外植体的种类及消毒方式对椰枣胚性愈伤组织诱导的影响

Effects of Explant Types and Disinfection Methods on Embryo Callus Induction of Date Palm

为了研究不同外植体及消毒方式对椰枣胚性愈伤组织诱导的影响,在MS培养基的基础上,以椰枣幼苗茎尖生长组织、幼嫩叶片、5年生大树的茎尖生长组织和种胚为外植体,对这4种材料分别用75%乙醇、2%次氯酸钠、2%山农一号(Ⅰ)型组合进行消毒。研究不同消毒方式对这4种典型取材部位的消毒效果及外植体后期的胚性愈伤诱导情况。结果表明:胚性愈伤组织诱导率,种胚>大树茎尖生长点>幼苗茎尖生长点>幼嫩叶片,其中种胚的诱导率最高,达到66.67%,幼嫩叶片的诱导率最低,为26.67%;最佳消毒方案是:75%乙醇+2%次氯酸钠+2%山农一号(Ⅰ)型,种胚、幼苗茎尖生长点、大树茎尖生长点、幼嫩叶片的成活率分别为86.67%、93.33%、83.33%、70.00%。愈伤组织最佳诱导培养基:MS+3.5 g/L植物凝胶+30 g/L蔗糖+1 g/L活性炭+150 mg/L谷氨酰胺+0.3 mg/L烟酸+0.5 mg/L盐酸吡哆醇+100 mg/L肌醇+0.7 g/L磷酸二氢钾+2 mg/L6-BA+3 mg/L NAA+5 mg/L 2,4-D+1 mg/L 2-ip。综上结果,种胚、茎尖生长点部位更适合作为胚性愈伤组织诱导的外植体。

In order to study the effects of different explants and disinfection methods on embryo callus induction of date palm.On the basis of MS medium,stem tip growth tissue of seedling,young leaves,stem tip growth tissue of 5-year old tree and seed embryo were used as explants.The four materials were disinfected with a combination of 75%alcohol,2%sodium hypochlorite and 2%Shannong No.1(Ⅰ).The disinfecting effects of different disinfecting methods on the five typical sampling parts and the embryogenic callus induction in the anaphasis stage of explants were studied.The results show that the induction rate of embryogenic callus was seed embryo>stem tip growth tissue of large tree>stem tip growth tissue of seedling>young leaves.The induction rate of seed embryo was the highest,reaching 66.67%.The induction rate of young leaves was the lowest,reaching 26.67%.The best disinfection method:75%alcohol+2%sodium hypochlorite+2%Shannong No.1(Ⅰ).The survival rate of seed embryo,stem tip growth tissue of seedling、stem tip growth tissue of large tree,young leaves were respectively reached 86.67%,93.33%,83.33%,70.00%.The optimal medium for callus induction:MS+3.5 g/L phytagel+30 g/L sucrose+1 g/L AC+150 mg/L Gln+0.3 mg/L nicotinic acid+0.5 mg/L pyridoxine HCl+100 mg/L inositol+0.7 g/L KH_(2)PO4+2 mg/L 6-BA+3 mg/L NAA+5 mg/L 2,4-D+1 mg/L 2-ip.In conclusion,seed embryos and shoot tips are more suitable for embryogenic callus induction.