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1.
J Agric Food Chem ; 67(32): 8905-8918, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31380641

RESUMO

NAC TFs play crucial roles in response to abiotic stresses in plants. Here, ZmNAC071 was identified as a nuclear located transcriptional repressor. Overexpression of ZmNAC071 in Arabidopsis enhanced sensitivity of transgenic plants to ABA and osmotic stress. The expression levels of SODs, PODs, P5CSs, and AtMYB61 were inhibited by ZmNAC071, which results in reduced ROS scavenging and proline content, increased ROS level, and water loss. Besides, the expression levels of some ABA or abiotic stress-related genes, like ABIs, RD29A, DREBs, and LEAs were also significantly inhibited by ZmNAC071. Yeast one-hybrid assay demonstrated that ZmNAC071 specifically bound to the cis-acting elements containing CGT[G/A] core sequences in the promoter of stress-related genes, suggesting that ZmNAC071 may participate in the regulation of transcription of these genes through recognizing the core sequences CGT[G/A]. These results will facilitate further studies concerning the cis-elements and downstream genes targeted by ZmNAC071 in maize.


Assuntos
Arabidopsis/efeitos dos fármacos , Arabidopsis/fisiologia , Ácido Ascórbico/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/fisiologia , Fatores de Transcrição/genética , Zea mays/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação para Baixo/efeitos dos fármacos , Pressão Osmótica , Plantas Geneticamente Modificadas/genética , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico , Fatores de Transcrição/metabolismo
2.
BMC Plant Biol ; 19(1): 231, 2019 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-31159735

RESUMO

BACKGROUND: Four-Coumarate:CoA ligase gene (4CL) plays multiple important roles in plant growth and development by catalyzing the formation of CoA ester. 4CL belongs to the plant phenylpropane derivative, which is related to the synthesis of flavonoids and lignin and is a key enzyme in the biosynthetic pathway. RESULTS: In this study, 12 4CL genes of Fraxinus mandschurica were identified and named Fm4CL1-Fm4CL12, respectively. The analysis of the expression pattern of Fm4CL genes indicate that Fm4CL-like 1 gene may play a role in the lignin synthesis pathway. Our study indicate that overexpression of Fm4CL-like 1 increases the lignin content of transgenic tobacco by 39.5% compared to WT, and the S/G ratio of transgenic tobacco increased by 19.7% compared with WT. The xylem cell layer of transgenic line is increased by 40% compared to WT, the xylem cell wall thickness increased by 21.6% compared to the WT. Under mannitol-simulated drought stress, the root length of transgenic tobacco is 64% longer than WT, and the seed germination rate of the transgenic lines is 47% higher than that of WT. In addition, the H2O2 content in the transgenic tobacco was 22% lower than that of WT, while the POD and SOD content was higher than WT by 30 and 24% respectively, which showed Fm4CL-like 1 affect the accumulation of reactive oxygen species (ROS). The MDA content and relative conductivity was 25 and 15% lower than WT, respectively. The water loss rate is 16.7% lower than that of WT. The relative expression levels of stress-related genes NtHAK, NtAPX, NtCAT, NtABF2, and NtZFP were higher than those of WT under stress treatment. The stomatal apertures of OE (Overexpression) were 30% smaller than those of WT, and the photosynthetic rate of OE was 48% higher than that of WT. These results showed that the overexpression line exhibited stronger adaptability to osmotic stress than WT. CONCLUSIONS: Our results indicate that Fm4CL-like 1 is involved in secondary cell wall development and lignin synthesis. Fm4CL-like 1 play an important role in osmotic stress by affecting cell wall and stomatal development.


Assuntos
Secas , Proteínas de Plantas/genética , Tabaco/fisiologia , Clonagem Molecular , Fraxinus/genética , Fraxinus/metabolismo , Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/genética , Tabaco/genética
3.
BMC Plant Biol ; 19(1): 237, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31170915

