgenic tobacco T0 plants were obtained by hygromycin resistance screening and PCR detection. PgF3′5′H showed a higher expression level in all PgF3′5′H transgenic tobacco plants than in control plants. Under the drive of the cauliflower mosaic virus (CaMV) 35S promoter, the over-expressed PgF3′5′H produced dihydromyricetin (DHM) and some new anthocyanin pigments (including delphinidin, petunidin, peonidin, and malvidin derivatives), and increased dihydrokaempferol (DHK), taxifolin, tridactyl, cyanidin derivatives, and pelargonidin derivatives in PgF3′5′H transgenic tobacco plants by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) analysis, resulting in a dramatic color alteration from light pink to magenta. These results indicate that PgF3′5′H products have F3′5′H enzyme activity. In addition, PgF3′5′H transfer alters flavonoid pigment synthesis and accumulation in tobacco. Thus, PgF3′5′H may be considered a candidate gene for gene engineering to enhance anthocyanin accumulation and the molecular breeding project for blue flowers. [ABSTRACT FROM AUTHOR]

Temperature affects the growth and yield of yam (Dioscorea opposite Thunb.), and calcium-dependent protein kinases (CDPKs) play an important role in the plant stress response. However, there has been a lack of system analyses of yam’s CDPK gene family. In this study, 29 CDPK transcriptome sequences with complete open reading frames (ORFs) were identified from yam RNA sequencing data. The sequences were classified into four groups (I–VI) using phylogenetic analysis. Two DoCDPK genes were randomly selected from each group and the gene patterns of yam leaves were determined using quantitative real-time PCR (qRT-PCR) under high and low temperature stress in order to show their unique functions in mediating specific responses. Among them, DoCDPK20 was significantly induced in high temperatures. The pPZP221-DoCDPK20 was transformed into tobacco leaves using an agrobacterium-mediated method. Under high temperature stress, DoCDPK20 overexpression reduced photosynthesis and improved heat tolerance in transgenic tobacco. Our research offers meaningful perspectives into CDPK genes and new avenues for the genetic engineering and molecular breeding of yam. [ABSTRACT FROM AUTHOR]

Pumpkin (Cucurbita moschata Duch.) productivity is severely hindered by powdery mildew (PM) worldwide. The causative agent of pumpkin PM is Podosphaera xanthii, a biotrophic fungus. Pathogenesis-related protein 1 (PR1) homolog was previously identified from transcriptomic analysis of a PM-resistant pumpkin. Here, we investigated the effects of CmPR1 gene from pumpkin for resistance to PM. Subcellular localization assay revealed that CmPR1 is a cytoplasmic protein in plants. The expression of CmPR1 gene was strongly induced by P. xanthii inoculation at 48 h and exogenous ethylene (ET), jasmonic acid (JA) and NaCl treatments, but repressed by H2O2 and salicylic acid (SA) treatments. Visual disease symptoms, histological observations of fungal growth and host cell death, and accumulation of H2O2 in transgenic tobacco plants indicated that CmPR1 overexpression significantly enhanced the resistance to Golovinomyces cichoracearum compared to wild type plants during PM pathogens infection, possibly due to inducing cell death and H2O2 accumulation near infected sites. The expression of PR1a was significantly induced in transgenic tobacco plants in response to G. cichoracearum, suggesting that CmPR1 overexpression positively modulates the resistance to PM via the SA signaling pathway. These findings indicate that CmPR1 is a defense response gene in C. moschata and can be exploited to develop disease-resistant crop varieties. [ABSTRACT FROM AUTHOR]

