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ABA inducible rice protein phosphatase 2C confers ABA insensitivity and abiotic stress tolerance in Arabidopsis.

Singh A, Jha SK, Bagri J, Pandey GK - PLoS ONE (2015)

Bottom Line: At adult stage, OsPP108 overexpression leads to high tolerance to salt, mannitol and drought stresses with far better physiological parameters such as water loss, fresh weight, chlorophyll content and photosynthetic potential (Fv/Fm) in transgenic Arabidopsis plants.Expression profile of various stress marker genes in OsPP108 overexpressing plants revealed interplay of ABA dependent and independent pathway for abiotic stress tolerance.Overall, this study has identified a potential rice group A PP2C, which regulates ABA signaling negatively and abiotic stress signaling positively.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi, India.

ABSTRACT
Arabidopsis PP2C belonging to group A have been extensively worked out and known to negatively regulate ABA signaling. However, rice (Oryza sativa) orthologs of Arabidopsis group A PP2C are scarcely characterized functionally. We have identified a group A PP2C from rice (OsPP108), which is highly inducible under ABA, salt and drought stresses and localized predominantly in the nucleus. Genetic analysis revealed that Arabidopsis plants overexpressing OsPP108 are highly insensitive to ABA and tolerant to high salt and mannitol stresses during seed germination, root growth and overall seedling growth. At adult stage, OsPP108 overexpression leads to high tolerance to salt, mannitol and drought stresses with far better physiological parameters such as water loss, fresh weight, chlorophyll content and photosynthetic potential (Fv/Fm) in transgenic Arabidopsis plants. Expression profile of various stress marker genes in OsPP108 overexpressing plants revealed interplay of ABA dependent and independent pathway for abiotic stress tolerance. Overall, this study has identified a potential rice group A PP2C, which regulates ABA signaling negatively and abiotic stress signaling positively. Transgenic rice plants overexpressing this gene might provide an answer to the problem of low crop yield and productivity during adverse environmental conditions.

No MeSH data available.


Related in: MedlinePlus

Sub-cellular localization of OsPP108 in Nicotiana benthamiana.Agrobacterium-infiltrated tobacco leaves expressing the GFP-OsPP108 fusion protein driven by the 2XCaMV35S promoter. Confocal images of fluorescence (green) for cell expressing OsPP108 (35S:OsPP108-GFP) are showing its distribution predominantly in the nucleus and partly in cytoplasm (upper panel). Fluorescence is distributed throughout the cells, transformed with vector only (35S: GFP) (middle panel). Co-localization of green (GFP) signal with blue DAPI organelle marker confirms the localization of OsPP108 in nucleus (lower panel). Detection of fluorescence was performed under a confocal laser-scanning microscope (wavelength: 488nm for GFP and 405nm for DAPI). Scale bar = 20 μm.
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pone.0125168.g003: Sub-cellular localization of OsPP108 in Nicotiana benthamiana.Agrobacterium-infiltrated tobacco leaves expressing the GFP-OsPP108 fusion protein driven by the 2XCaMV35S promoter. Confocal images of fluorescence (green) for cell expressing OsPP108 (35S:OsPP108-GFP) are showing its distribution predominantly in the nucleus and partly in cytoplasm (upper panel). Fluorescence is distributed throughout the cells, transformed with vector only (35S: GFP) (middle panel). Co-localization of green (GFP) signal with blue DAPI organelle marker confirms the localization of OsPP108 in nucleus (lower panel). Detection of fluorescence was performed under a confocal laser-scanning microscope (wavelength: 488nm for GFP and 405nm for DAPI). Scale bar = 20 μm.

Mentions: One of the key evidences for the function of a protein inside the cell is its spatial and temporal localization. To determine the sub-cellular localization of OsPP108, the coding sequence was fused with green fluorescent protein (GFP) tag and the expression was driven by 2XCaMV35S promoter. Agrobacterium carrying this construct was infiltrated into Nicotiana benthamiana epidermal peel cells to transiently express the fusion protein. Confocal microscopy analysis of Nicotiana epidermal cells revealed that OsPP108 protein predominantly resides in the nucleus with partial cytosolic distribution (Fig 3). Nuclear localization of OsPP108 was further supported by co-localization of DAPI with strong GFP signal in nucleus.


