沙地云杉体细胞胚胎发生及组织细胞学观察

发布时间：2018-09-06 12:40

【摘要】：本试验分别以沙地云杉未成熟合子胚与成熟合子胚为外植体,研究不同基本培养基、不同激素组合,不同株系对胚性愈伤诱导、增殖以及体胚分化成熟的影响,建立并优化了沙地云杉体细胞胚胎发生体系,结果表明:1.沙地云杉胚性愈伤组织的诱导:以未成熟合子胚为外植体诱导胚性愈伤组织最适诱导条件包括:最佳的激素组合为添加6-BA 1.2～1.8 mg/L、2,4-D 1.8～2.7 mg/L,胚性愈伤组织诱导率最高可达74.00%;最佳采集日期为8月5日前后,胚性愈伤组织诱导率最高为67.70%;不同株系胚性愈伤组织诱导率最低为15.91%,最高可达77.50%,且诱导率与其饱籽率呈显著正相关(r=0.865,P=0.006)关系;以成熟合子胚为外植体诱导胚性愈伤组织的培养条件为:初代培养采用LM +6-BA 2.0 mg/L + KT 1.0 mg/L + 2,4-D 4.0 mg/L,继代培养采用 1/2LM + 6-BA 1.0 mg/L + KT 0.5 mg/L + 2,4-D 2 mg/L,胚性愈伤组织诱导率为12.00%,其它处理没能诱导产生胚性愈伤组织。2.胚性愈伤组织增殖最佳激素组合为添加6-BA 0.8 mg/L、2,4-D 0.8～1.6 mg/L,在愈伤组织增殖过程中,2,4-D的作用更明显,最高增殖率为7.24倍。3.沙地云杉体细胞胚胎分化与成熟最适培养条件为:1/2LM+ABA 22.5mg/L +PEG4000 45 g/L,体胚分化数最高为362个/g,畸形胚数为108个/g,畸形胚率为29.83%。K-81株系体胚分化数最高为573.4个/g,其中畸形胚数为151个/g,畸形胚率为26.33%。4.沙地云杉体细胞胚胎发生发育过程组织细胞学观察表明:胚性愈伤组织是由椭圆形或长条形的胚性细胞以及由胚性细胞分裂所产生的胚性胚柄团组成;在愈伤组织成熟发育过程中,体细胞胚胎发育共经过早期原胚、球形胚,早期体胚及子叶胚四个阶段。
[Abstract]:In this experiment, immature zygotic embryos and mature zygotic embryos of Picea spruce were used as explants to study the effects of different basic media, different hormone combinations and different strains on embryogenic callus induction, proliferation and somatic embryo differentiation and maturation. The somatic embryogenesis system of Picea spruce was established and optimized. Embryogenic callus induction of Picea spruce: the optimum conditions for embryogenic callus induction with immature zygotic embryos as explants are as follows: the best hormone combination is the rate of embryogenic callus induction added with 6-BA 1.2n 1.8 mg/L,2,4-D 1.82.7 mg/L,. Up to 74.00; the best date of collection is around August 5, The highest rate of embryogenic callus induction was 67.70%, the lowest rate of embryogenic callus was 15.91, the highest was 77.50%, and there was a significant positive correlation between the induction rate and the full seed rate (r 0.865P0. 006). The culture conditions of embryogenic callus induced by mature zygote embryos were as follows: the primary culture was carried out with LM 6-BA 2.0 mg/L KT 1.0 mg/L 2G 4-D 4.0 mg/L, subculture and 1/2LM 6-BA 1.0 mg/L KT 0.5 mg/L 2N 4-D 2 mg/L, embryos. The rate of callus induction was 12.00%, while other treatments failed to induce embryogenic callus. The best hormone combination of embryogenic callus proliferation was that the addition of 6-BA 0.8 mg/L,2,4-D 0.81. 6 mg/L, had a more obvious effect on callus proliferation, and the highest proliferation rate was 7.24 times. 3. The optimum culture conditions for somatic embryo differentiation and maturation of Picea spruce are as follows: 1 / 2 LM ABA 22.5mg/L PEG4000 45 g / L, somatic embryo differentiation highest 362 / g, abnormal embryo number 108 / g, abnormal embryo rate of 29.83%.K-81 strain highest 573.4% / g. The number of embryos is 151 / g, and the rate of abnormal embryo is 26.33. 4. Histological observation of somatic embryogenesis and development of Picea spruce showed that embryogenic callus was composed of elliptical or long embryonic cells and embryogenic petiole produced by embryogenic cell division. During the process of callus maturation, somatic embryogenesis went through four stages: early proembryo, globular embryo, early somatic embryo and cotyledon embryo.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S791.18

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