scholarly journals Molecular Manipulation of the MiR396/GRF Expression Module Alters the Salt Stress Response of Arabidopsis thaliana

Agronomy ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1751
Author(s):  
Joseph L. Pegler ◽  
Duc Quan Nguyen ◽  
Jackson M.J. Oultram ◽  
Christopher P.L. Grof ◽  
Andrew L. Eamens
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Gene Family ◽  
Expression Regulation ◽  
Treatment Regime ◽  
Wild Type ◽  
Salt Stress Response ◽  
Stress Treatment ◽  
Plant Line ◽  
Expression Module

We previously demonstrated that microRNA396 (miR396) abundance is altered in 15-day-old Arabidopsis thaliana (Arabidopsis) whole seedlings following their exposure to a 7-day salt stress treatment regime. We, therefore, used a molecular modification approach to generate two new Arabidopsis transformant populations with reduced (MIM396 plants) and elevated (MIR396 plants) miR396 abundance. The exposure of 8-day-old wild-type Arabidopsis whole seedlings and a representative plant line of the MIM396 and MIR396 transformant populations to a 7-day salt stress treatment regime revealed unique phenotypic and physiological responses to the imposed stress by unmodified wild-type Arabidopsis plants and the MIM396 and MIR396 transformat lines. A quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) approach was, therefore, applied to demonstrate that the plant line specific responses to salt stress likely stemmed from the unique molecular profile of each of the GROWTH REGULATING FACTOR (GRF) transcription factor gene family members which form posttranscriptional targets of miR396-directed expression regulation. RT-qPCR additionally revealed that, in 15-day-old Arabidopsis whole seedlings, the three previously identified putative target genes of miR396 belonging to the NEUTRAL/ALKALINE NONLYSOSOMAL CERAMIDASE-LIKE (NCER) gene family, including NCER1, NCER2, and NCER3, do not form targets of miR396-directed expression regulation at the posttranscriptional level. Taken together, the phenotypic and molecular analyses presented here demonstrate that alteration of the miR396/GRF expression module is central to the molecular response of Arabidopsis to salt stress.

scholarly journals Molecular Manipulation of the miR399/PHO2 Expression Module Alters the Salt Stress Response of Arabidopsis thaliana

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 73
Author(s):  
Joseph L. Pegler ◽  
Jackson M.J. Oultram ◽  
Christopher P.L. Grof ◽  
Andrew L Eamens
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Target Gene ◽  
Molecular Level ◽  
Biological Processes ◽  
Wild Type ◽  
Salt Stress Response ◽  
Molecular Modification ◽  
Molecular Manipulation ◽  
Expression Module

In Arabidopsis thaliana (Arabidopsis), the microRNA399 (miR399)/PHOSPHATE2 (PHO2) expression module is central to the response of Arabidopsis to phosphate (PO4) stress. In addition, miR399 has been demonstrated to also alter in abundance in response to salt stress. We therefore used a molecular modification approach to alter miR399 abundance to investigate the requirement of altered miR399 abundance in Arabidopsis in response to salt stress. The generated transformant lines, MIM399 and MIR399 plants, with reduced and elevated miR399 abundance respectively, displayed differences in their phenotypic and physiological response to those of wild-type Arabidopsis (Col-0) plants following exposure to a 7-day period of salt stress. However, at the molecular level, elevated miR399 abundance, and therefore, altered PHO2 target gene expression in salt-stressed Col-0, MIM399 and MIR399 plants, resulted in significant changes to the expression level of the two PO4 transporter genes, PHOSPHATE TRANSPORTER1;4 (PHT1;4) and PHT1;9. Elevated PHT1;4 and PHT1;9 PO4 transporter levels in salt stressed Arabidopsis would enhance PO4 translocation from the root to the shoot tissue which would supply additional levels of this precious cellular resource that could be utilized by the aerial tissues of salt stressed Arabidopsis to either maintain essential biological processes or to mount an adaptive response to salt stress.

scholarly journals Molecular Manipulation of MicroRNA397 Abundance Influences the Development and Salt Stress Response of Arabidopsis thaliana

