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栓皮栎体胚再生与增殖能力的保持研究
贾小明1, 张焕玲1, 张存旭1
西北农林科技大学 林学院
摘要:
【目的】提高栓皮栎体胚的再生与增殖能力。【方法】将栓皮栎不同发育阶段合子胚诱导的胚性愈伤组织及不同发育时期的体胚,分别接种于含不同组合激素的MS培养基上,在不同光照条件下培养,探讨影响栓皮栎体胚再生与增殖能力保持的因素。【结果】随着继代次数的增加,胚性愈伤组织再生体胚能力呈下降趋势;6-BA与NAA配合使用可延缓继代过程中胚性愈伤组织再生体胚能力的下降,1.00 mg/L 6-BA与0.25 mg/L NAA是继代中保持胚性愈伤组织再生体胚能力的最佳培养基激素组合;胚性愈伤组织再生体胚需要自然散射光,强光及全程暗培养均不利于体胚再生;合子胚外植体发育程度影响胚性愈伤组织再生体胚能力,处于心形期及早子叶形期的幼嫩合子胚诱导的胚性愈伤组织,其再生体胚能力强于成熟子叶形合子胚诱导的胚性愈伤组织;体胚发育阶段越老,再生与增殖体胚能力越差,胚性愈伤组织及处于球形期的体胚,再生增殖体胚能力强于其他发育时期的体胚。【结论】选择处于心形期的幼嫩合子胚诱导胚性愈伤组织,然后在含1.00 mg/L 6-BA与0.25 mg/L NAA的MS培养基上于自然散射光下培养,可有效保持栓皮栎的体胚再生与增殖能力。
关键词:  栓皮栎  体细胞胚  增殖与再生
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基金项目:陕西省科技攻关项目(2007K01-11-04);西北农林科技大学唐仲英育种基金项目(09YZ082)
Studies on regeneration and proliferation of somatic embryos in Quercus variabilis
Abstract:
【Objective】The research was done to improve regeneration and proliferation abilities of somatic embryos of Quercus variabilis.【Method】Factors affecting regeneration and proliferation abilities of somatic embryos of Q.variabilis were discussed by culturing embryogenic calluses at different development stages of somatic embryos,which were induced from immature zygotic embryos on the MS medium supplied with different phytohormone combinations in variable light conditions.【Result】The embryoids regeneration ability of embryogenic callus presented downward trend with increasing times of subculture on medium,while match usage of 6-BA and NAA could delay this trend.The optimal medium for maintaining embryoids regeneration ability of embryogenic callus was MS medium containing 1.00 mg/L 6-BA and 0.25 mg/L NAA.Natural light was essential to maintain embryoids regeneration ability of embryogenic callus,while darkness and intensive light were inimical.Embryoids regeneration ability of embryogenic callus was also influenced by the development degree of zygotic embryos.Embryogenic callus from junior hart-shape or early cotyledonary zygotic embryos had higher embryoids regeneration ability than the one from mature cotyledonary zygotic embryos.The older was the development stage of somatic embryo,the weaker was the regeneration and proliferation ability.The regeneration and proliferation abilities of initial embryogenic calluses and globular-shape somatic embryos were higher than others.【Conclusion】Regeneration and proliferation abilities of somatic embryos of Q.variabilis can be maintained effectively by selecting embryogenic calluses from junior zygotic embryos and culturing them on the MS medium containing 1.00 mg/L 6-BA and 0.25 mg/L NAA under the natural light.
Key words:  Quercus variabilis  somatic embryos  proliferation and regeneration

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