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1.
Physiol Plant ; 176(4): e14492, 2024.
Article in English | MEDLINE | ID: mdl-39166265

ABSTRACT

Genomic DNA methylation patterns play a crucial role in the developmental processes of plants and mammals. In this study, we aimed to investigate the significant effects of epigenetic mechanisms on the development of soybean seedlings and metabolic pathways. Our analyses show that 5-azaC-treatment affects radicle development from two Days After Imbibition (DAI), as well as both shoot and root development. We examined the expression levels of key genes related to DNA methylation and demethylation pathways, such as DRM2, which encodes RNA-directed DNA Methylation (RdDM) pathway, SAM synthase, responsible for methyl group donation, and ROS1, a DNA demethylase. In treated seedling roots, we observed an increase in DRM2 expression and a decrease in ROS1 expression. Additionally, 5-azaC treatment altered protein accumulation, indicating epigenetic control over stress response while inhibiting nitrogen assimilation, urea cycle, and glycolysis-related proteins. Furthermore, it influenced the levels of various phytohormones and metabolites crucial for seedling growth, such as ABA, IAA, ethylene, polyamines (PUT and Cad), and free amino acids, suggesting that epigenetic changes may shape soybean responses to pathogens, abiotic stress, and nutrient absorption. Our results assist in understanding how hypomethylation shapes soybean responses to pathogens, abiotic stress, and nutrient absorption crucial for seedling growth, suggesting that the plant's assimilation of carbon and nitrogen, along with hormone pathways, may be influenced by epigenetic changes.


Subject(s)
DNA Methylation , Glycine max , Metabolic Networks and Pathways , Plant Growth Regulators , DNA Methylation/genetics , Glycine max/genetics , Glycine max/metabolism , Glycine max/growth & development , Plant Growth Regulators/metabolism , Metabolic Networks and Pathways/genetics , Metabolic Networks and Pathways/drug effects , Gene Expression Regulation, Plant/drug effects , Seedlings/genetics , Seedlings/growth & development , Seedlings/metabolism , Plant Roots/genetics , Plant Roots/metabolism , Plant Roots/growth & development , Epigenesis, Genetic , Plant Proteins/metabolism , Plant Proteins/genetics
2.
Plant Cell Environ ; 47(11): 4227-4245, 2024 Nov.
Article in English | MEDLINE | ID: mdl-38950037

ABSTRACT

Nitrate is a nutrient and signal that regulates gene expression. The nitrate response has been extensively characterized at the organism, organ, and cell-type-specific levels, but intracellular mRNA dynamics remain unexplored. To characterize nuclear and cytoplasmic transcriptome dynamics in response to nitrate, we performed a time-course expression analysis after nitrate treatment in isolated nuclei, cytoplasm, and whole roots. We identified 402 differentially localized transcripts (DLTs) in response to nitrate treatment. Induced DLT genes showed rapid and transient recruitment of the RNA polymerase II, together with an increase in the mRNA turnover rates. DLTs code for genes involved in metabolic processes, localization, and response to stimulus indicating DLTs include genes with relevant functions for the nitrate response that have not been previously identified. Using single-molecule RNA FISH, we observed early nuclear accumulation of the NITRATE REDUCTASE 1 (NIA1) transcripts in their transcription sites. We found that transcription of NIA1, a gene showing delayed cytoplasmic accumulation, is rapidly and transiently activated; however, its transcripts become unstable when they reach the cytoplasm. Our study reveals the dynamic localization of mRNAs between the nucleus and cytoplasm as an emerging feature in the temporal control of gene expression in response to nitrate treatment in Arabidopsis roots.


Subject(s)
Arabidopsis , Cell Nucleus , Cytoplasm , Gene Expression Regulation, Plant , Nitrates , Plant Roots , RNA, Messenger , Arabidopsis/genetics , Arabidopsis/metabolism , Nitrates/metabolism , Nitrates/pharmacology , Plant Roots/metabolism , Plant Roots/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Cell Nucleus/metabolism , Cytoplasm/metabolism , Gene Expression Regulation, Plant/drug effects , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Nitrate Reductase/metabolism , Nitrate Reductase/genetics
3.
New Phytol ; 243(5): 1810-1822, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38970467

