尾巨桉愈伤组织诱导与再生体系的建立  

Callus Induction and in vitro Regeneration for Eucalyptus urophylla×E. grandis

作　　者：孙长斌 郭棣 张兰英[2] 

机构地区：[1]普罗米绿色能源(深圳)有限公司,深圳518000 [2]中国科学院华南植物研究所,广州510650

出　　处：《基因组学与应用生物学》2012年第6期592-596,共5页Genomics and Applied Biology

摘　　要：以尾巨桉（Eucalyptus urophyllaxE. Grandis）白化苗茎段为外植体,进行愈伤诱导和芽诱导获得不定芽,建立了生产上广泛应用的桉树优良杂交品系 DH32-29高效离体再生体系.户外采集桉树2~3年生尾巨桉 DH32-29枝条进行无性系繁殖,生根苗去除顶芽在无激素 MS＋蔗糖5%培养基上黑暗培养获得白化苗,以此白化苗茎切段3~6 mm 为外植体,在 TDZ、CPPU、Zt 和 KT 分别与 NAA 组合诱导愈伤组织形成,结果表明 TDZ 和 CPPU 的效果好,愈伤诱导率达100%;所得愈伤组织在含0.5 mg/L BAA 和0.1 mg/L NAA 芽诱导培养基上诱导不定芽的形成,CPPU 诱导的愈伤组织不定芽诱导率最高,达83.42%;诱导芽经过伸长和生根阶段形成再生植株,生根率为92.17%.本体系所用材料为无性系无菌苗,易得到大量均一的材料,具有很高的再生率,为利用现代分子育种技术对桉树的遗传改良奠定了良好的基础.Whitened internodal stems were used as explants to establish a high efficient regeneration protocol for Eucalyptus urophyllaíE. Grandis elite hybrid line DH32-29 which are widely planted in China. Micro-propagation were established from a 2 to 3-year old DH32-29 and then rooted plant cut shoot apices were incubated on MS medium with 5% sugar in dark to obtain whitened plantlet. Calli were induced from internodal stems derived from whitened plantlet stems with 3 to 6 mm length incubated on a B5 medium, supplemented with different kinds and concentrations of plant growth regulating substances, at highest frequency to 100%. After 21 days,the calli ob-tained were transferred to B5 medium including 0.5 mg/L BAA and 0.1 mg/L NAA obtained highest frequency to 83.42%. Shoot elongation was then stimulated on MS medium supplemented with BA, NAA for 20 to 30 days and 20 mm long shoots were cultivated on root induction medium for 25 days and rooted frequency up to 92.17%. The merit of this method is that abundant resource and genetic stability explants can be used to construct regeneration of eucalyptus at high frequency system that will be an efficient tool for introducing interesting genes into eucalyp-tus which is still considered recalcitrant.

关 键 词：桉树 愈伤诱导 不定芽诱导 杂交品系 

分 类 号：S792.39[农业科学—林木遗传育种]