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1.
BMC Plant Biol ; 19(1): 368, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31429706

RESUMO

BACKGROUND: We previously reported the involvement of nitric oxide (NO) and cyclic nucleotide-gated ion channel 6 (CNGC6) in the responses of plants to heat shock (HS) exposure. To elucidate their relationship with heat tolerance in Arabidopsis thaliana, we examined the effects of HS on several groups of seedlings: wild type, cngc6, and cngc6 complementation and overexpression lines. RESULTS: After HS exposure, the level of NO was lower in cngc6 seedlings than in wild-type seedlings but significantly elevated in the transgenic lines depending on CNGC6 expression level. The treatment of seeds with calcium ions (Ca2+) enhanced the NO level in Arabidopsis seedlings under HS conditions, whereas treatment with EGTA (a Ca2+ chelator) reduced it, implicating that CNGC6 stimulates the accumulation of NO depending on an increase in cytosolic Ca2+ ([Ca2+]cyt). This idea was proved by phenotypic observations and thermotolerance testing of transgenic plants overexpressing NIA2 and NOA1, respectively, in a cngc6 background. Western blotting indicated that CNGC6 stimulated the accumulation of HS proteins via NO. CONCLUSION: These data indicate that CNGC6 acts upstream of NO in the HS pathway, which improves our insufficient knowledge of the initiation of plant responses to high temerature.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Cálcio/metabolismo , Canais de Cátion Regulados por Nucleotídeos Cíclicos/metabolismo , Óxido Nítrico/metabolismo , Termotolerância , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Canais de Cálcio/metabolismo , Canais de Cátion Regulados por Nucleotídeos Cíclicos/genética , Citosol/metabolismo , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Resposta ao Choque Térmico , Mutação , Nitrato Redutase/genética , Nitrato Redutase/metabolismo , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/metabolismo , Plântula/genética , Plântula/metabolismo
2.
BMC Plant Biol ; 19(1): 338, 2019 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-31375063

RESUMO

BACKGROUND: In native environments plants frequently experience simultaneous or sequential unfavourable abiotic and biotic stresses. The plant's response to combined stresses is usually not the sum of the individual responses. Here we investigated the impact of cold on plant defense against subsequent herbivory by a generalist and specialist insect. RESULTS: We determined transcriptional responses of Arabidopsis thaliana to low temperature stress (4 °C) and subsequent larval feeding damage by the lepidopteran herbivores Mamestra brassicae (generalist), Pieris brassicae (specialist) or artificial wounding. Furthermore, we compared the performance of larvae feeding upon cold-experienced or untreated plants. Prior experience of cold strongly affected the plant's transcriptional anti-herbivore and wounding response. Feeding by P. brassicae, M. brassicae and artificial wounding induced transcriptional changes of 1975, 1695, and 2239 genes, respectively. Of these, 125, 360, and 681 genes were differentially regulated when cold preceded the tissue damage. Overall, prior experience of cold mostly reduced the transcriptional response of genes to damage. The percentage of damage-responsive genes, which showed attenuated transcriptional regulation when cold preceded the tissue damage, was highest in M. brassicae damaged plants (98%), intermediate in artificially damaged plants (89%), and lowest in P. brassicae damaged plants (69%). Consistently, the generalist M. brassicae performed better on cold-treated than on untreated plants, whereas the performance of the specialist P. brassicae did not differ. CONCLUSIONS: The transcriptional defense response of Arabidopsis leaves to feeding by herbivorous insects and artificial wounding is attenuated by a prior exposure of the plant to cold. This attenuation correlates with improved performance of the generalist herbivore M. brassicae, but not the specialist P. brassicae, a herbivore of the same feeding guild.


Assuntos
Arabidopsis/fisiologia , Herbivoria , Animais , Arabidopsis/metabolismo , Borboletas/fisiologia , Resposta ao Choque Frio , Dieta , Regulação da Expressão Gênica de Plantas/fisiologia , Herbivoria/fisiologia , Larva , Mariposas/fisiologia , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Transcriptoma
3.
J Agric Food Chem ; 67(35): 9757-9771, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31373492