ABSTRACT

Shoot branching is determined by a balance between factors that promote axillary bud dormancy and factors that release buds from the quiescent state. The TCP family of transcription factors is classified into two classes, Class I and Class II, which usually play different roles. While the role of the Class II TCP BRANCHED1 (BRC1) in suppressing axillary bud development in Arabidopsis thaliana has been widely explored, the function of Class I TCPs in this process remains unknown. We analyzed the role of Class I TCP14 and TCP15 in axillary branch development in Arabidopsis through a series of genetic and molecular studies. In contrast to the increased branch number shown by brc1 mutants, tcp14 tcp15 plants exhibit a reduced number of branches compared with wild-type. Our findings provide evidence that TCP14 and TCP15 act by counteracting BRC1 function through two distinct mechanisms. First, they indirectly reduce BRC1 expression levels. Additionally, TCP15 directly interacts with BRC1 decoying it from chromatin and thereby preventing the transcriptional activation of a set of BRC1-dependent genes. We describe a molecular mechanism by which Class I TCPs physically antagonize the action of the Class II TCP BRC1, aligning with their opposite roles in axillary bud development.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Gene Expression Regulation, Plant , Transcription Factors , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/drug effects , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Transcription Factors/metabolism , Transcription Factors/genetics , Gene Expression Regulation, Plant/drug effects , Mutation/genetics , Protein Binding/drug effects , Chromatin/metabolism , Plant Shoots/growth & development , Plant Shoots/drug effects , Plant Shoots/genetics
4.
Plant Physiol Biochem ; 214: 108889, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38954945

ABSTRACT

Abscisic acid (ABA) is crucial for plant water deficit (WD) acclimation, but how the interplay between ABA and guard cell (GC) metabolism aids plant WD acclimation remains unclear. Here, we investigated how ABA regulates GC metabolism and how this contributes to plant WD acclimation using tomato wild type (WT) and the ABA-deficient sitiens mutant. These genotypes were characterized at physiological, metabolic, and transcriptional levels under recurring WD periods and were used to perform a13C-glucose labelling experiment using isolated guard cells following exogenously applied ABA. ABA deficiency altered the level of sugars and organic acids in GCs in both irrigated and WD plants and the dynamic of accumulation/degradation of these compounds in GCs during the dark-to-light transition. WD-induced metabolic changes were more pronounced in sitiens than WT GCs. Results from the 13C-labelling experiment indicate that ABA is required for the glycolytic fluxes toward malate and acts as a negative regulator of a putative sucrose substrate cycle. The expression of key ABA-biosynthetic genes was higher in WT than in sitiens GCs after two cycles of WD. Additionally, the intrinsic leaf water use efficiency increased only in WT after the second WD cycle, compared to sitiens. Our results highlight that ABA deficiency disrupts the homeostasis of GC primary metabolism and the WD memory, negatively affecting plant WD acclimation. Our study demonstrates which metabolic pathways are activated by WD and/or regulated by ABA in GCs, which improves our understanding of plant WD acclimation, with clear consequences for plant metabolic engineering in the future.


Subject(s)
Abscisic Acid , Solanum lycopersicum , Abscisic Acid/metabolism , Abscisic Acid/pharmacology , Solanum lycopersicum/metabolism , Solanum lycopersicum/genetics , Plant Stomata/metabolism , Plant Stomata/drug effects , Gene Expression Regulation, Plant/drug effects
5.
Braz J Biol ; 84: e280778, 2024.
Article in English | MEDLINE | ID: mdl-38922193

ABSTRACT

Despite being valuable for producing a natural sweetener Curculin, Curculigo latifolia has a low growth and difficult to domestificate. So, to solve this problem, propagation on in vitro culture will be an alternative method to propagated this spesies under different cytokinins and light condition. Cytokinins and light has major role in organogenesis, growth and gene expression of many species. Thus, in this study, we aimed to improve the Curculigo latifolia growth on in vitro condition and expression of curculin gene by combining cytokinins addition and different light exposure. Four weeks seedlings were sub-cultured into medium (MS free hormone) containing 3 mg/L benzyladenine (BA) and various concentrations of meta-Topolin (mT) including 0.1 mg/L, 0.5 mg/L, and 5 mg/L. The cultures then incubated under different light types (red, blue, white LED lights and white fluorescence light) with 16-h light/ 18-h dark photoperiod for 14 weeks at 25 ± 2°C. Several parameters, including plant height, leaf number, chlorophyll contents, stomatal structure, and density and curculin expression, were observed every week. Unexpectedly, our results showed that C. latifolia growth displayed significant improvement when it was treated under white LED light without any additional cytokinins. In sum, white LED light further improves plantlets phenotype, such as plant height, leaf number, chlorophyll production, and stomatal number and structure, whereas, red LED light lead to a decreased phenotypes but increase the curculin gene expression.


