Study On Cryopreservation Of Anthurium Andraeanum By Droplet-vitrification

Anthurium andraeanum Lind.is a perennial evergreen plant of the genus Anthurium of the family Araceae.It is the second largest tropical flower and one of the most-selling"noble"flowers in the world.It has extremely high ornamental value and economic value.Obtaining the genetic resources available in the gene bank is a key factor for the success of genetic improvement of Anthurium andraeanum breeding,but most traditional varieties are gradually lost during the promotion of new varieties.Therefore,the preservation of germplasm resources is extremely important for the production of Anthurium andraeanum and the breeding of new varieties.The cryopreservation procedure is suitable for any plant tissue with regenerative ability.Research the cryopreservation technology through the droplet-vitrification method in order to establish a method that is simple to operate and can stably preserve the germplasm resources of Anthurium andraeanum for a long time.Provide technical reference for the establishment of the gene bank of cryopreservation resources of Anthurium andraeanum and the cryopreservation of other tropical herbs.In this study,the sterile seedlings of Anthurium andraeanum"Pink Champion"（PC）,"Alabama"and"Xueyu"were used as materials,and the ultra-low temperature preservation technology of small drop vitrification was used.The main results are as follows:the highest regrowth of Anthurium（63.70%）was achieved after preculture for 4 days on MS solid medium containing 0.4 M sucrose and 2 M glycerol,dehydration with 80%PVS₂ at 0°C for 50 min,cooling in droplets of PVS₂ placed on aluminum foil strips,after immersing in liquid nitrogen for 2 s,directly transfer it to a cryotube filled with liquid nitrogen,and keep it in liquid nitrogen for at least 30 minutes;after rewarm them with MS liquid medium containing 1.2 M sucrose at room temperature for 20 minutes,transferred to recovery medium.Through ISSR and SSR molecular marker detection,the genetic stability of the regenerated plants of Anthurium andraeanum PC after cryopreservation was tested.The results showed that no specific band appeared in the regenerated plants,indicating that the genetic stability of Anthurium andraeanum PC was unchanged.Using the optimized solution obtained from the PC axillary buds of Anthurium andraeanum,the study on the cryopreservation of different varieties and different explant types by droplet vitrification method found that the effect of cryopreservation of different varieties of Anthurium andraeanum by the droplet vitrification method:"Pink Champion"（PC）>"Alabama">"Xueyu".The effect of ultra-low temperature preservation of different explant types of Anthurium andraeanum with droplet vitrification:stem segment（including axillary bud）>petiole>leaf.