两种中国原产百合体细胞胚发生发育的组织学初探

发布时间：2018-03-28 11:55

本文选题：百合　切入点：体细胞胚起源　出处：《沈阳农业大学》2017年硕士论文

【摘要】：体细胞胚发生对于百合种球繁殖、种质保存、遗传性状改良等方面具有重要意义。目前百合体细胞胚发生的研究主要集中在植物生长调节剂和不同培养基对体细胞胚诱导效果的影响,大部分研究只凭肉眼观察到的形态特征来判断是否属于体细胞胚,未进行组织细胞学验证,特别是对体细胞获得胚性能力这个关键阶段缺少必要的组织形态学研究,使得百合体细胞胚的形态建成研究受到限制。本试验利用细胞生物学技术,研究兰州百合(Lilium davidii var.unicor)和细叶百合(LiliumpumilumDC.Fisch)体细胞胚发生途径和起源方式,以期为明确百合体细胞胚发生机制奠定基础。试验结果如下:1.体细胞胚发生起源兰州百合和细叶百合体细胞胚起源均存在外起源和内起源两种方式,其中外起源方式所占的比例分别是91.45%和83.85%。胚性细胞均在诱导体细胞胚发生的第9 d以外起源方式开始分裂。外起源位置分为两种:表皮细胞和亚表皮细胞。兰州百合的内起源方式发生在诱导体细胞胚的第12 d,而细叶百合内起源发生时间为诱导体细胞胚的第15 d。两种百合体细胞胚发育过程中都存在导管细胞,均为多细胞起源发生方式。本试验中,体细胞胚均以间接途径发生,即先形成胚性愈伤组织,经脱分化再分化后形成体细胞胚,最终形成个体植株。2.体细胞胚发育过程中指示因子的差异两种百合胚性愈伤组织阶段的持续时间存在明显差异,细叶百合胚性愈伤组织持续时间长达42 d,而兰州百合仅为28 d。兰州百合在诱导体细胞胚的第9 d出现垂周分裂和平周分裂,而细叶百合出现垂周分裂和平周分裂时间为第15 d。体细胞胚发生过程中均发现表皮原细胞和维管束形成,兰州百合和细叶百合出现表皮原细胞时间分别为诱导体细胞胚的第24 d和第30 d,维管束形成分别是在鱼雷形胚时期和球形胚时期。兰州百合和细叶百合体细胞胚形成生理隔离的时间也存在差异,分别为诱导体细胞胚的第30 d 和第 33 d。3.体细胞胚发育过程中淀粉粒的变化在诱导体细胞胚培养9 d后,两种百合胚性细胞内充斥着大量淀粉粒,为淀粉粒积累第一次高峰期,在球形胚期出现第二次积累高峰,第三次积累高峰是在早期鱼雷形胚期,随着体细胞胚的发育逐渐减少。4.体细胞胚发生过程的导管细胞在体细胞胚发生过程中,发现在胚性细胞周围一直伴随着导管细胞的出现,并且在体细胞胚发育后期多个导管细胞基本形成一条直线。
[Abstract]:Somatic embryogenesis for bulb propagation, germplasm preservation, Studies on somatic embryogenesis in lily are mainly focused on the effects of plant growth regulators and different media on somatic embryogenesis. Most of the studies only depend on the morphological characteristics observed by the naked eye to determine whether the somatic embryo is a somatic embryo, and has not been verified by histocytology, especially in the critical stage of somatic embryogenic ability. The morphogenesis of somatic embryogenesis in Lilium davidii var. unicor. and Lilium pumilum DC.Fisch) was studied by using cell biology technique in this experiment, and the way of somatic embryogenesis and the origin of somatic embryogenesis in Lilium davidii var. unicor. and Lilium pumilum DC.Fischchwere studied in this experiment. The results are as follows: 1. The origin of somatic embryogenesis in lily, Lanzhou lily and Lilium lilium, has two ways: external origin and internal origin. The proportion of external origin was 91.45% and 83.85% respectively. Embryogenic cells began to divide outside the 9th day of somatic embryogenesis. There are two types of exogenetic sites: epidermal cells and subepidermal cells. The internal origin of lily occurred at the 12th day of somatic embryogenesis, while that of Lilium tenuifolia was at the 15th day of somatic embryogenesis. Ductal cells were present in the development of somatic embryogenesis in both lily and lily. In this experiment, somatic embryogenesis occurred indirectly, that is to say, embryogenic callus was formed first, and somatic embryos were formed after dedifferentiation and redifferentiation. The difference of indicator factors in somatic embryogenesis was significant difference in the duration of embryogenic callus between two species of lily. The duration of embryogenic callus of Lilium tenuifolia was 42 days, but that of Lanzhou lily was only 28 days. On the other hand, the time of vertical division and circumferential division was 15 days. Epidermal protocells and vascular bundles were found in somatic embryogenesis. The time of appearance of epidermal protocells in Lanzhou lily and fine leaf lily was the 24th day and the 30th day of inducing somatic embryo, respectively, and the formation of vascular bundle was at the stage of torpedo embryo and globular embryo, respectively, and the somatic embryo of Lanzhou lily and lily finely leafed lily was formed at the same time as the somatic embryo. There are also differences in the time when physiological isolation is formed, The changes of starch granules during somatic embryogenesis were observed on the 30th day and 33rd day of somatic embryogenesis. After 9 days of induced somatic embryogenesis, a large number of starch granules were found in the embryogenic cells of the two lily species, which was the first peak of starch accumulation. The second accumulation peak occurred in the globular embryo stage, and the third accumulation peak occurred in the early torpedo stage. With the development of somatic embryos, the number of ducted cells in the somatic embryogenesis process decreased gradually, and the third accumulation peak occurred during the somatic embryogenesis. It was found that ductal cells appeared all the time around embryogenic cells, and many ductal cells formed a straight line in the late stage of somatic embryogenesis.
【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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