土党参茎段的愈伤组织诱导与植株再生

摘要

为了建立土党参愈伤组织途径的高效再生体系,以无菌苗的带芽茎段为外植体,接种于含有不同植物激素的MS培养基中,探讨不同植物激素对愈伤组织诱导、再分化出芽以及生根的影响.结果表明:在不同处理中可获得2种类型的愈伤组织,黄色致密愈伤(类型Ⅰ)和杂夹有少量白色球形体细胞胚的黄色疏松愈伤(类型Ⅱ),细胞分裂素主导着愈伤组织类型的形成.米黄色致密状胚性愈伤(类型Ⅰ)的适宜配方为MS+1.0 mg·L-1 2,4-D+1.0 mg·L-1 6-BA+0.5 mg·L-1 NAA,米黄色疏松愈伤组织(类型Ⅱ)的适宜配方为MS+1.0 mg·L-1 2,4-D.致密愈伤组织在含有细胞分裂素(6-BA、KT、TDZ)的培养基上可分化出芽点进而形成不定芽块,转接到MS+0.5 mg·L-1 6-BA+0.5 mg·L-1 GA3+0.1 mg·L-1 NAA培养基后,芽伸长并增殖为丛生芽.单芽体经300 mg·L-1 IBA溶液浸泡15 min 后,转接于不含任何植物激素的MS培养基上,生根率为100%;将生根良好的植株移栽到泥炭土、蛭石、珍珠岩体积比1∶1∶1的基质中培养30 d,成活率为73.3%.通过愈伤组织途径建立了土党参的高效再生体系,为种苗繁育、多倍体育种、转基因操作和细胞突变育种提供了技术基础.%In order to estabilished an efficient plant regeneration system via callus from stem segments with bud of Campanumoea javanica B1.The aseptic stems were used as explants and then cultured on MS medium supplemented with different plant growth regulators (PGRs) to establish an effective protocol for callus induction, shoots induction and development, and roots induction.The results showed that two types of callus could be obtained under different treatments, of which type Ⅰ was yellow and compact while type Ⅱ was light yellow, fragile and mixed with a small number of white globular embryos.The type of callus could be regulated by cytokinin.Type I callus was induced on MS medium supplemented with 1.0 mg·L-1 2, 4-D+1.0 mg·L-1 6-BA + 0.5 mg·L-1 NAA, while type Ⅱ callus was induced on MS medium supplemented with 1.0 mg·L-1 2, 4-D.The compact callus could differentiated into adventitious buds on the medium supplementing cytokinin (6-BA, KT, TDZ).The adventitious buds further developed and proliferated after 30 d culture on MS medium containing 0.5 mg·L-1 6-BA+0.5 mg·L-1 GA3+0.1 mg·L-1 NAA.Excised shoots were treated with 300 mg·L-1 IBA for 15 min and then cultured on MS basal medium for 30 d, and the rooting rate reached 100%.When transplanted into a mixture of peat soil, vermiculite and perlite (volume ratios was 1∶1∶1), the survival rate of regenerated plantlets could reached 73.3% after 30 d.The experiment developed an effective protocol for C.javanica shoot organogenesis via callus, which might laid the foundation of researches on polyploidy, transgenic and cell mutation breeding.