Subject(s)
Curculigo , Cytokinins , Light , Plant Growth Regulators , Cytokinins/pharmacology , Curculigo/genetics , Plant Growth Regulators/pharmacology , Gene Expression Regulation, Plant/drug effects
6.
Int J Mol Sci ; 22(23)2021 Nov 24.
Article in English | MEDLINE | ID: mdl-34884520

ABSTRACT

Low temperature remarkably limits rubber tree (Hevea brasiliensis Muell. Arg.) growth, latex production, and geographical distribution, but the underlying mechanisms of Hevea brasiliensis cold stress response remain elusive. Here, we identified HbSnRK2.6 as a key component in ABA signaling functions in phytohormone abscisic acid (ABA)-regulated cold stress response in Hevea brasiliensis. Exogenous application of ABA enhances Hevea brasiliensis cold tolerance. Cold-regulated (COR) genes in the CBF pathway are upregulated by ABA. Transcript levels of all five HbSnRK2.6 members are significantly induced by cold, while HbSnRK2.6A, HbSnRK2.6B, and HbSnRK2.6C can be further activated by ABA under cold conditions. Additionally, HbSnRK2.6s are localized in the cytoplasm and nucleus, and can physically interact with HbICE2, a crucial positive regulator in the cold signaling pathway. Overexpression of HbSnRK2.6A or HbSnRK2.6B in Arabidopsis extensively enhances plant responses to ABA and expression of COR genes, leading to increased cold stress tolerance. Furthermore, HbSnRK2.6A and HbSnRK2.6B can promote transcriptional activity of HbICE2, thus, increasing the expression of HbCBF1. Taken together, we demonstrate that HbSnRK2.6s are involved in ABA-regulated cold stress response in Hevea brasiliensis by regulating transcriptional activity of HbICE2.


Subject(s)
Abscisic Acid/pharmacology , Cold-Shock Response , Gene Expression Regulation, Plant/drug effects , Hevea/metabolism , Plant Proteins/metabolism , Protein Kinases/metabolism , Transcription Factors/metabolism , Hevea/drug effects , Hevea/genetics , Plant Growth Regulators/pharmacology , Plant Proteins/genetics , Protein Kinases/genetics , Transcription Factors/genetics
7.
Sci Rep ; 11(1): 19644, 2021 10 04.
Article in English | MEDLINE | ID: mdl-34608228

ABSTRACT

To date, the investigation of genes involved in Al resistance has focused mainly on microarrays and short periods of Al exposure. We investigated genes involved in the global response under Al stress by tracking the expression profile of two inbred popcorn lines with different Al sensitivity during 72 h of Al stress. A total of 1003 differentially expressed genes were identified in the Al-sensitive line, and 1751 were identified in the Al-resistant line, of which 273 were shared in both lines. Genes in the category of "response to abiotic stress" were present in both lines, but there was a higher number in the Al-resistant line. Transcription factors, genes involved in fatty acid biosynthesis, and genes involved in cell wall modifications were also detected. In the Al-resistant line, GST6 was identified as one of the key hub genes by co-expression network analysis, and ABC6 may play a role in the downstream regulation of CASP-like 5. In addition, we suggest a class of SWEET transporters that might be involved in the regulation of vacuolar sugar storage and may serve as mechanisms for Al resistance. The results and conclusions expand our understanding of the complex mechanisms involved in Al toxicity and provide a platform for future functional analyses and genomic studies of Al stress in popcorn.


Subject(s)
Aluminum/metabolism , Gene Expression Profiling , Gene Expression Regulation, Plant , Transcriptome , Zea mays/genetics , Zea mays/metabolism , Aluminum/toxicity , Computational Biology/methods , Gene Expression Regulation, Plant/drug effects , Molecular Sequence Annotation , Plant Breeding
8.
Cells ; 10(9)2021 09 07.
Article in English | MEDLINE | ID: mdl-34571988

ABSTRACT

Papaya is a fleshy fruit that undergoes fast ethylene-induced modifications. The fruit becomes edible, but the fast pulp softening is the main factor that limits the post-harvest period. Papaya fast pulp softening occurs due to cell wall disassembling coordinated by ethylene triggering that massively expresses pectinases. In this work, RNA-seq analysis of ethylene-treated and non-treated papayas enabled a wide transcriptome overview that indicated the role of ethylene during ripening at the gene expression level. Several families of transcription factors (AP2/ERF, NAC, and MADS-box) were differentially expressed. ACO, ACS, and SAM-Mtase genes were upregulated, indicating a high rate of ethylene biosynthesis after ethylene treatment. The correlation among gene expression and physiological data demonstrated ethylene treatment can indeed simulate ripening, and regulation of changes in fruit color, aroma, and flavor could be attributed to the coordinated expression of several related genes. Especially about pulp firmness, the identification of 157 expressed genes related to cell wall metabolism demonstrated that pulp softening is accomplished by a coordinated action of several different cell wall-related enzymes. The mechanism is different from other commercially important fruits, such as strawberry, tomato, kiwifruit, and apple. The observed behavior of this new transcriptomic data confirms ethylene triggering is the main event that elicits fast pulp softening in papayas.


