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Spatial distribution pattern analysis of Dof1 transcription factor in different tissues of three Eleusine coracana genotypes differing in their grain colour, yield and photosynthetic efficiency

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Abstract

In the present study Dof1 gene of finger millet was cloned and sequenced. In silico analysis reveals 61% identity with the Sorghum bicolor and 57% identity with the Oryza sativa Dof1 sequence. A comparative analysis of gene sequences from different crops and three finger millet genotypes {Brown (PRM-1), Golden (PRM-701) and White (PRM-801)} differing in grain colour, yield and photosynthetic efficiency showed a high degree of sequence identity of Dof1 sequence gene ranging from 22 to 70% as evident from distance matrix of the built phylogenetic tree showing two major clusters. A total of five conserved motifs were observed in Dof1 sequences of different cereals. Motif 1 with multilevel consensus sequence CKNCRRYWTKGGAMRNVPVG contains zinc finger Dof domain. Motif 3 and motif 5 contains protein kinase phosphorylation site. Motif 2 contains Dof domain and zinc finger N-glycosylation site while motif 4 is involved in Zinc finger type profiling. Further, we studied the spatial distribution of Dof1 gene in three vegetative tissues (root, stem and flag leaf) as well as four stages of developing spikes (S1, S2, S3 and S4) of the three finger millet genotypes using qualitative and quantitative PCR based approaches. Physiological parameters (plant height, leaf area, chlorophyll content, SPAD value and photosynthetic efficiency) at the time of flowering was found to be highest in white (PRM-801) genotype followed by golden (PRM-701) and brown (PRM-1) genotype. Semi-quantitative RT-PCR and quantitative real-time PCR analysis revealed that the expression of Dof1 is highest in leaves and lowest in roots, which suggests its role in regulation of photosynthesis-related genes and carbon skeleton synthesis. Also at grain maturity stage, expression of Dof1 was higher in white (PRM-801) genotype followed by golden (PRM-701) and brown (PRM-1) genotype. The result is suggestive of Dof1 role in the accumulation of grain protein and yield attribute through regulation of key enzymes involved in source to sink relationship during grain filling stage.

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Abbreviations

Dof:

DNA binding with one finger only

RT PCR:

Reverse transcriptase polymerase chain reaction

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Acknowledgments

The authors wish to acknowledge the Department of Biotechnology, Govt. of India for providing financial support in the form of Programme Support for research and development in Agricultural Biotechnology at G.B. Pant University of Agriculture and Technology, Pantnagar (Grant No. BT/PR7849/AGR/02/374/2006). Nidhi Gupta’s and Atul Kumar Gupta’s work was supported by Junior Research Fellowship from the DBT and DST. Authors gratefully acknowledge the support provided from Bioinformatics Sub-DIC, Pantnagar and Ms. Anshita Goel, for in silico analysis. The support provided by Dean, College of Basic Sciences and Humanities and Director, Experiment Station, G.B. Pant University of Agriculture and Technology, Pantnagar is also thankfully acknowledged.

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  1. Department of Molecular Biology & Genetic Engineering, College of Basic Sciences & Humanities, G. B. Pant University of Agriculture & Technology, Pantnagar, U.S. Nagar, 263 145, India

    Nidhi Gupta, Atul Kumar Gupta & Anil Kumar

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  1. Nidhi Gupta
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  2. Atul Kumar Gupta
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  3. Anil Kumar
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Correspondence to Anil Kumar.

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Gupta, N., Gupta, A.K. & Kumar, A. Spatial distribution pattern analysis of Dof1 transcription factor in different tissues of three Eleusine coracana genotypes differing in their grain colour, yield and photosynthetic efficiency. Mol Biol Rep 39, 2089–2095 (2012). https://doi.org/10.1007/s11033-011-0956-2

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