2020 ◽  
Vol 21 (21) ◽  
pp. 7879
Author(s):  
Duc Quan Nguyen ◽  
Christopher W. Brown ◽  
Joseph L. Pegler ◽  
Andrew L. Eamens ◽  
Christopher P. L. Grof
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Lignin Content ◽  
Quantitative Polymerase Chain Reaction ◽  
Regulatory Rna ◽  
Wild Type ◽  
Salt Stress Response ◽  
Small Regulatory Rna ◽  
Heterologous System ◽  
Molecular Manipulation

Arabidopsis thaliana (Arabidopsis) has been used extensively as a heterologous system for molecular manipulation to genetically characterize both dicotyledonous and monocotyledonous plant species. Here, we report on Arabidopsis transformant lines molecularly manipulated to over-accumulate the small regulatory RNA microRNA397 (miR397) from the emerging C4 monocotyledonous grass model species Setaria viridis (S. viridis). The generated transformant lines, termed SvMIR397 plants, displayed a range of developmental phenotypes that ranged from a mild, wild-type-like phenotype, to a severe, full dwarfism phenotype. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR)-based profiling of the SvMIR397 transformant population revealed a strong correlation between the degree of miR397 over-accumulation, repressed LACCASE (LAC) target gene expression, reduced lignin content, and the severity of the developmental phenotype displayed by SvMIR397 transformants. Further, exposure of SvMIR397 transformants to a 7-day regime of salt stress revealed the SvMIR397 transformant lines to be more sensitive to the imposed stress than were wild-type Arabidopsis plants. Taken together, the findings reported here via the use of Arabidopsis as a heterologous system show that the S. viridis miR397 small regulatory RNA is able to repress the expression of three Arabidopsis LAC genes which led to reduced lignin content and increased salt stress sensitivity.

scholarly journals ROS-Scavengers, Osmoprotectants and Violaxanthin De-Epoxidation in Salt-Stressed Arabidopsis thaliana with Different Tocopherol Composition

2021 ◽  
Vol 22 (21) ◽  
pp. 11370
Author(s):  
Ewa Surówka ◽  
Dariusz Latowski ◽  
Michał Dziurka ◽  
Magdalena Rys ◽  
Anna Maksymowicz ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Transgenic Line ◽  
Wild Type ◽  
Carbon And Nitrogen ◽  
Nacl Concentration ◽  
Tocopherol Composition ◽  
Ros Scavengers ◽  
Excessive Accumulation

To determine the role of α- and γ-tocopherol (TC), this study compared the response to salt stress (200 mM NaCl) in wild type (WT) Arabidopsis thaliana (L.) Heynh. And its two mutants: (1) totally TC-deficient vte1; (2) vte4 accumulating γ-TC instead of α-TC; and (3) tmt transgenic line overaccumulating α-TC. Raman spectra revealed that salt-exposed α-TC accumulating plants were more flexible in regulating chlorophyll, carotenoid and polysaccharide levels than TC deficient mutants, while the plants overaccumulating γ-TC had the lowest levels of these biocompounds. Tocopherol composition and NaCl concentration affected xanthophyll cycle by changing the rate of violaxanthin de-epoxidation and zeaxanthin formation. NaCl treated plants with altered TC composition accumulated less oligosaccharides than WT plants. α-TC deficient plants increased their oligosaccharide levels and reduced maltose amount, while excessive accumulation of α-TC corresponded with enhanced amounts of maltose. Salt-stressed TC-deficient mutants and tmt transgenic line exhibited greater proline levels than WT plants, lower chlorogenic acid levels, and lower activity of catalase and peroxidases. α-TC accumulating plants produced more methylated proline- and glycine- betaines, and showed greater activity of superoxide dismutase than γ-TC deficient plants. Under salt stress, α-TC demonstrated a stronger regulatory effect on carbon- and nitrogen-related metabolites reorganization and modulation of antioxidant patterns than γ-TC. This suggested different links of α- and γ-TCs with various metabolic pathways via various functions and metabolic loops.

