Genetic engineering of indica rice with AtDREB1A gene for enhanced abiotic stress tolerance
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Drought and cold stresses are major abiotic stresses that affect rice productivity. Therefore, enhancing tolerance to these stresses is necessary for sustaining rice productivity. Overexpression of the AtDREB1A gene has been shown to confer tolerance to both drought and cold stresses in diverse plant species. Hence, we genetically engineered indica rice cv. Pusa Sugandh 2 with AtDREB1A gene under the transcriptional control of stress responsive AtRD29A promoter. The transformants were confirmed for the stable integration of transgene in the rice genome by using PCR, RT-PCR and Southern blot analyses. Two single copy transgenic events (T3) and non-transgenic (NT) plants grown in pots were subjected to drought and cold stresses for 14 days. Transgenic plants exhibited enhanced tolerance to both drought and cold stresses as compared with NT plants. Transgenic plants maintained significantly higher leaf relative water content (LRWC), chlorophyll content, total sugars and proteins, but lower canopy temperature as compared with NT plants. Microarray analysis of AtDREB1A transgenic rice line (TL4) subjected to drought stress at reproductive stage led to the identification of 256 differentially expressed genes (DEGs), of which 201 were upregulated under drought stress. Interestingly, ~ 38% of the up-regulated genes coded proteins for chloroplast structure and function, which is a unique finding of this study. About 47% of the DEGs were enriched with CRT/DRE cis-regulatory elements in their promoters. Of these CRT/DRE cis-element containing genes, 50% genes coded for chloroplast function suggesting that these genes might be the direct targets of DREB1A TF. Up-regulation of TFs ZFP179 and NF-YC1, which are positive regulators of stress tolerance and downregulated TFs HOX22 and OsNAP, which are negative regulators of stress tolerance might have contributed to the enhanced drought tolerance of AtDREB1A transgenic plants. In addition, AtDREB1A-mediated orchestration of genes for chloroplast function appeared to have played an important role in not only providing carbon requirements of plants for survival and growth, but also helped minimize photo-inhibition and ROS accumulation in chloroplast under drought stress. Thus, stress-inducible overexpression of AtDREB1A in rice is a useful strategy to enhance drought and cold tolerance in the major staple food crop of the world.
KeywordsAbiotic stress AtDREB1A Cold Drought Indica rice RD29A promoter
Pusa Sugandh 2
Dehydration responsive elements binding factor
Differentially expressed genes
Revese transcription polymerase chain reaction
The whole work was carried out at ICAR-NRCPB, IARI, New Delhi and funded by “ICAR Network Project on Transgenics in Crops”. The authors are thankful to Project Director, NRCPB, for all support and the laboratory facility for this work. National Phytotron Facility, ICAR-IARI, New Delhi, India is duly acknowledged for providing the facility to grow and phenotypes transgenics.
GML generated transgenic rice plants, conducted molecular and physiological experiments and drafted the manuscript, KVR assisted in transgenic development and molecular analysis, JML helped in generation and analysis of microarray data, DP participated in molecular data analysis, AKS took part in phenotyping, KCB and VC analyzed the data and helped in manuscript writing, KRSSR participated in work supervision and manuscript preparation and TM conceived the study, supervised the work and edited the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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