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1.
Plant Sci ; 313: 111085, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34763870

RESUMO

Soybean, a glycophyte that is sensitive to salt stress, is greatly affected by salinity at all growth stages. A mapping population derived from a cross between a salt-sensitive Korean cultivar, Cheongja 3, and a salt-tolerant landrace, IT162669, was used to identify quantitative trait loci (QTLs) conferring salt tolerance in soybean. Following treatment with 120 mM NaCl for 2 weeks, phenotypic traits representing physiological damage, leaf Na+ content, and K+/Na+ ratio were characterized. Among the QTLs mapped on a high-density genetic map harboring 2,630 single nucleotide polymorphism markers, we found two novel major loci, qST6, on chromosome 6, and qST10, on chromosome 10, which controlled traits related to ion toxicity and physiology in response to salinity, respectively. These loci were distinct from the previously known salt tolerance allele on chromosome 3. Other QTLs associated with abiotic stress overlapped with the genomic regions of qST6 and qST10, or with their paralogous regions. Based on the functional annotation and parental expression differences, we identified eight putative candidate genes, two in qST6 and six in qST10, which included a phosphoenolpyruvate carboxylase and an ethylene response factor. This study provides additional genetic resources to breed soybean cultivars with enhanced salt tolerance.


Assuntos
Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Locos de Características Quantitativas/genética , Tolerância ao Sal/genética , Tolerância ao Sal/fisiologia , Soja/genética , Soja/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/fisiologia , Variação Genética , Genótipo , Fenótipo
2.
Plant Sci ; 313: 111092, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34763876

RESUMO

WD40 transcription factors are an ancient protein family whose members play important roles in plant growth and stress resistance. In this study, a new WD40 gene was cloned from Ginkgo biloba L. via the rapid amplification of cDNA ends (RACE) technique. This gene was 824 bp in length and encoded 109 amino acids. Sequence alignment and phylogenetic analysis showed that this transcription factor was most similar to the LWD1 protein, and it was thus named GbLWD1-like. This gene was expressed mainly in the leaves, followed by the roots. Phenotypic analysis showed that the transgenic plants grew better, were taller, and had significantly more roots than the control check (CK) plants. Moreover, the transgenic plants were more tolerant to salt stress than the CK plants. After 11 days of salt treatment, all the leaves of the CK plants had dried up and fallen off, whereas in the transgenic lines, only the edges of the bottom leaves had turned yellow. Under salt stress, the expression levels of some genes related to salt tolerance were higher in the transgenic plants than in the CK plants. This study suggests that the GbLWD1-like gene may be related to the growth potential and improved salt tolerance of plants and may play an important role in the response to adversity.


Assuntos
Ginkgo biloba/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Populus/genética , Populus/fisiologia , Tolerância ao Sal/genética , Tolerância ao Sal/fisiologia , China , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Melhoramento Vegetal/métodos , Estresse Salino , Fatores de Transcrição
3.
Int J Mol Sci ; 22(19)2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34639173

RESUMO

C2H2 zinc finger proteins (ZFPs) play important roles in plant development and response to abiotic stresses, and have been studied extensively. However, there are few studies on ZFPs in mangroves and mangrove associates, which represent a unique plant community with robust stress tolerance. MpZFP1, which is highly induced by salt stress in the mangrove associate Millettia pinnata, was cloned and functionally characterized in this study. MpZFP1 protein contains two zinc finger domains with conserved QALGGH motifs and targets to the nucleus. The heterologous expression of MpZFP1 in Arabidopsis increased the seeds' germination rate, seedling survival rate, and biomass accumulation under salt stress. The transgenic plants also increased the expression of stress-responsive genes, including RD22 and RD29A, and reduced the accumulation of reactive oxygen species (ROS). These results indicate that MpZFP1 is a positive regulator of plant responses to salt stress due to its activation of gene expression and efficient scavenging of ROS.


