Abstract
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.
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Abbreviations
- BAP:
-
6-benzylaminopurine
- CH:
-
casein hydrolysate
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- DPD:
-
Durand, Potrykus and Donn (1973) medium
- Kn:
-
kinetin
- MES:
-
2[N-morpholino]ethane-sulfolic acid
- NAA:
-
α-naphthaleneacetic acid
- PCR:
-
polymerase chain reaction
- MS:
-
Murashige and Skoog (1962)medium
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Zhang, G.N., Jia, J.F., Hao, J.G. et al. Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoides . Biol Plant 52, 373–376 (2008). https://doi.org/10.1007/s10535-008-0078-4
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DOI: https://doi.org/10.1007/s10535-008-0078-4