Enhancement of crop productivity under various abiotic stresses is a major objective for researchers in the current scenario. Heat stress adversely affects yield of wheat (Triticum aestivum L.) plants. To cope up with stress conditions, plants respond by overexpressing their heat stress-related genes and transcription factors. Wheat is one of the world's most staple food crops which is highly sensitive to heat stress especially during anthesis stage, thereby affecting both yield and quality. During abiotic stress, plant heat shock factors (Hsfs) play a crucial role and confers stress tolerance. In this study, we have isolated highly heat stress-responsive transcription factor from wheat cultivar HD3086, after checking the gene expression using real-time PCR in contrasting wheat genotypes (HD2894 and HD3086). The candidate gene (TaMBF1c) was cloned in pJET1.2/blunt vector and then further into binary vector followed by transformation in tobacco (Nicotiana tabacum) via Agrobacterium mediated genetic transformation. The transgenic tobacco plants raised were validated for heat stress tolerance using different physiological and biochemical assays (RWC, MDA, proline content and chlorophyll content). The gene expression was checked in transgenic plants using qRT-PCR. At T1 generation, seeds of transgenic plants were germinated on MS selection media and a segregation inheritance of 3:1 (resistance: susceptible) ratio was obtained which, followed the Mendelian inheritance pattern. For future research work, TaMBF1c would be taken in different crop plants to develop heat stress tolerant crops for sustainable development under globally changing climate conditions. Key message: TaMBF1c was isolated from thermo tolerant wheat cultivar (HD3086), proved its role in providing heat stress tolerance to transgenic tobacco plants. [ABSTRACT FROM AUTHOR]

Abstract Background Jatropha curcas is a promising alternative bio-energy resource. However, underrun limited its broad application in the industry. Luckily, TAW1 is a high-productivity promoting gene that increases the lateral branches by prolonging the identification of inflorescence meristems to generate more spikes and flowers. Results In the current study, we introduced the Jatropha JcTAW1 gene into tobacco to depict its functional profile. Ectopically expressed JcTAW1 increased the lateral branches and ultimate yield of the transgenic tobacco plants. Moreover, the JcTAW1 lines had significantly higher plant height, longer roots, and better drought resistance than those of wild-type (W.T.). We performed RNA sequencing and weighted gene co-expression network analysis to determine which biological processes were affected by JcTAW1. The results showed that biological processes such as carbon metabolism, cell wall biosynthesis, and ionization transport were extensively promoted by the ectopic expression of JcTAW1. Seven hub genes were identified. Therein, two up-regulated genes affect glucose metabolism and cell wall biosynthesis, five down-regulated genes are involved in DNA repair and negative regulation of TOR (target-of-rapamycin) signaling which was identified as a central regulator to promote cell proliferation and growth. Conclusions Our study verified a new promising candidate for Jatropha productive breeding and discovered several new features of JcTAW1. Except for boosting flowering, JcTAW1 was found to promote stem and root growth. Additionally, transcriptome analysis indicated that JcTAW1 might promote glucose metabolism while suppressing the DNA repair system.

The expression of the soybean Bowman–Birk proteinase isoinhibitor DII (BBI-DII) gene and the inducible activity of its promoter were studied under salt, drought, low temperature, and abscisic acid (ABA) exposure conditions. The BBI-DII gene was induced by salt, drought, low temperature, and ABA, and the relative expression levels were 103.09-, 107.01-, 17.25- and 27.24-fold, respectively, compared with the untreated control. The putative promoter, designated BP1 (− 1255 to + 872 bp), located 5′-upstream of the BBI-DII gene was cloned. The expression of the GUS gene in pCAM-BP1 transgenic tobacco plants was highest at 5 h after treatment with salt, drought, low temperature and ABA, especially under salt and drought. Using histochemical staining and fluorescence analysis of GUS, BP1 activity under salt and drought conditions after 5 h was 1.03 and 1.07-fold, respectively, compared with that of the CaMV35S promoter. Based on a 5′ deletion analysis, the segment (+ 41 to + 474 bp) was the basal region that responded to salt and drought, whereas the segment (− 820 to + 41 bp) was the area that responded to increased salt and drought activity. The BP2 (− 820 to + 872) activities were 0.98- and 1.02-fold compared with that of BP1 under salt and drought conditions and was 435 bp shorter than BP1. The salt- and drought-inducible activities of the BP2 promoter in the roots, stems, and leaves of transgenic tobacco plants were stable. Taken together, BP2 is more suitable than the BP1 promoter for the study and molecular breeding of stress-resistant soybean plants. [ABSTRACT FROM AUTHOR]

