本文選題：百合　切入點：體細(xì)胞胚起源　出處：《沈陽農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：體細(xì)胞胚發(fā)生對于百合種球繁殖、種質(zhì)保存、遺傳性狀改良等方面具有重要意義。目前百合體細(xì)胞胚發(fā)生的研究主要集中在植物生長調(diào)節(jié)劑和不同培養(yǎng)基對體細(xì)胞胚誘導(dǎo)效果的影響,大部分研究只憑肉眼觀察到的形態(tài)特征來判斷是否屬于體細(xì)胞胚,未進(jìn)行組織細(xì)胞學(xué)驗證,特別是對體細(xì)胞獲得胚性能力這個關(guān)鍵階段缺少必要的組織形態(tài)學(xué)研究,使得百合體細(xì)胞胚的形態(tài)建成研究受到限制。本試驗利用細(xì)胞生物學(xué)技術(shù),研究蘭州百合(Lilium davidii var.unicor)和細(xì)葉百合(LiliumpumilumDC.Fisch)體細(xì)胞胚發(fā)生途徑和起源方式,以期為明確百合體細(xì)胞胚發(fā)生機制奠定基礎(chǔ)。試驗結(jié)果如下:1.體細(xì)胞胚發(fā)生起源蘭州百合和細(xì)葉百合體細(xì)胞胚起源均存在外起源和內(nèi)起源兩種方式,其中外起源方式所占的比例分別是91.45%和83.85%。胚性細(xì)胞均在誘導(dǎo)體細(xì)胞胚發(fā)生的第9 d以外起源方式開始分裂。外起源位置分為兩種:表皮細(xì)胞和亞表皮細(xì)胞。蘭州百合的內(nèi)起源方式發(fā)生在誘導(dǎo)體細(xì)胞胚的第12 d,而細(xì)葉百合內(nèi)起源發(fā)生時間為誘導(dǎo)體細(xì)胞胚的第15 d。兩種百合體細(xì)胞胚發(fā)育過程中都存在導(dǎo)管細(xì)胞,均為多細(xì)胞起源發(fā)生方式。本試驗中,體細(xì)胞胚均以間接途徑發(fā)生,即先形成胚性愈傷組織,經(jīng)脫分化再分化后形成體細(xì)胞胚,最終形成個體植株。2.體細(xì)胞胚發(fā)育過程中指示因子的差異兩種百合胚性愈傷組織階段的持續(xù)時間存在明顯差異,細(xì)葉百合胚性愈傷組織持續(xù)時間長達(dá)42 d,而蘭州百合僅為28 d。蘭州百合在誘導(dǎo)體細(xì)胞胚的第9 d出現(xiàn)垂周分裂和平周分裂,而細(xì)葉百合出現(xiàn)垂周分裂和平周分裂時間為第15 d。體細(xì)胞胚發(fā)生過程中均發(fā)現(xiàn)表皮原細(xì)胞和維管束形成,蘭州百合和細(xì)葉百合出現(xiàn)表皮原細(xì)胞時間分別為誘導(dǎo)體細(xì)胞胚的第24 d和第30 d,維管束形成分別是在魚雷形胚時期和球形胚時期。蘭州百合和細(xì)葉百合體細(xì)胞胚形成生理隔離的時間也存在差異,分別為誘導(dǎo)體細(xì)胞胚的第30 d 和第 33 d。3.體細(xì)胞胚發(fā)育過程中淀粉粒的變化在誘導(dǎo)體細(xì)胞胚培養(yǎng)9 d后,兩種百合胚性細(xì)胞內(nèi)充斥著大量淀粉粒,為淀粉粒積累第一次高峰期,在球形胚期出現(xiàn)第二次積累高峰,第三次積累高峰是在早期魚雷形胚期,隨著體細(xì)胞胚的發(fā)育逐漸減少。4.體細(xì)胞胚發(fā)生過程的導(dǎo)管細(xì)胞在體細(xì)胞胚發(fā)生過程中,發(fā)現(xiàn)在胚性細(xì)胞周圍一直伴隨著導(dǎo)管細(xì)胞的出現(xiàn),并且在體細(xì)胞胚發(fā)育后期多個導(dǎo)管細(xì)胞基本形成一條直線。
[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.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S682.29

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