scholarly journals AtSIBP1, a Novel BTB Domain-Containing Protein, Positively Regulates Salt Signaling in Arabidopsis thaliana

Plants ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 573 ◽  
Author(s):  
Xia Wan ◽  
Lu Peng ◽  
Jie Xiong ◽  
Xiaoyi Li ◽  
Jianmei Wang ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Marker Genes ◽  
Oxygen Species ◽  
Gus Gene ◽  
Wild Type ◽  
Btb Domain ◽  
Highly Sensitive ◽  
Sessile Organisms ◽  
Salt Signaling

Because they are sessile organisms, plants need rapid and finely tuned signaling pathways to adapt to adverse environments, including salt stress. In this study, we identified a gene named Arabidopsis thaliana stress-induced BTB protein 1 (AtSIBP1), which encodes a nucleus protein with a BTB domain in its C-terminal side and is induced by salt and other stresses. The expression of the β-glucuronidase (GUS) gene driven by the AtSIBP1 promoter was found to be significantly induced in the presence of NaCl. The sibp1 mutant that lost AtSIBP1 function was found to be highly sensitive to salt stress and more vulnerable to salt stress than the wild type WT, while the overexpression of AtSIBP1 transgenic plants exhibited more tolerance to salt stress. According to the DAB staining, the sibp1 mutant accumulated more reactive oxygen species (ROS) than the WT and AtSIBP1 overexpression plants after salt stress. In addition, the expression levels of stress-induced marker genes in AtSIBP1 overexpression plants were markedly higher than those in the WT and sibp1 mutant plants. Therefore, our results demonstrate that AtSIBP1 was a positive regulator in salinity responses in Arabidopsis.

The Structure of the Arabidopsis thaliana SOS3: Molecular Mechanism of Sensing Calcium for Salt Stress Response

2005 ◽  
Vol 345 (5) ◽  
pp. 1253-1264 ◽  
Author(s):  
María José Sánchez-Barrena ◽  
Martín Martínez-Ripoll ◽  
Jian-Kang Zhu ◽  
Armando Albert
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Stress Response ◽  
Molecular Mechanism ◽  
Salt Stress Response

scholarly journals REDUCED CHLOROPLAST COVERAGE genes from Arabidopsis thaliana help to establish the size of the chloroplast compartment

2016 ◽  
Vol 113 (8) ◽  
pp. E1116-E1125 ◽  
Author(s):  
Robert M. Larkin ◽  
Giovanni Stefano ◽  
Michael E. Ruckle ◽  
Andrea K. Stavoe ◽  
Christopher A. Sinkler ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Energy Metabolism ◽  
Gene Family ◽  
Unknown Function ◽  
Cell Expansion ◽  
Mutant Allele ◽  
Eukaryotic Cells ◽  
Wild Type ◽  
Photosynthetic Organisms ◽  
Cellular Volume

Eukaryotic cells require mechanisms to establish the proportion of cellular volume devoted to particular organelles. These mechanisms are poorly understood. From a screen for plastid-to-nucleus signaling mutants in Arabidopsis thaliana, we cloned a mutant allele of a gene that encodes a protein of unknown function that is homologous to two other Arabidopsis genes of unknown function and to FRIENDLY, which was previously shown to promote the normal distribution of mitochondria in Arabidopsis. In contrast to FRIENDLY, these three homologs of FRIENDLY are found only in photosynthetic organisms. Based on these data, we proposed that FRIENDLY expanded into a small gene family to help regulate the energy metabolism of cells that contain both mitochondria and chloroplasts. Indeed, we found that knocking out these genes caused a number of chloroplast phenotypes, including a reduction in the proportion of cellular volume devoted to chloroplasts to 50% of wild type. Thus, we refer to these genes as REDUCED CHLOROPLAST COVERAGE (REC). The size of the chloroplast compartment was reduced most in rec1 mutants. The REC1 protein accumulated in the cytosol and the nucleus. REC1 was excluded from the nucleus when plants were treated with amitrole, which inhibits cell expansion and chloroplast function. We conclude that REC1 is an extraplastidic protein that helps to establish the size of the chloroplast compartment, and that signals derived from cell expansion or chloroplasts may regulate REC1.

