biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 52:373-376, 2008 | DOI: 10.1007/s10535-008-0078-4

Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoides

G. N. Zhang1,2, J. F. Jia1,*, J. G. Hao1, X. R. Wang1, T. He1
1 School of Life Science, Northwest University, Xi'an, P.R. China
2 College of Agriculture of Henan University of Science and Technology, Luoyang, P.R. China

Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoides Pall. was here developed. The protoplasts were isolated directly from the leaves of the hairy root-induced plants. The highest yield of protoplasts was obtained from fully expanded leaves of young plants. Their viability was up to 72 ± 2.3 %. The highest division frequency (32.4 ± 0.13 %) and sustained divisions were obtained in Durand, Potrykus and Donn (DPD) medium supplemented with 2.0 mg dm-3 2,4-dichlorophenoxyacetic acid, 0.2 mg dm-3 6-benzylaminopurine, 0.3 M mannitol, 2 % sucrose and 500 mg dm-3 casein hydrolysate at the plating density of 3.0 × 105 cm-3. The frequency of shoot differentiation from protocalli reached to 91.75 ± 3.1 %. Opine synthesis and polymerase chain reaction analysis confirmed that T-DNA still existed in the protoplast regenerated plants.

Keywords: growth regulators; opine; protoplast culture; T-DNA; transformation
Subjects: Agrobacterium rhizogenes; Astragalus melilotoides; cytokinins; in vitro culture, regeneration; nutrient medium, Murashige and Skoog (MS); protoplasts; transgenic plants

Received: August 8, 2006; Accepted: March 21, 2007; Published: June 1, 2008  Show citation

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Zhang, G.N., Jia, J.F., Hao, J.G., Wang, X.R., & He, T. (2008). Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoides. Biologia plantarum, 52(2), 373-376. doi: 10.1007/s10535-008-0078-4
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