RESUMO

BAK1 effects on plant stress responses have been well documented, but little is known regarding its effects on plant growth. In this study, we functionally characterized MdBAK1. Overexpressing MdBAK1 in Arabidopsis thaliana and apple trees promoted growth. Longitudinal stem cells were longer in transgenic plants than in wild-type plants. The size and number of cells and the area of the transverse stem were greater in the transgenic lines than in the wild-type plants. Moreover, transgenic A. thaliana and apple plants were more sensitive to an exogenous brassinosteroid. A transcriptome analysis of wild-type and transgenic apple revealed that MdBAK1 overexpression activated the brassinosteroid and ethylene signals, xylem production, and stress responses. Trend and Venn analyses indicated that carbohydrate, energy, and hormone metabolic activities were greater in transgenic plants during different periods. Moreover, a weighted gene coexpression network analysis proved that carbohydrate, hormone, and xylem metabolism as well as cell growth may be critical for MdBAK1-mediated apple tree growth and development. Compared with the corresponding levels in wild-type plants, the endogenous brassinosteroid, cytokinin, starch, sucrose, trehalose, glucose, fructose, and total sugar contents were considerably different in transgenic plants. Our results imply that MdBAK1 helps to regulate the growth of apple tree through the above-mentioned pathways. These findings provide new information regarding the effects of MdBAK1 onplant growth and development.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Malus/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Proteínas Serina-Treonina Quinases/metabolismo , Arabidopsis/metabolismo , Brassinosteroides/metabolismo , Etilenos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Malus/genética , Malus/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteínas Serina-Treonina Quinases/genética , Transdução de Sinais
4.
J Agric Food Chem ; 67(35): 9877-9884, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31398030

RESUMO

Heavy metal contaminants and nutrient deficiencies in soil negatively affect crop growth and human health. The plant cadmium resistance (PCR) protein transports heavy metals. The abundance of PCR is correlated with that of cell number regulator (CNR) protein, and the two proteins have similar conserved domains. Hence, CNR might also participate in heavy metal transport. We isolated and analyzed TaCNR5 from wheat (Triticum aestivum). The expression level of TaCNR5 in the shoots of wheat increased under cadmium (Cd), zinc (Zn), or manganese (Mn) treatments. Transgenic plants expressing TaCNR5 showed enhanced tolerance to Zn and Mn. Overexpression of TaCNR5 in Arabidopsis increased Cd, Zn, and Mn translocation from roots to shoots. The concentrations of Zn and Mn in rice grains were increased in transgenic plants expressing TaCNR5. These roles of TaCNR5 in the translocation and distribution of heavy metals mean that it has potential as a genetic biofortification tool to fortify cereal grains with micronutrients.


Assuntos
Manganês/metabolismo , Oryza/genética , Oryza/metabolismo , Proteínas de Plantas/genética , Triticum/genética , Zinco/metabolismo , Arabidopsis/química , Arabidopsis/genética , Arabidopsis/metabolismo , Biofortificação , Transporte Biológico , Cádmio/análise , Cádmio/metabolismo , Manganês/análise , Oryza/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Triticum/química , Triticum/metabolismo , Zinco/análise
5.
Plant Cell Physiol ; 60(8): 1633-1645, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31292642

RESUMO

Plants respond to a rise in ambient temperature by increasing the growth of petioles and hypocotyls. In this work, we show that Arabidopsis thaliana class I TEOSINTE BRANCHED 1, CYCLOIDEA, PCF (TCP) transcription factors TCP14 and TCP15 are required for optimal petiole and hypocotyl elongation under high ambient temperature. These TCPs influence the levels of the DELLA protein RGA and the expression of growth-related genes, which are induced in response to an increase in temperature. However, the class I TCPs are not required for the induction of the auxin biosynthesis gene YUCCA8 or for auxin-dependent gene expression responses. TCP15 directly targets the gibberellin biosynthesis gene GA20ox1 and the growth regulatory genes HBI1 and PRE6. Several of the genes regulated by TCP15 are also targets of the growth regulator PIF4 and show an enrichment of PIF4- and TCP-binding motifs in their promoters. PIF4 binding to GA20ox1 and HBI1 is enhanced in the presence of the TCPs, indicating that TCP14 and TCP15 directly participate in the induction of genes involved in gibberellin biosynthesis and cell expansion by high temperature functionally interacting with PIF4. In addition, overexpression of HBI1 rescues the growth defects of tcp14 tcp15 double mutants, suggesting that this gene is a major outcome of regulation by both class I TCPs during thermomorphogenesis.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Giberelinas/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Regulação da Expressão Gênica de Plantas , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Complexo de Endopeptidases do Proteassoma/genética , Temperatura Ambiente , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
6.
Plant Sci ; 286: 1-6, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31300135

