Study On Tissue Culture And Establishment Of Regeneration System Of Salix Chaenomeloides ’Variegata’

salt and alkali. It has certain value on wetland restoration and landscaping. With tissue culture, S. chaenomeloides’Variegata’is in rapid multiplication without seasonal restriction. Establishment of regeneration system with callus to provide technical support for transfer insect-resistant genes to Salix chaenomeloides’Variegata’. Main results are listed as follows:Micropropagation with nodal segments: ① Suitable disinfection and sterilization method: The stems were collected from May to July, which was surface sterilized with 75% etha nol for 30-35s, followed by 15min in 2% NaClO; ②The optimum medium for lateral bud induction is WPM supplemented without any plant growth regulators; ③The optimum me dium for shoot multiplication is:WPM+6-BA1.0mg·L-1+NAA0.1mg·L-1, The optimum medi um for shoot elongation is:WPM+6-BA0.5mg·L-1+NAA0.1mg·L-1+active carbon1.0g·L-1, A dding active carbon, combined with using 1.Omg·L-16-BA and 0.5mg·L-16-BA alternately, w hich can maintain a high multiplication value, shoot elongation without apical necrosis, vit rification and browning. The appropriate subculture cycle is about 30d; ④The optimum m edium for plantlet rooting is:1/2WPM+NAA0.05mg·L-1+active carbon1.0g·L-1; ⑤ The opti mum time of opening sealing membrane and smelting plants is 5 days; Suitable transplanti ng medium is peat and perlite volume ratio 1:1.Establishment of regeneration system with callus: ① Suitable disinfection and sterilization method:The leave were collected from February to July, which was surface sterilized with 75% ethanol for 30-35s, followed by 10min in 2% NaClO; ②The optimum medium for callus induction is:MS+6-BA1.0mg·L-1+NAA0.5mg·L-1,7days for dark treatment; ③ The optimum medium for callus proliferation is:MS+6-BA1.0mg·L-1+NAA0.05mg·L-1, The appropriate subculture cycle is about 30d; ④ The optimum medium for callus differentiation is: MS+6-BA4.0mg·L-1.