Assuntos
Arabidopsis/fisiologia , Dedos de Zinco CYS2-HIS2 , Regulação da Expressão Gênica de Plantas , Millettia/fisiologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Tolerância ao Sal , Arabidopsis/genética , Arabidopsis/metabolismo , Secas , Millettia/genética , Millettia/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico
4.
Int J Mol Sci ; 22(19)2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34639192

RESUMO

As with other environmental stresses, cold stress limits plant growth, geographical distribution, and agricultural productivity. CBF/DREB (CRT-binding factors/DRE-binding proteins) regulate tolerance to cold/freezing stress across plant species. ICE (inducer of CBF expression) is regarded as the upstream inducer of CBF expression and plays a crucial role as a main regulator of cold acclimation. Snow lotus (Saussurea involucrata) is a well-known traditional Chinese herb. This herb is known to have greater tolerance to cold/freezing stress compared to other plants. According to transcriptome datasets, two putative ICE homologous genes, SiICE1 and SiICE2, were identified in snow lotus. The predicted SiICE1 cDNA contains an ORF of 1506 bp, encoding a protein of 501 amino acids, whereas SiICE2 cDNA has an ORF of 1482 bp, coding for a protein of 493 amino acids. Sequence alignment and structure analysis show SiICE1 and SiICE2 possess a S-rich motif at the N-terminal region, while the conserved ZIP-bHLH domain and ACT domain are at the C-terminus. Both SiICE1 and SiICE2 transcripts were cold-inducible. Subcellular localization and yeast one-hybrid assays revealed that SiICE1 and SiICE2 are transcriptional regulators. Overexpression of SiICE1 (35S::SiICE1) and SiICE2 (35S::SiICE2) in transgenic Arabidopsis increased the cold tolerance. In addition, the expression patterns of downstream stress-related genes, CBF1, CBF2, CBF3, COR15A, COR47, and KIN1, were up-regulated when compared to the wild type. These results thus provide evidence that SiICE1 and SiICE2 function in cold acclimation and this cold/freezing tolerance may be regulated through a CBF-controlling pathway.


Assuntos
Arabidopsis/fisiologia , Resposta ao Choque Frio , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Saussurea/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Saussurea/genética , Saussurea/metabolismo , Fatores de Transcrição/genética , Ativação Transcricional
5.
Plant Sci ; 312: 111034, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34620438

RESUMO

MicroRNAs (miRNAs) are small, non-coding regulatory RNAs that regulate gene expression by facilitating target mRNA cleavage in plants. They are crucial for responses to diverse stresses. The novel drought-responsive miRNA ZmmiR190 was previously identified during an analysis of the maize transcriptome. In this study, we revealed that transgenic Arabidopsis thaliana overexpressing ZmmiR190 is more sensitive to drought than the wild-type control. The transcript of a nuclear-localized gene, ZmCRP04, was identified as a likely target of ZmmiR190. Moreover, ZmmiR190 and ZmCRP04 had the opposite expression profiles following drought and salt treatments. Additionally, 5' RACE and coexpression analyses in A. thaliana provided evidence of the in vivo targeting of the ZmCRP04 transcript by ZmmiR190. Furthermore, the overexpression of ZmCRP04 in A. thaliana and rice significantly enhanced drought tolerance, with lower malonaldehyde contents and relative electrolyte leakage in the transgenic A. thaliana and rice plants than in the wild-type control. Transgenic plants overexpressing ZmmiR190 or ZmCRP04 were hypersensitive to abscisic acid. These results suggest that the ZmCRP04 transcript is targeted by ZmmiR190 and may encode a protein that positively regulates drought stress tolerance via an abscisic acid-dependent pathway. These findings may be relevant for future molecular breeding aimed at improving crop drought tolerance.


