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建立哈密大枣叶片离体再生体系,为遗传转化奠定基础。采用哈密大枣叶片为外植体,研究基本培养基、激素浓度、暗培养时间、AgNO3等因素对离体叶片不定芽诱导的影响,并获得再生植株。MS、NN69、WPM三种培养基相比较,NN69更适合做为诱导愈伤组织的基本培养基;TDZ诱导叶片再生不定芽的效果显著优于BA;再生培养基中添加1 mg/L AgNO3对不定芽的形成有显著的影响(P<0.05);培养初期经过2周避光培养更有利于提高再生效率;采用10 mg/L维生素C浸泡15 min可以防止褐化,并能提高不定芽再生率;培养基中添加聚乙烯吡咯烷酮(PVP)或者维生素C,不定芽再生率无显著提高(P>0.05);培养基添加活性炭(AC)会抑制外植体的分化。叶片在NN69+1.0 mg/L TDZ+0.20 mg/L IBA+AgNO31.0 mg/L培养基上,暗培养2周后转入光照培养,出芽外植体转入MS+1 mg/L 6-BA+0.20 mg/L IBA不定芽再生效果最好。不定芽生长至3 cm以上时,转至0.40 mg/L IBA的1/2MS培养基上进行诱导生根。
Establishment of Hami jujube leaves in vitro regeneration system, lay the foundation for genetic transformation. Hami jujube leaves were used as explants to study the effects of basic medium, hormone concentration, dark culture time, AgNO3 and other factors on the induction of adventitious buds in vitro, and to obtain regenerated plants. NN69 and WPM, NN69 was more suitable as a basic medium for inducing callus; TDZ induced the regeneration of adventitious buds significantly better than BA; 1mg / L AgNO3 was added to regeneration medium Adventitious bud formation was significantly affected (P <0.05). After 2 weeks of culture, darkening culture was more beneficial to improve regeneration efficiency. Soaking with 10 mg / L vitamin C for 15 minutes could prevent browning and increase the regeneration of adventitious buds (P> 0.05). Addition of activated carbon (AC) to the medium inhibited the differentiation of explants. The medium supplemented with polyvinylpyrrolidone (PVP) or vitamin C did not significantly improve the regeneration rate of adventitious buds. The leaves were transferred to light culture medium NN69 + 1.0 mg / L TDZ + 0.20 mg / L IBA + AgNO 31.0 mg / L for 2 weeks and the explants were transferred to MS + 1 mg / L 6- BA + 0.20 mg / L IBA adventitious bud regeneration best. When adventitious buds grew to more than 3 cm, they were transferred to 1/2 MS medium containing 0.40 mg / L IBA to induce rooting.