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    沙棘优良抗旱品种离体再生体系的建立和优化

    In vitro plant regeneration from a droughtresistant filial generation of Hippophae rhamnoides L

    • 摘要: 本文在沙棘杂交育种工作和抗旱性综合评价研究的基础上,获得了优良抗旱品种雄性植株无性系,并较系统地研究了该无性系通过器官发生途径实现离体培养,建立和优化再生体系所需的条件。结果表明:春季采集生长旺盛的健康新梢,建立无菌培养体系效果最好,试管苗再生可以通过两条途径实现:1)采用培养基WPM+BA0.5 mg/L+2,4-D0.5 mg/L可以诱导无菌叶片和茎段切口处产生愈伤组织,采用培养基WPM+BA0.4~0.6+IBA0.02~0.03,可以诱导茎段来源的愈伤组织再分化形成大量绿色不定芽点;经继代培养后,不定芽最终可以生长成为具有一定高度和粗度的健壮新梢;在生根培养基1/2WPM+IBA0.5 mg/L+NAA0.5 mg/L上培养20 d后,生根率可以达到100%。2)不定芽也可以直接从茎段或叶片的切口处发生,采用最佳的培养基(WPM+TDZ0.01~0.02 mg/L+IBA0.01~0.02 mg/L)和适宜的试材类型(从无根组培苗上剪取的茎段和从有根组培苗上剪取的叶盘),不定芽发生率可以达到100%,平均每个茎段上产生的不定芽数为4.45个,叶片上产生325个;不定芽再经过增殖、伸长、壮苗(培养基为WPM+BA0.2 mg/L+IBA0.05 mg/L)后,在生根培养基1/2WPM+IBA0.5 mg/L+NAA0.5 mg/L上培养20 d后亦可100%生根。

       

      Abstract: Breeding efforts to obtain more droughtresistant seabuckthorn (Hippophae rhammoides) materials have resulted in a good filial generation, in which a Mongolia seabuckthorn‘Wulanshalin’ was used as female parent and one of the improved Chinese seabuckthorn ‘Fengning’ supplied pollen. In this work, the authors tested the in vitro regenerability of the seabuckthorn through organogenesis. New shoots derived from healthy and vigorously growing plants of spring in the greenhouse are the most appropriate explants to the whole plant regeneration. Regenerated plantlets can be efficiently obtained by two medium systems. System I consisted of callus induction medium of WPM (wood plant medium) supplemented with 0.5 mg/L BA and 0.5 mg/L 2,4-D and shoot induction medium of WPM with 0.4-0.6 mg/L BA and 0.02-0.03 mg/L IBA. SystemⅡincluded WPM medium supplemented with 0.01-0.02 mg/L TDZ and 0.01-0.02 mg/L IBA and WPM medium with 0.2 mg/L BA and 0.05 mg/L IBA. Stem segments from unrooted plantlets and leaf discs from rooted plantlets are appropriate explants and 100% efficiency can be achieved. A total of 4.45 buds on each stem segment and 3.25 buds on each leaf disc were attained. The media of both systems were adopted in this research to receive adventitious buds either regenerated from calli or directly derived from stem segments and leaves. The length of buds increased after several times of subculture. The regenerated shoots from the effective culture systems could root with nearly 100% efficiency in half strength WPM medium supplemented with 0.5 mg/L IBA and 0.5 mg/L NAA after 20 days. 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