ABA inducible rice protein phosphatase 2C confers ABA insensitivity and abiotic stress tolerance in Arabidopsis.

Singh A, Jha SK, Bagri J, Pandey GK - PLoS ONE (2015)

Sub-cellular localization of OsPP108 in Nicotiana benthamiana.Agrobacterium-infiltrated tobacco leaves expressing the GFP-OsPP108 fusion protein driven by the 2XCaMV35S promoter. Confocal images of fluorescence (green) for cell expressing OsPP108 (35S:OsPP108-GFP) are showing its distribution predominantly in the nucleus and partly in cytoplasm (upper panel). Fluorescence is distributed throughout the cells, transformed with vector only (35S: GFP) (middle panel). Co-localization of green (GFP) signal with blue DAPI organelle marker confirms the localization of OsPP108 in nucleus (lower panel). Detection of fluorescence was performed under a confocal laser-scanning microscope (wavelength: 488nm for GFP and 405nm for DAPI). Scale bar = 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401787&req=5

pone.0125168.g003: Sub-cellular localization of OsPP108 in Nicotiana benthamiana.Agrobacterium-infiltrated tobacco leaves expressing the GFP-OsPP108 fusion protein driven by the 2XCaMV35S promoter. Confocal images of fluorescence (green) for cell expressing OsPP108 (35S:OsPP108-GFP) are showing its distribution predominantly in the nucleus and partly in cytoplasm (upper panel). Fluorescence is distributed throughout the cells, transformed with vector only (35S: GFP) (middle panel). Co-localization of green (GFP) signal with blue DAPI organelle marker confirms the localization of OsPP108 in nucleus (lower panel). Detection of fluorescence was performed under a confocal laser-scanning microscope (wavelength: 488nm for GFP and 405nm for DAPI). Scale bar = 20 μm.
Mentions: One of the key evidences for the function of a protein inside the cell is its spatial and temporal localization. To determine the sub-cellular localization of OsPP108, the coding sequence was fused with green fluorescent protein (GFP) tag and the expression was driven by 2XCaMV35S promoter. Agrobacterium carrying this construct was infiltrated into Nicotiana benthamiana epidermal peel cells to transiently express the fusion protein. Confocal microscopy analysis of Nicotiana epidermal cells revealed that OsPP108 protein predominantly resides in the nucleus with partial cytosolic distribution (Fig 3). Nuclear localization of OsPP108 was further supported by co-localization of DAPI with strong GFP signal in nucleus.

Bottom Line: At adult stage, OsPP108 overexpression leads to high tolerance to salt, mannitol and drought stresses with far better physiological parameters such as water loss, fresh weight, chlorophyll content and photosynthetic potential (Fv/Fm) in transgenic Arabidopsis plants.Expression profile of various stress marker genes in OsPP108 overexpressing plants revealed interplay of ABA dependent and independent pathway for abiotic stress tolerance.Overall, this study has identified a potential rice group A PP2C, which regulates ABA signaling negatively and abiotic stress signaling positively.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi, India.

ABSTRACT
Arabidopsis PP2C belonging to group A have been extensively worked out and known to negatively regulate ABA signaling. However, rice (Oryza sativa) orthologs of Arabidopsis group A PP2C are scarcely characterized functionally. We have identified a group A PP2C from rice (OsPP108), which is highly inducible under ABA, salt and drought stresses and localized predominantly in the nucleus. Genetic analysis revealed that Arabidopsis plants overexpressing OsPP108 are highly insensitive to ABA and tolerant to high salt and mannitol stresses during seed germination, root growth and overall seedling growth. At adult stage, OsPP108 overexpression leads to high tolerance to salt, mannitol and drought stresses with far better physiological parameters such as water loss, fresh weight, chlorophyll content and photosynthetic potential (Fv/Fm) in transgenic Arabidopsis plants. Expression profile of various stress marker genes in OsPP108 overexpressing plants revealed interplay of ABA dependent and independent pathway for abiotic stress tolerance. Overall, this study has identified a potential rice group A PP2C, which regulates ABA signaling negatively and abiotic stress signaling positively. Transgenic rice plants overexpressing this gene might provide an answer to the problem of low crop yield and productivity during adverse environmental conditions.

No MeSH data available.


Related in: MedlinePlus