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Lipoxygenase (LOX, EC1.13.11.12) catalyzes the oxygenation of polyunsaturated fatty acids and further generates oxylipins via enzymatic or non-enzymatic pathways, thus participating in the regulation of growth and development, stress response, and other processes. The objective of this research was to characterize the function and evaluate the mechanism of lipoxygenase gene AfLOX4's role in enhancing drought tolerance. Transcriptomic data were generated and analyzed for the LOX gene, under different concentrations of drought (polyethylene glycol-6000) (PEG6000) and abscisic acid (ABA) stress treatment, to determine their effects on the expression of the AfLOX4 gene and its role in plant physiology. The localization of the AfLOX4 protein was detected through protoplast transformation. The expression of AfLOX4 in Amorpha fruticosa tissues was determined by quantitative real-time PCR (qRT-PCR). Six LOX genes of A. fruticosa that showed an upregulated pattern under different concentrations of drought (PEG6000) stress were found via transcriptomic analysis and were named AfLOX1 ~ 6. The AfLOX4 protein was localized in the cytoplasm. AfLOX4 was expressed in the stems, roots, flowers, and leaves of Amorpha fruticosa L., and its expression level was the highest in leaves. The expression of AfLOX4 increased with the prolongation of stress time under different stresses. After ABA treatment, the germination rate of tobacco lines overexpressing AfLOX4 was significantly increased, while under NaHCO3 stress, the tolerance of tobacco plants overexpressing AfLOX4 was higher than that of wild-type tobacco. The tobacco with overexpression of the AfLOX4 gene detected by the natural drought method had a higher survival rate in the drought stress recovery experiment, and improved the drought tolerance of tobacco. This study reveals the function of the AfLOX4 gene in resisting drought stress and lays a solid theoretical foundation for the genetic improvement of Amorpha fruticosa L. plant varieties, as well as improvement of resource utilization. [ABSTRACT FROM AUTHOR]

The NAC transcription factor family is well known to play vital roles in plant development and stress responses. For this research, a salt-inducible NAC gene, PsnNAC090 (Po-tri.016G076100.1), was successfully isolated from Populus simonii × Populus nigra. PsnNAC090 contains the same motifs at the N-terminal end of the highly conserved NAM structural domain. The promoter region of this gene is rich in phytohormone-related and stress response elements. Transient transformation of the gene in the epidermal cells of both tobacco and onion showed that the protein was targeted to the whole cell including the cell membrane, cytoplasm and nucleus. A yeast two-hybrid assay demonstrated that PsnNAC090 has transcriptional activation activity with the activation structural domain located at 167–256aa. A yeast one-hybrid experiment showed that PsnNAC090 protein can bind to ABA-responsive elements (ABREs). The spatial and temporal expression patterns of PsnNAC090 under salt and osmotic stresses indicated that the gene was tissue-specific, with the highest expression level in the roots of Populus simonii × Populus nigra. We successfully obtained a total of six transgenic tobacco lines overexpressing PsnNAC090. The physiological indicators including peroxidase (POD) activity, superoxide dismutase (SOD) activity, chlorophyll content, proline content, malondialdehyde (MDA) content and hydrogen peroxide (H2O2) content were measured in three transgenic tobacco lines under NaCl and polyethylene glycol (PEG) 6000 stresses. The findings reveal that PsnNAC090 improves salt and osmotic tolerance by enhancing reactive oxygen species (ROS) scavenging and reducing membrane lipid peroxide content in transgenic tobacco. All the results suggest that the PsnNAC090 gene is a potential candidate gene playing an important role in stress response. [ABSTRACT FROM AUTHOR]

Copyright of Bulletin of Botanical Research is the property of Bulletin of Botanical Research Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Pinus massoniana (P. massoniana) is an important fast-growing tree species in China. To explore photosynthesis-related gene resources and find possible targets for high photosynthetic-efficiency breeding of P. massoniana, we cloned two full-length cDNAs with conserved sequences of RubisCO activase (RCA) genes from P. massoniana. The RCA enzyme is important in maintaining the activity of RubisCO in vivo. Two genes, named PmRCA1 and PmRCA2, showed high similarity to RCA genes from other species and were identical to each other except for a 137 bp intervening sequence in the open reading frame (ORF) of PmRCA1, which led to different protein products, herein referred to as PmRCA1 and PmRCA2, respectively. PmRCA1 and PmRCA2 showed high homology to RCA of Pinus halepensis, and PmRCA1 had two cysteine residues in its carboxyl-terminal extension that were unique to the large RCA isoform. To characterize the function of PmRCA1 and PmRCA2, plant expression vectors driven by the constitutive 35S promoter were transformed in tobacco plants. Transgenic plants of PmRCA2 were grown better than wild tobacco plants and had significantly more leaves. Overexpression of PmRCA2 increased the maximum net photosynthetic rate and dark respiration rate of tobacco by 16.49% and 81.85%, respectively. Overexpression of PmRCA1 increased the maximum net photosynthetic rate and dark respiration rate of tobacco more than overexpression of PmRCA2, but it simultaneously enhanced photoinhibition of photosynthesis. [ABSTRACT FROM AUTHOR]

