樟子松无性系(GS1)胚性愈伤组织的初步诱导

Preliminary Study of Embryogenic Callus Induction of Pinus sylvestris var.mongolica Clone(GS1)

本研究以樟子松无性系(GS1)为研究对象,探讨合子胚不同发育时期愈伤组织的诱导情况,探索不同培养基类型及不同激素配比对其愈伤组织诱导的影响,初步建立体胚发生技术体系,为樟子松(GS1)体胚发生技术建立和获得植株再生提供一定基础。本研究对樟子松无性系(GS1)合子胚不同发育时期进行观察并进行不同时期、不同激素体胚诱导。樟子松无性系(GS1)合子胚形态变化可划分为9个阶段,将这9个阶段的合子胚分别接种到不同培养基上进行诱导。结果显示,DCR培养基比MS和LM培养基更适合GS1愈伤组织的诱导和生长。第7阶段的合子胚在DCR+2,4-D 5.0 mg/L+NAA 1.0 mg/L+6-BA 0.5 mg/L培养基上愈伤组织诱导率最高,在DCR+2,4-D 3.0 mg/L+NAA 1.0 mg/L+6-BA 0.5 mg/L培养基上胚性愈伤组织诱导率最高。诱导过程会产生两种类型的愈伤组织,分别为白色透明状的愈伤组织和淡黄褐色愈伤组织。经显微观察有两种结构愈伤组织,一种是近球形的薄壁细胞结构,为非胚性愈伤组织;另一类是由胚性细胞和胚柄细胞组成的胚性细胞团。樟子松无性系(GS1)合子胚第7阶段的合子胚诱导愈伤组织效率高,低盐培养基DCR适合GS1愈伤诱导,高浓度的2,4-D能够促进GS1产生大量的愈伤组织。胚性愈伤组织诱导最佳培养基为DCR+2,4-D 3.0 mg/L+NAA 1.0 mg/L+6-BA 0.5 mg/L。

In this study,Pinus sylvestris var.mongolica clone(GS1)was used as the research object to investigate the callus induction of zygotic embryo at different developmental stages,to explore the effects of different medium types and different hormone ratios on callus induction,and to preliminarily establish the somatic embryogenesis technology system,which provided a certain basis for the establishment of somatic embryogenesis technology and plant regeneration of Pinus sylvestris var.mongolica(GS1).In this study,we observed the zygotic embryos of Pi-nus sylvestris var.mongolica clonal(GS1)at different developmental stages and induced somatic embryos at differ-ent stages with different hormones.Morphological changes of the zygotic embryos of Pine sylvestris clones(GS1)could be divided into 9 stages.The zygotic embryos of these 9 stages were inoculated on different media for in-duction.The results showed that DCR medium was more suitable for the induction and growth of GS1 callus than MS and LM medium.The zygous embryos at stage 7 had the highest callus induction rate on the medium of DCR+2,4-D 5.0 mg/L+NAA 1.0 mg/L+6-BA 0.5 mg/L,and the highest embryogenic callus induction rate on the medi-um of DCR+2,4-D 3.0 mg/L+NAA 1.0 mg/L+6-BA 0.5 mg/L.Two types of callus were produced during induc-tion,namely,white transparent callus and light yellow brown callus.The two kinds of callus showed two kinds of structure by microscopic observation,one is the near-spherical parenchyma cell structure,which is non-embryonic callus;The other type is the group of embryonic cells composed of embryonic cells and stem cells.Pinus sylvestris var.mongolica clones(GS1)zygote embryo morphology changes can be divided into nine stages,including 7 stages of zygote embryo callus induction of high efficiency and low salt medium DCR suits GS1 callus induction,a high concentration of 2,4-D can promote GS1 to produce large amounts of callus.The optimal medium for em-bryogenic callus induction was DCR+2,4-D 3.0 mg/L+NAA 1.0 mg/L+6-BA 0.5 mg/L.