RESUMO

BACKGROUND: The Agrobacterium-mediated transient transformation is a versatile and indispensable way of rapid analyzing gene function in plants. Despite this transient expression system has been successfully applied in a number of plant species, it is poorly developed in Caragana intermedia. RESULTS: In this study, we established an Agrobacterium-mediated transient expression system in C. intermedia leaves and optimized the effect of different Agrobacterial strains, several surfactants and the concentration of Silwet L-77, which would affect transient expression efficiency. Among the 5 Agrobacterial strains examined, GV3101 produced the highest GUS expression level. Besides, higher level of transient expression was observed in plants infiltrated with Silwet L-77 than with Triton X-100 or Tween-20. Silwet L-77 at a concentration of 0.001% greatly improved the level of GUS transient expression. Real-time PCR showed that expression of CiDREB1C was highly up-regulated in transiently expressed plants and reached the highest level at the 2nd day after infiltration. Based on this optimized transient transformation method, we characterized CiDREB1C function in response to drought, salt and ABA treatment. The results showed that transiently expressed CiDREB1C in C. intermedia leaves could enhance the survival rate and chlorophyll content, and reduce the lodging rate compared with the control seedlings under drought, salt and ABA treatments. Furthermore, the rate of leaf shedding of CiDREB1C transient expression seedlings was lower than that of the control under ABA treatment. CONCLUSIONS: The optimized transient expression condition in C. intermedia leaves were infiltrated with Agrobacterial strains GV3101 plus Silwet L-77 at a concentration of 0.001% added into the infiltration medium. Transiently expressed CiDREB1C enhanced drought, salt and ABA stress tolerance, indicated that it was a suitable and effective tool to determine gene function involved in abiotic stress response in C. intermedia.


Assuntos
Agrobacterium/genética , Caragana/fisiologia , Regulação da Expressão Gênica de Plantas , Engenharia Genética/métodos , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Fatores de Transcrição/genética , Proteínas de Arabidopsis , Caragana/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Fatores de Transcrição/metabolismo
4.
Plant Physiol Biochem ; 141: 164-171, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31170640

RESUMO

For a comprehensive understanding of gene expression, enzyme activity and sugar concentrations in response to short-term water deficit in apple (Greensleeves), sugar-modulated gene expression and enzyme activities were analyzed. Water stress resulted in the accumulation of sorbitol, glucose, fructose, galactose and starch, accompanied by a significant reduction in photosynthesis and sucrose concentration. In response to short-term water deficits, the activities of aldose-6-phosphate reductase (A6PR; EC 1.1.1.200), sorbitol dehydrogenase (SDH; EC 1.1.1.14), neutral invertase (NINV; EC 3.2.1.26), sucrose synthase (SUSY; EC 2.4.1.13), and fructokinase (FK; EC 2.7.1.4) were higher, whereas cell wall invertase (CWINV; EC 3.2.1.26) and hexokinase (HK; EC 2.7.1.1) activities were lower. In addition, sucrose phosphate synthase (SPS; EC 2.4.1.14) activity increased during the initial stages of dehydration and then decreased as the drought strengthened. Transcript levels of MdA6PR, MdSDH1/2, MdNINV1/2, MdSUSY3, MdFK1/2/4, MdSOT1/2, MdSUC1-3, MdTMT2/3, MdvGT1, MdpGlcT1-4 were upregulated, whereas transcript levels of MdCWINV1/2, MdHK1/2/3/5, and MdTMT1 were downregulated after 6 days of water stress. These findings suggest that the sorbitol metabolism pathway is induced and high levels of hexose derived from photosynthetic products are transported into vacuoles for adjustment to the water deficit. Our results provide insights into the relationships between sugar levels and sugar-modulated gene and enzyme activity in response to the imposition of short-term water stress.


Assuntos
Metabolismo dos Carboidratos/genética , Secas , Regulação da Expressão Gênica de Plantas , Malus/fisiologia , Folhas de Planta/fisiologia , Parede Celular/metabolismo , Frutas/metabolismo , Glucosiltransferases/metabolismo , Malus/genética , Fotossíntese , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Sorbitol/metabolismo , Estresse Fisiológico , Sacarose/metabolismo , beta-Frutofuranosidase/metabolismo
5.
Plant Physiol Biochem ; 141: 300-305, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31202194

RESUMO

MYB-type transcription factors are known to participate in the response of plants to a number of stress agents. MsMYB2L is an alfalfa member of this large gene family. Its transcription in alfalfa seedlings was found to be rapidly and strongly induced by salinity, moisture deficiency and exogenously supplied abscisic acid. An analysis based on a yeast one hybrid assay indicated that its product is able to activate transcription, consistent with its function as a transcription factor. When the gene was constitutively expressed in Arabidopsis thaliana, both germination and seedling growth were more sensitive to ABA treatment than wild type, and growth was less strongly compromised by salinity and moisture deficiency stress, presumably as a result of the induction of certain stress-related genes active in ABA-dependent pathways. The transgenic seedlings' enhanced the synthesis of many osmotic regulatory substances such as proline and soluble sugar, and decreased the lipid peroxidation. In all, MsMYB2L represents a potential candidate gene for manipulating the salinity and drought tolerance of alfalfa.