Subject(s)
Carica/metabolism , Ethylenes/metabolism , Fruit/metabolism , Carica/enzymology , Carica/genetics , Cell Wall/metabolism , Ethylenes/pharmacology , Fruit/drug effects , Fruit/enzymology , Gene Expression/genetics , Gene Expression Regulation, Plant/drug effects , Genes, Plant , Plant Proteins/metabolism , Systems Biology/methods , Transcription Factors/metabolism , Transcriptome/drug effects
9.
BMC Plant Biol ; 21(1): 420, 2021 Sep 13.
Article in English | MEDLINE | ID: mdl-34517831

ABSTRACT

BACKGROUND: Natural rubber (cis-1,4-polyioprene, NR) is an indispensable industrial raw material obtained from the Pará rubber tree (H. brasiliensis). Natural rubber cannot be replaced by synthetic rubber compounds because of the superior resilience, elasticity, abrasion resistance, efficient heat dispersion, and impact resistance of NR. In NR production, latex is harvested by periodical tapping of the trunk bark. Ethylene enhances and prolongs latex flow and latex regeneration. Ethephon, which is an ethylene-releasing compound, applied to the trunk before tapping usually results in a 1.5- to 2-fold increase in latex yield. However, intense mechanical damage to bark tissues by excessive tapping and/or over-stimulation with ethephon induces severe oxidative stress in laticifer cells, which often causes tapping panel dryness (TPD) syndrome. To enhance NR production without causing TPD, an improved understanding of the molecular mechanism of the ethylene response in the Pará rubber tree is required. Therefore, we investigated gene expression in response to ethephon treatment using Pará rubber tree seedlings as a model system. RESULTS: After ethephon treatment, 3270 genes showed significant differences in expression compared with the mock treatment. Genes associated with carotenoids, flavonoids, and abscisic acid biosynthesis were significantly upregulated by ethephon treatment, which might contribute to an increase in latex flow. Genes associated with secondary cell wall formation were downregulated, which might be because of the reduced sugar supply. Given that sucrose is an important molecule for NR production, a trade-off may arise between NR production and cell wall formation for plant growth and for wound healing at the tapping panel. CONCLUSIONS: Dynamic changes in gene expression occur specifically in response to ethephon treatment. Certain genes identified may potentially contribute to latex production or TPD suppression. These data provide valuable information to understand the mechanism of ethylene stimulation, and will contribute to improved management practices and/or molecular breeding to attain higher yields of latex from Pará rubber trees.


Subject(s)
Ethylenes/metabolism , Gene Expression Profiling , Gene Expression Regulation, Plant/drug effects , Hevea/genetics , Hevea/metabolism , Latex/metabolism , Seedlings/genetics , Seedlings/metabolism , Crops, Agricultural/genetics , Crops, Agricultural/metabolism , Genes, Plant , Indonesia
10.
Int J Mol Sci ; 22(12)2021 Jun 11.
Article in English | MEDLINE | ID: mdl-34208198

ABSTRACT

The role of auxin in the fruit-ripening process during the early developmental stages of commercial strawberry fruits (Fragaria x ananassa) has been previously described, with auxin production occurring in achenes and moving to the receptacle. Additionally, fruit softening is a consequence of the depolymerization and solubilization of cell wall components produced by the action of a group of proteins and enzymes. The aim of this study was to compare the effect of exogenous auxin treatment on the physiological properties of the cell wall-associated polysaccharide contents of strawberry fruits. We combined thermogravimetric (TG) analysis with analyses of the mRNA abundance, enzymatic activity, and physiological characteristics related to the cell wall. The samples did not show a change in fruit firmness at 48 h post-treatment; by contrast, we showed changes in the cell wall stability based on TG and differential thermogravimetric (DTG) analysis curves. Less degradation of the cell wall polymers was observed after auxin treatment at 48 h post-treatment. The results of our study indicate that auxin treatment delays the cell wall disassembly process in strawberries.


Subject(s)
Biopolymers/metabolism , Cell Wall/metabolism , Fragaria/metabolism , Fruit/metabolism , Indoleacetic Acids/pharmacology , Cell Wall/drug effects , Cell Wall/genetics , Fragaria/drug effects , Fragaria/genetics , Fruit/drug effects , Fruit/genetics , Gene Expression Regulation, Plant/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Temperature , Thermogravimetry , Transcription, Genetic/drug effects , Triiodobenzoic Acids/pharmacology
11.
BMC Plant Biol ; 21(1): 259, 2021 Jun 05.
Article in English | MEDLINE | ID: mdl-34090337

