Cryopreservation Of In Vitro Shoot Tips Of Haskaop By Vitrification

The Japanese Hokkaido nutrition fruit tree HASKAOP (Lonicera caerulea L. var. emphyllacaryx Nakai) which belongs to honeysuckle family has the features of drought resistance, cold resistance and other characteristics.Its fruit are rich in special nutrients, It can be used as a kind of fruit or medicine.It’s a very precious resource of special local product and also can be used as the raw material of fruit tree’s breeding.In order to preserved this kind of nature resource successfully,several main factors of cryopresservat-ion were researched in this experiment.The cryopreservation developed in recent decades.The principle of cryopreservation is that live cells’ metabolism almost stopped in liquid nitrogen conditions-196degrees, the plant were in a relatively stable biological state,because of the material saved can greatly reduce its metabolism,so as to the purpose of long-term preservation of germplasm. Common methods of cryopreservation are vitrification, progressive freezing methods, drying dehydration, etc. This experiment was researched in Japanese small fruit HASKAO with vitrification method(encapsulation-vitrification),designed to discuss the effects of HASKAOP in vitro cryopreservation of shoot tips’relationship between the optimal combination through the orthogonal test.And set up a more simple cryopreservat-ion procedures,so as to lay a foundation for large-scale planting in China. The results are as follows:1.Established a relatively simple cryopreservation procedure of HASKAOP in vitro Shoot-tips by vitrification.The2-3mm length shoot-tips were precultured for Id on MS medium with0.7mol/L sucrose at4℃.After pretreating in loading solution(LS) for60min,the shoot tips were treated with PVS2at0℃for60min.Then put them into liquid nitrogen directly and conserved for at least1h.After being rapidly thawed in a water bath at40℃for60s,the shoot tips were washed twice by sucrose solution(1.2mol/L).Then the shoot tips were plated on MS medium contained0.3mol/L sucrose for48h in darkness. Then transferred the materials onto MS medium with0.5mg/L6-BA,0.1mg/L NAA.After culturing in darkness for7days, the shoot tips were cultivated again in normal light. The survival rate of the shoot tips reached to66.67%.The recycle rate up to33.33%. 2.Screened the recovery media.The HASKAOP stem apexes cryopreserved in MS+6-BA0.5mg/L+NAA0.1mg/L+sucrose30g/L+agar7g/L are recoved better and growed faster,the appearance of the leaves was fine.