山杏不同外植体愈伤组织的诱导及其体细胞胚胎发生

发布时间：2018-05-06 21:40

本文选题：山杏 + 外植体　；参考：《内蒙古农业大学》2017年硕士论文

【摘要】：山杏是北方地区主要的经济树种和生态绿化树种其用途广泛和实用价值,山杏常规繁殖速度慢,并且山杏种子数量受气候影响极大,存在大小年现象,妨碍优良品种的推广与应用。运用植物组织培养技术,具有繁殖系数大,可获得温度优良性状的山杏苗木,是解决山杏繁殖问题的有效途径,本文以叶片、种子、和茎尖为外植体,研究不同外植体愈伤组织的诱导和分化,通过愈伤组织途径建立了再生植株体系、通过诱导胚性愈伤组织分化研究其体细胞胚胎发生,获得了山杏体细胞胚。结果如下:(1)试验筛选结果得出:成熟胚和茎尖分别是山杏愈伤组织诱导和体细胞胚胎发生的最佳外植体。(2)种子随着消毒时间的增加,感染率降低,诱导率下降,75%酒精30s→2%NaC1O 4min处理种子,感染率为6.6%,但诱导率高达90%,是种子最佳消毒方法;而茎尖在75%酒精30s→0.2%升汞6min →2%NaC10 4min下消毒效果最好。(3)成熟胚在MS+6-BA(0.5mg/L)+NAA(0.5mg/L)培养基中愈伤组织诱导率达100%,当6-BA浓度为2.0mg/L,NAA浓度为0.2mg/L时,愈伤组织的分化率最高为5.88;而成熟胚在光照培养下直接诱导丛生芽,当6-BA(1.0 mg/L)和NAA(0.2 mg/L)组合中芽丛长势旺盛,叶片浓绿,生长良好且分化率高达10.75,是成熟胚诱导丛生芽的最佳培养基;最后将芽丛切下在NAA和IBA作用下诱导生根,NAA配合IBA使用时,芽丛的生根率较低甚至没有生根,基部有大量愈伤,单独使用NAA时,随着NAA的浓度增加,芽丛生根率也随之增加,浓度为0.5mg/L生根率最高为 75%。(4)对胚性愈伤组织增殖试验添加附加物水解乳蛋白、水解酪蛋白和肌醇,添加水解酪蛋白愈伤增殖率为92.2%最高;而胚性愈伤在1.Omg/L 6-BA和0.05mmg/LNAA的激素组合中诱导率最高为40%,作为胚性愈伤适宜分化培养基。
[Abstract]:Apricot is the main economic tree species and ecological greening tree species in the north of China, which is widely used and practical value. The rate of conventional propagation of almonds is slow, and the number of apricot seeds is greatly affected by climate, and there is a phenomenon of large and small years. It hinders the popularization and application of fine varieties. It is an effective way to solve the problem of propagation of apricot by using plant tissue culture technique, which has high reproduction coefficient and good temperature character. In this paper, leaves, seeds and stem tips are used as explants. The callus induction and differentiation of different explants were studied. The regenerated plant system was established through callus pathway. Somatic embryogenesis was studied by inducing embryogenic callus, and somatic embryogenesis of apricot was obtained. The results showed that mature embryo and stem tip were the best explants for callus induction and somatic embryogenesis, respectively. The infection rate decreased with the increase of disinfection time. The induction rate decreased by 75% alcohol for 30s and 2%NaC1O 4min was used to treat seeds, the infection rate was 6.6%, but the induction rate was as high as 90%, so it was the best way to sterilize the seeds. The callus induction rate of mature embryo in MS 6-BAN 0.5 mg / L NAA 0.5 mg / L medium was 100 when the 6-BA concentration was 2.0 mg / L 0.2mg/L, while the stem tip was sterilized at 75% alcohol for 30 s or 0.2% mercuric acid (2%NaC10 4min), and the rate of callus induction was 100% in MS 6-BAN 0.5 mg / L NAA / L medium. When the 6-BA concentration was 2.0 mg / L, the concentration of NAA was 0.2mg/L. The highest differentiation rate of callus was 5.88, while mature embryos induced cluster buds directly under light culture. When 6-BA(1.0 mg / L) and NAA(0.2 mg / L) combinations, buds grew vigorously and leaves were thick green. It was the best medium for mature embryos to induce tufted buds. Finally, when NAA and IBA were used to induce rooting with IBA, the rooting rate of bud plexus was lower or not, and a large number of calli were found at the base. When NAA was used alone, the rooting rate of bud tufts increased with the increase of NAA concentration. The highest rooting rate of 0.5mg/L was 75. 4) the addition of hydrolactin, casein and inositol was added to embryogenic callus proliferation test. The highest rate of callus proliferation was 92. 2% in addition of hydrolysate casein, and the highest rate of embryogenic callus was 40 in the combination of 1.Omg/L 6-BA and 0.05mmg/LNAA, which was used as the suitable medium for differentiation of embryogenic callus.
【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】：S662.2

【参考文献】