REGENERATION SYSTEM AND AGROBACTERIUM-MEDIATED TRANSFORMATION OF SWEET CLOVER WITH HERBICIDE-RESISTANCE GENE

In order to set up a transformation system for sweet clover (Melilotus suaveclens Ledeb), the regeneration of shoots and roots was compared from two different explants, hypocotyls and cotyledons, the affect of MS culture medium in combination with plant growth regulators kinetin, benzyladenine, indole acetic acid, or naphthaleneacetic acid at different concentrations (0.2-8 mg/L). The percent of shoot regeneration was significantly higher on hypocotyl explants than on cotyledon explants, and increased with higher concentrations of BA. Transformation of sweet clover hypocotyl explants was conducted using Agrobacterium-mediated transformation.
The tissues were co-cultivated for two days with Agrobacterium tumefaciens disarmed strain C58 carrying a binary vector pCB 302-3, which contains bar gene conferring resistance to glufosinate herbicide.
The hypocotyl explants, regenerated shoots, regenerated roots, and all plantlets were cultured and selected at 3 mg/L glufosinate.
Successful genetic transformation was confirmed by PCR and Southern hybridization.

VI International Symposium on In Vitro Culture and Horticultural Breeding

Yanjun Liu, Jinghui Yang, Junxuan Huang, Jianke Li, Shuangyue Li, Yudong Liu, Chunxia Wu, Enqin Yang

Melilotus suaveclens Ledeb, plant growth regulators, explants, bar gene, transgenic plant, Agrobacterium tumefaciens, genetic transformation, regeneration

https://doi.org/10.17660/ActaHortic.2009.829.50