EXPRESSION OF PORPHYRA YEZOENSIS TPS GENE IN TRANSGENIC RICE ENHANCED THE SALT TOLENRANCE

Bao-Tai Guo, Bin Wang, Man-Li Weng, Li-Xian Qiao, Yan-Bin Feng, Li Wang, Peng-Yan Zhang, Xiu-Liang Wang, Jiong-Ming Sui, Tao Liu, De-Lin Duan, Bin Wang

Abstract


The trehalose-6-phosphate synthase gene of Porphyra yezoensis (PyTPS) was isolated and cloned into a plant gene expression vector pCAMBIA2300-35S-OCS, and the resulting construct pCAMBIA2300-PyTPS was transformed into Agrobacterium tumefaciens (A. tumefaciens ) strain AGL1. Genetic transformation of rice variety TP309 was performed with the A. tumefaciens containing pCAMBIA2300-PyTPS. After antibiotic G418 screening and PCR analysis, one hundred T0 transgenic plants were seclected and transplanted into the trial field in the greenhouse and used for further study. Ninety-five of these 100 T0 transgenic cultivaries produced their seeds, which were harvested and stored separately. All of the 95 potential T1 transgenic lines were re-identified by PCR analysis, and their salt-tolerance was tested with 3‰ and 5‰ NaCl solutions. Results indicated that 78 of the 95 T1 transgenic lines were PCR- positive and resistant to 5‰ NaCl solution. Salt-tolerance of these 78 T1 transgenic lines was further tested with higher concentration of NaCl solutions. Of which, three lines (H155, H191 and Y308) showed resistance to 8‰ NaCl in the test. These 3 lines were comprehensively analyzed by PCR, Southern hybridization, northern hybridization and RT-PCR analyses. In addition, trehalose content measurement and preliminary yield evaluation were carried out, results indicated that the PyTPS gene was integrated into the genomic DNA sequences of these 3 transgenic lines and expressed indeed in the transgenic plants. Detection of the transformed PyTPS gene in these 3 transgenic lines was performed in plants from T1 to T6 generations; results indicated that the transformed PyTPS gene was present in transgenic plants from T1 to T6 generations.


Keywords


Agrobacterium-mediated transformation, Porphyra yezoensis Salt-tolerance, Transgenic rice, Trehalose-6- phosphate synthase (TPS)

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References


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Copyright (c) 2014 Bao-Tai Guo, Bin Wang, Man-Li Weng, Li-Xian Qiao, Yan-Bin Feng, Li Wang, Peng-Yan Zhang, Xiu-Liang Wang, Jiong-Ming Sui, Tao Liu, De-Lin Duan, Bin Wang

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Journal of Plant Breeding and Genetics
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