Full Length Research Paper

Characterization of a C₄ maize pyruvate orthophosphate dikinase expressed in C₃ transgenic rice plants

Jianfu Zhang^1,2,3,4, Bandyopadhyay A.³, Krisnan Sellappan³, Guoying Wang¹, Hua’an Xie², K. Datta^3,4 and Swapan K. Datta^3,4*

¹National Key Laboratory of Agrobiotechnology and college of biological sciences, China Agricultural University, Beijing, 100094, China.

²Subcenter of Fuzhou, National Rice Improvement Center of China and Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, 350018, Fujian province, China.

³Plant Breeding, Genetics and Biotechnology division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.

⁴University of Calcutta, Plant Molecular Biology and Biotechnology Lab., Botany Department, Ballygunge Circular Road, Kolkata-700019, India.

*Corresponding author. E-mail: swpndatta@yahoo.com. Tel: 91-9830668907. Fax: 91-33-2461-4848.

Accepted 26 May, 2009

   Abstract

Pyruvate orthophosphate dikinase (PPDK) is a key enzyme in plants that utilize the C₄ photosynthetic pathway to fix CO₂. The enzymatic reaction catalyzed by PPDK is critically controlled by light and is one of the rate-limiting steps of the C4 pathway. The intact maize (Zea mays) C₄-PPDK gene, containing its own promoter, terminator sequences and exon/intron structure was introduced into rice (Oryza sativa L. Indica “IR64”), a C₃ plant. Expression of C₄-PPDK in most transgenic rice lines resulted in increased CO₂ assimilation rates compared to untransformed control plants. Most of the transformants showed higher photosynthetic activities than that of wild-type plant. Total nitrogen in the flag leaves of C₄-PPDK transgenics was analyzed. Results showed an increase in total nitrogen compared to untransformed control plants suggesting that C₄-PPDK expression in rice promoted nitrogen absorption from the soil. In addition, the photosynthesis rate of some transgenic IR64 lines was also increased in the greenhouse. Molecular analysis revealed that the intact PPDK gene integrated in the rice genome and affected the phenotypes of plants particularly tillers and enhanced yield of transgenic IR64 rice plants in the greenhouse.

Key words: Indica rice IR64, Maize (Zea mays), photosynthesis, PPDK (pyruvate orthophosphate dikinase), C4 rice.