Assuntos
Ácido Abscísico/metabolismo , Arabidopsis/fisiologia , Proteínas de Ligação a DNA/genética , Secas , Medicago sativa/genética , Proteínas de Plantas/genética , Salinidade , Fatores de Transcrição/genética , Regulação da Expressão Gênica de Plantas , Técnicas Genéticas , Germinação , Filogenia , Plantas Geneticamente Modificadas/fisiologia , Polietilenoglicóis/química , Plântula/fisiologia , Estresse Fisiológico , Açúcares/química , Transcrição Genética , Ativação Transcricional
6.
Plant Physiol Biochem ; 141: 343-352, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31207495

RESUMO

Plant U-box (PUB) E3 ubiquitin ligases play crucial roles in the plant response to abiotic stress and the phytohormone abscisic acid (ABA) signaling, but little is known about them in bryophytes. Here, a representative U-box armadillo repeat (PUB-ARM) ubiquitin E3 ligase from Antarctic moss Pohlia nutans (PnSAG1), was explored for its role in abiotic stress response in Arabidopsis thaliana and Physcomitrella patens. The expression of PnSAG1 was rapidly induced by exogenous abscisic acid (ABA), salt, cold and drought stresses. PnSAG1 was localized to the cytoplasm and showed E3 ubiquitin ligase activity by in vitro ubiquitination assay. The PnSAG1-overexpressing Arabidopsis enhanced the sensitivity with respect to ABA and salt stress during seed germination and early root growth. Similarly, heterogeneous overexpression of PnSAG1 in P. patens was more sensitive to the salinity and ABA in their gametophyte growth. The analysis by RT-qPCR revealed that the expression of salt stress/ABA-related genes were downregulated in PnSAG1-overexpressing plants after salt treatment. Taken together, our results indicated that PnSAG1 plays a negative role in plant response to ABA and salt stress.


Assuntos
Ácido Abscísico/farmacologia , Arabidopsis/fisiologia , Briófitas/enzimologia , Bryopsida/fisiologia , Estresse Salino , Ubiquitina-Proteína Ligases/genética , Regiões Antárticas , Arabidopsis/genética , Briófitas/genética , Bryopsida/genética , Biologia Computacional , Secas , Regulação da Expressão Gênica de Plantas , Células Germinativas Vegetais/metabolismo , Germinação , Raízes de Plantas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/fisiologia , Transdução de Sinais
7.
Plant Physiol Biochem ; 141: 446-455, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31247427

RESUMO

Coumarin plays a pivotal role in plant response to biotic stress, as well as in the mediation of nutrient acquisition. However, its functions in response to abiotic stresses are largely unknown. In this work, a homologous gene, GmF6'H1, of AtF6'H1, which encodes the enzyme catalyzing the final rate-limiting step in the biosynthesis pathway of coumarin, was isolated from soybean. GmF6'H1 protein shares very high amino acid identity with AtF6'H1, and expression of GmF6'H1 in atf6'h1 can successfully restore the decreased coumarin production in the T-DNA insertion mutant. Further study revealed that the expression of GmF6'H1 in soybean was remarkably induced by salt stress. Constitutive expression of GmF6'H1 in Arabidopsis, driven by 35S promoter, significantly enhanced the resistance to salt of transgenic Arabidopsis. All these results suggest that GmF6'H1 can be used as a potential candidate gene for the engineering of plants with improved resistance to both biotic and abiotic stresses.


Assuntos
Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Tolerância ao Sal , Soja/enzimologia , Arabidopsis/genética , Clorofila/química , Clonagem Molecular , Cumarínicos/química , Perfilação da Expressão Gênica , Germinação , Fenótipo , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Regiões Promotoras Genéticas , Soja/genética
8.
BMC Plant Biol ; 19(1): 214, 2019 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-31122194

