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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

Expression profile of stress marker genes.qPCR analysis was done to generate expression profile of different stress marker genes such as RD29A, RD29B, RAB18 and KIN1 in WT (Col-0) and two OsPP108 overexpression lines L1 and L2 after ABA (50μM), salt (300mM) and mannitol (400mM) stress treatment. Various time points of stress treatment are indicated at X-axis and relative expression value (fold change) is indicated on Y-axis. Data from mean of two replicates is presented as columns and standard deviation among the samples is denoted by error bar. *p-value < 0.05 and **p-value- < 0.01 indicate statistically significant expression change of transgenic lines w.r.t. respective WT in different treatment and different time points.
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pone.0125168.g008: Expression profile of stress marker genes.qPCR analysis was done to generate expression profile of different stress marker genes such as RD29A, RD29B, RAB18 and KIN1 in WT (Col-0) and two OsPP108 overexpression lines L1 and L2 after ABA (50μM), salt (300mM) and mannitol (400mM) stress treatment. Various time points of stress treatment are indicated at X-axis and relative expression value (fold change) is indicated on Y-axis. Data from mean of two replicates is presented as columns and standard deviation among the samples is denoted by error bar. *p-value < 0.05 and **p-value- < 0.01 indicate statistically significant expression change of transgenic lines w.r.t. respective WT in different treatment and different time points.

Mentions: One of the important parameters to assess the response to stress is by monitoring the expression pattern of crucial stress marker genes. Since OsPP108OX plants exhibited enhanced ABA and abiotic stress tolerance, it is quite imperative to analyse the expression level of various stress marker genes in WT and transgenic plants under normal and stress conditions. Therefore, we performed qPCR based expression analysis for selected and well established stress marker genes, including RD29A, RD29B, RAB18 and KIN1, using RNA from 3 week old Arabidopsis plants grown on MS media, and treated with different stresses (NaCl, mannitol and ABA) for 0hrs, 6hrs and 12hrs time periods. We observed that transcript level of most of these stress marker genes was higher in OsPP108OX plants than WT even without any stress (Fig 8). Interestingly, RD29A was found to be induced at several fold higher level in WT than in OsPP108OX transgenic plants, under exogenous ABA treatment. It expressed at almost similar level in WT and transgenic plants under salt stress, however expression level was slightly higher in transgenic plants under mannitol treatment. Expression level of RD29B was significantly higher in OsPP108OX lines under ABA, salt and mannitol stresses. Notably, expression level of RAB18 was found to decline more in transgenic plants than in WT plants under ABA, salt and mannitol stresses. Expression level of KIN1 was consistently higher in transgenic lines under ABA and mannitol treatments, and during salt stress induction was higher in OsPP108OX lines than WT after 6 hrs but higher expression observed in WT than OsPP108OX lines after 12hrs of treatment. Overall, overexpression of OsPP108 in Arabidopsis plants enhanced the expression level of stress responsive genes without ABA and stress treatment. But under ABA and stress conditions, variable expression pattern was observed, especially for ABA related genes (RD29A and RAB18), which might be contributing to stress adaptation in overexpressing transgenic plants in ABA dependent or independent manner.


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)

Expression profile of stress marker genes.qPCR analysis was done to generate expression profile of different stress marker genes such as RD29A, RD29B, RAB18 and KIN1 in WT (Col-0) and two OsPP108 overexpression lines L1 and L2 after ABA (50μM), salt (300mM) and mannitol (400mM) stress treatment. Various time points of stress treatment are indicated at X-axis and relative expression value (fold change) is indicated on Y-axis. Data from mean of two replicates is presented as columns and standard deviation among the samples is denoted by error bar. *p-value < 0.05 and **p-value- < 0.01 indicate statistically significant expression change of transgenic lines w.r.t. respective WT in different treatment and different time points.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0125168.g008: Expression profile of stress marker genes.qPCR analysis was done to generate expression profile of different stress marker genes such as RD29A, RD29B, RAB18 and KIN1 in WT (Col-0) and two OsPP108 overexpression lines L1 and L2 after ABA (50μM), salt (300mM) and mannitol (400mM) stress treatment. Various time points of stress treatment are indicated at X-axis and relative expression value (fold change) is indicated on Y-axis. Data from mean of two replicates is presented as columns and standard deviation among the samples is denoted by error bar. *p-value < 0.05 and **p-value- < 0.01 indicate statistically significant expression change of transgenic lines w.r.t. respective WT in different treatment and different time points.
Mentions: One of the important parameters to assess the response to stress is by monitoring the expression pattern of crucial stress marker genes. Since OsPP108OX plants exhibited enhanced ABA and abiotic stress tolerance, it is quite imperative to analyse the expression level of various stress marker genes in WT and transgenic plants under normal and stress conditions. Therefore, we performed qPCR based expression analysis for selected and well established stress marker genes, including RD29A, RD29B, RAB18 and KIN1, using RNA from 3 week old Arabidopsis plants grown on MS media, and treated with different stresses (NaCl, mannitol and ABA) for 0hrs, 6hrs and 12hrs time periods. We observed that transcript level of most of these stress marker genes was higher in OsPP108OX plants than WT even without any stress (Fig 8). Interestingly, RD29A was found to be induced at several fold higher level in WT than in OsPP108OX transgenic plants, under exogenous ABA treatment. It expressed at almost similar level in WT and transgenic plants under salt stress, however expression level was slightly higher in transgenic plants under mannitol treatment. Expression level of RD29B was significantly higher in OsPP108OX lines under ABA, salt and mannitol stresses. Notably, expression level of RAB18 was found to decline more in transgenic plants than in WT plants under ABA, salt and mannitol stresses. Expression level of KIN1 was consistently higher in transgenic lines under ABA and mannitol treatments, and during salt stress induction was higher in OsPP108OX lines than WT after 6 hrs but higher expression observed in WT than OsPP108OX lines after 12hrs of treatment. Overall, overexpression of OsPP108 in Arabidopsis plants enhanced the expression level of stress responsive genes without ABA and stress treatment. But under ABA and stress conditions, variable expression pattern was observed, especially for ABA related genes (RD29A and RAB18), which might be contributing to stress adaptation in overexpressing transgenic plants in ABA dependent or independent manner.

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