ABSTRACT

BACKGROUND: Nitrogen (N) and phosphorus (P) are macronutrients essential for crop growth and productivity. In cultivated fields, N and P levels are rarely sufficient, contributing to the gap between realized and potential production. Fertilizer application increases nutrient availability, but is not available to all farmers, nor are current rates of application sustainable or environmentally desirable. Transcriptomic studies of cereal crops have revealed dramatic responses to either low N or low P single stress treatments. In the field, however, levels of both N and P may be suboptimal. The interaction between N and P starvation responses remains to be fully characterized. RESULTS: We characterized growth and root and leaf transcriptomes of young maize plants under nutrient replete, low N, low P or combined low NP conditions. We identified 1555 genes to respond to our nutrient treatments, in one or both tissues. A large group of genes, including many classical P starvation response genes, were regulated antagonistically between low N and P conditions. An additional experiment over a range of N availability indicated that a mild reduction in N levels was sufficient to repress the low P induction of P starvation genes. Although expression of P transporter genes was repressed under low N or low NP, we confirmed earlier reports of P hyper accumulation under N limitation. CONCLUSIONS: Transcriptional responses to low N or P were distinct, with few genes responding in a similar way to the two single stress treatments. In combined NP stress, the low N response dominated, and the P starvation response was largely suppressed. A mild reduction in N availability was sufficient to repress the induction of P starvation associated genes. We conclude that activation of the transcriptional response to P starvation in maize is contingent on N availability.


Subject(s)
Nitrogen/pharmacology , Phosphorus/pharmacology , Zea mays/drug effects , Zea mays/growth & development , Gene Expression Profiling , Gene Expression Regulation, Plant/drug effects , Nitrogen/administration & dosage , Phosphorus/administration & dosage , Plant Leaves/growth & development , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/growth & development , Plant Roots/metabolism , Seedlings/growth & development , Stress, Physiological/drug effects , Zea mays/metabolism
12.
Plant Mol Biol ; 107(4-5): 387-404, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34189708

ABSTRACT

KEY MESSAGE: The moss Pseudocrossidium replicatum is a desiccation-tolerant species that uses an inducible system to withstand severe abiotic stress in both protonemal and gametophore tissues. Desiccation tolerance (DT) is the ability of cells to recover from an air-dried state. Here, the moss Pseudocrossidium replicatum was identified as a fully desiccation-tolerant (FDT) species. Its gametophores rapidly lost more than 90% of their water content when exposed to a low-humidity atmosphere [23% relative humidity (RH)], but abscisic acid (ABA) pretreatment diminished the final water loss after equilibrium was reached. P. replicatum gametophores maintained good maximum photosystem II (PSII) efficiency (Fv/Fm) for up to two hours during slow dehydration; however, ABA pretreatment induced a faster decrease in the Fv/Fm. ABA also induced a faster recovery of the Fv/Fm after rehydration. Protein synthesis inhibitor treatment before dehydration hampered the recovery of the Fv/Fm when the gametophores were rehydrated after desiccation, suggesting the presence of an inducible protective mechanism that is activated in response to abiotic stress. This observation was also supported by accumulation of soluble sugars in gametophores exposed to ABA or NaCl. Exogenous ABA treatment delayed the germination of P. replicatum spores and induced morphological changes in protonemal cells that resembled brachycytes. Transcriptome analyses revealed the presence of an inducible molecular mechanism in P. replicatum protonemata that was activated in response to dehydration. This study is the first RNA-Seq study of the protonemal tissues of an FDT moss. Our results suggest that P. replicatum is an FDT moss equipped with an inducible molecular response that prepares this species for severe abiotic stress and that ABA plays an important role in this response.


Subject(s)
Adaptation, Physiological/genetics , Bryopsida/genetics , Gene Expression Profiling/methods , Gene Expression Regulation, Plant/genetics , Abscisic Acid/pharmacology , Adaptation, Physiological/drug effects , Alpha-Amanitin/pharmacology , Bryopsida/metabolism , Cycloheximide/pharmacology , Dehydration , Gene Expression Regulation, Plant/drug effects , Geography , Mexico , Nucleic Acid Synthesis Inhibitors/pharmacology , Plant Growth Regulators/pharmacology , Protein Synthesis Inhibitors/pharmacology , RNA-Seq/methods , Stress, Physiological , Time Factors
13.
Int J Mol Sci ; 22(9)2021 May 06.
Article in English | MEDLINE | ID: mdl-34066536