RESUMO

BACKGROUND: MicroRNA319 (miR319) acts as an essential regulator of gene expression during plant development and under stress conditions. Although the role of miR319a in regulating leaf development has been well studied in tomato (Solanum lycopersicum), the function of the recently discovered wild tomato Solanum habrochaites miRNA319d (sha-miR319d) remains poorly understood. In this study, we overexpressed sha-miR319d in cultivated tomato 'Micro-Tom' to further investigate its role in tomato temperature stress responses. RESULTS: Under chilling or heat stress, sha-miR319d-overexpressing plants showed enhanced stress tolerance, including lower relative electrolyte leakage (REL), malondialdehyde (MDA) concentration, O2- generation and H2O2 concentration and higher chlorophyll contents and Fv/Fm values than wild-type (WT) plants. Overexpression of sha-miR319d enhanced the activities of superoxide dismutase (SOD) and catalase (CAT), with possible correlation with elevated expression levels of the genes FeSOD, CuZnSOD and CAT. Moreover, different expression levels of key genes involved in chilling (MYB83 and CBF1), heat (HsfA1a, HsfA1b and Hsp90), and reactive oxygen species (ROS) (ZAT12 and ZAT10) signaling in transgenic plants and WT were determined, suggesting a role for sha-miR319d in regulating tomato temperature stress via chilling, heat and ROS signaling. Silencing GAMYB-like1 increased tomato chilling tolerance as well as the expression levels of CBF1, CuZnSOD, CAT, APX1, APX2, ZAT12 and ZAT10. Additionally, overexpression of sha-miR319d in tomato caused plant leaf crinkling and reduced height. CONCLUSIONS: Overexpression of sha-miR319d confers chilling and heat stress tolerance in tomato. Sha-miR319d regulates tomato chilling tolerance, possibly by inhibiting expression of GAMYB-like1 and further alters chilling, heat and ROS signal transduction. Our research provides insight for further study of the role of sha-miR319d in tomato growth and stress regulation and lays a foundation for the genetic improvement of tomato.


Assuntos
Resposta ao Choque Frio/genética , Regulação da Expressão Gênica de Plantas , Resposta ao Choque Térmico/genética , Lycopersicon esculentum/fisiologia , MicroRNAs/genética , RNA de Plantas/genética , Solanum/fisiologia , Lycopersicon esculentum/genética , MicroRNAs/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , RNA de Plantas/metabolismo , Solanum/genética , Termotolerância/genética
9.
BMC Plant Biol ; 19(1): 218, 2019 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-31133003

RESUMO

BACKGROUND: An eukaryotic translation elongation factor-2 (eEF-2) plays an important role in protein synthesis, however, investigation on its role in abiotic stress responses is limited. A cold responsive eEF2 named as MfEF2 was isolated from yellow-flowered alfalfa [Medicago sativa subsp. falcata (L.) Arcang, thereafter M. falcata], a forage legume with great cold tolerance, and transgenic tobacco (Nicotiana tabacum L.) plants overexpressing MfEF2 were analyzed in cold tolerance and proteomic profiling was conducted under low temperature in this study. RESULTS: MfEF2 transcript was induced and peaked at 24 h and remained at the high level during cold treatment up to 96 h. Overexpression of MfEF2 in trasngenic tobacco plants resulted in enhanced cold tolerance. Compared to the wild type, transgenic plants showed higher survival rate after freezing treatment, higher levels of net photosynthetic rate (A), maximum photochemical efciency of photosystem (PS) II (Fv/Fm) and nonphotochemical quenching (NPQ) and lower levels of ion leakage and reactive oxygen species (ROS) production after chilling treatment. iTRAQ-based quantitative proteomic analysis identified 336 differentially expressed proteins (DEPs) from leaves of one transgenic line versus the wild type after chilling treatment for 48 h. GO and KEGG enrichment were conducted for analysis of the major biological process, cellular component, molecular function, and pathways of the DEPs involving in. It is interesting that many down-regulated DEPs were grouped into "photosynthesis" and "photosynthesis-antenna", such as subunits of PSI and PSII as well as light harvesting chlorophyll protein complex (LHC), while many up-regulated DEPs were grouped into "spliceosome". CONCLUSIONS: The results suggest that MfEF2 confers cold tolerance through regulating hundreds of proteins synthesis under low temperature conditions. The elevated cold tolerance in MfEF2 transgenic plants was associated with downregulation of the subunits of PSI and PSII as well as LHC, which leads to reduced capacity for capturing sunlight and ROS production for protection of plants, and upregulation of proteins involving in splicesome, which promotes alternative splicing of pre-mRNA under low temperature.


