| 摘要: |
| 红锥（Castanopsis hystrix ）为壳斗科栲属常绿乔木，种皮坚硬、渗透性强，这是阻碍以种子
为外植体启动组织培养的主要原因之一。为成功建立红锥组培快繁体系，系统研究了红锥种子灭菌方案、
种子萌发与丛生芽诱导培养基。结果表明，红锥种子最佳的灭菌方案为：先将带种皮种子用 75% 酒精浸
泡 40 s，用无菌水冲洗 1 次，然后用 0.1% 升汞浸泡 12 min，无菌水冲洗 5~6 次，接着剥除种皮，再用
0.1% 升汞浸泡 6 min，无菌水冲洗 5~6 次，污染率最低，为 26.7%，发芽率最高，为 73.3%；较好的萌发
培养基为改良 MS+GA3 3.0 mg · L-1，接种后第 2 天种子即开始萌动，萌发率 93.3%，生长旺盛；较好的丛
芽诱导培养基为改良 MS +6-BA2.0~3.0 mg · L-1+NAA 0.2 mg · L-1+GA3 3.0 mg · L-1, 丛芽数量平均 6.01 个。
外植体诱导效果与灭菌方案、基本培养基、培养基中激素种类和浓度等有显著的关系。较低盐浓度的改
良 MS 培养基、较高水平的 6-BA 有利于丛芽的诱导。 |
| 关键词: 红锥 种子 灭菌方案 初代培养 |
| DOI： |
| 投稿时间：2018-10-13修订日期：2018-11-09 |
| 基金项目:广东省省级科技计划项目（项目编号：2016B020233001）；广东省林业科技创新专项资金项目（项目编号：2014KJCX002） |
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| Sterilization and Primary Culture of Seeds from Castanopsis hystrix |
| wuqiaona,liruiping,guomengqing,liaohuanqin,dengxiaomei |
| (College of Forestry and Landscape Architecture, South China Agricultural University;Guangdong Academy Of Forestry) |
| Abstract: |
| Castanopsis hystrix is a genus of superior, valuable evergreen trees. Its seed coat were of
strong permeability but very hard, which leads to difficulty in primary culture of seeds. Disinfection procedure,
germination and shoot-inducing medium were systematically studied. The results showed that the best disinfection
procedure for C. hystrix seed was that, the seeds with the outer seed coat and subsequent tissue were disinfected
with 75% (v/v) ethanol for 40 s followed by one rinse (1 min ) with sterile distilled water, and then surface
disinfected with an aqueous solution of 0.1% of mercuric chloride (w/v) for 12 min followed by five or six rinses
(1 min each) with sterile distilled water. Subsequently, the outer seed coat and subsequent tissue were removed
to expose intact embryos, and then the embryos were disinfected with an aqueous solution of 0.1% of mercuric
chloride (w/v) for 6 min followed by five or six rinses (1 min each) with sterile distilled water, yielding 73.3%
contamination-free germinated buds with 26.7% contamination rate. Modified-MS supplemented with 3.0 mg · L-1gibberellic acid (GA3) was found to be best for seed germination, seed began to germinate the next day after being
put into the medium, and the germination frequency reached up to 93.3%. Modified-MS containing 2.0-3.0 mg · L-1
6-benzyladenine(6-BA), 0.2 mg · L-1 1-napthyl acetic acid(NAA), 3.0 mg · L-1 GA3 was proved to be optimal
for adventitious shoot induction with average 6.01 buds per explant.The induction effect of the explant was
significantly related to the sterilization protocol, the basic medium, and the hormone type and concentration in the
medium. The improved MS medium with lower salt concentration and higher level of 6-BA were beneficial to the
induction of the shoot. |
| Key words: Castanopsis hystrix seeds disinfection procedure primary culture |
