甘蔗胚性细胞团诱导影响因子研究

Factors affecting induction of callus and embryogenic cell cluster derived from young leaf sheath of sugarcane

以不同基因型甘蔗品种新台糖22号、新台糖16号、桂糖21号及拔地拉为供试材料,以近生长点0～3 cm的嫩叶鞘组织为外植体、MS固体培养基为基本培养基,研究诱导初期不同分化刺激培养时间和不同激素组合(2,4-D 1.0～7.0 mg/L、ABA 0～2.0 mg/L)对愈伤组织胚性细胞团诱导或分化的影响。结果表明,在MS+6-BA 1.5 mg/L+NAA 0.5mg/L培养基中对嫩叶鞘愈伤组织进行分化刺激培养24～72 h,可显著提高胚性细胞团的诱导率,以培养48 h的诱导率最高,平均诱导率为44.4%,比对照增加15.7个百分点;在不同激素组合培养基中,以在培养基MS+2,4-D 3.0 mg/L+ABA0.5 mg/L中培养的不同甘蔗品种胚性细胞团诱导率较高、生长较好,其次是MS+2,4-D 3.0 mg/L;在分化培养基MS+6-BA 1.5 mg/L+NAA 0.1 mg/L中对来自不同诱导培养基的胚性细胞团进行分化培养,结果发现在MS+2,4-D 3.0 mg/L+ABA 0.5 mg/L和MS+2,4-D 3.0 mg/L中诱导的胚性细胞团的绿苗分化率较高,不同甘蔗品种平均绿苗分化率分别为65.0%、64.9%,但绿苗生根率较低。

In the present study,different sugarcane varieties,viz.,ROC22,ROC16,GT21 and Badila were used for investigating the effects of different differentiation incentive culture time at early induction stage,and hormone combinations（2,4-D 1.0-7.0 mg/L,ABA 0-2.0 mg/L） on induction or differentiation of embryogenic cell cluster.The 0-3 cm of young leaf sheath（near to growing point） was used as explants,and solid MS was used as the basic culture medium.The induction rate of embyrogenic cell cluster derived from callus of young leaf sheath increased significantly through incentive culture for 24-72 h in medium MS ＋6-BA 1.5 mg/L ＋NAA 0.5 mg/L,the highest induction rate（44.4%） was found when the plantlets were cultured for 48 h and it increased by 15.7 percent as compared to the control.It was found that the induction rate and growth of embryogenic cell cluster in different sugarcane varieties were better when cultured in medium MS＋ 2,4-D 3.0 mg/L＋ ABA 0.5 mg/L,followed by MS＋2,4-D 3.0 mg/L.After differentiation culture in medium MS＋6-BA 1.5 mg/L＋NAA 0.1 mg/L,the differentiation rate of green plantlets derived from embryogenic cell cluster（cultured in media MS＋2,4-D 3.0 mg/L＋ ABA 0.5 mg/L and MS＋2,4-D 3.0 mg/L） was higher than that of the other media by 65.0 and 64.9% of average differentiation rate,respectively,while the rooting rate of green plantlets was found to be lower.