Assuntos
Adaptação Fisiológica/genética , Regulação da Expressão Gênica de Plantas , Medicago/fisiologia , Fator 2 de Elongação de Peptídeos/genética , Proteínas de Plantas/genética , Tabaco/fisiologia , Temperatura Baixa , Medicago/genética , Fator 2 de Elongação de Peptídeos/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/genética , Tabaco/genética
10.
BMC Plant Biol ; 19(1): 208, 2019 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-31109298

RESUMO

BACKGROUND: Cotton fiber is a single cell that arises from the epidermis of ovule. It is not only a main economic product of cotton, but an ideal material for studying on the growth and development of plant cell. Our previous study indicated that phytosterol content and the ratio of campesterol to sitosterol fluctuated regularly in cotton fiber development. However, what effects of modified phytosterol content and composition on the growth and development of cotton fiber cell is unknown. In this study, we overexpressed the GhSMT2-1, a cotton homologue of sterol C-24 methyltransferase 2 gene in transgenic upland cotton plants to modify phytosterol content and composition in fiber cells and investigated the changes on fiber elongation and secondary cell wall deposition. RESULTS: GhSMT2-1 overexpression led to changes of phytosterol content and the ratio of campesterol to sitosterol in fiber cell. At the rapid elongation stage of fiber cell, total phytosterol and sitosterol contents were increased while campesterol content was decreased in transgenic fibers when compared to control fibers. Accordingly, the ratio of campesterol to sitosterol declined strikingly. Simultaneously, the transgenic fibers were shorter and thicker than control fibers. Exogenous application of sitosterol or campesterol separately inhibited control fiber cell elongation in cotton ovule culture system in vitro. In addition, campesterol treatment partially rescued transgenic fiber elongation. CONCLUSION: These results elucidated that modification of phytosterol content and composition influenced fiber cell elongation and secondary cell wall formation. High sitosterol or low ratio of campesterol to sitosterol suppresses fiber elongation and/or promote secondary cell wall deposition. The roles of sitosterol and campesterol were discussed in fiber cell development. There might be a specific ratio of campesterol to sitosterol in different developmental stage of cotton fibers, in which GhSMT2-1 play an important role. Our study, at a certain degree, provides novel insights into the regulatory mechanisms of fiber cell development.


Assuntos
Gossypium/química , Gossypium/fisiologia , Fitosteróis/química , Proteínas Arqueais/genética , Proteínas Arqueais/metabolismo , Crescimento Celular , Parede Celular , Fibra de Algodão , Gossypium/genética , Metiltransferases/genética , Metiltransferases/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia
11.
J Zhejiang Univ Sci B ; 20(4): 322-331, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30932377

RESUMO

We investigated the microRNA172 (miR172)-mediated regulatory network for the perception of changes in external and endogenous signals to identify a universally applicable floral regulation system in ornamental plants, manipulation of which could be economically beneficial. Transgenic gloxinia plants, in which miR172 was either overexpressed or suppressed, were generated using Agrobacterium-mediated transformation. They were used to study the effect of altering the expression of this miRNA on time of flowering and to identify its mRNA target. Early or late flowering was observed in transgenic plants in which miR172 was overexpressed or suppressed, respectively. A full-length complementary DNA (cDNA) of gloxinia (Sinningia speciosa) APETALA2-like (SsAP2-like) was identified as a target of miR172. The altered expression levels of miR172 caused up- or down-regulation of SsAP2-like during flower development, which affected the time of flowering. Quantitative real-time reverse transcription PCR analysis of different gloxinia tissues revealed that the accumulation of SsAP2-like was negatively correlated with the expression of miR172a, whereas the expression pattern of miR172a was negatively correlated with that of miR156a. Our results suggest that transgenic manipulation of miR172 could be used as a universal strategy for regulating time of flowering in ornamental plants.


Assuntos
Proteínas de Arabidopsis/genética , Flores/fisiologia , Proteínas de Homeodomínio/genética , Lamiales/fisiologia , MicroRNAs/genética , Proteínas Nucleares/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Clonagem Molecular , DNA Complementar/metabolismo , Flores/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteínas de Homeodomínio/metabolismo , Lamiales/genética , MicroRNAs/metabolismo , Proteínas Nucleares/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Transgenes
12.
Int J Mol Sci ; 20(8)2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-31018553

RESUMO

Dehydrins (DHNs), as a sub-family of group two late embryogenesis-abundant (LEA) proteins, have attracted considerable interest owing to their functions in enhancing abiotic stress tolerance in plants. Our previous study showed that the expression of CaDHN5 (a dehydrin gene from pepper) is strongly induced by salt and osmotic stresses, but its function was not clear. To understand the function of CaDHN5 in the abiotic stress responses, we produced pepper (Capsicum annuum L.) plants, in which CaDHN5 expression was down-regulated using VIGS (Virus-induced Gene Silencing), and transgenic Arabidopsis plants overexpressing CaDHN5. We found that knock-down of CaDHN5 suppressed the expression of manganese superoxide dismutase (MnSOD) and peroxidase (POD) genes. These changes caused more reactive oxygen species accumulation in the VIGS lines than control pepper plants under stress conditions. CaDHN5-overexpressing plants exhibited enhanced tolerance to salt and osmotic stresses as compared to the wild type and also showed increased expression of salt and osmotic stress-related genes. Interestingly, our results showed that many salt-related genes were upregulated in our transgenic Arabidopsis lines under salt or osmotic stress. Taken together, our results suggest that CaDHN5 functions as a positive regulator in the salt and osmotic stress signaling pathways.


