表油菜素内酯对粗枝云杉胚性组织诱导的影响

发布时间：2018-03-15 07:09

本文选题：粗枝云杉　切入点：表油菜素内酯　出处：《中国林业科学研究院》2017年硕士论文　论文类型：学位论文

【摘要】：粗枝云杉(Picea asperata Mast.)体细胞胚胎发生技术为解决其无性繁殖和品种选育的问题提供了新途径,但目前仍存在胚性组织诱导率低、增殖后期胚性组织的高胚性难以维持、胚性组织分化能力低等问题。本研究以2015年粗枝云杉3个基因型的未成熟胚为试材,在已有体胚发生技术体系的基础上,在胚性组织诱导时,培养基中添加不同浓度EBR,提高了胚性组织的诱导率,优化了体胚发生的诱导体系,并进一步探讨EBR对同一增殖周期第0 d和第10 d时胚性组织中生理生化的变化,以及对体细胞胚分化的影响。结果表明:粗枝云杉未成熟合子胚在诱导培养基中,添加低浓度的EBR,能够有效地提高粗枝云杉胚性组织的诱导率和生长量。粗枝云杉基因型A、B和C未成熟胚在添加EBR诱导后,胚性组织诱导率的变化分别为:15%~47.5%,12.5%~23.3%和50%~77.5%。粗枝云杉3个基因型未成熟胚在不同浓度EBR诱导后的胚性组织中,内源激素含量最多的是IAA和ABA。在同一增殖周期的第0 d和第10 d时,实验组CTK/IAA的值均比对照组的值低,而CTK/GA的值,基因型A和C的实验组均比对照组的值高,而基因型B的实验组却比对照组的值低。各胚性组织内源激素含量的比值变化均不同,各激素之间通过联合或拮抗共同调节维持平衡。诱导后的胚性组织在同一增殖周期的第0 d和第10 d时,共检测到16种游离氨基酸的含量和3种游离糖的含量,实验组中游离氨基酸的总含量均低于对照组;基因型A和基因型B实验组的胚性组织,在同一增殖周期第0 d时,游离糖的总含量高于对照组,但在第10 d时,基因型A实验组胚性组织的游离糖总含量低于对照组,而基因型B实验组与对照组无显著差异;基因型C未成熟胚在诱导后形成的胚性组织,在第0 d和第10 d,实验组中游离糖的总含量以及多糖、葡萄糖和果糖的含量均低于对照组。胚性组织中抗氧化酶活性、MDA以及可溶性蛋白含量的检测发现:实验组与对照组相比,在第0 d和第10 d时,CAT活性以及可溶性蛋白含量在升高,而SOD与POD活性的变化则相反,MDA无显著差异,它们共同维持着胚性组织中活性氧的代谢平衡。粗枝云杉基因型A未成熟胚在添加0.5μM EBR诱导后的胚性组织有利于体细胞胚的分化,而基因型B和基因型C未成熟胚在添加1.0μM EBR诱导后的胚性组织有利于体细胞胚的分化。分化6周后,子叶胚的数量分别为54.01、52.36和74.85个/克,较未添加EBR诱导后的胚性组织分化的子叶胚数量分别高209.87%、238.90%和105.57%。本研究首次通过在诱导培养基中添加EBR,提高了粗枝云杉胚性组织的诱导率,优化了粗枝云杉胚性组织诱导体系。对同一增殖周期第0 d和第10 d的胚性组织,首次在生理生化水平上进行了探究,并且揭示了在EBR诱导后的胚性组织,体细胞胚分化的差异。
[Abstract]:Somatic embryogenesis of Picea asperata Mast.provides a new way to solve the problem of asexual reproduction and variety selection. However, there is still a low rate of embryogenic tissue induction, and it is difficult to maintain the high embryogenesis of embryogenic tissue in the late stage of proliferation. In this study, immature embryos of three genotypes of spruce spruce on 2015 were used as materials, on the basis of the technique system of somatic embryogenesis, when embryogenic tissue was induced, The induction rate of embryogenic tissue was improved and the induction system of somatic embryogenesis was optimized. The physiological and biochemical changes of embryogenic tissue at 0 and 10 days of the same proliferating cycle were further studied by adding different concentrations of EBR to the medium. The results showed that immature zygote embryos of Picea crassifolia were induced in culture medium. The induction rate and growth of embryogenic tissue of spruce were increased by adding low concentration of EBR.The immature embryos were induced by EBR. The percentage of embryogenic tissue induction was 23.3% and 5077.5%, respectively. The contents of endogenous hormones in embryogenic tissues of three genotypes of spruce spruce induced by different concentrations of EBR were the highest in IAA and ABA. at the 0 and 10 days of the same proliferating cycle, the contents of endogenous hormones in the immature embryos of the three genotypes of spruce spruce were higher than that of the control group at the 10th day and the 10th day of the same proliferative cycle, and the contents of endogenous hormones were the highest in the embryogenic tissues induced by different concentrations of EBR. The values of CTK/IAA in the experimental group were lower than those in the control group, while the values of CTK/GA, genotype A and C in the experimental group were higher than those in the control group, while the values of the experimental group with genotype B were lower than those in the control group. The levels of 16 kinds of free amino acids and 3 kinds of free sugar were detected at the 0 and 10 days of the same proliferating cycle in the embryogenic tissue after induction. The total content of free amino acids in the experimental group was lower than that in the control group, and the total content of free sugar in the embryogenic tissue of genotype A and genotype B groups was higher than that in the control group at the 0th day of the same proliferating cycle, but on the 10th day, the total content of free sugar in the experimental group was higher than that. The total content of free sugar in embryogenic tissue of genotype A group was lower than that of control group, but there was no significant difference between genotype B group and control group. The contents of free sugar, polysaccharides, glucose and fructose in the experimental group were lower than those in the control group at day 0 and day 10. The activity of cat and the content of soluble protein increased on day 0 and day 10, but there was no significant difference between SOD and POD. They maintained the metabolic balance of reactive oxygen species in embryogenic tissues, and the embryogenic tissues induced by the addition of 0.5 渭 M EBR from immature embryos of spruce genotype A were beneficial to the differentiation of somatic embryos. The embryogenic tissue of genotype B and genotype C immature embryos induced by 1.0 渭 M EBR was beneficial to the differentiation of somatic embryos. After 6 weeks of differentiation, the number of cotyledon embryos was 54.01 and 74.85 / g, respectively. The number of cotyledon embryos induced by EBR was 209.87% and 105.57% higher than that of embryogenic tissue induced by EBR for the first time, the induction rate of embryogenic tissue of Picea crassifolia was increased by adding EBR to the culture medium for the first time. The embryogenic tissue induction system of Picea crassifolia was optimized. The embryogenic tissue of the same proliferating cycle on day 0 and day 10 was studied at the physiological and biochemical level for the first time, and the embryogenic tissue induced by EBR was revealed. Differences in somatic embryogenesis.
【学位授予单位】：中国林业科学研究院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S791.18

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