RESUMO

The excessive and harmful light energy absorbed by the photosystem (PS) II of higher plants is dissipated as heat through a protective mechanism termed non-photochemical quenching (NPQ) of chlorophyll fluorescence. PsbS-knock-out (KO) mutants lack the trans-thylakoid proton gradient (ΔpH)-dependent part of NPQ. To elucidate the molecular mechanism of NPQ, we investigated its dependency on oxygen. The development of NPQ in wild-type (WT) rice under low-oxygen (LO) conditions was reduced to more than 50% of its original value. However, under high-oxygen (HO) conditions, the NPQ of both WT and PsbS-KO mutants recovered. Moreover, WT and PsbS-KO mutant leaves infiltrated with the ΔpH dissipating uncoupler nigericin showed increased NPQ values under HO conditions. The experiments using intact chloroplasts and protoplasts of Arabidopsis thaliana supported that the LO effects observed in rice leaves were not due to carbon dioxide deficiency. There was a noticeable 90% reduction in the half-time of P700 oxidation rate in LO-treated leaves compared with that of WT control leaves, but the HO treatment did not significantly change the half-time of P700 oxidation rate. Overall, the results obtained here indicate that the stroma of the PsbS-KO plants could be potentially under O2 deficiency. Because the functions of PsbS in rice leaves are likely to be similar to those in other higher plants, our findings offer novel insights into the role of oxygen in the development of NPQ.


Assuntos
Adaptação Fisiológica/efeitos da radiação , Arabidopsis/metabolismo , Oryza/metabolismo , Oxigênio/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Arabidopsis/efeitos da radiação , Cloroplastos/metabolismo , Cloroplastos/efeitos da radiação , Oryza/efeitos da radiação , Complexo de Proteína do Fotossistema II/genética , Folhas de Planta/genética , Folhas de Planta/fisiologia , Folhas de Planta/efeitos da radiação , Protoplastos/metabolismo , Protoplastos/efeitos da radiação
7.
Plant Sci ; 286: 28-36, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31300139

RESUMO

MYB family genes act as important regulators modulating the response to abiotic stress in plants. However, much less is known about MYB proteins in cotton. Here, we found that a cotton MYB gene, GhMYB73, was induced by NaCl and abscisic acid (ABA). Silencing GhMYB73 expression in cotton increased sensitivity to salt stress. The cotyledon greening rate of Arabidopsis thaliana over-expressing GhMYB73 under NaCl or mannitol treatment was significantly enhanced during the seedling germination stage. What's more, several osmotic stress-induced genes, such as AtNHX1, AtSOS3 and AtP5CS1, were more highly induced in the over-expression lines than in wild type under salt treatment, supporting the hypothesis that GhMYB73 contributes to salinity tolerance by improving osmotic stress resistance. Arabidopsis lines over-expressing GhMYB73 had superior germination and cotyledon greening under ABA treatment, and some abiotic stress-induced genes involved in ABA pathways (AtPYL8, AtABF3, AtRD29B and AtABI5), had increased transcription levels under salt-stress conditions in these lines. Furthermore, we found that GhMYB73 physically interacts with GhPYL8 and AtPYL8, suggesting that GhMYB73 regulates ABA signaling during salinity stress response. Taken together, over-expression of GhMYB73 significantly increases tolerance to salt and ABA stress, indicating that it can potentially be used in transgenic technology approaches to improve cotton salt tolerance.


Assuntos
Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Gossypium/fisiologia , Proteínas de Plantas/genética , Estresse Salino/genética , Fatores de Transcrição/genética , Arabidopsis/genética , Inativação Gênica , Genes myb , Gossypium/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Tolerância ao Sal/genética , Fatores de Transcrição/metabolismo
8.
Plant Sci ; 286: 78-88, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31300145

RESUMO

Chloroplastic Cpn60 proteins are type I chaperonins comprising of Cpn60α and Cpn60ß subunits. Arabidopsis genome contains six entries in Cpn60 family, out of which two are for Cpn60α subunit and four for Cpn60ß subunit. We noted that the cpn60ß4 knockout mutant plants (T-DNA insertion salk_064887 line) differed from the wild type Col-0 plants in the developmental programming. cpn60ß4 mutant plants showed early seed germination. Radical emergence, hypocotyl emergence and cotyledons opening were faster in cpn60ß4 mutant plants than WT. Importantly, cpn60ß4 mutant plants showed early-flowering phenotype. The number of flowers and siliques as well as weight of the seeds were higher in cpn60ß4 mutant plants as compared to Col-0 plants. These effects were reverted to wild type like growth and developmental patterns when genomic fragment of Arabidopsis encompassing Cpn60ß4 gene was complemented in the mutant background. The overexpression of Cpn60ß4 gene using CaMV35 promoter in wild type background (OE-Cpn60ß4) delayed the floral transition as against wild type plants. The plastid division were affected in cpn60ß4 mutant plants compared to Col-0. The results of this study suggest that Cpn60ß4 plays important role(s) in chloroplast development and is a key factor in plant growth, development and flowering in Arabidopsis.


