海南白花油茶花药再生体系的初步建立

摘要

Lack of suitable seedlings for local cultivation is one of the key factors restricting sustainable development of Camellia oleifera industry in Hainan.In order to obtain somatic cell regenerated tissue culture plantlets with main roots through the anther somatic cell embryogenesis way, we observed pollen development phases, selected primary callus induction medium and culture methods, studied regeneration and transplantation of anther somatic cell embryo plants, and preliminarily established regeneration system of anther somatic cell embryo in C.oleifera in Hainan.The results show that,when flower bud diameter was 1.19-1.30 cm,pollen development was at the mononuclear side phase,and the anthers at this phase were the optimal explants for inducing anther embryogenic callus. The suitable primary medium for inducing anther embryogenic callus in C.oleifera was modified MS added with NAA and KT hormone combination, and the suitable way was dark culture. Anthers embryogenic calli could re-differentiate somatic cell embryos and induce maturation of somatic cell embryos in the medium added with NAA, BA and Zt hormone combination.%适合本地种植的良种壮苗匮乏,是制约海南省油茶产业可持续发展的关键因素之一.为了通过花药体细胞胚发生途径来获得具主根的体细胞再生植株组培苗,从花粉发育时期的鉴别、初代愈伤组织诱导培养基和培养方式的筛选以及花药体胚植株的再生和移栽等方面进行探索,初步建立了海南白花油茶花药体细胞胚再生体系.结果表明:海南白花油茶未开花蕾的横径为1.19~1.30 cm时,花粉发育多处于单核靠边期,此期的花药是诱导花药胚性愈伤组织的合适外植体;适用于海南油茶花药胚性愈伤组织诱导的初代培养基为改良MS配以NAA和KT激素组合的培养基,适宜的培养方式为暗培养;花药胚性愈伤组织在NAA、BA和Zt激素组合的培养基上可再分化出体细胞胚和诱导体胚成熟.