Metabolic and Morphological Analysis of DOF1 Transgenic T2 Wheat Lines under Nitrogen Stress
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Background. Over the last century, the increased crop production has largely been attributed to rampant input of nitrogen fertilizers causing environmental deterioration. There is a need to engineer crops that require minimal fertilizer input. A transcription factor ‘Triticum aestivum Dof1 (TaDof1)’ is known to improve the nitrogen use efficiency (NUE) of crop plants, by regulating the activity of multiple genes, involved in carbon and nitrogen metabolism, when plants are grown under nitrogen-limiting conditions. Previously, transgenic wheat plants, overexpressing TaDof1, were developed.
Methods. The main objective of the current study was to compare the T2 generation of six different transgenic wheat lines transformed with TaDof1-namely, F1, G1, G2, G3, G4, and G5-with respect to their metabolic, biochemical, and morphological traits under normal and nitrogen-deficient conditions. BASTA assay and conventional PCR were used to screen out the positive plants. The expression of TaDof1 in aforementioned transgenic lines along with the four genes (Glutamine synthetase, nitrite reductase, phosphoenolpyruvate carboxylase and pyruvate kinase) associated with TaDof1 in carbon and nitrogen metabolism were quantified through RT-PCR and real-time PCR.
Results. The T2 generation of TaDof1 transgenic wheat lines overexpressed the transcription factor TaDof1 along with the other regulated genes. The expression of TaDof1 gene ranged from 0.68-7.61 folds, with the highest fold recorded in line G2. Protein, soluble sugars, phosphorous, chlorophyll, and relative water content were enhanced in almost all transgenic lines.
Conclusion. Overall, galaxy transgenic lines, specifically-G1, showed better metabolic profile as compared to Faisalabad transgenic line under nitrogen stress.
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