本文選題：粗枝云杉　切入點(diǎn)：表油菜素內(nèi)酯　出處：《中國林業(yè)科學(xué)研究院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：粗枝云杉(Picea asperata Mast.)體細(xì)胞胚胎發(fā)生技術(shù)為解決其無性繁殖和品種選育的問題提供了新途徑,但目前仍存在胚性組織誘導(dǎo)率低、增殖后期胚性組織的高胚性難以維持、胚性組織分化能力低等問題。本研究以2015年粗枝云杉3個(gè)基因型的未成熟胚為試材,在已有體胚發(fā)生技術(shù)體系的基礎(chǔ)上,在胚性組織誘導(dǎo)時(shí),培養(yǎng)基中添加不同濃度EBR,提高了胚性組織的誘導(dǎo)率,優(yōu)化了體胚發(fā)生的誘導(dǎo)體系,并進(jìn)一步探討EBR對(duì)同一增殖周期第0 d和第10 d時(shí)胚性組織中生理生化的變化,以及對(duì)體細(xì)胞胚分化的影響。結(jié)果表明:粗枝云杉未成熟合子胚在誘導(dǎo)培養(yǎng)基中,添加低濃度的EBR,能夠有效地提高粗枝云杉胚性組織的誘導(dǎo)率和生長量。粗枝云杉基因型A、B和C未成熟胚在添加EBR誘導(dǎo)后,胚性組織誘導(dǎo)率的變化分別為:15%~47.5%,12.5%~23.3%和50%~77.5%。粗枝云杉3個(gè)基因型未成熟胚在不同濃度EBR誘導(dǎo)后的胚性組織中,內(nèi)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時(shí),實(shí)驗(yàn)組CTK/IAA的值均比對(duì)照組的值低,而CTK/GA的值,基因型A和C的實(shí)驗(yàn)組均比對(duì)照組的值高,而基因型B的實(shí)驗(yàn)組卻比對(duì)照組的值低。各胚性組織內(nèi)源激素含量的比值變化均不同,各激素之間通過聯(lián)合或拮抗共同調(diào)節(jié)維持平衡。誘導(dǎo)后的胚性組織在同一增殖周期的第0 d和第10 d時(shí),共檢測到16種游離氨基酸的含量和3種游離糖的含量,實(shí)驗(yàn)組中游離氨基酸的總含量均低于對(duì)照組;基因型A和基因型B實(shí)驗(yàn)組的胚性組織,在同一增殖周期第0 d時(shí),游離糖的總含量高于對(duì)照組,但在第10 d時(shí),基因型A實(shí)驗(yàn)組胚性組織的游離糖總含量低于對(duì)照組,而基因型B實(shí)驗(yàn)組與對(duì)照組無顯著差異;基因型C未成熟胚在誘導(dǎo)后形成的胚性組織,在第0 d和第10 d,實(shí)驗(yàn)組中游離糖的總含量以及多糖、葡萄糖和果糖的含量均低于對(duì)照組。胚性組織中抗氧化酶活性、MDA以及可溶性蛋白含量的檢測發(fā)現(xiàn):實(shí)驗(yàn)組與對(duì)照組相比,在第0 d和第10 d時(shí),CAT活性以及可溶性蛋白含量在升高,而SOD與POD活性的變化則相反,MDA無顯著差異,它們共同維持著胚性組織中活性氧的代謝平衡。粗枝云杉基因型A未成熟胚在添加0.5μM EBR誘導(dǎo)后的胚性組織有利于體細(xì)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誘導(dǎo)后的胚性組織有利于體細(xì)胞胚的分化。分化6周后,子葉胚的數(shù)量分別為54.01、52.36和74.85個(gè)/克,較未添加EBR誘導(dǎo)后的胚性組織分化的子葉胚數(shù)量分別高209.87%、238.90%和105.57%。本研究首次通過在誘導(dǎo)培養(yǎng)基中添加EBR,提高了粗枝云杉胚性組織的誘導(dǎo)率,優(yōu)化了粗枝云杉胚性組織誘導(dǎo)體系。對(duì)同一增殖周期第0 d和第10 d的胚性組織,首次在生理生化水平上進(jìn)行了探究,并且揭示了在EBR誘導(dǎo)后的胚性組織,體細(xì)胞胚分化的差異。
[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.
【學(xué)位授予單位】：中國林業(yè)科學(xué)研究院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S791.18

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