ABSTRACT

Plant food production is severely affected by fungi; to cope with this problem, farmers use synthetic fungicides. However, the need to reduce fungicide application has led to a search for alternatives, such as biostimulants. Rare-earth elements (REEs) are widely used as biostimulants, but their mode of action and their potential as an alternative to synthetic fungicides have not been fully studied. Here, the biostimulant effect of gadolinium (Gd) is explored using the plant-pathosystem Arabidopsis thaliana-Botrytis cinerea. We determine that Gd induces local, systemic, and long-lasting plant defense responses to B. cinerea, without affecting fungal development. The physiological changes induced by Gd have been related to its structural resemblance to calcium. However, our results show that the calcium-induced defense response is not sufficient to protect plants against B. cinerea, compared to Gd. Furthermore, a genome-wide transcriptomic analysis shows that Gd induces plant defenses and modifies early and late defense responses. However, the resistance to B. cinerea is dependent on JA/ET-induced responses. These data support the conclusion that Gd can be used as a biocontrol agent for B. cinerea. These results are a valuable tool to uncover the molecular mechanisms induced by REEs.


Subject(s)
Arabidopsis/immunology , Arabidopsis/microbiology , Botrytis/physiology , Cyclopentanes/metabolism , Ethylenes/metabolism , Gadolinium/pharmacology , Oxylipins/metabolism , Protective Agents/pharmacology , Arabidopsis/drug effects , Arabidopsis/genetics , Botrytis/drug effects , Botrytis/growth & development , Gene Expression Regulation, Plant/drug effects , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Roots/drug effects , Plant Roots/growth & development , Reactive Oxygen Species/metabolism , Salicylic Acid/metabolism , Stress, Physiological/drug effects , Transcriptional Activation/drug effects , Transcriptional Activation/genetics
14.
Sci Rep ; 11(1): 11257, 2021 05 27.
Article in English | MEDLINE | ID: mdl-34045631

ABSTRACT

Soil contamination with heavy metals is a major problem worldwide, due to the increasing impact mainly caused by anthropogenic activities. This research evaluated the phytoremediation capacity of, Lolium perenne for heavy metals such as cadmium (Cd2+) and mercury (Hg2+), and the effects of these metals on morphology, biomass production, and the changes on gene expression. Seeds of L. perenne were exposed to six concentrations of Cd2+ and Hg2+ in the range of 0 to 25 mg L-1, and two mixtures of Cd2+-Hg2. The Non-Observed Effect Level (NOEL) was established with dose response curves and the expression of specific genes was evaluated applying a commercially available quantitative reverse transcription (RT-qPCR) assay. There was no significant effect when exposing the seeds to Hg2+, for Cd2+ the maximum concentration was established in 0.1 mg L-1, and for the two concentrations of mixtures, there was a negative effect. An increase of expression of genes that regulate antioxidant activity and stress was found when the plant was exposed to heavy metals. Given the high tolerance to metals analyzed that was reflected both, the development of the plant and in its molecular response, these results highlight that L. perenne is a plant with phytoremediator potential.


Subject(s)
Cadmium/pharmacology , Gene Expression Regulation, Plant/drug effects , Lolium/drug effects , Mercury/pharmacology , Soil Pollutants/pharmacology , Lolium/genetics , Lolium/growth & development , Stress, Physiological/drug effects
15.
Plant J ; 106(6): 1791-1806, 2021 06.
Article in English | MEDLINE | ID: mdl-33797826

ABSTRACT

Low-molecular-weight organic acid (OA) extrusion by plant roots is critical for plant nutrition, tolerance to cations toxicity, and plant-microbe interactions. Therefore, methodologies for the rapid and precise quantification of OAs are necessary to be incorporated in the analysis of roots and their exudates. The spatial location of root exudates is also important to understand the molecular mechanisms directing OA production and release into the rhizosphere. Here, we report the development of two complementary methodologies for OA determination, which were employed to evaluate the effect of inorganic ortho-phosphate (Pi) deficiency and aluminum toxicity on OA excretion by Arabidopsis roots. OA exudation by roots is considered a core response to different types of abiotic stress and for the interaction of roots with soil microbes, and for decades has been a target trait to produce plant varieties with increased capacities of Pi uptake and Al tolerance. Using targeted ultra-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS), we achieved the quantification of six OAs in root exudates at sub-micromolar detection limits with an analysis time of less than 5 min per sample. We also employed targeted (MS/MS) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) to detect the spatial location of citric and malic acid with high specificity in roots and exudates. Using these methods, we studied OA exudation in response to Al toxicity and Pi deficiency in Arabidopsis seedlings overexpressing genes involved in OA excretion. Finally, we show the transferability of the MALDI-MSI method by analyzing OA excretion in Marchantia polymorpha gemmalings subjected to Pi deficiency.