Alpha-galactosidase seed imbibition protein (VvSIP) isolated from Vitis vinifera is up-regulated upon salt stress and mediates osmotic stress responses in a tolerant grapevine cultivar. So far, little is known about the putative role of this stress-responsive gene. In the present study, VvSIP function was investigated in model tobacco plants via Agrobacterium-mediated genetic transformation. Our results showed that overexpression of VvSIP exhibited increased tolerance to salinity at germination and late vegetative stage in transgenic Nicotiana benthamiana compared to the nontransgenic plants based on the measurement of the germination rate and biomass production. High salt concentrations of 200 and 400 mM NaCl in greenhouse-grown pot assay resulted in better relative water content, higher leaf osmotic potential, and leaf water potential in transgenic lines when compared to the wild-type (WT) plants. These physiological changes attributed to efficient osmotic adjustment improved plant performance and tolerance to salinity compared to the WT. Moreover, the VvSIP-expressing lines SIP1 and SIP2 showed elevated amounts of chlorophyll with lower malondialdehyde content indicating a reduced lipid peroxidation required to maintain membrane stability. When subjected to high salinity conditions, the transgenic tobacco VvSIP exhibited higher soluble sugar content, which may suggest an enhancement of the carbohydrate metabolism. Our findings indicate that the VvSIP is involved in plant salt tolerance by functioning as a positive regulator of osmotic adjustment and sugar metabolism, both of which are responsible for stress mitigation. Such a candidate gene is highly suitable to alleviate environmental stresses and thus could be a promising candidate for crop improvement. [ABSTRACT FROM AUTHOR]

Background: Jatropha curcas is a promising alternative bio-energy resource. However, underrun limited its broad application in the industry. Luckily, TAW1 is a high-productivity promoting gene that increases the lateral branches by prolonging the identification of inflorescence meristems to generate more spikes and flowers. Results: In the current study, we introduced the Jatropha JcTAW1 gene into tobacco to depict its functional profile. Ectopically expressed JcTAW1 increased the lateral branches and ultimate yield of the transgenic tobacco plants. Moreover, the JcTAW1 lines had significantly higher plant height, longer roots, and better drought resistance than those of wild-type (W.T.). We performed RNA sequencing and weighted gene co-expression network analysis to determine which biological processes were affected by JcTAW1. The results showed that biological processes such as carbon metabolism, cell wall biosynthesis, and ionization transport were extensively promoted by the ectopic expression of JcTAW1. Seven hub genes were identified. Therein, two up-regulated genes affect glucose metabolism and cell wall biosynthesis, five down-regulated genes are involved in DNA repair and negative regulation of TOR (target-of-rapamycin) signaling which was identified as a central regulator to promote cell proliferation and growth. Conclusions: Our study verified a new promising candidate for Jatropha productive breeding and discovered several new features of JcTAW1. Except for boosting flowering, JcTAW1 was found to promote stem and root growth. Additionally, transcriptome analysis indicated that JcTAW1 might promote glucose metabolism while suppressing the DNA repair system. [ABSTRACT FROM AUTHOR]

DNA binding with one finger (Dof) is a plant-specific transcription factor (TF) that has a key role in seed development and germination, light-mediated control, and plant responses to biotic and abiotic stressors. Herein, we aimed to evaluate the role of the soybean Dof -TFs in plant tolerance breeding. In this study, GmDof4.2 was isolated from the soybean. GmDof4.2 in soybean seedlings responded to drought, salt, heat, and cold stresses, according to the results of real-time fluorescence quantitative PCR (qPCR). GmDof4.2 protein had a 59-amino-acid Dof domain with a CX2CX21CX2C motif and four conserved cysteine (Cys) residues, according to the predicted amino acid sequence. The GmDof4.2 protein shared the most genetic similarities with the Cajanus cajan CcDof4 protein. Seven cis-elements associated with stress or hormone induction, as well as two TF binding sites, were found in the GmDof4.2 promoter region. GmDof4.2 protein bound specifically to the AAAG motif in vivo according to the results of yeast one-hybrid. Moreover, the overexpression of GmDof4.2 resulted in higher soluble carbohydrates content and proline content and lower relative electrolyte leakage and malondialdehyde content in transgenic tobacco than in wild-type (WT) tobacco under drought stress. Furthermore, the expression levels of nine stress-related genes were considerably greater in transgenic tobacco than in WT tobacco. The underlined data suggested that the GmDof4.2 improved drought tolerance in transgenic tobacco. Key Message: A Dof transcription factor, GmDof4.2, was isolated from soybean. The expression of GmDof4.2 could be induced by abiotic stress. Overexpression of GmDof4.2 improved drought tolerance in transgenic tobacco. [ABSTRACT FROM AUTHOR]