Assuntos
Arabidopsis/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Cloroplastos/metabolismo , Flores/genética , Regulação da Expressão Gênica no Desenvolvimento , Reprodução
9.
Nat Commun ; 10(1): 2904, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31266952

RESUMO

Plant survival necessitates constant monitoring of fluctuating light and balancing growth demands with adaptive responses, tasks mediated via interconnected sensing and signaling networks. Photoreceptor phytochrome B (phyB) and plastidial retrograde signaling metabolite methylerythritol cyclodiphosphate (MEcPP) are evolutionarily conserved sensing and signaling components eliciting responses through unknown connection(s). Here, via a suppressor screen, we identify two phyB mutant alleles that revert the dwarf and high salicylic acid phenotypes of the high MEcPP containing mutant ceh1. Biochemical analyses show high phyB protein levels in MEcPP-accumulating plants resulting from reduced expression of phyB antagonists and decreased auxin levels. We show that auxin treatment negatively regulates phyB abundance. Additional studies identify CAMTA3, a MEcPP-activated calcium-dependent transcriptional regulator, as critical for maintaining phyB abundance. These studies provide insights into biological organization fundamentals whereby a signal from a single plastidial metabolite is transduced into an ensemble of regulatory networks controlling the abundance of phyB, positioning plastids at the information apex directing adaptive responses.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Fitocromo B/metabolismo , Plastídeos/metabolismo , Arabidopsis/genética , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Eritritol/análogos & derivados , Eritritol/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Ácidos Indolacéticos/metabolismo , Luz , Fitocromo B/genética , Plastídeos/genética , Transdução de Sinais/efeitos da radiação , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
10.
BMC Plant Biol ; 19(1): 320, 2019 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-31319813

RESUMO

BACKGROUND: Plant cell walls participate in all plant-environment interactions. Maintaining cell wall integrity (CWI) during these interactions is essential. This realization led to increased interest in CWI and resulted in knowledge regarding early perception and signalling mechanisms active during CWI maintenance. By contrast, knowledge regarding processes mediating changes in cell wall metabolism upon CWI impairment is very limited. RESULTS: To identify genes involved and to investigate their contributions to the processes we selected 23 genes with altered expression in response to CWI impairment and characterized the impact of T-DNA insertions in these genes on cell wall composition using Fourier-Transform Infrared Spectroscopy (FTIR) in Arabidopsis thaliana seedlings. Insertions in 14 genes led to cell wall phenotypes detectable by FTIR. A detailed analysis of four genes found that their altered expression upon CWI impairment is dependent on THE1 activity, a key component of CWI maintenance. Phenotypic characterizations of insertion lines suggest that the four genes are required for particular aspects of CWI maintenance, cell wall composition or resistance to Plectosphaerella cucumerina infection in adult plants. CONCLUSION: Taken together, the results implicate the genes in responses to CWI impairment, cell wall metabolism and/or pathogen defence, thus identifying new molecular components and processes relevant for CWI maintenance.


Assuntos
Arabidopsis/genética , Parede Celular/metabolismo , Genes de Plantas/fisiologia , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Ascomicetos , Parede Celular/fisiologia , Resistência à Doença/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Técnicas de Silenciamento de Genes , Interações Hospedeiro-Patógeno , Doenças das Plantas/imunologia , Plântula/metabolismo , Plântula/fisiologia , Espectroscopia de Infravermelho com Transformada de Fourier
11.
J Photochem Photobiol B ; 197: 111535, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31319267

