红皮云杉体胚发生体系优化和超低温保存技术研究

[目的]通过对红皮云杉(Picea koraiensis)体细胞胚胎发生条件的筛选和超低温保存技术研究,建立其完整的体胚发生体系及超低温保存条件,为红皮云杉优良种质资源的大量繁殖及保存提供基础.[方法]以红皮云杉合子胚为外植体,以两种基本培养基、6种植物生长调节剂组合为培养条件,筛选胚性愈伤组织诱导的适宜条件.并将获得...

Optimization of somatic embryogenesis system and cryopreservation of Picea koraiensis

【Objective】This study aimed to establish a complete somatic embryogenesis system and optimal cryopreservation conditions to provide a basis for the breeding and preservation of Picea koraiensis (Korean spruce) planting resources. We screened somatic embryogenesis conditions and the cryopreservation of Korean spruce. 【Method】We explanted zygote embryos in two basal culture media with six plant growth regulators to define the optimal culture conditions in which the embryogenic mass was introduced. The embryogenic mass was cryopreserved, and the somatic embryo maturation and germination were assessed. We screened three abscisic acid (ABA) and Gelrite concentrations and four basic culture media to assess the optimal conditions for the somatic embryo maturation and germination. 【Result】①The two basal culture media were compared, and the modified induction ratio in RJW medium was 30.00% which was greater than that of 1/2 LV (induction ratio, 22.50%) basal culture medium. The highest induction ratio was 50.00% in the proliferation medium containing 3.0 mg/L of 1-naphthalacetic acid (NAA) and 0.5 mg/L of N6-benzyladenine (6-BA). ②The embryogenic mass was cultured for 3 months, then cryopreservation was assessed. The cryopreserved embryogenic mass from three cell lines survived after 10 days of restoration. ③The numbers of somatic embryos in the three cell lines were higher, but the ratio of abnormal HY-1 somatic embryos was 77.14% in the maturation medium containing Gelrite (6.0 g/L) and ABA (20 mg/L). Increasing the Gelrite concentration to 8.0 g/L decreased the ratio of abnormal somatic embryos in all three cell lines; the number of HY-2 somatic embryos was 396.00 per gram, and the ratio of abnormal somatic embryos was 10.33%. ④The somatic embryos of Korean spruce matured for two months in the germination medium. The amount of somatic embryo germination was the lowest on 1/2 LM, followed by on LM medium and was the highest on RJW medium. The germination ability of the cell lines differed, with a somatic germination ratio of 65.00%.【Conclusion】①The optimal basal culture medium for the embryogenic mass induction was modified RJW, and the appropriate plant growth regulator concentration was NAA 3.0 mg/L + 6-BA 0.5 mg/L. ②The embryogenic mass obtained survived cryopreservation. ③The optimal ABA and Gelrite concentrations for somatic embryo maturation were 20 mg/L and 8.0 g/L, respectively. ④Also, the optimal basal culture medium for somatic embryo germination was modified RJW. We established the ideal conditions for the regeneration, cryopreservation, and somatic embryogenesis of Korean spruce. This technique can be applied to propagate the best planting resources and to establish multi-varietal forestry by using Korean spruce.