Assuntos
Ácido Abscísico/metabolismo , Arabidopsis/genética , Secas , MicroRNAs/genética , MicroRNAs/metabolismo , Oryza/genética , Estresse Fisiológico/genética , Zea mays/genética , Arabidopsis/metabolismo , Produtos Agrícolas/genética , Produtos Agrícolas/fisiologia , Regulação da Expressão Gênica de Plantas , Variação Genética , Genótipo , Oryza/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Zea mays/metabolismo
6.
Int J Mol Sci ; 22(19)2021 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-34638606

RESUMO

Drought has become a major threat to food security, because it affects crop growth and development. Drought tolerance is an important quantitative trait, which is regulated by hundreds of genes in crop plants. In recent decades, scientists have made considerable progress to uncover the genetic and molecular mechanisms of drought tolerance, especially in model plants. This review summarizes the evaluation criteria for drought tolerance, methods for gene mining, characterization of genes related to drought tolerance, and explores the approaches to enhance crop drought tolerance. Collectively, this review illustrates the application prospect of these genes in improving the drought tolerance breeding of crop plants.


Assuntos
Produtos Agrícolas/genética , Produtos Agrícolas/fisiologia , Secas , Genes de Plantas , Melhoramento Vegetal/métodos , Aclimatação/genética , Aclimatação/fisiologia , Fenótipo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Locos de Características Quantitativas
7.
Plant Sci ; 311: 111016, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34482919

RESUMO

The yields of cereal crops grown on acidic soils are often reduced by aluminum (Al) toxicity because the prevalence of toxic Al3+ cations increases as pH falls below 5.0. The Al-dependent release of citrate from resistant lines of maize is controlled by ZmMATE1 which encodes a multidrug and toxic compound extrusion (MATE) transporter protein. ZmMATE6 is another member of this family in maize whose expression is also increased by Al treatment. We investigated the function of this gene in more detail to determine whether it also contributes to Al resistance. Quantitative RT-PCR measurements found that ZmMATE6 was expressed in the roots and leaves of Al-resistant and sensitive inbred lines. Treatment with Al induced ZmMATE6 expression in all tissues but several other divalent or trivalent cations tested had no effect on expression. This expression pattern and the induction by Al treatment was confirmed in ZmMATE6 promoter-ß-glucuronidase fusion lines. Heterogeneous expression of ZmMATE6 displayed a greater Al-activated release of citrate from the roots and was significantly resistant to Al toxicity than controls. This was associated with reduced accumulation of Al in the root tissues. Our results demonstrated that ZmMATE6 expression is induced by Al and functions as a citrate transporter.


Assuntos
Adaptação Fisiológica/genética , Adaptação Fisiológica/fisiologia , Alumínio/efeitos adversos , Arabidopsis/genética , Proteínas de Transporte/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Zea mays/genética , Arabidopsis/fisiologia , Proteínas de Transporte/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/fisiologia , Genes de Plantas , Variação Genética , Genótipo , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Análise de Sequência de Proteína , Zea mays/fisiologia
8.
Plant Sci ; 311: 111021, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34482922

RESUMO

HD-Zip is a plant-specific HB transcription factor, which participates in plant development and stress response. In this study, we identified 63 poplar HD-Zip transcription factors, which were randomly distributed on 19 chromosomes of poplar. Based on the gene structure and phylogenetic relationship, these members are divided into four groups, which have a variety of collinear evolutionary relationships. They also have rich segmental replication events and experienced strong purification selection. Based on RNA-seq analysis, we profiled the expression pattern of the 63 HD-Zip members under salt stress. Subsequently, we carried out in-depth study on the significantly up-regulated PsnHDZ63 in the stems and leaves. The transgenic Populus simonii × P.nigra plants over-expressing PsnHDZ63 displayed better morphological and physiological indexes than WT under salt stress. In addition, PsnHDZ63 enhanced salt stress tolerance of transgenic lines by combining effective stress-resistant elements to improve reactive oxygen species scavenging ability. These studies laid a foundation for a comprehensive understanding of poplar HD-Zip family members, and revealed the important role of PsnHDZ63 in plant salt tolerance.