RESUMO

Measurement of Pulse-Amplitude-Modulated (PAM) chlorophyll a fluorescence is widely used method for obtaining information on the functional state of photosystem II (PSII). Recently, it has been shown that some of long-established fluorescence parameters must be interpreted with caution, when the light-induced chloroplast movements occur. In our work we have analyzed the effect of chloroplast movements on these parameters. We have derived new parameters that are independent of the change in PSII absorption occurring during measurement. To verify whether there is a need for new parameters or the difference between the parameters commonly used and the newly derived ones is insignificant, we conducted an experiment with Arabidopsis thaliana wild type plants and its phot1 phot2 mutant defective in chloroplast movement. Plants were exposed to light of different qualities (450, 470, 550 or 660 nm) and quantities (100, 400 or 1200 µmol m-2 s-1) for up to 40 min. Since the blue light-induced chloroplast avoidance reaction is a photoprotective mechanism, we expected that phot1 phot2 mutant will compensate the lack of this mechanism by increasing non-photochemical quenching. However, using the light at both 450 and 470 nm, the calculation of commonly used parameter, ΦNPQ (quantum yield of regulated light-induced thermal energy dissipation in PSII) based on Hendrickson et al. [L. Hendrickson, R.T. Furbank, W.S. Chow, Photosynth. Res. 82 (2004) 73-81] showed the opposite. On the other hand, the results obtained using our newly proposed formulae to determine quantum yield of PSII thermal energy dissipation were in line with our assumption. Thus, the experimental data showed that some formulae of fluorescence parameters are dependent on the change in PSII absorption and need to be interpreted carefully. On the contrary, the formulae introduced by us can remove the effect of changes in PSII absorption that occur during measurement, without additional measurements, and give the real estimate of light-induced non-photochemical quenching.


Assuntos
Proteínas de Arabidopsis/metabolismo , Clorofila A/química , Complexo de Proteína do Fotossistema II/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Cloroplastos/fisiologia , Luz , Modelos Teóricos , Mutagênese , Complexo de Proteína do Fotossistema II/química , Complexo de Proteína do Fotossistema II/genética , Folhas de Planta/química , Teoria Quântica , Termodinâmica
12.
BMC Plant Biol ; 19(1): 304, 2019 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-31291882

RESUMO

BACKGROUND: In flowering plants, proper seed development is achieved through the constant interplay of fertilization products, embryo and endosperm, and maternal tissues. Communication between these compartments is supposed to be tightly regulated at their interfaces. Here, we characterize the deposition pattern of an apoplastic lipid barrier between the maternal inner integument and fertilization products in Arabidopsis thaliana seeds. RESULTS: We demonstrate that an apoplastic lipid barrier is first deposited by the ovule inner integument and undergoes de novo cutin deposition following central cell fertilization and relief of the FERTILIZATION INDEPENDENT SEED Polycomb group repressive mechanism. In addition, we show that the WIP zinc-finger TRANSPARENT TESTA 1 and the MADS-Box TRANSPARENT TESTA 16 transcription factors act maternally to promote its deposition by regulating cuticle biosynthetic pathways. Finally, mutant analyses indicate that this apoplastic barrier allows correct embryo sliding along the seed coat. CONCLUSIONS: Our results revealed that the deposition of a cutin apoplastic barrier between seed maternal and zygotic tissues is part of the seed coat developmental program.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Lipídeos de Membrana/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Óvulo Vegetal/genética , Óvulo Vegetal/crescimento & desenvolvimento , Óvulo Vegetal/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo
13.
Molecules ; 24(13)2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31288414

RESUMO

Due to their sedentary lifestyle, plants are constantly exposed to different stress stimuli. Stress comes in variety of forms where factors like radiation, free radicals, "replication errors, polymerase slippage", and chemical mutagens result in genotoxic or cytotoxic damage. In order to face "the base oxidation or DNA replication stress", plants have developed many sophisticated mechanisms. One of them is the DNA mismatch repair (MMR) pathway. The main part of the MMR is the MutS homologue (MSH) protein family. The genome of Arabidopsis thaliana encodes at least seven homologues of the MSH family: AtMSH1, AtMSH2, AtMSH3, AtMSH4, AtMSH5, AtMSH6, and AtMSH7. Despite their importance, the functions of AtMSH homologs have not been investigated. In this work, bioinformatics tools were used to obtain a better understanding of MSH-mediated DNA repair mechanisms in Arabidopsis thaliana and to understand the additional biological roles of AtMSH family members. In silico analysis, including phylogeny tracking, prediction of 3D structure, interactome analysis, and docking site prediction, suggested interactions with proteins were important for physiological development of A. thaliana. The MSH homologs extensively interacted with both TIL1 and TIL2 (DNA polymerase epsilon catalytic subunit), proteins involved in cell fate determination during plant embryogenesis and involved in flowering time repression. Additionally, interactions with the RECQ protein family (helicase enzymes) and proteins of nucleotide excision repair pathway were detected. Taken together, the results presented here confirm the important role of AtMSH proteins in mismatch repair and suggest important new physiological roles.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Simulação de Acoplamento Molecular , Proteínas MutS/metabolismo , Proteínas de Arabidopsis/química , Sítios de Ligação , Simulação por Computador , Dano ao DNA , Reparo de Erro de Pareamento de DNA , Reparo do DNA , Replicação do DNA , Proteínas MutS/química , Conformação Proteica
14.
Plant Mol Biol ; 101(3): 257-268, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31302867

