biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 53:578-582, 2009 | DOI: 10.1007/s10535-009-0105-0

Regeneration of transgenic citrus plants from the trimmed shoot/root region of etiolated seedlings

D. L. Li1, B. Tan1, Y. X. Duan1, W. W. Guo1,*
1 National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, Wuhan, P.R. China

Transformation and high efficient regeneration of transgenic plants from the trimmed etiolated shoot/root region (TESRR) of Anliucheng sweet orange [Citrus sinensis (L.) Osb.] seedling was reported. A visual green fluorescent protein (GFP) marker gene was introduced to evaluate transformation efficiency by using the explants from TESRR and epicotyls. The transformation protocol was: infection 20 min, co-culture 3 d, selection culture 30 d, and rooting 15 d. Out of a total of 288 sprouted shoots obtained from TESRR, 34 shoots (11.8 %) yielded GFP expression. In contrast, only 2 (3.0 %) of the 67 sprouted shoots from epicotyl transformation yielded GFP expression. In all plants showing the green fluorescence an expected 500 bp GFP fragment was proved by PCR analysis. Southern blot analysis further confirmed the integration of GFP gene into citrus genome. Transgenic plantlets were obtained within 80 d using the TESRR, compared within 150 d by using epicotyls.

Keywords: Agrobacterium tumefaciens; sweet orange; GFP; plantlet regeneration; transformation
Subjects: Agrobacterium tumefaciens; Citrus sinensis; ex vitro transfer; green fluorescent protein (GFP); in vitro culture, direct regeneration; Southern blot analysis; sweet orange; transgenic plants

Received: September 28, 2007; Accepted: February 26, 2008; Published: September 1, 2009  Show citation

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Li, D.L., Tan, B., Duan, Y.X., & Guo, W.W. (2009). Regeneration of transgenic citrus plants from the trimmed shoot/root region of etiolated seedlings. Biologia plantarum, 53(3), 578-582. doi: 10.1007/s10535-009-0105-0
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