| Diling is Chinese unique half-wild herb. Its plummy and wartery rhizome contains abundant stachyose. The last research of carbohydrate showed the stachyose has important health protection function and big marketing potential. In production, the rhizome sowing is the main propagation way of Diling; The generation rate is lower and seed resource is fairly deficient. Meanwhile, Asexaul propagation in the long time can make Diling to carry and accumulate virus, lead variety feature to degenerate and bring much more risks to production. It is improtant for breeding popularization and purification rejuvenation of good varity of Diling to establish a rapid propagation system which can be used to big scale production of Diling by founfing and excelling various conditions of the test tube propagation. The study results show as follow.The rhizome is the best explant for Diling in vitro regeneration under the way of terminal bud or lateral bud. The rhizomes of Diling disinfected respectively for 25-30 seconds and 16-18 minutes by 75% alcohol and 0.1% mercuric chloride can control efficiently contamination.The single or mingle usage of hormone can cause of the different effects of seedling proliferation. The simutaneous usage of 6-BA and KT can induce rhizoma of Diling to produce lots of adventitious buds, but it is disadvantageous for father differentiation, growth and developing of the adventitious bud. The stem of Diling can get efficient proliferation under lower concentration of 6-BA or KT. The usage effect of KT which is milder and has not vitrification superiors to that of 6-BA. The mingle usage of cytokinins and auxins can improve growth of seedling. The generation rate is 4.20-4.67 on the MB medium containing 0.5mg L-1 NAA supplemented with 1.0-1.5 mg L-1KT. The hormone concentration ratio is efficient for seedling proliferation of Diling.The seedling has many ways for root inducation. In general test tuber root inducation, the number difference of root induced is not big between 1/2MB and 1/4 MB basic medium, butquality of the root induced is rised on the 1/2MB medium. The best concentration of NAA for root inducation is 0.5-1.0 mg L-1. The higher concentration of NAA is disadvantages for root growth. The circumstance of root inducation is diferent on the different media. The root inducation rate is 100% on the cotton, pearl rock and agar medium, but the number of root induced is maximum. The average number is 5.47. The young root possess many multistage branchs. The transplant effect also showed pearl rock is ideal substitute for agar. The cuttage of stem is also a efficient way of root inducation, but the general effect is worse than test tube root inducation. The root number and root length of water culture cuttage are maximum in all kands of cuttage, but the young root is too slender to provide high transplant survival rate. The soak treatment of the stem bottom by 1.0 mg L-1 NAA solution before cuttage can improve the inducation and growth of root. The stem of Diling after 4-5 times continous transfer can form seedling through one-step culture on the MS medium containing 0.5 mg L-1 6-BA, 0.5 mg L-1 KT and 0.1 mg L-1 NAA. The seedling propagation period shorten 10-15 days on the way.The iducation of Diling microtuber are affected by many factors. On the basic medium aspect, the MS medium is superior to MB on the microtuber number and wet weight, but the MB medium is more advantage of inducing and producing big microtuber under high concentration level (10%)of sucrose. The efficient sucrose concentration for inducing microtuber is 8%-10%. The lower sucrose concentration can not induce the stem of Diling to form microtuber. The affect of external stimulus on microtubers induction is obvious. The optimum effect comes from the treatment of 8.0 mgL-16-BA, the next is 10 mg L-1 SA and 5.0 mg L-1 B9, the effect of IAA and CCC is worst. The CCC is not suited to induce microtuber of the Diling.
|
PDF Full Text Request