Assuntos
Arabidopsis/genética , Plantas Geneticamente Modificadas/fisiologia , Populus/crescimento & desenvolvimento , Populus/genética , Tolerância ao Sal/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Estudo de Associação Genômica Ampla , Genótipo , Análise de Sequência de Proteína
9.
Int J Mol Sci ; 22(18)2021 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-34576101

RESUMO

Near-infrared spectroscopy (NIRS) has become a more popular approach for quantitative and qualitative analysis of feeds, foods and medicine in conjunction with an arsenal of chemometric tools. This was the foundation for the increased importance of NIRS in other fields, like genetics and transgenic monitoring. A considerable number of studies have utilized NIRS for the effective identification and discrimination of plants and foods, especially for the identification of genetically modified crops. Few previous reviews have elaborated on the applications of NIRS in agriculture and food, but there is no comprehensive review that compares the use of NIRS in the detection of genetically modified organisms (GMOs). This is particularly important because, in comparison to previous technologies such as PCR and ELISA, NIRS offers several advantages, such as speed (eliminating time-consuming procedures), non-destructive/non-invasive analysis, and is inexpensive in terms of cost and maintenance. More importantly, this technique has the potential to measure multiple quality components in GMOs with reliable accuracy. In this review, we brief about the fundamentals and versatile applications of NIRS for the effective identification of GMOs in the agricultural and food systems.


Assuntos
Plantas Geneticamente Modificadas/fisiologia , Espectroscopia de Luz Próxima ao Infravermelho , Produtos Agrícolas/fisiologia , Alimentos
10.
Plant Physiol Biochem ; 167: 198-209, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34365290

RESUMO

Two pepper methionine sulfoxide reductase B2 (CaMsrB2) gene expressing transgenic rice lines (L-8 and L-23) were interrogated with respect to their physiological and photochemical attributes along with control (WT, Ilmi) as a standard against varying levels of salt concentration which are 75 mM, 150 mM and 225 mM. Against various levels of salt (NaCl) concentration, recurring detrimental effects of extreme salt stress was observed and more pronounced in the wild type plants as compared to our transgenic lines. As the exacerbated effects of salinity is responsible for pushing the plants to their ecological tolerance, our transgenic lines performed well uplifted in different realms of physiology and photochemistry such as relative water content (RWC = 60-75%), stomatal conductance (gs = 70-190 mmolm-2s-1), performance index (PIABS = 1.0-4.5), maximal photochemical yield of PSII (FV/FM = 0.48-0.72) and chlorophyll content index (CCI = 5-7.2 au) in comparison to the control. Relative gene expression, ion analysis and antioxidants activity were analyzed in all treatments to ensure the hypothesis obtained from data of physiology and photochemistry. Photosynthetic apparatus is known to lose energy in various forms such as NPQ, DIO/CS, damages of reaction center (FV/FO) which are the markers of poor health were clearly decreased in the L-23 line as compared to L-8 and WT. Present study revealed the protruding tolerance of L-23 and L-8 transgenic lines with L-23 line in the lead in comparison to control and L-8 transgenic lines.


Assuntos
Metionina Sulfóxido Redutases , Oryza , Capsicum/enzimologia , Clorofila , Ecossistema , Metionina Sulfóxido Redutases/genética , Oryza/genética , Oryza/fisiologia , Fotossíntese , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico
11.
Int J Biol Macromol ; 184: 967-980, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34197850

RESUMO

Soil salinization is a vital factor that restricts the efficient and sustainable development of global agriculture. Studies enlightened that the C2H2 zinc finger proteins (C2H2-ZFP) were involved in regulating the stress response in plants. However, knowledge of the C2H2-ZFP subfamily C1 (ZAT; Zinc finger of Arabidopsis thaliana) in cotton is still a mystery. In this study, 47, 45, 94, and 88 ZAT genes were obtained from diploid A2, D5 and tetraploid AD1, AD2 cotton genomes, respectively. The function of hybridization and allopolyploidy in the evolutionary linkage of allotetraploid cotton was explained by the family of ZAT gene in 4 species. Duplication of gene activities indicates that the family of ZAT gene of cotton evolution was under strong purifying selection. The integration of previous transcriptome data related to NaCl stress, strongly suggests the GhZAT34 and GhZAT79 may interact with salt resistance in upland cotton. The expression level of certain ZAT genes, higher seed germination rate of transgenic Arabidopsis and gene- silenced cotton revealed that both genes were involved in the salt tolerance of upland cotton. This study may pave the substantial understandings into the role of ZATs genes in plants as well as suggest appropriate candidate genes for breeding of cotton varieties against salinity tolerance.


