The Preliminary Establishment Of Plant Regenerration System About Paeonia Lactiflora ‘hang Bai’ Embryo Direct Oganogenesis By The Way Of Multiple Shoot Clumps Indution

Herbaceous peony(Paeonia lactiflora Pall.) as China’s traditional precious flowers is loved by people deeply. However its low propagation coefficient and long breeding cycle limit the peony breeding process and industry development for a long period, which urges people to to build a peony’s tissue culture regeneration system to achieve fast peony’s propagation as for there are many basic problems, none of efficient has been presented.This study choosed P. lactiflora ‘Hang Bai’, which has both medicinal and oil use value for research object. Different raties of plant growth regulators were used to induced embryo explants into multiple shoot clumps, and proliferated the multiple shoot clumps then poduce roots and form complete plants with roots at last. Our study preliminarily established a new plant regenerration system for P.lactiflora ‘Hang Bai’ by means of embryo direct organogenesis based on multiple shoot clumps indution. This systerm avoids the phase of de-differentiation and re-differentiation process, reduce the regeneration cycle greatly. An embryo establish sterile system was intrduced to overcome these fundermengtal problerms for peony the these,such as explant culture pollution at the initial stage and rooting in later period. The experimental materials were taken from Caozhou Peony Garden in Heze in late July at which peony pods began to split and the seeds turned from green to dark brown, which were washed and stored in the refrigerator at 4℃ befor being tested for study. Experimental study results are as follows:1.At the experimental materials processing soaking the P.lactiflora ‘Hang Bai’ seeds in distilled water for 52 hours, then using 70% ethanol sterilized 30 s combined with 2.0% sodium hypochlorite sterilization for 15~20min befor stripping the embryo out for inoculation, the sterilization effect is good, explant material contamination is low at 7.89% and the inoculation loss rate is low at 6.6%.2.At the embryo culture conditions controlling process get the result of culture treated gradient of dark stars P. lactiflora ‘Hang Bai’ embryo germination rate and embryo growth and development of embryo are not directly related to dark culture by setting different darkness.3.The appropriate embryo culture medium is MS+6-BA1.0mg/L+GA31.0mg/L, the embryo germinated at 7 days and the germination rate is above 90%, the treatment has greatly increased the peony embryo germination rate, at the same time shorted the breeding cycle by means of breaking the peony species of embryo dormancy, which is higher than the germination rate under natural conditions whose germination rate is about 40% and artificial seeding process whose germination rate is 60%~80% for peony.4.The study concluded that the active carbon will promote embryo radicle development priorities, with the increase of activated carbon content, radicle development and rapid root growth slender, really slow the leaf germination and ultimately it is difficult to form a robust seedling. The embryo can grow into a seedling at the hinght of 4cm after 90 days, which greatly reduced the peony breeding cycle, after 130~160days the seeding is 8cm high, the seedlings grew well by adjusting the 6-BA(2.0~3.0mg/L) concentrations, thus the study established a stable embryo seedlings of peony cultivation system.5.This experiment researched the case of P. lactiflora ‘Hang Bai’ embryo germination and growth at different culture medium conditions and find out that the embryo germinated earlier and growed faster and is easy to induce multiple shoot clumps on the medium MS+6-BA1.5mg/L+GA31.0mg/L, then divert the whole explants to the proliferation medium of MS+6-BA3.0mg/L+GA31.0mg/L for the next indution of multiple shoot clumps after being cultured for 30 days, the inducing effect was good, the average embryo proliferation was 1.27. The experiment at last inducted the explant to poduced root and researched that the appropriate rooting medium was 1/2MS+IBA1.0mg/L+NAA0.5mg/L+PVP1.0g/L+AC0.4g/L, the rooting medium with PVP1.0g/L was good for reduceing the explant browning at which the root-inducing rate was 48.72% after the explants being cultured for 45 days, a P. lactiflora ‘Hang Bai’ embryo grows to be 1.27 seedings after a proliferation of propagation, the hole cultivating cycle is 120 days. The multiple shoot clumps proliferation medium was MS+6-BA2.0mg/L+GA31.0mg/L, after 40 days’ cultivating the average proliferation was 2.53, then inducing the seedlings to poduce roots, the root-inducing is 23.77% after 45 days, the original one seed can be induced to 3.21 seedlings with twice proliferation of propagation of embryo multiple shoot clumps, what’s more the the hole cultivating cycle was 160 days.