Biologia plantarum 55:634, 2011 | DOI: 10.1007/s10535-011-0161-0

Heterologous expression of P5CS gene in chickpea enhances salt tolerance without affecting yield

S. Kiran Kumar Ghanti1,6,*, K. G. Sujata2, B. M. Vijay Kumar3, N. Nataraja Karba2, K. Janardhan Reddy4, M. Srinath Rao1, P. B. Kavi Kishor5
1 Department of Botany, Gulbarga University, Gulbarga, India
2 Department of Crop Physiology, University of Agricultural Science, Bengaluru, India
3 Indian Institute of Science, Bengaluru, India
4 Department of Botany, Osmania University, Hyderabad, India
5 Department of Genetics, Osmania University, Hyderabad, India
6 Department of Plant Physiology, University of Agricultural Science, Bengaluru, India

Vigna Δ1-pyrroline-5-carboxylate synthetase (P5CS) cDNA was transferred to chickpea (Cicer arietinum L.) cultivar Annigeri via Agrobacterium tumefaciens mediated transformation. Following selection on hygromycin and regeneration, 60 hygromycin-resistant plants were recovered. Southern blot analysis of five fertile independent lines of T0 and T1 generation revealed single and multiple insertions of the transgene. RT-PCR and Western blot analysis of T0 and T1 progeny demonstrated that the P5CS gene is expressed and produced functional protein in chickpea. T1 transgenic lines accumulated higher amount of proline under 250 mM NaCl compared to untransformed controls. Higher accumulation of Na+ was noticed in the older leaves but negligible accumulation in seeds of T1 transgenic lines as compared to the controls. Chlorophyll stability and electrolyte leakage indicated that proline overproduction helps in alleviating salt stress in transgenic chickpea plants. The T1 transgenics lines were grown to maturity and set normal viable seeds under continuous salinity stress (250 mM) without any reduction in plant yield in terms of seed mass.

Keywords: Agrobacterium tumefaciens; Cicer arietinum; Δ1-pyrroline-5-carboxylate synthetase; NaCl; proline accumulation; RT-PCR; Southern blot; Western blot

Received: November 30, 2009; Accepted: June 1, 2010; Published: December 1, 2011Show citation

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Kiran Kumar Ghanti, S., Sujata, K.G., Kumar, B.M., Karba, N., Reddy, K., Rao, M., & Kishor, P.B. (2011). Heterologous expression of P5CS gene in chickpea enhances salt tolerance without affecting yield. Biologia plantarum, 55(4), 634. doi: 10.1007/s10535-011-0161-0.
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