Assuntos
Arabidopsis/fisiologia , Gossypium/fisiologia , Tolerância ao Sal , Fatores de Transcrição/genética , Arabidopsis/genética , Dedos de Zinco CYS2-HIS2 , Diploide , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Família Multigênica , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Seleção Genética , Tetraploidia , Fatores de Transcrição/química
12.
Int J Mol Sci ; 22(11)2021 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-34073070

RESUMO

Although recent studies suggest that the plant cytoskeleton is associated with plant stress responses, such as salt, cold, and drought, the molecular mechanism underlying microtubule function in plant salt stress response remains unclear. We performed a comparative proteomic analysis between control suspension-cultured cells (A0) and salt-adapted cells (A120) established from Arabidopsis root callus to investigate plant adaptation mechanisms to long-term salt stress. We identified 50 differentially expressed proteins (45 up- and 5 down-regulated proteins) in A120 cells compared with A0 cells. Gene ontology enrichment and protein network analyses indicated that differentially expressed proteins in A120 cells were strongly associated with cell structure-associated clusters, including cytoskeleton and cell wall biogenesis. Gene expression analysis revealed that expressions of cytoskeleton-related genes, such as FBA8, TUB3, TUB4, TUB7, TUB9, and ACT7, and a cell wall biogenesis-related gene, CCoAOMT1, were induced in salt-adapted A120 cells. Moreover, the loss-of-function mutant of Arabidopsis TUB9 gene, tub9, showed a hypersensitive phenotype to salt stress. Consistent overexpression of Arabidopsis TUB9 gene in rice transgenic plants enhanced tolerance to salt stress. Our results suggest that microtubules play crucial roles in plant adaptation and tolerance to salt stress. The modulation of microtubule-related gene expression can be an effective strategy for developing salt-tolerant crops.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis , Microtúbulos/fisiologia , Oryza , Tolerância ao Sal , Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Oryza/fisiologia , Plantas Geneticamente Modificadas/fisiologia
13.
Nat Plants ; 7(7): 914-922, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34155371

RESUMO

The activities of cold-responsive C-repeat-binding transcription factors (CBFs) are tightly controlled as they not only induce cold tolerance but also regulate normal plant growth under temperate conditions1-4. Thioredoxin h2 (Trx-h2)-a cytosolic redox protein identified as an interacting partner of CBF1-is normally anchored to cytoplasmic endomembranes through myristoylation at the second glycine residue5,6. However, after exposure to cold conditions, the demyristoylated Trx-h2 is translocated to the nucleus, where it reduces the oxidized (inactive) CBF oligomers and monomers. The reduced (active) monomers activate cold-regulated gene expression. Thus, in contrast to the Arabidopsis trx-h2 (AT5G39950) null mutant, Trx-h2 overexpression lines are highly cold tolerant. Our findings reveal the mechanism by which cold-mediated redox changes induce the structural switching and functional activation of CBFs, therefore conferring plant cold tolerance.


Assuntos
Arabidopsis/genética , Arabidopsis/fisiologia , Temperatura Baixa , Resposta ao Choque Frio/genética , Resposta ao Choque Frio/fisiologia , Oxirredução , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia
14.
Plant Signal Behav ; 16(8): 1913306, 2021 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-34134596

