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以马铃薯试管苗为试材,对其茎尖小滴玻璃化法超低温保存的影响因素进行了研究,并对再生植株进行了遗传稳定性检测。结果表明,马铃薯茎尖依次在含有0.3mol·L-1和0.5mol·L-1蔗糖的液体MS培养基中预培养各1d后,在0℃下PVS2处理30min,转到铝箔条上PVS2小滴上(约15μL),将粘有茎尖的铝箔条在液氮里蘸一下,然后直接装入盛满液氮的冷冻管中,投入液氮至少保持1h。室温下用含有1.2mol·L-1蔗糖的MS液体培养基解冻并洗涤30min后,接种到MS+0.5mg·L-1Zeatin+0.1mg·L-1NAA+1.0mg·L-1GA3恢复培养基上,存活率和再生率最高达79.91%和62.52%。通过SSR分子标记检测,再生植株的遗传稳定性没有发生改变。
Potato plantlets were used as experimental materials to study the influence factors of the cryogenic preservation of the shoot tip droplets by vitrification, and the genetic stability of the regenerated plants was tested. The results showed that the shoot tips of potato were pre-incubated for 1 day in liquid MS medium containing 0.3 mol·L-1 and 0.5 mol·L-1 sucrose, respectively, and then treated with PVS2 at 0 ° C for 30 min, Drop on (about 15 μL). Dip the tip of aluminum foil into liquid nitrogen and dip directly into a cryovial filled with liquid nitrogen. Maintain liquid nitrogen for at least 1 h. After thawing and washing with MS liquid medium containing 1.2 mol·L -1 sucrose for 30 min at room temperature, the medium was inoculated into MS + 0.5 mg · L-1 Zeatin + 0.1 mg · L -1 NAA + 1.0 mg · L -1 GA 3 recovery medium , The survival rate and regeneration rate reached as high as 79.91% and 62.52%. The genetic stability of regenerated plants did not change by SSR molecular markers.