红掌遗传转化受体系统的建立Ⅰ离体高效培养体系的建立

Establishment of Genetic Transformation Recepient System in Anthurium andraeanumⅠEstablishment of High Efficient Culture System in Vitro

以红掌叶片和叶柄为材料,研究了影响红掌愈伤组织的诱导和分化的因素。结果表明,基因型对愈伤组织诱导有显著的影响,Pink Champion愈伤组织诱导率最高,为90%。基本培养基对愈伤组织诱导影响显著。在改良MS培养基上,愈伤组织诱导率为85%,出愈伤多,质量高。消毒时间和接种方式对愈伤组织诱导率亦有显著影响。初展开叶片用氯化汞消毒8-10 min,背面向下接入培养基,愈伤组织诱导率高。外源激素对愈伤组织的诱导和芽分化影响显著。单独使用BA不能诱导红掌叶片产生愈伤组织,高浓度BA、低浓度2,4-D时,愈伤组织诱导率高。在MS+BA0.5 mg·L-1+NAA 0.5 mg·L-1+CM 5%培养基上,芽分化率为97.5%,平均芽分化数为4.5个/块,芽粗壮。将分化出的芽转入1/2 MS+NAA 0.2 mg·L-1+AC 1 g·L-1培养基上诱导生根,生根率达100%。通过上述研究建立了红掌离体高效培养系统。

Leaf and petiole were employed to investigate factors influencing on the induction and dedifferentiation of callus in Anthurium andraeanum.The induction rates of callus among different varieties were significantly different.The callus induction of 'Pink champion'was the highest(90%).Basic medium significantly influenced the callus formation.The suitable medium for callus induction was the improved MS medium on which the callus induction rate was the highest and the callus was in high quality.Sterilization time and inoculation mode significantly influenced the induction rate of callus.The high induction rate of callus was obtained when young leaves were sterilized in a 0.1%w/v mercuric chloride solution for 8～10 min and then inoculated on improved MS medium with the back of leaves downwards.Exogenous hormones also significantly affected callus formation and proliferation.High concentration of BA and low concentration of 2,4-D promoted callus formation.The frequency of bud differentiation on MS medium with BA 0.5 mg? L^(-1)+NAA 0.5 mg·L^(-1)+ CM 5%was 97.5%,and the average amount of differentiated bud per callus was 4.5.The root differentiation rate on 1/2 MS culture medium supplemented with NAA 0.2 mg·L^(-1)+ AC 1.0 g·L^(-1) was 100%.