RESUMO

Abiotic stresses are significant environmental issues that restrict plant growth, productivity, and survival while also posing a threat to global food production and security. Plants produce compatible solutes known as osmolytes to adapt themselves in such changing environment. Osmolytes contribute to homeostasis maintenance, provide the driving gradient for water uptake, maintain cell turgor by osmotic adjustment, and redox metabolism to remove excess level of reactive oxygen species (ROS) and reestablish the cellular redox balance as well as protect cellular machinery from osmotic stress and oxidative damage. Perceiving the mechanisms how plants interpret environmental signals and transmit them to cellular machinery to activate adaptive responses is important for crop improvement programs to get stress-tolerant varieties. A large number of studies conducted in the last few decades have shown that osmolytes accumulate in plants and have strong associations with abiotic stress tolerance. Production of abundant osmolytes is needed for tolerance in many plant species. In addition, transgenic plants overexpressing genes for different osmolytes showed enhanced tolerance to various abiotic stresses. Many important aspects of their mechanisms of action are yet to be largely identified, especially regarding the relevance and relative contribution of specific osmolytes to the stress tolerance of a given species. Therefore, more efforts and resources should be invested in the study of the abiotic stress responses of plants in their natural habitats. The present review focuses on the possible roles and mechanisms of osmolytes and their association toward abiotic stress tolerance in plants. This review would help the readers in learning more about osmolytes and how they behave in changing environments as well as getting an idea of how this knowledge could be applied to develop stress tolerance in plants.


Assuntos
Aclimatação , Aminoácidos/biossíntese , Carboidratos/biossíntese , Pressão Osmótica , Plantas/metabolismo , Poliaminas/metabolismo , Estresse Fisiológico , Produtos Agrícolas/metabolismo , Produtos Agrícolas/fisiologia , Citoproteção , Secas , Osmorregulação , Osmose , Oxirredução , Estresse Oxidativo , Desenvolvimento Vegetal , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Salinidade , Álcoois Açúcares/metabolismo , Açúcares/metabolismo , Água
15.
Plant Sci ; 308: 110928, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34034876

RESUMO

Cell death is an inevitably cryo-injury in cell and tissue cryopreservation. The research on programmed cell death (PCD) in plant cryopreservation is still in its infancy. In this study, the survival rate of Agapanthus praecox embryogenic callus was significantly improved when the vitrification solution was added with 20 µM E-64, which is an inhibitor of cathepsin B. For further investigating the relation between cathepsin B and cryo-injury, the coding gene of cathepsin B, ApCathB was isolated and characterized. A subcellular localization assay showed that ApCathB was located in cytomembrane. Heterologous overexpression of ApCathB reduced the recovery rate during Arabidopsis seedlings cryopreservation from 29.56 % to 16.46 %. Transgenic seedlings lost most of cell viability in hypocotyl after dehydration and lead to aggravated cryo-injury. The reduced survival rate of ApCathB-overexpressing embryogenic callus of A. praecox further confirmed its negatively function in cryo-injury tolerance. In addition, the survival of ApCathB-overexpressing lines was almost rescued by E-64. TUNEL detection showed intensified signal and ROS was burst, especially for H2O2. Furthermore, VPE, Metacaspase 1, Cyp15a and AIF genes related to cell death regulation were remarkably up-regulated in ApCathB-overexpressing embryogenic callus during cryopreservation. Additionally, the expression level of genes regulating cell degradation was also elevated, indicating accelerated cell death caused by ApCathB-overexpressing. Taken together, this work verified that ApCathB negatively regulated the cryo-injury tolerance and cell viability through mediating the PCD event in plant cryopreservation. Significantly, cathepsin B has potential to be a target to improve survival rate after cryopreservation.


Assuntos
Amaryllidaceae/fisiologia , Arabidopsis/fisiologia , Catepsina B/genética , Resposta ao Choque Frio , Proteínas de Plantas/genética , Amaryllidaceae/genética , Sequência de Aminoácidos , Arabidopsis/genética , Catepsina B/química , Catepsina B/metabolismo , Resposta ao Choque Frio/genética , Congelamento , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Alinhamento de Sequência
16.
Plant Cell Rep ; 40(7): 1155-1170, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33950277