Assuntos
Capsicum/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/fisiologia , Capsicum/fisiologia , Secas , Genes de Plantas , Osmorregulação , Pressão Osmótica , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Tolerância ao Sal
13.
Int J Mol Sci ; 20(8)2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-31018555

RESUMO

Polyamines play an important role in plant growth and development, and response to abiotic stresses. Previously, differentially expressed proteins in sugar beet M14 (BvM14) under salt stress were identified by iTRAQ-based quantitative proteomics. One of the proteins was an S-adenosylmethionine decarboxylase (SAMDC), a key rate-limiting enzyme involved in the biosynthesis of polyamines. In this study, the BvM14-SAMDC gene was cloned from the sugar beet M14. The full-length BvM14-SAMDC was 1960 bp, and its ORF contained 1119 bp encoding the SAMDC of 372 amino acids. In addition, we expressed the coding sequence of BvM14-SAMDC in Escherichia coli and purified the ~40 kD BvM14-SAMDC with high enzymatic activity. Quantitative real-time PCR analysis revealed that the BvM14-SAMDC was up-regulated in the BvM14 roots and leaves under salt stress. To investigate the functions of the BvM14-SAMDC, it was constitutively expressed in Arabidopsis thaliana. The transgenic plants exhibited greater salt stress tolerance, as evidenced by longer root length and higher fresh weight and chlorophyll content than wild type (WT) under salt treatment. The levels of spermidine (Spd) and spermin (Spm) concentrations were increased in the transgenic plants as compared with the WT. Furthermore, the overexpression plants showed higher activities of antioxidant enzymes and decreased cell membrane damage. Compared with WT, they also had low expression levels of RbohD and RbohF, which are involved in reactive oxygen species (ROS) production. Together, these results suggest that the BvM14-SAMDC mediated biosynthesis of Spm and Spd contributes to plant salt stress tolerance through enhancing antioxidant enzymes and decreasing ROS generation.


Assuntos
Adenosilmetionina Descarboxilase/genética , Beta vulgaris/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Tolerância ao Sal , Regulação para Cima , Arabidopsis/genética , Arabidopsis/fisiologia , Beta vulgaris/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico
14.
Plant Physiol Biochem ; 139: 504-512, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31015089

RESUMO

Plant NAC proteins constitute one of the largest transcription factor families. They play pivotal functions during responses to various abiotic stresses. However, knowledge on roles of NAC proteins in abiotic stress tolerance as well as corresponding mechanisms has not been fully studied in perennial woody plants, including domesticated apple (Malus domestica). In the present study, we characterized the role of apple MdNAC1 transcription factor in response to drought stress. Apple plants overexpressing MdNAC1 gene exhibited promoted tolerance to drought stress, as evident by reduced water loss and electrolyte leakage in leaves, and maintenance of photosynthesis and photosynthetic pigments content under drought conditions. In addition, the levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were significantly lower for transgenic apple lines than those for nontransgenic plants under drought conditions. This was accompanied by higher activities of several antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as increased expression of the associated genes in transgenic lines. Together, our results indicate that overexpression of the apple MdNAC1 gene enhances drought stress tolerance in apple plants by promoting higher photosynthesis and activities of ROS-scavenging enzymes.


Assuntos
Secas , Malus/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Fatores de Transcrição/metabolismo , Catalase/genética , Catalase/metabolismo , Regulação da Expressão Gênica de Plantas , Malus/genética , Malus/fisiologia , Peroxidase/genética , Peroxidase/metabolismo , Fotossíntese/genética , Fotossíntese/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Fatores de Transcrição/genética
15.
Tree Physiol ; 39(3): 345-355, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30917196

