Genetic Transformation of McCHIT1 Gene into Rice Shoot Tips via Agrobacterium-mediated Method via Agrobacterium-mediated Method

doi:10.11924/j.issn.1000-6850.casb16010036

Abstract

Abstract: To improve the efficiency of genetic transformation of rice shoot tips via Agrobacterium-mediated method, a glutinous rice variety“Pingtang Heinuo”was used as the research material, the vector pCambia-Ubi-McCHIT1 with Ubi promotor driving bitter melon chitinase gene McCHIT1 was used as the transformation vector, we investigated systematically the influence of Agrobacterium concentration, vacuum treatment time and vacuum osmotic pressure on the efficiency of Agrobacterium-mediated genetic transformation of rice shoot tips by orthogonal experimental design method. The results showed that, among the three factors, the vacuum treatment time was the key factor which influenced the transformation efficiency and death rate. The optimum Agrobacterium concentration was OD600=1.0, the optimum vacuum treatment time was 7 min and the optimum vacuum osmotic pressure was 15 kPa. The histochemical staining of GUS activity and the identifying of PCR amplification confirmed that McCHIT1 gene was integrated into the rice genome. It demonstrated that we had created a novel transgenic rice germplasm with McCHIT1, and developed a high efficiency genetic transformation system for rice shoot tips by Agrobacterium-mediated method.

CLC Number:

Q785

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