RESUMO

KEY MESSAGE: The drought and salt tolerances of wheat were enhanced by ectopic expression of the Arabidopsis ornithine aminotransferase (AtOAT) encoded gene. The OAT was confirmed to play a role in proline biosynthesis in wheat. Proline (Pro) accumulation is a common response to both abiotic and biotic stresses in plants. Ornithine aminotransferase (OAT) is pyridoxal-5-phosphate dependent enzyme involved in plant proline biosynthesis. During stress condition, proline is synthesized via glutamate and ornithine pathways. The OAT is the key enzyme in ornithine pathway. In this study, an OAT gene AtOAT from Arabidopsis was expressed in wheat for its functional characterization under drought, salinity, and heat stress conditions. We found that the expression of AtOAT enhanced the drought and salt stress tolerances of wheat by increasing the proline content and peroxidase activity. In addition, it was confirmed that the expression of AtOAT also played a partial tolerance to heat stress in the transgenic wheat plants. Moreover, quantitative real-time PCR (qRT-PCR) analysis showed that the transformation of AtOAT up-regulated the expression of the proline biosynthesis associated genes TaOAT, TaP5CS, and TaP5CR, and down-regulated that of the proline catabolism related gene TaP5CDH in the transgenic plants under stress conditions. Moreover, the genes involved in ornithine pathway (Orn-OAT-P5C/GSA-P5CR-Pro) were up-regulated along with the up-regulation of those genes involved in glutamate pathway (Glu-P5CS-P5C/GSA-P5CR-Pro). Therefore, we concluded that the expression of AtOAT enhanced wheat abiotic tolerance via modifying the proline biosynthesis by up-regulating the expression of the proline biosynthesis-associated genes and down-regulating that of the proline catabolic gene under stresses condition.


Assuntos
Proteínas de Arabidopsis/genética , Ornitina-Oxo-Ácido Transaminase/genética , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/genética , Triticum/fisiologia , Secas , Regulação da Expressão Gênica de Plantas , Resposta ao Choque Térmico/genética , Plantas Geneticamente Modificadas/genética , Prolina/genética , Prolina/metabolismo , Tolerância ao Sal/genética , Estresse Fisiológico/fisiologia , Triticum/genética
17.
Int J Mol Sci ; 22(7)2021 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-33806243

RESUMO

Arachis hypogaea abscisic acid transporter like-1 (AhATL1) modulates abscisic acid (ABA) sensitivity by specifically influencing the importing of ABA into cells, and is a key player in plant stress responses. However, there is limited information on ABA transporters in crops. In this study, we found that the level of AhATL1 expression and AhATL1 distribution increased more rapidly in the second drought (D2) compared with in the first drought (D1). Compared with the first recovery (R1), the AhATL1 expression level and ABA content remained at a higher level during the second recovery (R2). The heterologous overexpression of AhATL1 in Arabidopsis changed the expression pattern of certain memory genes and changed the post response gene type into the memory gene type. Regarding the proline and water content of Col (Arabidopsis thaliana L. Heynh., Col-0), atabcg22, and AhATL1-OX during drought training, the second drought (D2) was more severe than the first drought (D1), which was more conducive to maintaining the cell osmotic balance and resisting drought. In summary, drought stress memory resulted in a rapid increase in the AhATL1 expression and AhATL1 distribution level, and then raised the endogenous ABA content and changed the post response gene type into the memory gene type, which enhanced the drought resistance and recovery ability.


Assuntos
Arabidopsis/fisiologia , Arachis/genética , Secas , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ácido Abscísico/química , Arabidopsis/genética , Perfilação da Expressão Gênica , Proteínas de Fluorescência Verde/metabolismo , Malondialdeído/química , Osmose , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Prolina/química , Ubiquitina-Proteína Ligases/genética
18.
Genes (Basel) ; 12(3)2021 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-33808710

RESUMO

In mango (Mangifera indica L.), fruitlet abscission limits productivity. The INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) peptide acts as a key component controlling abscission events in Arabidopsis. IDA-like peptides may assume similar roles in fruit trees. In this study, we isolated two mango IDA-like encoding-genes, MiIDA1 and MiIDA2. We used mango fruitlet-bearing explants and fruitlet-bearing trees, in which fruitlets abscission was induced using ethephon. We monitored the expression profiles of the two MiIDA-like genes in control and treated fruitlet abscission zones (AZs). In both systems, qRT-PCR showed that, within 24 h, both MiIDA-like genes were induced by ethephon, and that changes in their expression profiles were associated with upregulation of different ethylene signaling-related and cell-wall modifying genes. Furthermore, ectopic expression of both genes in Arabidopsis promoted floral-organ abscission, and was accompanied by an early increase in the cytosolic pH of floral AZ cells-a phenomenon known to be linked with abscission, and by activation of cell separation in vestigial AZs. Finally, overexpression of both genes in an Atida mutant restored its abscission ability. Our results suggest roles for MiIDA1 and MiIDA2 in affecting mango fruitlet abscission. Based on our results, we propose new possible modes of action for IDA-like proteins in regulating organ abscission.