Identification of novel candidate phosphatidic acid-binding proteins involved in the salt-stress response of Arabidopsis thaliana roots

2013 ◽  
Vol 450 (3) ◽  
pp. 573-581 ◽  
Author(s):  
Fionn McLoughlin ◽  
Steven A. Arisz ◽  
Henk L. Dekker ◽  
Gertjan Kramer ◽  
Chris G. de Koster ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Salt Stress ◽  
Phosphatidic Acid ◽  
Stress Responses ◽  
Ribosomal Proteins ◽  
Binding Proteins ◽  
Affinity Purification ◽  
Lipid Binding ◽  
Biotic And Abiotic Stress ◽  
Salt Stress Response

PA (phosphatidic acid) is a lipid second messenger involved in an array of processes occurring during a plant's life cycle. These include development, metabolism, and both biotic and abiotic stress responses. PA levels increase in response to salt, but little is known about its function in the earliest responses to salt stress. In the present study we have combined an approach to isolate peripheral membrane proteins of Arabidopsis thaliana roots with lipid-affinity purification, to identify putative proteins that interact with PA and are recruited to the membrane in response to salt stress. Of the 42 putative PA-binding proteins identified by MS, a set of eight new candidate PA-binding proteins accumulated at the membrane fraction after 7 min of salt stress. Among these were CHC (clathrin heavy chain) isoforms, ANTH (AP180 N-terminal homology) domain clathrin-assembly proteins, a putative regulator of potassium transport, two ribosomal proteins, GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and a PI (phosphatidylinositol) 4-kinase. PA binding and salt-induced membrane recruitment of GAPDH and CHC were confirmed by Western blot analysis of the cellular fractions. In conclusion, the approach of the present study is an effective way to isolate biologically relevant lipid-binding proteins and provides new leads in the study of PA-mediated salt-stress responses in roots.

scholarly journals SPL36 Encodes a Receptor-like Protein Kinase that Regulates Programmed Cell Death and Defense Responses in Rice

Rice ◽  
2021 ◽  
Vol 14 (1) ◽  
Author(s):  
R. A. O. Yuchun ◽  
J. I. A. O. Ran ◽  
W. A. N. G. Sheng ◽  
W. U. Xianmei ◽  
Y. E. Hanfei ◽  
...  
Keyword(s):  
Cell Death ◽  
Salt Stress ◽  
Protein Kinase ◽  
Programmed Cell Death ◽  
Stress Responses ◽  
Defense Responses ◽  
Base Substitution ◽  
Wild Type ◽  
Salt Stress Response ◽  
Lesion Mimic

AbstractLesion mimic mutants spontaneously produce disease spots in the absence of biotic or abiotic stresses. Analyzing lesion mimic mutants’ sheds light on the mechanisms underlying programmed cell death and defense-related responses in plants. Here, we isolated and characterized the rice (Oryza sativa) spotted leaf 36 (spl36) mutant, which was identified from an ethyl methanesulfonate-mutagenized japonica cultivar Yundao population. spl36 displayed spontaneous cell death and enhanced resistance to rice bacterial pathogens. Gene expression analysis suggested that spl36 functions in the disease response by upregulating the expression of defense-related genes. Physiological and biochemical experiments indicated that more cell death occurred in spl36 than the wild type and that plant growth and development were affected in this mutant. We isolated SPL36 by map-based cloning. A single base substitution was detected in spl36, which results in a cysteine-to-arginine substitution in SPL36. SPL36 is predicted to encode a receptor-like protein kinase containing leucine-rich domains that may be involved in stress responses in rice. spl36 was more sensitive to salt stress than the wild type, suggesting that SPL36 also negatively regulates the salt-stress response. These findings suggest that SPL36 regulates the disease resistance response in rice by affecting the expression of defense- and stress-related genes.

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