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Construction and utilization of plant expression vectors with PMI gene as selection marker to recover transgenic plants of Citrus sinensis L. Osbeck

Published online by Cambridge University Press:  24 April 2009

Zeng Li-Hui*
Affiliation:
College of Horticulture, Fujian Agricultural and Forestry University, Fuzhou 350002, China
Xu Hai-Feng
Affiliation:
College of Horticulture, Fujian Agricultural and Forestry University, Fuzhou 350002, China
Wang Hui-Quang
Affiliation:
College of Horticulture, Fujian Agricultural and Forestry University, Fuzhou 350002, China
Wu Shao-Hua
Affiliation:
College of Horticulture, Fujian Agricultural and Forestry University, Fuzhou 350002, China
Zhu Yi-Xuan
Affiliation:
College of Horticulture, Fujian Agricultural and Forestry University, Fuzhou 350002, China
*
*Corresponding author. E-mail: [email protected]

Abstract

The plant expression vectors pCAMBIA1301PMI and pBIPMI were constructed by substituting the Escherichia coli phosphomannose-isomerase (PMI) gene for the hpt gene of pCAMBIA1301 and gus gene of pBI121, respectively. Epicotyl explants of the Xuegan sweet orange (Citrus sinensis L. Osbeck) were inoculated with Agrobacterium tumefaciens EHA105- pCAMBIA1301PMI and EHA105-pBIPMI and subsequently selected in a medium supplemented with a combination of 25 g/l mannose and 5 g/l sucrose as the carbon source. The transformation efficiency rate was 27.7% when transformed by pCAMBIA1301PMI and 12.7% by pBIPMI. Genetic transformation was confirmed by chlorophenol red assay and polymerase chain reaction (PCR). A new method for obtaining transgenic Xuegan sweet orange plants was developed using the PMI/mannose selection system.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2009

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