Potassium (K+) is the most abundant cation in plants, which is absorbed by roots and distributed throughout the plants and within plant cells, and is involved in various cellular processes. Shaker K+ channel plays crucial roles in the absorption and distribution of K+ and in the response to abiotic stress in plants. Herein, a total of six shaker K+ channel genes, BvKAT1 , BvKAT3 , BvAKT1 , BvAKT2 , BvAKT5 , and BvSKOR , were identified in the genome of sugar beet (Beta vulgaris L.). The coding domain sequences (CDS) of these genes ranged from 2232 to 2739 bp, and protein lengths were varied from 743 to 912 aa. The shaker K+ channel genes contained hormone-related and light responsiveness cis -acting regulatory elements. The phylogenetic analysis showed that BvSKOR was highly conserved and contained six transmembrane structures. The expression patterns of BvSKOR under salt and osmotic stress were analyzed by qRT-PCR, and found that the expression level of BvSKOR under low concentration salt and osmotic stress at short period of treatment were significantly higher than that of the control group. The function of BvSKOR was further verified in tobacco (Nicotiana tabacum), and the results showed that under salt and osmotic stress, the roots of transgenic plants were significantly stronger than those of wild type (WT) plants, and the relative water content (RWC), chlorophyll, proline, soluble sugar, soluble proteins contents and antioxidant enzyme activity were significantly higher than those of WT plants. These results indicated that overexpression of BvSKOR can significantly enhance the salt and drought tolerance in transgenic tobacco plants. This study could provide theoretical support and genetic resources for genetic improvement of crops stress resistance. • A total of six genes of the shaker K+ channel family were identified in the genome of sugar beet. • The expression level of BvSKOR was significantly higher under salt and drought stresses compared with control. • Overexpression of BvSKOR significantly enhanced the tolerance of transgenic tobacco to salt and drought stresses. • BvSKOR has research potential in improving the genetic improvement of crop stress resistance by genetic engineering. [ABSTRACT FROM AUTHOR]

Background. Soil salinity is one of the limiting factors for plant growth and productivity. The areas of saline lands increase annually, so it is important to study the mechanisms of plant resistance to salt stress.Material and methods. We studied the effect of salt stress on tobacco plants (Nicotiana tabacum L.) of the wild type (‘Samsun’) and the transgenic line Cod 38 obtained by introducing the сodA gene, encoding bacterial choline oxidase, from Arthrobacter globiformis. Salt tolerance of the compared genotypes was assessed according to the growth indicators and the ability to preserve the pool of photosynthetic pigments under model salt stress conditions (150 mМ NaCl). The sensitivity of plants to salt stress was analyzed using biochemical tests that reflected the intensity of peroxidation processes and the activity of antioxidant enzymes (superoxide dismutase, and peroxidase).Results. The survival rates and biometric characteristics of transformants under salt stress were significantly higher than in wild-type plants. Under the impact of salt stress, the content of chlorophylls and carotenoids in the leaves of ‘Samsun’ plants decreased 1.5 and 1.3 times, respectively. Contrastingly, transformants under the same conditions showed a tendency to increase the pool of plastid pigments. A peculiarity of transgenic plants was also the reduced malondialdehyde content in their leaves, which indicates a low intensity of lipid peroxidation during salinization and can be explained by the functioning of endogenous glycine betaine as a compound with a multifunctional effect.Conclusions. It was shown that the transformation of plants with the bacterial gene of choline oxidase, followed by the accumulation of the protein product of the codA gene – glycine betaine, even in a minimal amount, was accompanied by positive effects on tobacco plants under salt stress conditions.

