天山云杉成熟与未成熟合子胚的胚性愈伤组织诱导及增殖研究

Study on the Induction and Proliferation of Somatic Embryogenesis from the Mature and Immature Zygote Embryos of Picea schrenkiana

【目的】研究天山云杉成熟与未成熟合子胚的胚性愈伤组织诱导及增殖条件技术,对该树种的胚胎发生机理和建立相关的快繁技术体系奠定了基础。【方法】以带胚乳和不带胚乳的未成熟合子胚及成熟合子胚为外植体,在不同生长调节剂浓度的DCR培养基上进行诱导培养。对不同生长时期合子胚的胚性愈伤组织诱导率和增殖量进行研究。【结果】(1)添加不同生长调节剂(2,4-D、6-BA、TDZ)浓度组合的DCR培养基上诱导成熟合子胚时,20μM 2,4-D和5μM 6-BA为最佳组合浓度,胚性愈伤组织诱导率最高,为55%。(2)对不同采样时间的三种合子胚胚性愈伤组织诱导率进行分析表明,带胚乳的未成熟合子胚(6月)为32.6%,成熟合子胚(9月)为60.6%,不带胚乳的未成熟合子胚(7月)为83.1%,7月为天山云杉胚性愈伤组织诱导的最佳采摘期。(3)胚性愈伤组织在不同5种培养基上增殖培养表明,培养基之间增殖量差异显著(P<0.05)。BM2和SH培养基的增殖效果明显优于MSG,1/2DCR,DCR,以SH的综合增殖效果最好,其增殖量为4.55 g,增殖率为600%。(4)5种个体胚性愈伤组织在SH培养基上增殖培养表明,个体之间增殖量变化不明显(P>0.05)。【结论】建立了天山云杉胚性愈伤组织发生技术平台,为天山云杉遗传改良种质创新、缩短育种周期奠定了研究基础。

[ Objective] To study the induction and proliferation of somatic embryogenesis from the mature and immature zygote embryos of Picea schrenkiana, which might have great significance for the understanding of the mechanism of embryonic development and the establishment of rapid propagation system. [ Method ] Immature embryo and mature embryo were used as explants to induce cuhure in DCR medium with different growth regulator concentrations. The induction rates of zygotic embryos in different growth stages and its prolif- eration were studied using immature zygotic embryos with endosperm, immature zygotic embryos without endo- sperm and mature embryos as explants. [ Result] （ 1 ） When DCR medium added with different concentrations of 2,4 - D and 6 - BA was used to induce mature zygotic embryos, the combination of 20 tzM 2,4 - D and 5 bLM 6 - BA was the best and somatic embryogenesis induction rate was the highest, up to 55 %. （2） The in- duction rate of three zygotic embryos of somatic embryogenesis collected in different periods showed that the immature zygotic embryos with endosperm （June） was up to 32.6%, the mature zygotic embryos （ September） was up to 60.6%, Immature embryo without endosperm （July） was 83.1%, which indicated that July was the best picking period of P. schrenkiana cones. （3） The somatic embryogenesis in the five different proliferation medium showed the the proliferation rate was significant （ P 〈 0.05 ）. The proliferation results of somatic embryogenesis in different proliferation medium showed that the effect of BM2 and SH were better than the MSG, 1/2DCR, DCR, and the comprehensive proliferation effect of SH was the best. The proliferation rate was 4.55 g and the rate was up 600%. ） （4） The five different somatic embryogenesis in SH proliferation medium showed that the difference between somatic embryogenesis was not significant （ P 〉 0.05 ）. [ Conclusion ] This study has established the somatic embryogenesis technology platform of P. schrenkiana for the first time, which has provided a scientific basis for the genetic improvement of germplasm and shortened the breed- ing cycle of P. Schrenkiana.