马尾松幼胚离体培养器官发生植株再生研究

Micropropagation in immature embryos of Pinus massoniana in vitro

以马尾松幼胚为外植体,经丛生芽诱导、增殖和伸长培养、生根诱导,获得了再生植株。对影响马尾松幼胚器官发生植株再生的关键因子,如减轻外植体褐变的处理方式、基本培养基类型、最适诱导培养基激素种类和浓度、增殖培养条件以及生根诱导影响因子(激素、活性炭和蔗糖)等,分别进行了随机区组试验设计。试验数据经SPSS16方差分析,LSD多重比较,结果表明:1)选择WPM培养基,添加抗坏血酸Vc 0.15 g/L,并且每2天转接1次能有效地减轻外植体接种处理初期出现的褐化现象;2)WPM+NAA 0.15 mg/L+6—BA 1.00 mg/L+KT 0.50 mg/L为较好的马尾松幼胚丛芽诱导分化培养基,丛芽诱导率可达98.15%;3)马尾松幼胚丛芽增殖和伸长培养的最适培养基为WPM+NAA 0.15 mg/L,增殖系数可达5.43;4)生根培养中,生根质量最好的培养基组合为WPM+IBA2.0 mg/L+NAA 0.15 mg/L+Ac 0.50 g/L,且生根培养基中添加的蔗糖浓度以20.0 g/L为宜,生根率达到66.67%。本研究结果可为利用马尾松幼胚建立组培快繁体系奠定研究基础。

An in vitro procedure for micropropagation in immature embryos of Pinus massoniana,consisting of the following four steps has been established： adventitious shoot induction,adventitious shoot growth,root induction,and root growth.Influence of certain selected factors on each of these steps was determined with random block experiments,including treatments on controlling brown,basal medium（GD,DCR and WPM）,plant hormones（6-BA,NAA and IBA）,active carbon（Ac） and sucrose.SPSS 16.0 statistics software were used to analyze those data.Results were as followed.1） For controlling brown,WPM containing ascorbic acid（0.15g/L） and subculture the explant every 2days were suitable.2） Maximum adventitious shoot initiation and growth（98.15% at the 4th weeks and 5.43 times at the 6th weeks） were obtained on WPM with NAA（0.15mg/L）,6-BA（1.00mg/L） and KT（0.50mg/L）.3） Adventitious shoot elongation（2.5cm at the 3th weeks） was achieved on WPM medium with NAA（0.15 mg/L）.4） Four rooting experiments involving IBA,NAA,sucrose and Ac concentration were carried out.Best root induction and growth occurred when shoots were put on a medium of WPM containing 2% sucrose with IBA（2.0mg/L）,NAA（0.15mg/L） and Ac（0.50g/L）,the rooting rate reached 66.67%.These findings provide foundation for setting up a tissue culture and rapid propagation system with immature embryos of Pinus massoniana.