以花曲柳合子胚的单片子叶为外植体成功诱导出体胚并获得再生植株。未成熟合子胚的子叶在添加400 mg·L-1水解酪蛋白、0.25 mg·L-1 6-BA、1.5 mg·L- 1 NAA、70 g·L-1蔗糖和6 g·L-1琼脂的MS1/2培养基上可以成功诱导产生体胚,诱导率达到34.7%,每个外植体上体胚数量为2~9个。成熟合子胚的子叶在添加0.25 mg·L-1 6-BA、2 mg·L- 1 NAA的MS1/2培养基(其他成分同上)上可以成功诱导产生体胚,诱导率为10.0%。体胚在MS1/2培养基上经过成熟培养后可以正常萌发,萌发率87.6%。萌发的体胚植株在MS1/2+0.01 mg·L-1NAA培养基上生长较好,具备实生幼苗的外观特征。经炼苗后的体胚苗移植到栽培基质(草炭土 ∶蛭石 ∶珍珠岩体积比为5 ∶4 ∶1)中可以正常生长,成活率为75.0%。
Somatic embryogenesis and plant regeneration of Fraxinus rhynchophylla were obtained by using single cotyledons of zygotic embryos as explants. Cotyledons of immature zygotic embryos, cultured on half strength MS medium supplemented with 400 mg·L-1 hydrolyzed casein, 70 g·L-1 sugar, 6 g·L-1 agar, 0.25 mg·L-1 6-BA and 1.5 mg·L-1 NAA, were able to be successfully induced somatic embryos. The somatic embryo (SE) induction rate could reach 34.7% and about 2-9 SEs were produced per each explant. However, cotyledons from mature zygotic embryos, culture on a similar medium with modification of adding extra 0.5 mg·L-1 NAA, and the SE induction rate could reach 10.0%. Somatic embryos after in vitro maturation on a MS1/2 medium could normally germinate, and the germination percentage rate reached to 87.6%. The germinated somatic plantlets grew well on MS1/2 medium containing 0.01 mg·L-1 NAA and had the similar morphological characteristics of zygotic seedlings. Acclimated somatic plantlets grew normally after transplanted in a medium with 50% peat moss, 40% vermiculite and 10% polite, with 75.0% survival rate.