Limits...
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

Germination and seedling growth analysis of OsPP108 overexpression Arabidopsis lines on salt and mannitol containing media.(A) Germination and vertical growth of WT (Col-0) and three OsPP108OX transgenic lines L1, L2 and L4 on 1/2MS (control) and different concentrations of NaCl (150 and 175mM) and mannitol (375 and 400mM) after 7 days. (B) Horizontal germination and growth pattern and seedling establishment of approximately 100 seeds from WT and transgenic lines on different MS media supplemented with different concentrations of NaCl (150 and 175mM) and mannitol (350 and 375mM) after 7 days growth. (C) Seed germination percentage of WT and three transgenic lines on 150mM, 175mM NaCl and 350mM, 375mM mannitol supplemented MS media. Germination percentage was counted for approximately 100 seeds for each genotype till 5 days of growth. Experiments were repeated three times and mean value ± SD is plotted on the graph. p-value < 0.05 was observed for all the transgenic lines (L1, L2 and L4) w.r.t. WT on different stress media, at different time points.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4401787&req=5

pone.0125168.g006: Germination and seedling growth analysis of OsPP108 overexpression Arabidopsis lines on salt and mannitol containing media.(A) Germination and vertical growth of WT (Col-0) and three OsPP108OX transgenic lines L1, L2 and L4 on 1/2MS (control) and different concentrations of NaCl (150 and 175mM) and mannitol (375 and 400mM) after 7 days. (B) Horizontal germination and growth pattern and seedling establishment of approximately 100 seeds from WT and transgenic lines on different MS media supplemented with different concentrations of NaCl (150 and 175mM) and mannitol (350 and 375mM) after 7 days growth. (C) Seed germination percentage of WT and three transgenic lines on 150mM, 175mM NaCl and 350mM, 375mM mannitol supplemented MS media. Germination percentage was counted for approximately 100 seeds for each genotype till 5 days of growth. Experiments were repeated three times and mean value ± SD is plotted on the graph. p-value < 0.05 was observed for all the transgenic lines (L1, L2 and L4) w.r.t. WT on different stress media, at different time points.

Mentions: Induction of OsPP108 transcript level under abiotic stresses such as drought and salt, and insensitivity of OsPP108OX transgenic plants towards ABA during seed germination enticed us to investigate the transgenic plant behaviour under abiotic stresses. Therefore, we performed seed germination and growth assays on MS media supplemented with NaCl and mannitol. OsPP108OX lines could germinate and survive on high salt stress (upto 175mM) and osmotic stress (upto 400mM mannitol), and hence showed tolerance to these abiotic stresses, when compared to untransformed WT (Fig 6A). Seed germination rate was much better for OsPP108OX lines than WT after 2 days of growth and more than 70% transgenic seeds could germinate on 175mM NaCl and 375mM mannitol while, ~ 45% and ~60% germination observed for WT on these stresses, respectively (Fig 6B and 6C). Analysis for cotyledon emergence after 4 days revealed that >50% transgenic seeds and only ~6% WT have emerged cotyledons on 175mM NaCl. Similarly, ~80% transgenic but only ~20% WT seeds had fully emerged cotyledon at 375mM mannitol (Fig 7A and 7B). Stress tolerance of transgenic plants was also assessed in term of fresh weight of full grown seedlings. This analysis showed that all the transgenic plants have much more weight at high salt and drought stress conditions than the WT seedlings (Fig 7C and 7D). Taken together, all these results indicated that OsPP108 overexpression confers tolerance to abiotic stresses such as salt and drought/osmotic stress in transgenic Arabidopsis.


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)

Germination and seedling growth analysis of OsPP108 overexpression Arabidopsis lines on salt and mannitol containing media.(A) Germination and vertical growth of WT (Col-0) and three OsPP108OX transgenic lines L1, L2 and L4 on 1/2MS (control) and different concentrations of NaCl (150 and 175mM) and mannitol (375 and 400mM) after 7 days. (B) Horizontal germination and growth pattern and seedling establishment of approximately 100 seeds from WT and transgenic lines on different MS media supplemented with different concentrations of NaCl (150 and 175mM) and mannitol (350 and 375mM) after 7 days growth. (C) Seed germination percentage of WT and three transgenic lines on 150mM, 175mM NaCl and 350mM, 375mM mannitol supplemented MS media. Germination percentage was counted for approximately 100 seeds for each genotype till 5 days of growth. Experiments were repeated three times and mean value ± SD is plotted on the graph. p-value < 0.05 was observed for all the transgenic lines (L1, L2 and L4) w.r.t. WT on different stress media, at different time points.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0125168.g006: Germination and seedling growth analysis of OsPP108 overexpression Arabidopsis lines on salt and mannitol containing media.(A) Germination and vertical growth of WT (Col-0) and three OsPP108OX transgenic lines L1, L2 and L4 on 1/2MS (control) and different concentrations of NaCl (150 and 175mM) and mannitol (375 and 400mM) after 7 days. (B) Horizontal germination and growth pattern and seedling establishment of approximately 100 seeds from WT and transgenic lines on different MS media supplemented with different concentrations of NaCl (150 and 175mM) and mannitol (350 and 375mM) after 7 days growth. (C) Seed germination percentage of WT and three transgenic lines on 150mM, 175mM NaCl and 350mM, 375mM mannitol supplemented MS media. Germination percentage was counted for approximately 100 seeds for each genotype till 5 days of growth. Experiments were repeated three times and mean value ± SD is plotted on the graph. p-value < 0.05 was observed for all the transgenic lines (L1, L2 and L4) w.r.t. WT on different stress media, at different time points.
Mentions: Induction of OsPP108 transcript level under abiotic stresses such as drought and salt, and insensitivity of OsPP108OX transgenic plants towards ABA during seed germination enticed us to investigate the transgenic plant behaviour under abiotic stresses. Therefore, we performed seed germination and growth assays on MS media supplemented with NaCl and mannitol. OsPP108OX lines could germinate and survive on high salt stress (upto 175mM) and osmotic stress (upto 400mM mannitol), and hence showed tolerance to these abiotic stresses, when compared to untransformed WT (Fig 6A). Seed germination rate was much better for OsPP108OX lines than WT after 2 days of growth and more than 70% transgenic seeds could germinate on 175mM NaCl and 375mM mannitol while, ~ 45% and ~60% germination observed for WT on these stresses, respectively (Fig 6B and 6C). Analysis for cotyledon emergence after 4 days revealed that >50% transgenic seeds and only ~6% WT have emerged cotyledons on 175mM NaCl. Similarly, ~80% transgenic but only ~20% WT seeds had fully emerged cotyledon at 375mM mannitol (Fig 7A and 7B). Stress tolerance of transgenic plants was also assessed in term of fresh weight of full grown seedlings. This analysis showed that all the transgenic plants have much more weight at high salt and drought stress conditions than the WT seedlings (Fig 7C and 7D). Taken together, all these results indicated that OsPP108 overexpression confers tolerance to abiotic stresses such as salt and drought/osmotic stress in transgenic Arabidopsis.

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