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Transgenic indica rice cv. ADT 43 expressing a Δ1-pyrroline-5-carboxylate synthetase (P5CS) gene from Vigna aconitifolia demonstrates salt tolerance

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Abstract

To develop salt tolerant rice, the P5CS gene of Vigna aconitifolia, encoding for proline synthesis, was introduced into the popular indica rice cultivar ADT 43. Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pCAMBIA 1301/P5CS, carrying the proline synthesis encoding gene P5CS, was co-cultivated with embryogenic callus of rice. Adding 100 μM acetosyringone to the Linsmaier and Skoog (LS) liquid and solid co-culture medium, along with 30 mg/l hygromycin and 250 mg/l timentin, contributed to significantly higher efficiency of transformation. Southern blot analysis of T1 independent transformants revealed that the copy number of transgene varied between one and three. When transgenic plants were subjected to salt stress, these plants grew well in the presence of up to 200 mM NaCl, while control plants died within 10 days under these treatment conditions. These transgenic plants grew under salt stress for a period of 4 weeks, and were capable of flowering and set seed. T1 plants segregated into 3:1 ratio suggesting Mendelian segregation pattern of inheritance of the P5CS transgene.

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Abbreviations

P5CS:

Δ1-pyrroline-5-carboxylate synthetase

BA:

N6-Benzyladenine

LS:

Linsmaier and Skoog medium

MS:

Murashige and Skoog medium

NAA:

Naphthalene acetic acid

(HPT):

Hygromycin phosphotransferase

2,4-D:

2,4- Dichlorophenoxyacetic acid

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Acknowledgements

This work was supported by a research grant from University Grants Commission, Govt. of India, New Delhi [F.No.34-251/2008(SR)]. A. Karthikeyan is grateful to University Grants Commission for the award of UGC Research Fellowship in Sciences for Meritorious Students (RFSMS) (F.4-3/2007(BSR)/11-61/2008). The authors gratefully acknowledge the use of the Bioinformatics Infrastructure facility, Alagappa University funded by the Department of Biotechnology, Ministry of Science and technology, Government of India (No. BT/BI/25/001/2006). The strains DH5α and Agrobacterium strain EHA 105 harbouring the binary plasmid pCAMBIA 1301and plasmid pUbP5CS were kindly provided by Dr. K. Veluthambi and Dr. A. K. Gupta (Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India), respectively. The strain LBA 4404 was kindly provided by Dr. P. Balasubramanian, Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India. We also thank Prof. A. Ganapathi for allowing us to perform non-radioactive Southern blot analysis and K. Subramanyam for technical support at Department of Biotechnology, Bharathidasan University, Tiruchirapalli, Tamil Nadu, India.

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  1. Department of Biotechnology, Alagappa University, Karaikudi, Tamil Nadu, 630 003, India

    Alagarsamy Karthikeyan, Shumugiah Karutha Pandian & Manikandan Ramesh

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  1. Alagarsamy Karthikeyan
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  2. Shumugiah Karutha Pandian
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Karthikeyan, A., Pandian, S.K. & Ramesh, M. Transgenic indica rice cv. ADT 43 expressing a Δ1-pyrroline-5-carboxylate synthetase (P5CS) gene from Vigna aconitifolia demonstrates salt tolerance. Plant Cell Tiss Organ Cult 107, 383–395 (2011). https://doi.org/10.1007/s11240-011-9989-4

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