Copyright of Chinese Journal of Oil Crop Sciences is the property of Oil Crops Research Institute of Chinese Academy of Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Chitinase plays a vital role in plant defense against insect pests and fungal pathogens due to its ability to hydrolyse chitin, which forms the major portion of the lining of gut membrane of insects and the cell wall of fungi. Earlier we reported a ~ 76 kDa chitinase protein isolated from Xenorhabdus nematophila strain (ATCC 19061). Purified chitinase protein was tested against the larvae of Helicoverpa armigera,a major crop pest and when fed orally to the larvae, it proved to be toxic. In the present study, we report the generation of transgenic tobacco plants expressing the chitinase gene of X. nematophila. The stable integration of the chitinase gene was confirmed by Southern blot while the expression of the gene was checked by RT-PCR analysis and quantification of chitinolytic activity. The transgenic lines were phenotypically normal and healthy. The result of bioassays revealed that the tobacco lines expressing increased level of chitinase protein showed 93–100% mortality of larvae of H. armigera when fed on the transgenic leaves expressing chitinase. A considerable reduction in leaf damage (55–76%) was also observed in transgenic lines as compared to the damage in untransformed control plants when the larvae were allowed to feed on the transgenic leaves. The larvae feeding on transgenic leaves showed adverse effect on their growth, development and survival. Thus we propose, the chitinase gene can prove to be a potent candidate for pest control in crop plants in future. Key message: Overexpression of insecticidal chitinase from Xenorhabdus nematophila protects transgenic tobacco plants against Helicoverpa armigera. Our results indicate that the gene encoding chitinase is a promising candidate for crop protection. [ABSTRACT FROM AUTHOR]

Main conclusion: HaNAC3 is a transcriptional activator located in the nucleus that may be involved in the response to high temperature, high salt and drought stresses as well as phytohormone IAA and ABA treatments. Our study demonstrated that HaNAC3 increased the tolerance of transgenic tobacco to abiotic stress and was involved in the regulation of a range of downstream genes and metabolic pathways. This also indicates the potential application of HaNAC3 as a plant tolerance gene. NAC transcription factors play a key role in plant growth and development and plant responses to biotic and abiotic stresses. However, the biological functions of NAC transcription factors in the desert plant Haloxylon ammodendron are still poorly understood. In this study, the NAC transcription factor HaNAC3 was isolated and cloned from a typical desert plant H. ammodendron, and its possible biological functions were investigated. Bioinformatics analysis showed that HaNAC3 has the unique N-terminal NAC structural domain of NAC transcription factor. Quantitative real-time fluorescence analysis showed that HaNAC3 was able to participate in the response to simulated drought, high temperature, high salt, and phytohormone IAA and ABA treatments, and was very sensitive to simulated high temperature and phytohormone ABA treatments. Subcellular localization analysis showed that the GFP-HaNAC3 fusion protein was localized in the nucleus of tobacco epidermal cells. The transcriptional self-activation assay showed that HaNAC3 had transcriptional self-activation activity, and the truncation assay confirmed that the transcriptional activation activity was located at the C-terminus. HaNAC3 gene was expressed exogenously in wild-type Nicotiana benthamiana, and the physiological function of HaNAC3 was verified by simulating drought and other abiotic stresses. The results indicated that transgenic tobacco had better resistance to abiotic stresses than wild-type B. fuminata. Further transcriptome analysis showed that HaNAC3 was involved in the regulation of a range of downstream resistance genes, wax biosynthesis and other metabolic pathways. These results suggest that HaNAC3 may have a stress resistance role in H. ammodendron and has potential applications in plant molecular breeding. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to clone the transcription factor gene BoWRKY10 of kale (Brassica oleracea var. acephala DC) and study the drought tolerance regulation mechanism of the gene.【Method】The gene BoWRKY10 was amplified from the cDNA of kale leaves by RT-PCR and transferred into tobacco. The expression pattern of the gene under drought stress was detected by real-time fluorescence quantitative PCR.【Result】The survival rate of transgenic lines were significantly higher than that of non-GMO wild types (WT) after 10 days of drought. There was no obvious difference between transgenic lines (OE3, OE4 and OE6) and WT in terms of germination rate in 1/2 MS culture medium, however, in medium containing 15% PEG4000, germination rate of OE3, OE4 and OE6 were 65.9%, 75.4% and 84.3%, respectively, while the one of WT was 22.5%. In medium containing 15% PEG4000, the root length of transgenic lines was significantly higher than that of WT after 14 days. Meanwhile, ABA content, proline content, soluble sugar content, POD, CAT and SOD activities were higher, while MDA and H2O2 content were significantly lower than those of WT. The expression level of drought responsive gene in OE3, like NtSOD, NtAPX, NtCAT, NtGST, NtSPSA and NtNCED1, was significantly higher than that of WT.【Conclusion】Expression of BoWRKY10 from kale remarkably improved the tolerance of tobacco to drought stresses in which the pleiotropy could be beneficial gene resource of gene engineering about resistance to stress in plant. [ABSTRACT FROM AUTHOR]