Subject(s)
Acids/chemistry , Aluminum/toxicity , Phosphorus/administration & dosage , Plant Exudates/chemistry , Plant Roots/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Arabidopsis/chemistry , Arabidopsis/drug effects , Arabidopsis/metabolism , Gene Expression Regulation, Plant/drug effects , Marchantia/chemistry , Marchantia/drug effects , Marchantia/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified
16.
Plant Cell Environ ; 44(6): 1961-1976, 2021 06.
Article in English | MEDLINE | ID: mdl-33529396

ABSTRACT

Plants host a diverse microbiome and differentially react to the fungal species living as endophytes or around their roots through emission of volatiles. Here, using divided Petri plates for Arabidopsis-T. atroviride co-cultivation, we show that fungal volatiles increase endogenous sugar levels in shoots, roots and root exudates, which improve Arabidopsis root growth and branching and strengthen the symbiosis. Tissue-specific expression of three sucrose phosphate synthase-encoding genes (AtSPS1F, AtSPS2F and AtSPS3F), and AtSUC2 and SWEET transporters revealed that the gene expression signatures differ from those of the fungal pathogens Fusarium oxysporum and Alternaria alternata and that AtSUC2 is largely repressed either by increasing carbon availability or by perception of the fungal volatile 6-pentyl-2H-pyran-2-one. Our data point to Trichoderma volatiles as chemical signatures for sugar biosynthesis and exudation and unveil specific modulation of a critical, long-distance sucrose transporter in the plant.


Subject(s)
Arabidopsis/growth & development , Hypocreales/chemistry , Sucrose/metabolism , Volatile Organic Compounds/pharmacology , Arabidopsis/drug effects , Arabidopsis/genetics , Arabidopsis/metabolism , Biological Transport , Gene Expression Regulation, Plant/drug effects , Glucose/metabolism , Glucosyltransferases/genetics , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Plant Exudates/metabolism , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/growth & development , Plants, Genetically Modified , Pyrones/pharmacology , Seedlings/growth & development , Seedlings/metabolism , Sucrose/pharmacology
17.
Int J Mol Sci ; 21(21)2020 Nov 07.
Article in English | MEDLINE | ID: mdl-33171770

ABSTRACT

Phosphate (Pi) is a pivotal nutrient that constraints plant development and productivity in natural ecosystems. Land colonization by plants, more than 470 million years ago, evolved adaptive mechanisms to conquer Pi-scarce environments. However, little is known about the molecular basis underlying such adaptations at early branches of plant phylogeny. To shed light on how early divergent plants respond to Pi limitation, we analyzed the morpho-physiological and transcriptional dynamics of Marchantia polymorpha upon Pi starvation. Our phylogenomic analysis highlights some gene networks present since the Chlorophytes and others established in the Streptophytes (e.g., PHR1-SPX1 and STOP1-ALMT1, respectively). At the morpho-physiological level, the response is characterized by the induction of phosphatase activity, media acidification, accumulation of auronidins, reduction of internal Pi concentration, and developmental modifications of rhizoids. The transcriptional response involves the induction of MpPHR1, Pi transporters, lipid turnover enzymes, and MpMYB14, which is an essential transcription factor for auronidins biosynthesis. MpSTOP2 up-regulation correlates with expression changes in genes related to organic acid biosynthesis and transport, suggesting a preference for citrate exudation. An analysis of MpPHR1 binding sequences (P1BS) shows an enrichment of this cis regulatory element in differentially expressed genes. Our study unravels the strategies, at diverse levels of organization, exerted by M. polymorpha to cope with low Pi availability.


Subject(s)
Marchantia/genetics , Marchantia/metabolism , Phosphates/metabolism , Ecosystem , Gene Expression Regulation, Plant/drug effects , Gene Expression Regulation, Plant/genetics , Gene Regulatory Networks/drug effects , Gene Regulatory Networks/genetics , Hepatophyta/metabolism , Phylogeny , Transcription Factors/metabolism
18.
Sci Rep ; 10(1): 19437, 2020 11 10.
Article in English | MEDLINE | ID: mdl-33173093

ABSTRACT

We tested two sources of lanthanum (La), LaCl3 and La(NO3)3 × 6H2O at a concentration of 40 µM each, in the treatment solution of cut flowers of 15 tulip (Tulipa gesneriana L.) cultivars. Ascorbic acid (AsA; 0.2 g/L) was used as a reference solution, while distilled water was evaluated as an absolute control. With both La sources, bud length and diameter, and stem length were increased; as a result, stem curvature was also significantly increased with La treatments. The cultivars Laura Fygi and Rosario registered the highest relative stem elongation. Lalibela and Acropolis displayed the greatest stem curvature on the last day in vase. At 3, 5, 7, 9 and 11 days after cutting, the highest solution uptake was recorded in flower stems treated with LaCl3, surpassing the control by 5, 11, 15, 18 and 24%, respectively. The relative stem elongations observed were 21.3, 27.4, 35.2 and 35.5% in the control, AsA, LaCl3 and La(NO3)3, respectively. The mean solution uptake per gram of stem fresh biomass weight was 1.44, 1.44, 1.71 and 1.54 mL in the control, AsA, LaCl3 and La(NO3)3, respectively. LaCl3 significantly increased the bud length and solution uptake of flower stems, while La(NO3)3 × 6H2O increased stem fresh weight.


