摘要
用正交实验设计对前期影响水仙胚性愈伤诱导和分化关联性大的因素进行综合分析,结果显示,培养基类型、激素及处理方式、不同添加物等对水仙外植体胚性愈伤诱导及分化的影响是基于培养基的多因素协同效应。水仙适宜胚性愈伤诱导培养基是:改良MB+NAA(0.2 mg/L)+6-BA(2 mg/L)+ABA(2 mg/L)+AgNO3(5 mg/L);适宜的分化培养基是:改良N6+2,4-D(0.5 mg/L)+6-BA(2.5 mg/L)+NAA(0.2 mg/L)+ABA(2 mg/L)。实验中观察到水仙体细胞胚的发生,在胚性愈伤诱导和分化体系优化基础上实现农杆菌介导GUS基因转化,稳定表达抗性愈伤GUS染色显示最佳胚性愈伤诱导培养基能促进水仙外植体感受态的形成,有利于外源基因的接受,PCR检测表明实验中获得3株转GUS基因阳性植株。
Based on single-factor screening experiments of embryogenic callus induction and differentiation of Chinese narcissus, comprehensive survey has been carried out with orthogonal experimental design. The results showed that embryonic callus induction and differentiation were under synergistic effects of mutil-factors, including medium type, hormonal concentration and different supplements, the best embryogenic callus induction medium is： improved MB ＋ NAA（0.2 mg/L） ＋ 6- BA （2 mg/L） ＋ABA （2 mg/L） ＋AgNO3（5 mg/L） ; the best differentiation medium is： improved N6 ＋2,4-D（0.5 mg/L） ＋ 6- BA（2.5 mg/L） ＋ NAA（0. 2 mg/L） ＋ ABA（2 mg/L） ; and embryogenesis was observed in embryogenic callus induction. GUS staining showed that improved Narcissus callus could accept foreign genes, and 3 stable expression transgenic plants test- ed by PCR was obtained via embryogenic callus in vitro. This study will lead to the broadening application of fast propagation, virus-free, cell engineering, e.g. , flower color gene engineering modification.
出处
《南京林业大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第4期33-37,共5页
Journal of Nanjing Forestry University：Natural Sciences Edition
基金
福建省自然科学基金资助项目(B0810040)
福建省教育厅A类科技项目(JA08154)
关键词
水仙
胚性愈伤
农杆菌介导法
遗传转化
GUS基因
Chinese narcissus
embryogenic callus
Agrobacterium-mediated tumefaciens
genetic transformation
GUS gene
