鹤望兰组织培养技术体系研究
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摘要
本研究以鹤望兰(Strelitzia reginae)种子切段为外植体,探索鹤望兰组织培养快速繁殖的技术体系,着重研究丛生芽和原球茎两种再生途径各阶段的最佳培养基配方及培养程序,以及培养过程中防止褐变的措施。结果表明:
     1)外植体以种子切段为好,此类型外植体褐变较轻。
     2)胚芽的启动培养:最适培养基为H+BA1.5mg/L+ZT1.0mg/L+NAA0.8mg/L,培养30天后可获得无菌芽苗。
     3)原球茎的启动培养:最适培养基为2H+BA2.0mg/L+ZT0.5mg/L+NAA0.6mg/L。
     4)丛芽增殖途径:最适培养基为H+BA1.5mg/L+ZT1.0mg/L+NAA0.6mg/L。褐变较严重,采用固液交替培养可有效减轻褐变。继代多次后,有愈伤组织发生,增殖加快,第六代进入增殖盛期,平均增殖系数可达5.0。
     5)原球茎增殖途径:最适培养基为2H+BA2.0mg/L+ZT1.0mg/L+NAA0.4mg/L。褐变相对较轻,但其增殖系数较丛芽途径低,平均为3.9。
     6)壮苗培养:最适培养基为H+NAA0.1mg/L,可促进苗的生长并能有效地促进根的诱导。
     7)生根培养:最适培养基为H+IBA0.5mg/L+NAA0.1mg/L+AC0.5g/L。基部可见白色根尖时将小苗转入无激素培养基中促根生长。
     8)炼苗移栽:竹林下的腐叶土可促进须根的萌发,以此为基质最好。
The seed segments were used as explants to establish a rapid technique system of tissue culture in Strelitzia reginae, and especially to find out some effective measures to control the browning. Two pathways of regeneration were observed: one was regeneration directly from clustered buds, and the other was adventitious shoots induction from protocorm. The results were as follows.
    1) The seed segments were used as explants because of their slighter browning.
    2)The initiation of embryo buds: H + BA1.5 mg/L + ZT1.0 mg/L + NAA0.8 mg/L was the best medium. Had been cultured 30 days, the buds or shoots could be got.
    3)The initiation of protocorms: the best medium was 2H+BA2.0mg/L + ZT0.5mg/L + NAA0.6mg/L.
    4)The multiplication of clustered buds: the best medium was H+BA1.5 mg/L+ ZT 1.0 mg/L + NAA0. 6 mg/L. The explants were cultured alternately on the solid medium and on the liquid medium with filter paper, which was the effective measure to prevent the explants from the browning to certain extent. Had been subcultured for the 6th generation, the multiplication was flourishing, and the average rate reached 5.0 per generation.
    5)The multiplication of protocorms: the best medium was 2H + BA2.0mg/L + ZT1.0mg/L + NAA0.4mg/L. The browning of explants was
    
    
    
    relatively slighter in this pathway. However, the rate of multiplication was lower, averaging 3.9 per generation. Had been cultured on the same medium for 30~60 days, the protocorms differentiated to small bud clusters.
    6) The suitable medium for the growth of clustered plantlets was H+ NAA0.1mg/L, which could significantly conduce the induction of roots.
    7) The best medium for the rooting of plantlets was H+IBA0.5mg/L + NAA0.1mg/L+AC0.5g/L. After 30 days, the very small white root tips emerged from most of the plantlets. Just then, the plantlets were transferred on H medium without any growth regulator, and the roots would grow up quickly.
    8)For nursling of plantlets, the soil of the rotten leaves in the bamboo groves was the best materials, which could conduce germination of fibrous roots effectively.
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