RESUMO

KEY MESSAGE: The C-terminal cysteine-rich motif of NYE1/SGR1 affects chlorophyll degradation likely by mediating its self-interaction and conformational change, and somehow altering its Mg-dechelating activity in response to the changing redox potential. During green organ senescence in plants, the most prominent phenomenon is the degreening caused by net chlorophyll (Chl) loss. NON-YELLOWING1/STAY-GREEN1 (NYE1/SGR1) was recently reported to be able to dechelates magnesium (Mg) from Chl a to initiate its degradation, but little is known about the domain/motif basis of its functionality. In this study, we carried out a protein truncation assay and identified a conserved cysteine-rich motif (CRM, P-X3-C-X3-C-X-C2-F-P-X5-P) at its C terminus, which is essential for its function. Genetic analysis showed that all four cysteines in the CRM were irreplaceable, and enzymatic assays demonstrated that the mutation of each of the four cysteines affected its Mg-dechelating activity. The CRM plays a critical role in the conformational change and self-interaction of NYE1 via the formation of inter- and intra-molecular disulfide bonds. Our results may provide insight into how NYE1 responds to rapid redox changes during leaf senescence and in response to various environmental stresses.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Clorofila/química , Proteínas de Cloroplastos/metabolismo , Motivos de Aminoácidos , Quelantes/química , DNA Complementar/metabolismo , Dissulfetos , Regulação da Expressão Gênica de Plantas , Magnésio/química , Oxirredução , Fenótipo , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Conformação Proteica , Domínios Proteicos , Estresse Fisiológico
15.
World J Microbiol Biotechnol ; 35(6): 90, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-31147784

RESUMO

The ability of plant growth promoting rhizobacteria (PGPR) for imparting abiotic stress tolerance to plants has been widely explored in recent years; however, the diversity and potential of these microbes have not been maximally exploited. In this study, we characterized four bacterial strains, namely, Pseudomonas aeruginosa PM389, Pseudomonas aeruginosa ZNP1, Bacillus endophyticus J13 and Bacillus tequilensis J12, for potential plant growth promoting (PGP) traits under osmotic-stress, induced by 25% polyethylene glycol (PEG) in the growth medium. Growth curve analysis was performed in LB medium with or without PEG, in order to understand the growth patterns of these bacteria under osmotic-stress. All strains were able to grow and proliferate under osmotic-stress, although their growth rate was slower than that under non-stressed conditions (LB without PEG). Bacterial secretions were analyzed for the presence of exopolysaccharides and phytohormones and it was observed that all four strains released these compounds into the media, both, under stressed and non-stressed conditions. In the Pseudomonas strains, osmotic stress caused a decrease in the levels of auxin (IAA) and cytokinin (tZ), but an increase in the levels of gibberellic acid. The Bacillus strains on the other hand showed a stress-induced increase in the levels of all three phytohormones. P. aeruginosa ZNP1 and B. endophyticus J13 exhibited increased EPS production under osmotic-stress. While osmotic stress caused a decrease in the levels of EPS in P. aeruginosa PM389, B. tequilensis J12 showed no change in EPS quantities released into the media under osmotic stress when compared to non-stressed conditions. Upon inoculating Arabidopsis thaliana seedlings with these strains individually, it was observed that all four strains were able to ameliorate the adverse effects of osmotic-stress (induced by 25% PEG in MS-Agar medium) in the plants, as evidenced by their enhanced fresh weight, dry weight and plant water content, as opposed to osmotic-stressed, non-inoculated plants.


Assuntos
Arabidopsis/microbiologia , Fenômenos Fisiológicos Bacterianos , Secas , Pressão Osmótica , Desenvolvimento Vegetal , Reguladores de Crescimento de Planta/metabolismo , Polissacarídeos Bacterianos/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Bacillus/crescimento & desenvolvimento , Bacillus/fisiologia , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Citocininas/metabolismo , Giberelinas/metabolismo , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/microbiologia , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/fisiologia , Rizosfera , Plântula/crescimento & desenvolvimento , Microbiologia do Solo , Estresse Fisiológico/fisiologia
16.
Gene ; 710: 210-217, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31176733

