【作者】 谢杰；

【导师】 余沛涛； 王全喜；

【作者基本信息】 上海师范大学 ， 水生生物学， 2007， 硕士

【摘要】 宜兴百合是我国四大食用百合之一,具有良好的食用、药用价值。本文从小鳞茎的增殖、小鳞茎的膨大、愈伤组织诱导再生小鳞茎、环境因子对小鳞茎生长的影响四个方面探索了组织培养快速繁殖宜兴百合的方法,大大提高了宜兴百合的生长量;并通过组培百合与栽培百合进行有效成分的对比分析,以此保证组培百合的品质。实验结果如下:小鳞茎增殖:小鳞茎在固态培养基MS+0.15mg/LNAA+2.0mg/L6-BA中诱导系数最高,为858%,液体培养基MS+0.15mg/LNAA+2.0mg/L6-BA+0.1mg/LCCC中小鳞茎增殖系数最高,60d质量增重是对照组的3倍,是同激素浓度固态培养基的1.44倍,且小鳞茎发生早,周期短,芽长势旺盛。小鳞茎膨大增重:小鳞茎增重膨大的最适蔗糖浓度为6-8%;最适的CCC(矮壮素)浓度为0.1mg/L, 2个月平均每个小鳞茎质量增重28.85克,比栽培百合单个小鳞茎重12.18克。2,4-D对小鳞茎的增重无显著作用,但低浓度的2,4-D对出芽和出苗有一定的促进作用,当2,4-D浓度为0.5mg/L时长根较好,根较长且须根较多。加了活性炭的小鳞茎重量略微有增大,出芽率和长根率都比对照组稍高,但出苗率不如对照组。通过诱导愈伤进行快繁:在适当的2,4-D浓度下(0.01~0.1mg/L),从愈伤组织上分化出的芽的数量远远大于相同条件下直接从鳞茎再生的小鳞茎数,60d增长率达1826%。2,4-D能积极促进愈伤组织的生长,但过高浓度的2,4-D会使小鳞茎停止生长。当2, 4-D的质量浓度在0.01～0.1mg/L的范围内时,愈伤组织增长量与2,4-D浓度呈正比。小鳞茎增殖的最佳环境因子:(1)pH=5.8-6.2之间时小鳞茎生长情况最好,低于5.0或者高于6.2时小鳞茎的生长都会受到一定的阻碍,尤其是长时间在pH<5.0的环境下生长,鳞片会逐渐由绿色变黑,枯萎以致死亡。pH>6.2时,小鳞茎可以维持一段时间的生命力,但生长极为缓慢。(2)温度也是影响小鳞茎生长的一个重要因素,最适宜小鳞茎增殖的温度范围是22-28℃。(3)在500lx~3000lx范围内,光照强弱对试管鳞茎增殖的影响差异不显著,较高强度的光照能略微提高小鳞茎的增殖率。(4)50mg/L Asp反而对小鳞茎的增殖有抑制作用,长根无明显差异。组培百合与栽培百合主要成分的比较:组培百合与栽培百合中都含有总皂甙、生物碱及总蛋白,组培百合中各类主要成分的含量均高于栽培百合。更多还原

【Abstract】 Lilium lancifolium Thunb. is one of the best edible lily in our country with great edible and officinal value.Rapid propagation of Lilium lancifolium Thunb. was introduced in five aspects: the reproduction of bulblets,the extantion of bulblets, the redifferentiation of bulblets through callus, the environment factors on bulblets propagation. In addition,the valuable components were analogized.The results were showed:The propagation of bulblets: The MS+0.15mg/L NAA+2.0mg/L 6-BA was the best solid medium for the scale of lily to reproduction and extention, propagation rate was 858%, The medium MS+0.15mg/LNAA+2.0mg/L6-BA+0.1mg/LCCC was the best liquid medium for bulblets propagation, adding weights by three times than control,and 1.44 times than the one with the same hormone concentration. The bulblets occured earlier and grew rapider.The weights and extention of bulblets: The appropriate sucrose concentration ranged 6% to 8%,the CCC concentration was 0.1 mg/L. After 2 months, the bulblet adding weights 28.85g, 12.18g more than the wild one separatly in each bulblet. There was no obvious effect on bulblets extention for 2,4-D,but low concentration of 2,4-D could accelerate buding rates and seedling rates. Roots were grown better when 2,4-D was 0.5mg/L,with longer and more fibres. There was a little process for bulblets when adding 400mg/L active carbon.The propagation of bulblets through callus inducing: The number of bulblets from callus were more than that directly from previous bulbs.The most suitable concentration for bulblets to induce callus was 0.01-0.1mg/L.The propagation rate through callus could reach 1826% in 60 days. 2,4-D could accelerate callus growing activity,but higher concentration of CCC could confine bulbs grow. When the concentration of 2,4-D ranged 0.01mg/L - 0.1mg/L, there was direct ratio growth of callus issues.The suitable environmental factors for bulblets repropagation:(1).The bulblets would grow fairly when pH was 5.8 to 6.2,The bulblets would be under normal condition when pH was below 5.0 or above 6.2.It would be bad when pH was under 5.0 for a long time, with scales becoming black and die. When pH is above 6.2, bulblets could keep a short life and growth was quite slow.(2).Temperature was also an important factor for bulblets growth,the appropriate temperature was 22-28℃.(3).Illumination was another condition for bulblets growth while under 500lx to 3000lx condition, there was no prominence effect on bulblets reproduction and higher illumination can elevate bulblets reproduction a little.(4).50mg/L Asp could inhibit bulblets reproduction,and there was no difference on rooting and differentiation.Contrast analysis major components of tissue culture lily and wild lily: There were saponin, alkaloid and protein in tissue culture lily and wild lily,and these three major components in tissue culture lily was higher than in wild lily.更多还原