外植体采集时期与冷藏处理对马尾松愈伤组织诱导的影响  

Effects of Explant Collecting Time and Storage Duration on Callus Induction of Pinus massoniana

作　　者：伊书亮[1] 张冬林[1,2] 杨模华[1] 李志辉[1] 王茜[1] 史昆[1] 

机构地区：[1]中南林业科技大学林学院,湖南长沙410004 [2]乔治亚大学园艺系,美国乔治亚州雅典市30602

出　　处：《广西林业科学》2013年第1期8-13,共6页Guangxi Forestry Science

基　　金：教育部博士点基金博导类联合资助项目(20104321110002)

摘　　要：马尾松(Pinus massoniana)无性繁殖困难,研究常采用体细胞胚胎发生和器官发生的方式繁殖,外植体采集时期是实验成功的关键。本实验采用2011年10月至2012年10月期间(每周或每两周采集1次)采集的马尾松枝梢顶端分生组织为外植体,并将外植体置于4℃冰箱冷藏做不同冷藏时间处理(4～114天)。处理后的外植体接种在添加不同激素浓度的改良GD培养基中,24～25℃条件下暗培养。实验结果表明:冷藏时间对马尾松愈伤组织诱导没有显著影响;2012年3月中旬至4月底采集的马尾松外植体,愈伤组织诱导率最高达89.8%,其平均诱导率达到80%以上;外植体经流水冲洗2～3 h、75%的酒精处理20～30 s、0.1%的升汞处理9～10 min并去除鳞片和针叶,能有效降低外植体污染率;外植体经流水冲洗2～3 h、去除鳞片和针叶后切成长约0.2～0.5 cm的薄片后接种,能大大降低培养过程中外植体的褐化率。Pinus massoniana （ masson pine ） is heard to be propagated vegetatively and somatic embry- ogenesis and organogenesis are usually applied by researchers. However, when to collect explants is the key to successfully micro-propagated it. During October 2011 to October 2012, shoot apical meristem of Pinus massoniana was collected monthly, bi-weekly, or weekly and refrigerated at 4℃ for 4 to 114 days. Explants were inoculated onto improved GD medium which contained various hormones, then cultured in the dark condition at 24 - 25 ℃. The findings were that ： Storage duration had no significant influence on callus induction of Pinus massoniana; Explants collected from middle of March 2012 to late April 2012, had a highest callus induction rate of 89.8%, and the average rate was above 80% ; The procedures of flushing in pure water 2 - 3 h, peeling off the scales, removing needles, rinsing in 75 % alcohol for 2030 s, and soaking in 0. 1% of mercuric chloride for 9 - 10 rain, could effectively solve the pollution problem in the early period ; Cleaned explants significantly reduced browning after sling into 0. 2 -0. 5 cm discs for all experiments.

关 键 词：马尾松 组织培养 愈伤组织 冷藏处理 采集时期 

分 类 号：S791.248[农业科学—林木遗传育种]