大叶紫花苜蓿愈伤组织原生质体再生植株

PLANT REGENERATION FROM CALLUS PROTOPLASTS OF MEDICAGO SATIVA SSP. SATIVA

摘要: 大叶紫花苜蓿下胚轴诱导的愈伤组织在继代培养基上生长快速, 易于分散。继代第12d的愈伤组织原生质体的得率为6.5×10⁷/g鲜重。原生质体培养基为SH基本培养基, 含有1.0mg/L2, 4-0、0.5mg/LBA、2.0g/LCH、2%蔗糖、6%葡萄糖、5mmol/LMES, 培养密度为1.0×10⁵/mL.培养至第12d时的原生质体再生细胞植板率为3.7%.由原生质体形成的小愈伤组织在含2.0mg/L2, 4-D的MS固体培养基上大量增殖。增殖的愈伤组织转移至2.0mg/L2-ip+0.1mg/LNAA的B₅培养基上, 形成体细胞胚并发育成完整植株。
- 大叶紫花苜蓿 /
- 愈伤组织 /
- 原生质体培养
Abstract: Calli derived from hypocotyledon of Medicago sativa ssp. sativa grew rapidly and dispersed easily on subculture media, Protoplasts isolated from calli of l2 days after subculture at a high yield of 6.5×l0⁷/g fresh weigh. Protoplasts were cultured on modified SHmedium containing l.0mgl/L 2, 4-D, 0.5mg/L BA, 2.0g/L CH, 2%sucrose, 6% glucose, 5 mmol/L 2-(N-morpholino)-ethanesulfonic acid with plating density of 1.0×10⁵/mL. Protoplast plating efficiency was 3.7%after 12 days in culture. Microcalli derived from protoplasts multiplicated on solid MSmedium containing 2.0mg/L 2, 4-D. Somatic embryos and plant regeneration were obtained when these calli were transferred to B₅ medium containing 2.0mg/L 2-ip and 0.1mg/L NAA.
- Medicago sativa ssp. sativa /
- Callus /
- Protoplast culture