RESUMO

Condensed tannins (CTs) have been studied extensively as potential defenses against pests and pathogens, and for their beneficial effects on human health. They are known to possess high in vitro antioxidant capacity, but whether they can function as in planta antioxidants for protection against oxidative stress has not been previously tested. Here, we show that stress induction of CTs in poplar (Populus) is matched closely by an increase in antioxidant activity under both high light and nitrogen deficiency. We also investigate the effects of CTs as in vivo antioxidants directly, using transgenic poplar plants which overexpress poplar MYB transcription factors that regulate the CT pathway. These transgenics have 50-fold higher CT concentrations than controls, and and also have dramatically higher antioxidant activity. High-CT and control poplar leaves were exposed to methyl viologen for 24 h. Chlorophyll fluorescence was used to measure maximum quantum efficiency of photosystem II photochemistry (Fv/Fm), and leaf discs were stained with 3,3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) to assess hydrogen peroxide and superoxide levels. After methyl viologen exposure, high-CT transgenics retained higher Fv/Fm ratios and accumulated less hydrogen peroxide and superoxide than the controls. Our findings indicate that high-CT concentrations protect poplar against methyl viologen-induced oxidative stress and suggest a broader function of CTs than previously supposed.


Assuntos
Antioxidantes/metabolismo , Nitrogênio/deficiência , Estresse Oxidativo , Populus/fisiologia , Proantocianidinas/metabolismo , Plantas Geneticamente Modificadas/fisiologia
16.
Plant Sci ; 280: 1-11, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30823987

RESUMO

Abscisic acid (ABA) and jasmonates (JAs) are the primary plant hormones involved in mediating salt tolerance. In addition, these two plant hormones exert a synergistic effect to inhibit seed germination. However, the molecular mechanism of the interaction between ABA signalling and JA signalling is still not well documented. Here, a moss jasmonate ZIM-domain gene (PnJAZ1), which encodes a nucleus-localized protein with conserved ZIM and Jas domains, was cloned from Pohlia nutans. PnJAZ1 expression was rapidly induced by various abiotic stresses. The PnJAZ1 protein physically interacted with MYC2 and was degraded by exogenous 12-oxo-phytodienoic acid (OPDA) treatment, implying that the JAZ protein-mediated signalling pathway is conserved in plants. Transgenic Arabidopsis and Physcomitrella plants overexpressing PnJAZ1 showed increased tolerance to salt stress and decreased ABA sensitivity during seed germination and early development. The overexpression of PnJAZ1 inhibited the expression of ABA pathway genes related to seed germination and seedling growth. Moreover, the transgenic Arabidopsis lines exhibited enhanced tolerance to auxin (IAA) and glucose, mimicking the phenotypes of abi4 or abi5 mutants. These results suggest that PnJAZ1 acts as a repressor, mediates JA-ABA synergistic crosstalk and enhances plant growth under salt stress.


Assuntos
Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Proteínas de Plantas/metabolismo , Tolerância ao Sal/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Tolerância ao Sal/genética , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
17.
Int J Mol Sci ; 20(5)2019 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-30857203

RESUMO

APETALA2/ethylene-responsive factor superfamily (AP2/ERF) is a transcription factor involved in abiotic stresses, for instance, cold, drought, and low oxygen. In this study, a novel ethylene-responsive transcription factor named AdRAP2.3 was isolated from Actinidia deliciosa 'Jinkui'. AdRAP2.3 transcription levels in other reproductive organs except for the pistil were higher than those in the vegetative organs (root, stem, and leaf) in kiwi fruit. Plant hormones (Salicylic acid (SA), Methyl-jasmonate acid (MeJA), 1-Aminocyclopropanecarboxylic Acid (ACC), Abscisic acid (ABA)), abiotic stresses (waterlogging, heat, 4 °C and NaCl) and biotic stress (Pseudomonas Syringae pv. Actinidiae, Psa) could induce the expression of AdRAP2.3 gene in kiwi fruit. Overexpression of the AdRAP2.3 gene conferred waterlogging stress tolerance in transgenic tobacco plants. When completely submerged, the survival rate, fresh weight, and dry weight of transgenic tobacco lines were significantly higher than those of wile type (WT). Upon the roots being submerged, transgenic tobacco lines grew aerial roots earlier. Overexpression of AdRAP2.3 in transgenic tobacco improved the pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) enzyme activities, and improved the expression levels of waterlogging mark genes NtPDC, NtADH, NtHB1, NtHB2, NtPCO1, and NtPCO2 in roots under waterlogging treatment. Overall, these results demonstrated that AdRAP2.3 might play an important role in resistance to waterlogging through regulation of PDC and ADH genes in kiwi fruit.