RESUMO

Low temperature is a key stress factor for the growth and development of wheat (Triticum aestivum L.), and glycometabolism plays an important role in plant cold tolerance. Our previous study identified trehalose 6-phosphate synthase 11 gene (TaTPS11), which had a significantly different expression pattern between a high freezing-tolerant wheat cultivar and a low freezing-tolerant wheat cultivar. In this study, TaTPS11 was isolated from a winter-hardy wheat cultivar (D1) and overexpressed in Arabidopsis thaliana to study its effect on cold tolerance in plants. Transgenic plants expressing TaTPS11 had lower sucrose content, higher starch content, and higher activity of key enzyme (sucrose phosphate synthase, sucrose synthase, and invertase) involved in sucrose metabolism. In addition, the expression level of sucrose non-fermenting 1-related kinase 1 (SnRK1), which catalyzes the sucrose in plants, increased in the TaTPS11-overexpressed plants. These results indicated that heterologous expression of TaTPS11 influenced carbohydrate metabolism in Arabidopsis plants. The resultant plants had a significantly higher survival rate after -5 °C treatment for 2 h and exhibited enhanced cold tolerance without unfavorable phenotypes compared to wild-type. Our findings indicated that manipulation of TaTPS11 improved cold tolerance in plants and TaTPS11 had potential values in wheat cold-tolerance breeding.


Assuntos
Arabidopsis/genética , Resposta ao Choque Frio , Monoéster Fosfórico Hidrolases/genética , Triticum/enzimologia , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Metabolismo dos Carboidratos , Regulação da Expressão Gênica de Plantas , Monoéster Fosfórico Hidrolases/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Proteínas Serina-Treonina Quinases/genética , Reação em Cadeia da Polimerase em Tempo Real , Sacarose/metabolismo , Triticum/genética
17.
BMC Plant Biol ; 19(1): 241, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31170927

RESUMO

BACKGROUND: Plant chloroplasts and mitochondria utilize nuclear encoded proteins to replicate their DNA. These proteins are purposely built for replication in the organelle environment and are distinct from those involved in replication of the nuclear genome. These organelle-localized proteins have ancestral roots in bacterial and bacteriophage genes, supporting the endosymbiotic theory of their origin. We examined the interactions between three of these proteins from Arabidopsis thaliana: a DNA helicase-primase similar to bacteriophage T7 gp4 protein and animal mitochondrial Twinkle, and two DNA polymerases, Pol1A and Pol1B. We used a three-pronged approach to analyze the interactions, including Yeast-two-hybrid analysis, Direct Coupling Analysis (DCA), and thermophoresis. RESULTS: Yeast-two-hybrid analysis reveals residues 120-295 of Twinkle as the minimal region that can still interact with Pol1A or Pol1B. This region is a part of the primase domain of the protein and slightly overlaps the zinc-finger and RNA polymerase subdomains located within. Additionally, we observed that Arabidopsis Twinkle interacts much more strongly with Pol1A versus Pol1B. Thermophoresis also confirms that the primase domain of Twinkle has higher binding affinity than any other region of the protein. Direct-Coupling-Analysis identified specific residues in Twinkle and the DNA polymerases critical to positive interaction between the two proteins. CONCLUSIONS: The interaction of Twinkle with Pol1A or Pol1B mimics the minimal DNA replisomes of T7 phage and those present in mammalian mitochondria. However, while T7 and mammals absolutely require their homolog of Twinkle DNA helicase-primase, Arabidopsis Twinkle mutants are seemingly unaffected by this loss. This implies that while Arabidopsis mitochondria mimic minimal replisomes from T7 and mammalian mitochondria, there is an extra level of redundancy specific to loss of Twinkle function.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Bacteriófago T7/genética , DNA Polimerase Dirigida por DNA/genética , Complexos Multienzimáticos/genética , Enzimas Multifuncionais/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , DNA Polimerase Dirigida por DNA/metabolismo , Mitocôndrias/metabolismo , Enzimas Multifuncionais/metabolismo
18.
J Agric Food Chem ; 67(24): 6736-6747, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31184154

RESUMO

Nitrogen is essential for plant growth and crop productivity; however, nitrogen use efficiency (NUE) decreases with increasing N supply, resulting in a waste of resources. Molecular mechanisms underlying low-nitrogen (LN)-mediated enhancement of NUE are not clear. We used high-NUE Brassica napus genotype H (Xiangyou 15), low-NUE B. napus genotype L (814), and Arabidopsis mutant aux1 to elucidate the mechanism underlying the changes in NUE under different rates of N fertilizer application. NUE of B. napus increased under LN, which enhanced N uptake ability by regulating root system architecture and plasma membrane H+-ATPase activity; AUX1 was involved in this process. Additionally, BnNRT1.5 was upregulated and BnNRT1.8 was downregulated under LN, whereby more N was transferred to the shoot through enhanced N transport. Observed changes in photosynthesis under LN were associated with N assimilation efficiency. Our study provides new insights into the mechanisms of plant adaptation to the environment.


