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Expression of Rice CYP450-Like Gene (Os08g01480) in Arabidopsis Modulates Regulatory Network Leading to Heavy Metal and Other Abiotic Stress Tolerance.

Rai A, Singh R, Shirke PA, Tripathi RD, Trivedi PK, Chakrabarty D - PLoS ONE (2015)

Bottom Line: Heterologous expression of Os08g01480 in Arabidopsis provided significant tolerance towards abiotic stresses.Transgenic lines expressing reporter gene under control of Os08g01480 promoter demonstrated differential promoter activity in different tissues during environmental stresses.These studies indicated that differential expression of Os08g01480 might be modulating response of plants towards environmental stresses as well as in different developmental stages.

View Article: PubMed Central - PubMed

Affiliation: Council of Scientific and Industrial Research-National Botanical Research Institute (CSIR-NBRI), Rana Pratap Marg, Lucknow, 226001, India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2 Rafi Marg, New Delhi, 110 001, India.

ABSTRACT
Heavy metal (HM) toxicity has become a grave problem in the world since it leads to hazardous effects on living organisms. Transcriptomic/proteomic studies in plants have identified a large number of metal-responsive gene families. Of these, cytochrome-P450 (CYPs) family members are composed of enzymes carrying out detoxification of exogenous molecules. Here, we report a CYP-like protein encoded by Os08g01480 locus in rice that helps the plant to combat HM and other abiotic stresses. To functionally characterize CYP-like gene, cDNA and promoter were isolated from rice to develop Arabidopsis transgenic lines. Heterologous expression of Os08g01480 in Arabidopsis provided significant tolerance towards abiotic stresses. In silico analysis reveals that Os08g01480 might help plants to combat environmental stress via modulating auxin metabolism. Transgenic lines expressing reporter gene under control of Os08g01480 promoter demonstrated differential promoter activity in different tissues during environmental stresses. These studies indicated that differential expression of Os08g01480 might be modulating response of plants towards environmental stresses as well as in different developmental stages.

No MeSH data available.


Related in: MedlinePlus

Promoter activity in Arabidopsis line expressing ProOs08g01480:uidA.(A) Histochemical GUS staining (B) Relative expression level of uid-A gene of transgenic seedling (10 d old), grown in ½ MS media supplemented with 5 and 25 μM As(III), 50 and 100 μM As(V), 30 and 50 μM Cd, 50 and 100 μM Cr(VI), 150 and 300 mM mannitol, 100 and 150 mM NaCl. For temperature stress seeds of transgenic line grown in ½ MS media plate and exposed to -20°C, -80°C, 37°C and 45°C for 4 h then allowed to grow in control conditions (10 d). NT represents for non treated transgenic line.
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pone.0138574.g007: Promoter activity in Arabidopsis line expressing ProOs08g01480:uidA.(A) Histochemical GUS staining (B) Relative expression level of uid-A gene of transgenic seedling (10 d old), grown in ½ MS media supplemented with 5 and 25 μM As(III), 50 and 100 μM As(V), 30 and 50 μM Cd, 50 and 100 μM Cr(VI), 150 and 300 mM mannitol, 100 and 150 mM NaCl. For temperature stress seeds of transgenic line grown in ½ MS media plate and exposed to -20°C, -80°C, 37°C and 45°C for 4 h then allowed to grow in control conditions (10 d). NT represents for non treated transgenic line.

Mentions: GUS activity was examined in transgenic seedling grown in ½ MS media supplemented with different concentration of metals. Histochemical GUS staining showed significant activity of promoter in Arabidopsis seedlings during As(III), As(V), and Cd stress at different concentrations. Interestingly, no significant activity was observed during Cr(VI) exposure in comparison to other HMs stresses at lower concentration (Fig 7a). GUS activity was observed in flowers and roots of Arabidopsis plants exposed to different HMs. While in leaf tissue, GUS activity was observed only in the plants exposed to As(III), As(V), Cd stresses. In case of Cr(VI), GUS activity was observed in leaves only in higher concentration. GUS activity was also observed in shoot of Arabidopsis during As(III), As(V), Cd and Cr(VI) exposure at higher and lower concentration (Fig L and M in S1 File). Enhanced GUS activity was also found in other abiotic stresses such as temperature, osmotic and salt stress (Fig 7a). Relative expression of uid-A gene agreed with the GUS staining patterns (Fig 7b).