Assuntos
Perfilação da Expressão Gênica/métodos , Mangifera/fisiologia , Compostos Organofosforados/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Arabidopsis/genética , Arabidopsis/fisiologia , Citosol , Flores/genética , Flores/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Mangifera/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Análise de Sequência de RNA , Regulação para Cima
19.
Int J Mol Sci ; 22(9)2021 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-33919137

RESUMO

Introduction of C4 photosynthetic traits into C3 crops is an important strategy for improving photosynthetic capacity and productivity. Here, we report the research results of a variant line of sorghum-rice (SR) plant with big panicle and high spikelet density by introducing sorghum genome DNA into rice by spike-stalk injection. The whole-genome resequencing showed that a few sorghum genes could be integrated into the rice genome. Gene expression was confirmed for two C4 photosynthetic enzymes containing pyruvate, orthophosphate dikinase and phosphoenolpyruvate carboxykinase. Exogenous sorghum DNA integration induced a series of key traits associated with the C4 pathway called "proto-Kranz" anatomy, including leaf thickness, bundle sheath number and size, and chloroplast size in bundle sheath cells. Significantly, transgenic plants exhibited enhanced photosynthetic capacity resulting from both photosynthetic CO2-concentrating effect and improved energy balance, which led to an increase in carbohydrate levels and productivity. Furthermore, such rice plant exhibited delayed leaf senescence. In summary, this study provides a proof for the feasibility of inducing the transition from C3 leaf anatomy to proto-Kranz by spike-stalk injection to achieve efficient photosynthesis and increase productivity.


Assuntos
Oryza/fisiologia , Fotossíntese , Folhas de Planta/fisiologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Sorghum/fisiologia , Dióxido de Carbono/metabolismo , Genoma de Planta , Oryza/anatomia & histologia , Oryza/genética , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/anatomia & histologia , Plantas Geneticamente Modificadas/genética , Sorghum/anatomia & histologia , Sorghum/genética
20.
Int J Mol Sci ; 22(5)2021 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-33673599

RESUMO

Saline-alkali soil has become an important environmental problem for crop productivity. One of the most effective approaches is to cultivate new stress-tolerant plants through genetic engineering. Through RNA-seq analysis and RT-PCR validation, a novel bZIP transcription factor ChbZIP1, which is significantly upregulated at alkali conditions, was obtained from alkaliphilic microalgae Chlorella sp. BLD. Overexpression of ChbZIP1 in Saccharomyces cerevisiae and Arabidopsis increased their alkali resistance, indicating ChbZIP1 may play important roles in alkali stress response. Through subcellular localization and transcriptional activation activity analyses, we found that ChbZIP1 is a nuclear-localized bZIP TF with transactivation activity to bind with the motif of G-box 2 (TGACGT). Functional analysis found that genes such as GPX1, DOX1, CAT2, and EMB, which contained G-box 2 and were associated with oxidative stress, were significantly upregulated in Arabidopsis with ChbZIP1 overexpression. The antioxidant ability was also enhanced in transgenic Arabidopsis. These results indicate that ChbZIP1 might mediate plant adaptation to alkali stress through the active oxygen detoxification pathway. Thus, ChbZIP1 may contribute to genetically improving plants' tolerance to alkali stress.


Assuntos
Álcalis/toxicidade , Arabidopsis/fisiologia , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Chlorella/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico , Adaptação Fisiológica/genética , Arabidopsis/efeitos dos fármacos , Fatores de Transcrição de Zíper de Leucina Básica/genética , Chlorella/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Tolerantes a Sal/efeitos dos fármacos , Plantas Tolerantes a Sal/fisiologia
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