基于体细胞胚发生的五种切花百合高效再生体系建立

发布时间：2018-01-14 01:11

本文关键词：基于体细胞胚发生的五种切花百合高效再生体系建立　出处：《沈阳农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：体细胞胚发生对于百合种球繁殖、种质资源保存、遗传性状改良等具有重要意义。本试验以五个不同系列的百合切花品种为试材,研究了不同外植体、不同PIC浓度及不同培养条件对体细胞胚诱导以及成熟体胚萌发的影响,分别建立了 5种百合体细胞胚发生的高效再生体系,为百合种球繁育和基于体胚发生的遗传转化体系建立奠定了基础。主要研究结果如下:1.以叶片为外植体诱导体细胞胚:体细胞胚以间接途径发生,即首先形成胚性愈伤组织。'Strawberry and cream'的最适诱导培养基为 MS+2.0 mg·L~(-1) PIC+0.2 mg·L~(-1)NAA,其余四个品种的最适培养基为MS+1.5 mg·L~(-1) PIC+0.2 mg·L~(-1) NAA,在暗培养条件下诱导效果更佳。铁炮百合'White heaven'与LO系列百合'Triumphator'的诱导率均可达95.83%,LA 系列百合'Eyeliner'的诱导率可达 95.00%,亚洲百合'Strawberry and cream,与东方百合'Double surprise'的诱导率均为93.33%。2.以无菌苗鳞片为外植体诱导体细胞胚:不同百合品种的发生途径不同。结果表明,'Eyeliner'与'Double surprise'诱导体细胞胚的最适培养基为 MS+2.5 mg·L~(-1) PIC+0.2 mg·L~(-1)NAA,培养条件为暗培养,诱导率均可达90.83%。'White heaven'最佳培养基与'Eyeliner'相同,但在光培养条件下诱导效果更佳,可达96.67%。'Strawberry and cream'与'Triumphator'的最适培养基为 MS+2.0 mg,L~(-1) PIC+0.2 mg·L~(-1) NAA,其中'Strawberry and cream'在光培养条件下诱导效果更好,诱导率可达88.33%,而'Triumphator'在暗培养条件下诱导效果较好,诱导率为93.33%。3.体细胞胚的分化:将胚性愈伤组织转接到体胚分化培养基中,结果表明,暗培养更有利于胚性愈伤分化及体细胞胚的形成。'Triumphator'分化率达97.5%,'Eyeliner'与'Strawberry and cream'分化率均为 95%,'Double surprise' 分化率为 92.5%,'White heaven'分化率为90%。4.体细胞胚的萌发:将长有不同形态体胚的胚性愈伤组织转入MS+0.5 mg·L~(-1) BA,结果发现,光培养更有利于体胚的萌发。'Eyeliner'和'Triumphator'萌发率达到97.5%。'White heaven'和'Double surprise'萌发率均达到 95%。'Strawberry and cream'体细胞胚的萌发率达到92.5%。5.炼苗移栽:将生长状态良好的小苗移栽至草炭:蛭石:珍珠岩为1:1:1的基质中,幼苗成活率相均可达到90%以上。
[Abstract]:Somatic embryogenesis plays an important role in bulb propagation, germplasm conservation and genetic improvement of lily. In this study, five different series of lily cut flower varieties were used as test materials to study different explants. The effects of different PIC concentration and culture conditions on somatic embryogenesis and mature somatic embryo germination were studied. Five efficient regeneration systems of somatic embryogenesis of lily were established. The main results are as follows: 1. Somatic embryogenesis induces somatic embryogenesis by indirect pathway with leaf as explant. The optimal medium for inducing embryogenic callus. Strawberry and cream' was MS 2.0 mg 路L ~ (-1). PIC 0.2 mg 路L ~ (-1). The optimum medium for the other four varieties was MS 1.5 mg 路L ~ (-1) PIC 0.2 mg 路L ~ (-1) NAA. The induction rate of White Haven 'and Lo series lily Triumphator' was 95.83%. The induction rate of LA series lily Eyeliner' was 95.00, and that of Asian lily was Strawberry and cream. The inductive rates of double surprise' were 93.33.2.The somatic embryos were induced by the scales of aseptic plantlets as explants. The results showed that different varieties of lily had different paths of occurrence. The most suitable medium for inducing somatic embryos was MS 2.5 mg 路L ~ (-1). PIC 0.2 mg 路L ~ (-1). The culture conditions were dark culture, the induction rate could reach 90.83. the best medium of White Heaven' was the same as that of Eyeliner', but the effect was better under the condition of light culture. The optimum medium for Strawberry and cream' and Triumphator' was MS 2.0 mg. PIC 0.2 mg 路L ~ (-1) 路L ~ (-1) NAA, in which Strawberry and cream' was better induced under light culture. The induction rate was 88.33 and Triumphator' was better in dark culture. The induction rate was 93.330.The differentiation of somatic embryogenesis: the embryogenic callus was transferred to the somatic embryo differentiation medium. Dark culture was more favorable to embryogenic callus differentiation and somatic embryogenesis. The differentiation rate of Triumphator' was 97.5%. The differentiation rates of Eyeliner' and Strawberry and cream' were both 95%. The differentiation rate of double multiple 'was 92.5%. The differentiation rate of White Heaven' was 90.4.The germination of somatic embryos: the embryogenic calli with different somatic embryos were transferred to MS 0.5 mg 路L ~ (-1) BA. Turns out. Light culture is more favorable to the germination of somatic embryos. Eyeler 'and Triumphator' germinate to 97.5. White. Both Heaven' and double Surprise' germinated to 95% Strawberry and. The germinating rate of cream' somatic embryos was 92.5%. 5. Transplanting the plantlets in good growing condition into a medium with vermiculite: perlite = 1: 1: 1. The survival rate of seedlings can reach more than 90%.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S682.29

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