华重楼离体胚培养及植株再生初探

In vitro embryos culture and growth of regenerated plants in Paris polyphylla var. chinensis

关键词:  胚培养, 离体幼胚, 成熟胚, 光照, 植物生长调节剂

Key words:  embryos culture, immature in vitro embryos, mature embryos, light, plant-growth regulator

摘要:

为探明华重楼离体胚培养及植株再生的基本体系,该文以华重楼离体幼胚为试验材料,以MS培养基为基本培养基,研究不同光照、不同浓度梯度组合的植物生长调节剂对华重楼离体幼胚萌发、成苗的影响。结果表明:培养到60 d时,暗培养条件下华重楼离体幼胚的生长率和萌发率分别比光培养条件下高45.25%、19.17%,故暗培养比光培养更有利于华重楼离体幼胚生长发育。当GA₃浓度相同时,离体幼胚萌发所需时间随IAA浓度增加而延长。不同浓度的GA₃都可促进离体幼胚萌发,促进作用由强到弱依次为5 mg·L^-1 GA₃>1 mg·L^-1 GA₃> 10 mg·L^-1 GA₃。在黑暗条件下,优选得到最适华重楼离体幼胚生长发育配方为1/2 MS+30 g·L^-1 蔗糖+7 g·L^-1琼脂+0.5 g·L^-1 活性炭+5 mg·L^-1GA₃+1 mg·L^-1 IAA。此配方可诱导华重楼离体幼胚在2个月左右萌发,萌发率达50%需80 d左右。在华重楼成熟胚出苗培养时发现,高浓度的植物生长调节剂会抑制华重楼成熟胚生长发育,高浓度GA₃甚至会导致华重楼成熟胚死亡。优选得到最适华重楼成熟胚出苗配方为MS+30 g·L^-1 蔗糖+6.2 mg·L^-1 硼酸+7 g·L^-1 琼脂+0.7 g·L^-1 活性炭+0.5 mg·L^-1 2,4-D+1.5 mg·L^-1 IAA + 1.5 mg·L^-1 ZT+5mg·L^-1 GA₃,诱导成熟胚出苗需43 d左右,75 d左右可形成真叶。

Abstract:

Light and plant-growth regulators have a great influence on the regeneration efficiency of somatic embryos and the growth of regenerated plants. In this study, the effects of light and plant-growth regulators on the regeneration of in vitro embryos and the growth of regenerated plants were examined in immature in vitro embryos from Paris polyphylla var. chinensis seeds. The main purpose was to explore the basic system of in vitro embryos culture and growth of regenerated plants in P. polyphylla var. chinensis. In order to study the effects of plant-growth regulators on the growth and development of immature in vitro embryos and the growth of regenerated plants, supplemented with plant-growth regulators with different kinds and concentrations to MS mediums. In order to study the effect of light on the growth and development of immature in vitro embryos of P. polyphylla var. chinensis, the immature in vitro embryos were cultured in light and dark conditions. The results showed that dark conditions were more conducive to the growth and development of immature in vitro embryos in P. polyphylla var. chinensis than light conditions. When cultured for 60 d, the growth rate and germination rate of immature in vitro embryos of P. polyphylla var. chinensis under dark culture conditions were 45.25%, 19.17% higher than those under light culture conditions, respectively. When the concentration of GA₃ was the same, the germination time of immature in vitro embryos was prolonged as the concentration of IAA increased. Different concentrations of GA₃ promoted the germination of immature in vitro embryos. The promoting effect was 5 mg·L^-1 GA₃>1 mg·L^-1 GA₃> 10 mg·L^-1 GA₃. Under dark conditions, the best formula for immature in vitro embryos of P. polyphylla var. chinensis germination was 1/2 MS + 30 g·L^-1 sucrose + 7 g·L^-1 agar + 0.5 g·L^-1 activated carbon + 5 mg·L^-1 GA₃ + 1 mg·L^-1 IAA. This induced in vitro embryo P. polyphylla var. chinensis germination in about two months. The germination rate reached 50% after about 80 d. In the mature embryos emergence experiment, a high concentration of plant-growth regulators inhibited the growth and development of mature embryos in P. polyphylla var. chinensis. A high concentration of GA₃ in particular led to the death of mature embryos. The best formula for seedlings of mature embryos was MS + 30 g·L^-1 sucrose + 6.2 mg·L^-1 boric acid + 7 g·L^-1 agar + 0.7 g·L^-1 activated carbon + 0.5 mg·L^-1 2,4-D + 1.5 mg·L^-1 IAA + 1.5 mg·L^-1 ZT + 5 mg·L^-1 GA₃. Mature embryos could produce seedlings within only 43 d. True leaves were formed after around 75 days.