Isolation and characterization of drought and ABA responsive promoter of a transcription factor encoding gene from rice

Vaibhav Kumar; Amresh Kumar; Kalpana Tewari; Nitin Kumar Garg; Sushil S Changan; Aruna Tyagi

doi:10.1007/s12298-022-01246-9

Isolation and characterization of drought and ABA responsive promoter of a transcription factor encoding gene from rice

Physiol Mol Biol Plants. 2022 Oct;28(10):1813-1831. doi: 10.1007/s12298-022-01246-9. Epub 2022 Nov 3.

Authors

Vaibhav Kumar^{1

2}, Amresh Kumar^{1

3}, Kalpana Tewari^{1

2}, Nitin Kumar Garg^{1

4}, Sushil S Changan^{1

5}, Aruna Tyagi¹

Affiliations

¹ Division of Biochemistry, Indian Council of Agricultural Research-Indian Agricultural Research Institute, New Delhi, India.
² Basic Science Division, Indian Council of Agricultural Research-Indian Institute of Pulses Research, Kanpur, Uttar Pradesh India.
³ Indian Council of Agricultural Research-National Institute for Plant Biotechnology, New Delhi, India.
⁴ Rajasthan Agricultural Research Institute (SKNAU Jobner), Durgapura, Jaipur India.
⁵ Division of CPB and PHT, Indian Council of Agricultural Research-Central Potato Research Institute, Shimla, India.

Abstract

Water deficit is a significant impediment to enhancing rice yield. Genetic engineering tools have enabled agriculture researchers to develop drought-tolerant cultivars of rice. A common strategy to achieve this involves expressing drought-tolerant genes driven by constitutive promoters such as CaMV35S. However, the use of constitutive promoters is often limited by the adverse effects it has on the growth and development of the plant. Additionally, it has been observed that monocot-derived promoters are more successful in driving gene expression in monocots than in dicots. Substitution of constitutive promoters with stress-inducible promoters is the currently used strategy to overcome this limitation. In the present study, a 1514 bp AP2/ERF promoter that drives the expression of a transcription factor was cloned and characterized from drought-tolerant Indian rice genotype N22. The AP2/ERF promoter was fused to the GUS gene (uidA) and transformed in Arabidopsis and rice plants. Histochemical GUS staining of transgenic Arabidopsis plants showed AP2/ERF promoter activity in roots, stems, and leaves. Water deficit stress and ABA upregulate promoter activity in transformed Arabidopsis and rice. Quantitative PCR for uidA expression confirmed induced GUS activity in Arabidopsis and rice. This study showed that water deficit inducible Os-AP2/ERF-N22 promoter can be used to overcome the limitations of constitutive promoters. Transformants overexpressing Os-AP2/ERF-N22 showed higher relative water content, membrane stability index, total chlorophyll content, chlorophyll stability index, wax content, osmotic potential, stomatal conductance, transpiration rate, photosynthetic rate and radical scavenging activity. Drought tolerant (N22) showed higher expression of Os-AP2/ERF-N22 than the susceptible (MTU1010) cultivar.

Supplementary information: The online version contains supplementary material available at 10.1007/s12298-022-01246-9.

Keywords: Abiotic stress; Agrobacterium; Arabidopsis; Transgenic; Water deficit stress.

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