Expression of Rice CYP450-Like Gene (Os08g01480) in Arabidopsis Modulates Regulatory Network Leading to Heavy Metal and Other Abiotic Stress Tolerance.

Rai A, Singh R, Shirke PA, Tripathi RD, Trivedi PK, Chakrabarty D - PLoS ONE (2015)

Promoter activity in Arabidopsis line expressing ProOs08g01480:uidA.(A) Histochemical GUS staining (B) Relative expression level of uid-A gene of transgenic seedling (10 d old), grown in ½ MS media supplemented with 5 and 25 μM As(III), 50 and 100 μM As(V), 30 and 50 μM Cd, 50 and 100 μM Cr(VI), 150 and 300 mM mannitol, 100 and 150 mM NaCl. For temperature stress seeds of transgenic line grown in ½ MS media plate and exposed to -20°C, -80°C, 37°C and 45°C for 4 h then allowed to grow in control conditions (10 d). NT represents for non treated transgenic line.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581635&req=5

pone.0138574.g007: Promoter activity in Arabidopsis line expressing ProOs08g01480:uidA.(A) Histochemical GUS staining (B) Relative expression level of uid-A gene of transgenic seedling (10 d old), grown in ½ MS media supplemented with 5 and 25 μM As(III), 50 and 100 μM As(V), 30 and 50 μM Cd, 50 and 100 μM Cr(VI), 150 and 300 mM mannitol, 100 and 150 mM NaCl. For temperature stress seeds of transgenic line grown in ½ MS media plate and exposed to -20°C, -80°C, 37°C and 45°C for 4 h then allowed to grow in control conditions (10 d). NT represents for non treated transgenic line.
Mentions: GUS activity was examined in transgenic seedling grown in ½ MS media supplemented with different concentration of metals. Histochemical GUS staining showed significant activity of promoter in Arabidopsis seedlings during As(III), As(V), and Cd stress at different concentrations. Interestingly, no significant activity was observed during Cr(VI) exposure in comparison to other HMs stresses at lower concentration (Fig 7a). GUS activity was observed in flowers and roots of Arabidopsis plants exposed to different HMs. While in leaf tissue, GUS activity was observed only in the plants exposed to As(III), As(V), Cd stresses. In case of Cr(VI), GUS activity was observed in leaves only in higher concentration. GUS activity was also observed in shoot of Arabidopsis during As(III), As(V), Cd and Cr(VI) exposure at higher and lower concentration (Fig L and M in S1 File). Enhanced GUS activity was also found in other abiotic stresses such as temperature, osmotic and salt stress (Fig 7a). Relative expression of uid-A gene agreed with the GUS staining patterns (Fig 7b).

Bottom Line: Heterologous expression of Os08g01480 in Arabidopsis provided significant tolerance towards abiotic stresses.Transgenic lines expressing reporter gene under control of Os08g01480 promoter demonstrated differential promoter activity in different tissues during environmental stresses.These studies indicated that differential expression of Os08g01480 might be modulating response of plants towards environmental stresses as well as in different developmental stages.

View Article: PubMed Central - PubMed

Affiliation: Council of Scientific and Industrial Research-National Botanical Research Institute (CSIR-NBRI), Rana Pratap Marg, Lucknow, 226001, India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2 Rafi Marg, New Delhi, 110 001, India.

ABSTRACT
Heavy metal (HM) toxicity has become a grave problem in the world since it leads to hazardous effects on living organisms. Transcriptomic/proteomic studies in plants have identified a large number of metal-responsive gene families. Of these, cytochrome-P450 (CYPs) family members are composed of enzymes carrying out detoxification of exogenous molecules. Here, we report a CYP-like protein encoded by Os08g01480 locus in rice that helps the plant to combat HM and other abiotic stresses. To functionally characterize CYP-like gene, cDNA and promoter were isolated from rice to develop Arabidopsis transgenic lines. Heterologous expression of Os08g01480 in Arabidopsis provided significant tolerance towards abiotic stresses. In silico analysis reveals that Os08g01480 might help plants to combat environmental stress via modulating auxin metabolism. Transgenic lines expressing reporter gene under control of Os08g01480 promoter demonstrated differential promoter activity in different tissues during environmental stresses. These studies indicated that differential expression of Os08g01480 might be modulating response of plants towards environmental stresses as well as in different developmental stages.

No MeSH data available.


Related in: MedlinePlus