Assuntos
Arabidopsis/metabolismo , Brassica napus/metabolismo , Nitratos/metabolismo , Nitrogênio/metabolismo , Arabidopsis/genética , Transporte Biológico , Brassica napus/genética , Fertilizantes/análise , Regulação da Expressão Gênica de Plantas , Nitrogênio/análise , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo
19.
Plant Sci ; 285: 1-13, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31203874

RESUMO

Bioactive gibberellins (GAs) play multiple roles in plant development and stress responses. GA2-oxidases (GA2oxs) are a class of 2-oxoglutarate-dependent dioxygenases that regulate the deactivation of bioactive GAs. In this study, we investigated the phylogeny and domain structures of the seven GA2ox genes present in the Arabidopsis thaliana genome. Comprehensive expression analysis using translational reporter lines showed that the seven GA2ox genes are differentially expressed during Arabidopsis growth and development: GA2ox1 is specifically expressed in the hypocotyl and lateral root primordium; GA2ox2 is highly expressed in aboveground tissues; GA2ox3 is expressed in the chalazal endosperm of the early embryo sac and inflorescences; GA2ox4 is expressed in the shoot apical meristem and during lateral root initiation; GA2ox6 is expressed in the maturation zone, but not in the meristem or elongating zone of the root; GA2ox7 is constitutively expressed during almost all developmental stages; and GA2ox8 is exclusively expressed in stomatal cells. Overexpression of each of these GA2ox genes inhibited high temperature-induced hypocotyl elongation in both wild-type and elongated hypocotyl 5 plants, which have an elongated hypocotyl phenotype, suggesting that these genes negatively regulate hypocotyl elongation by reducing bioactive GA levels. This study provides a valuable resource for further elucidating the roles of GA2ox genes during different stages of development.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Genes de Plantas/fisiologia , Giberelinas/metabolismo , Oxirredutases/genética , Arabidopsis/enzimologia , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Giberelinas/fisiologia , Hipocótilo/crescimento & desenvolvimento , Hipocótilo/metabolismo , Oxirredutases/metabolismo , Oxirredutases/fisiologia , Filogenia , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Transcriptoma
20.
Plant Sci ; 285: 34-43, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31203892

RESUMO

Seed germination is a critical stage during the initiation of the plant lifecycle and is strongly affected by endogenous phytohormones and environmental stress. High temperature (HT) upregulates endogenous abscisic acid (ABA) to suppress seed germination, and ABA-INSENSITIVE 5 (ABI5) is the key positive regulator in the ABA signal-mediated modulation of seed germination. In plants, hydrogen sulfide (H2S) is a small gas messenger that participates in multiple physiological processes, but its role in seed germination thermotolerance has not been thoroughly elucidated to date. In this study, we found that H2S enhanced the seed germination rate under HT. Moreover, HT accelerates the efflux of the E3 ligase CONSTITUTIVE PHOTOMORPHOGENESIS 1 (COP1) from the nucleus to the cytoplasm, which results in increased nuclear accumulation of ELONG HYPCOTYL 5 (HY5) to activate the expression of ABI5 and thereby suppress seed germination. However, the H2S signal reversed the HT effect, as characterized by increased COP1 in the nucleus, which resulted in increased degradation of HY5 and reduced expression of ABI5 and thereby enhanced the seed germination thermotolerance. Thus, our findings reveal a novel role for the H2S signal in the modulation of seed germination thermotolerance through the nucleocytoplasmic partitioning of COP1 and the downstream HY5 and ABI5 pathways.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Germinação/fisiologia , Sulfeto de Hidrogênio/metabolismo , Proteínas Nucleares/metabolismo , Sementes/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ácido Abscísico/metabolismo , Ácido Abscísico/fisiologia , Arabidopsis/fisiologia , Proteínas de Arabidopsis/fisiologia , Fatores de Transcrição de Zíper de Leucina Básica/fisiologia , Núcleo Celular/enzimologia , Núcleo Celular/metabolismo , Giberelinas/metabolismo , Giberelinas/fisiologia , Temperatura Alta , Proteínas Nucleares/fisiologia , Reguladores de Crescimento de Planta/fisiologia , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase em Tempo Real , Sementes/fisiologia , Transdução de Sinais/fisiologia , Termotolerância , Ubiquitina-Proteína Ligases/fisiologia
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