The multidrug and toxic compound extrusion (MATE) protein family has been implicated in the transport of a diverse range of molecules, including specialized metabolites. In tobacco (Nicotiana tabacum), only a limited number of MATE transporters have been functionally characterized, and no MATE transporter has been studied in the context of flavonoid transport in this plant species so far. In the present study, we characterize two homeologous tobacco MATE genes, NtMATE21 and NtMATE22 , and demonstrate their role in flavonol transport and in plant growth and development. The expression of these two genes was reported to be up-regulated in trichomes as compared with the trichome-free leaf. The transcript levels of NtMATE21 and NtMATE22 were found to be higher in flavonol overproducing tobacco transgenic lines as compared with wild type tobacco. The two transporters were demonstrated to be localized to the plasma membrane. Genetic manipulation of NtMATE21 and NtMATE22 led to altered growth phenotypes and modulated flavonol contents in N. tabacum. The β-glucuronidase and green fluorescent protein fusion transgenic lines of promoter regions suggested that NtMATE21 and NtMATE22 are exclusively expressed in the trichome heads in the leaf tissue and petals. Moreover, in a transient transactivation assay, NtMYB12, a flavonol-specific MYB transcription factor, was found to transactivate the expression of NtMATE21 and NtMATE22 genes. Together, our results strongly suggest the involvement of NtMATE21 and NtMATE22 in flavonol transport as well as in the regulation of plant growth and development. [ABSTRACT FROM AUTHOR]

Breeding for flower cold resistance is a priority for flower breeding research in northern China. The identification of cold resistance genes will not only provide genetic resources for cold resistance breeding, but also form a basis for the study of plant cold resistance mechanisms.Based on the flower transcriptome of Iris laevigata, 20 R2R3‐MYBs were identified and comprehensive analysis, including conservative domain, phylogenetic analyses and functional distribution, were performed for R2R3‐MYBs. Expression patterns of the abiotic stress genes under cold stress were detected, the upregulated gene was genetically transformed into tobacco, and the related physiological indicators of the transgenic tobacco were measured.A novel cold resistance gene, IlMYB306, was obtained. qRT‐PCR indicated that IlMYB306 was dramatically induced by cold stress and was significantly upregulated in roots. The free proline content, MDA, SOD and POD activity of the transgenic tobacco improved after cold stress, and the chlorophyll content decreased slowly. In addition, overexpression of IlMYB306 improved cold resistance of the seeds. SEM results showed leaves of transgenic tobacco had obvious folds, more grooves and bulges on the lower leaf surface.Overall, we report a novel cold resistance R2R3‐MYB gene, IlMYB306, in the flower of I. laevigata, which could improve tobacco cold stress tolerance by thickening the waxy layer, increasing antioxidant activity and the content of proline. [ABSTRACT FROM AUTHOR]

Phosphatidylinositol-specific phospholipase C (PI-PLC) catalyzes the hydrolysis of phosphatidylinositol-4,5-bisphosphate into diacylglycerol and inositol 1,4,5-trisphosphate. It can play an essential role in plant stress response and signaling. However, the functions of PLCs remain unclear in wooden plants. This study carried out the bioinformatics analysis of the PLC gene family in poplar. The expression pattern analysis suggested the transient up-regulation of PsnPLC in salt stress conditions. The transgenic tobacco plants overexpressing PsnPLC were generated by the Agrobacterium-mediated leaf disc method. The transgenic lines showed a significant increase in plant height, SOD and POD activity, and proline content under salt stress. By contrast, transgenic lines showed a decreased malondialdehyde content under salt stress. Comparative transcriptome analysis indicated that the overexpression of PsnPLC mainly affects membrane-associated GO terms and signaling pathways, and the differential expression of salt-responsive genes may contribute to the enhanced salt tolerance. Overall, the results indicated that PsnPLC plays an essential role in modulating salt tolerance in transgenic tobacco. [ABSTRACT FROM AUTHOR]