Subject(s)
Flowers/drug effects , Flowers/metabolism , Lanthanum/pharmacology , Tulipa/drug effects , Tulipa/metabolism , Gene Expression Regulation, Plant/drug effects , Plant Proteins/metabolism
19.
Int J Mol Sci ; 21(21)2020 Oct 26.
Article in English | MEDLINE | ID: mdl-33114621

ABSTRACT

Some metals are beneficial to plants and contribute to critical physiological processes. Some metals, however, are not. The presence of aluminum ions (Al3+) can be very toxic, especially in acidic soils. Considerable parts of the world's arable land are acidic in nature; mechanistically elucidating a plant's response to aluminum stress is critical to mitigating this stress and improving the quality of plants. To identify the genes involved in sugarcane response to aluminum stress, we generated 372 million paired-end RNA sequencing reads from the roots of CTC-2 and RB855453, which are two contrasting cultivars. Data normalization resulted in 162,161 contigs (contiguous sequences) and 97,335 genes from a de novo transcriptome assembly (trinity genes). A total of 4858 and 1307 differently expressed genes (DEGs) for treatment versus control were identified for the CTC-2 and RB855453 cultivars, respectively. The DEGs were annotated into 34 functional categories. The majority of the genes were upregulated in the CTC-2 (tolerant cultivar) and downregulated in RB855453 (sensitive cultivar). Here, we present the first root transcriptome of sugarcane under aluminum stress. The results and conclusions of this study are a crucial launch pad for future genetic and genomic studies of sugarcane. The transcriptome analysis shows that sugarcane tolerance to aluminum may be explained by an efficient detoxification mechanism combined with lateral root formation and activation of redox enzymes. We also present a hypothetical model for aluminum tolerance in the CTC-2 cultivar.


Subject(s)
Aluminum/adverse effects , Gene Expression Profiling/methods , Plant Proteins/genetics , Saccharum/growth & development , Gene Expression Regulation, Plant/drug effects , High-Throughput Nucleotide Sequencing , Plant Roots/drug effects , Plant Roots/genetics , Plant Roots/growth & development , Saccharum/drug effects , Saccharum/genetics , Sequence Analysis, RNA , Signal Transduction/drug effects , Soil/chemistry , Stress, Physiological
20.
J Plant Physiol ; 253: 153261, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32947244

ABSTRACT

MicroRNAs (miRNAs) are small non-coding molecules that modulate gene expression through targeting mRNA by specific-sequence cleavage, translation inhibition, or transcriptional regulation. miRNAs are key molecules in regulatory networks in abiotic stresses such as salt stress and water deficit in plants. Throughout the world, soybean is a critical crop, the production of which is affected by environmental stress conditions. In this study, RNA-Seq libraries from leaves of soybean under salt treatment were analyzed. 17 miRNAs and 31 putative target genes were identified with inverse differential expression patterns, indicating miRNA-target interaction. The differential expression of six miRNAs, including miR482bd-5p, and their potential targets, were confirmed by RT-qPCR. The miR482bd-5p expression was repressed, while its potential HEC1 and BAK1 targets were increased. Polyethylene glycol experiment was used to simulate drought stress, and miR482bd-5p, HEC1, and BAK1 presented a similar expression pattern, as found in salt stress. Histone modifications occur in response to abiotic stress, where histone deacetylases (HDACs) can lead to gene repression and silencing. The miR482bd-5p epigenetic regulation by histone deacetylation was evaluated by using the SAHA-HDAC inhibitor. The miR482bd-5p was up-regulated, and HEC1 was down-regulated under SAHA-salt treatment. It suggests an epigenetic regulation, where the miRNA gene is repressed by HDAC under salt stress, reducing its transcription, with an associated increase in the HEC1 target expression.


Subject(s)
Epigenesis, Genetic , Gene Expression Regulation, Plant/drug effects , Glycine max/drug effects , Histone Deacetylase Inhibitors/pharmacology , MicroRNAs/genetics , Stress, Physiological/drug effects , Gene Library , Osmotic Pressure/drug effects , Plant Leaves/drug effects , Plant Leaves/genetics , Plant Leaves/physiology , RNA, Messenger/genetics , RNA, Plant/genetics , RNA-Seq , Salt Stress/drug effects , Glycine max/genetics , Glycine max/physiology
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