Assuntos
Actinidia/fisiologia , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico , Tabaco/fisiologia , Fatores de Transcrição/metabolismo , Actinidia/genética , Álcool Desidrogenase/genética , Álcool Desidrogenase/metabolismo , Genes de Plantas , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Piruvato Descarboxilase/genética , Piruvato Descarboxilase/metabolismo , Tabaco/genética , Fatores de Transcrição/genética
18.
Proc Natl Acad Sci U S A ; 116(8): 3006-3011, 2019 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-30723147

RESUMO

Monarch butterfly (Danaus plexippus) decline over the past 25 years has received considerable public and scientific attention, in large part because its decline, and that of its milkweed (Asclepias spp.) host plant, have been linked to genetically modified (GM) crops and associated herbicide use. Here, we use museum and herbaria specimens to extend our knowledge of the dynamics of both monarchs and milkweeds in the United States to more than a century, from 1900 to 2016. We show that both monarchs and milkweeds increased during the early 20th century and that recent declines are actually part of a much longer-term decline in both monarchs and milkweed beginning around 1950. Herbicide-resistant crops, therefore, are clearly not the only culprit and, likely, not even the primary culprit: Not only did monarch and milkweed declines begin decades before GM crops were introduced, but other variables, particularly a decline in the number of farms, predict common milkweed trends more strongly over the period studied here.


Assuntos
Asclepias/genética , Borboletas/fisiologia , Ecossistema , Plantas Geneticamente Modificadas/genética , Migração Animal/fisiologia , Animais , Asclepias/crescimento & desenvolvimento , Borboletas/genética , Resistência a Herbicidas/genética , Interações Hospedeiro-Parasita , Plantas Geneticamente Modificadas/fisiologia , Dinâmica Populacional , Estados Unidos
19.
Environ Sci Pollut Res Int ; 26(11): 10537-10551, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30762181

RESUMO

The group 5 LEA (late embryogenesis abundant) proteins are an atypical LEA protein group, which is associated with resistance to multiple stresses. In this study, OsLea14-A gene was isolated from Oryza sativa L., which encodes a 5C LEA protein with 151 amino acids. The qPCR analysis showed that OsLea14-A expressed in all tissues and organs at all times. The expression of OsLea14-A in the panicles of plumping stage were dramatically increased. The heterologous expression of OsLea14-A in Escherichia coli improved its growth performance under salinity, desiccation, high temperature, and freeze-thaw stresses. The purified OsLea14-A protein can protect LDH activity from freeze-thaw-, heat-, and desiccation-induced inactivation. The overexpression of OsLea14-A in rice improved tolerance to dehydration, high salinity, CuSO4, and HgCl2, but excluding K2Cr2O7. The analysis of metal contents showed that the accumulation of OsLea14-A protein in transgenic rice could increase the accumulation of Hg, but could not increase the accumulation of Na, Cr, and Cu after HgCl2, NaCl, K2Cr2O7, and CuSO4 treatment, respectively. These results suggested that OsLea14-A conferred multiple stress tolerance and Hg accumulation, which made it a possible gene in genetic improvement for plants to acclimatize itself to multiple stresses and remediate Hg-contaminated soil.


Assuntos
Mercúrio/metabolismo , Oryza/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Tolerância a Medicamentos , Escherichia coli/genética , Escherichia coli/metabolismo , Regulação da Expressão Gênica de Plantas , Temperatura Alta , Oryza/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Salinidade , Cloreto de Sódio/metabolismo , Estresse Fisiológico
20.
Plant Cell Rep ; 38(5): 577-586, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30758711

RESUMO

KEY MESSAGE: We developed a novel Agrobacterium-mediated anther transformation for Wucai in planta, and in this procedure, the male germ line was the predominant target. Wucai (Brassica campestris L.), a variant of non-heading Chinese cabbage, is widely cultured in China and only improved by classic breeding methods. Here, a novel and efficient in planta Agrobacterium-mediated anther transformation method is developed based on the optimization of several factors that affect anther transformation. After optimization, transformation with the manual pollination application led to increased transient GUS expression in anthers (reaching 91.59%) and the transformation efficacies in planta (0.59-1.56% for four commercial cultivars). The stable integration and inheritance of the transgenes were further examined by molecular and genetic analyses. Three T2 transgenic lines presented a segregation ratio of 3:1, which was consistent with the Mendelian feature of a single dominant gene. In addition, the GUS histochemical assay and genetic crossing analysis revealed that the male germ line was the predominant target in this transformation. This optimized transformation system could provide a useful tool for both the improvement of cultivar qualities and investigation of functional genes in Wucai.


Assuntos
Agrobacterium tumefaciens/genética , Brassica/genética , Brassica/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Transformação Genética/genética , Brassica/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Polinização/genética , Polinização/fisiologia
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