Flammutoxin (FTX), as well as its precursor TDP, is a protein from Flammulina velutipes with antiviral activity. Transgenic tobacco with the FTX271 (gene of FTX or TDP) can not only delay the onset time of symptoms but also alleviate the symptoms caused by tobacco mosaic virus (TMV), but the mechanism is still unclear. In this study, FTX271 was introduced into Nicotiana benthamiana, and the disease resistance mechanism activated by FTX271 was speculated by transcriptomic and proteomic techniques. The results showed that TDP was detected, and some genes, proteins and pathways were significant upregulated or enriched in transgenic tobacco, including the mitogen-activated protein kinase (MAPK) cascade signal transduction pathway, the expression of hypersensitive response (HR) marker genes H1N1 and HSR203J, pathogenesis-related (PR) genes, and the key genes COI1 and lipoxygenase gene LOX2 of the jasmonic acid (JA) signaling pathway, indicating FTX271 may activate the MAPK pathway and increase the content of reactive oxygen species (ROS) and JA, which promoted the HR and inducible systemic resistance (ISR). ISR caused increased expression of peroxidase (POD) and other proteins involved in pathogen defense. In addition, transgenic tobacco may use sHSP-assisted photoreparation to alleviate the symptoms of TMV. In conclusion, JA-mediated ISR and sHSP-assisted photoreparation are activated by FTX271 to protect tobacco from TMV infection and alleviate the symptoms caused by the virus. The study provided a theoretical basis for the TMV resistance mechanism of FTX271, which may represent a potential gene resource for plant antiviral transgenic breeding. [ABSTRACT FROM AUTHOR]

Melatonin is an important, multifunctional protective agent against a variety of abiotic and biotic stressors in plants. Caffeic acid O-methyltransferase (COMT) catalyzes the last step of melatonin synthesis in plants and reportedly participates in the regulation of stress response and tolerance. However, few studies have reported its function in melatonin-mediated drought resistance. In this study, CrCOMT was identified and was strongly induced by drought stress in Carex rigescens. CrCOMT overexpression in transgenic tobacco increased tolerance to drought stress with high levels of seed germination, relative water content, and survival rates. CrCOMT overexpression in tobacco improved membrane stability, and plants exhibited lower relative electrolytic leakage and malondialdehyde content, as well as higher photochemical efficiency than the wildtype (WT) under drought stress. The transgenic plants also had higher levels of proline accumulation and antioxidant enzyme activity, which decreased oxidative stress damage due to reactive oxygen species (ROS) hyperaccumulation under drought stress. The transcription of drought stress response and ROS scavenging genes was significantly higher in the CrCOMT overexpression plants than in the WT plants. In addition, CrCOMT transgenic tobacco plants exhibited higher melatonin content under drought stress conditions. Exogenous melatonin was applied to C. rigescens under drought stress to confirm the function of melatonin in mediating drought tolerance; the relative water content and proline content were higher, and the relative electrolytic leakage was lower in melatonin-treated C. rigescens than in the untreated plants. In summary, these results show that CrCOMT plays a positive role in plant drought stress tolerance by regulating endogenous melatonin content. [ABSTRACT FROM AUTHOR]

Summary: Effectors synthesised in the pharyngeal glands are important in the successful invasion of root-knot nematodes. Meloidogyne javanica is among the nematodes that cause the most damage to various crops. In this study, an effector named MJ-10A08 of M. javanica was identified and investigated. Mj-10A08 was exclusively expressed in the dorsal pharyngeal gland cell and highly expressed in the parasitic second-juvenile stage of M. javanica. Transgenic tobaccos that over-expressed Mj-10A08 were more susceptible to M. javanica ; however, host delivered RNAi of Mj-10A08 in tobacco significantly decreased the expression level of Mj-10A08 and the infection efficiency of M. javanica. Transient expression in tobacco leaves demonstrated that MJ-10A08 suppressed programmed cell death caused by BAX and Gpa2/RBP-1. Our results indicated that MJ-10A08 is implicated in the suppression of plant defence response during nematode infection and plays an important role in the parasitism of M. javanica. [ABSTRACT FROM AUTHOR]

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