本文選題：珙桐　切入點(diǎn)：苞片發(fā)育　出處：《中南林業(yè)科技大學(xué)》2017年碩士論文

【摘要】：珙桐(Davidia involucrata Baill.)是珙桐科孓遺植物,在地球上生活了近6000萬(wàn)年,是極珍貴的樹(shù)種,具有很高的研究?jī)r(jià)值,西方對(duì)珙桐在園林上的應(yīng)用有很高的評(píng)價(jià)。珙桐最具代表性的即是其圓形的頭狀花序和成對(duì)的白色苞片。苞片是植物在生殖發(fā)育中出現(xiàn)的一類特殊的器官,廣義上將其定義為與花或花序相關(guān)的葉片,珙桐是研究苞片發(fā)育的理想材料,珙桐苞片不僅表型十分獨(dú)特,而且相較于苞片和葉片同時(shí)期生長(zhǎng),苞片的生長(zhǎng)速度遠(yuǎn)遠(yuǎn)超過(guò)同時(shí)萌發(fā)的葉片。本研究主要從苞片發(fā)育過(guò)程中的生理生化分析及形態(tài)學(xué)觀察:采集苞片不同發(fā)育階段的樣品,記錄苞片顏色,大小苞片生長(zhǎng)數(shù)據(jù)和苞片生長(zhǎng)過(guò)程中的葉綠素含量等相關(guān)生理指標(biāo),將珙桐苞片的生長(zhǎng)發(fā)育階段進(jìn)行分段分析,重點(diǎn)分析各發(fā)育階段珙桐苞片內(nèi)源激素含量變化。參考實(shí)際觀測(cè)結(jié)果和激素變化情況,結(jié)合本實(shí)驗(yàn)組已有的珙桐苞片轉(zhuǎn)錄組數(shù)據(jù)庫(kù)挑選與珙桐苞片生長(zhǎng)發(fā)育有關(guān)的ASR基因,提取總RNA,克隆相關(guān)基因并用原核蛋白表達(dá)其特異性蛋白。使用適合于珙桐苞片發(fā)育定量分析的內(nèi)參基因,根據(jù)表達(dá)模式對(duì)葉片與苞片,及苞片不同發(fā)育階段的樣品進(jìn)qRT-PCR定量分析,利用qRT-PCR驗(yàn)證相關(guān)基因的表達(dá)規(guī)律。實(shí)驗(yàn)結(jié)果如下:1珙桐苞片形態(tài)發(fā)育規(guī)律珙桐苞片的生長(zhǎng)發(fā)育階段可以主要分為Ⅳ個(gè)時(shí)期:在苞片Ⅰ期,珙桐苞片在形成初期呈綠色,苞片與葉片同在混合芽中,芽鱗未完全脫落,與普通葉片形態(tài)差別很小,色差很小。苞片Ⅱ期,珙桐苞片顏色逐漸與葉片在色彩上出現(xiàn)差異,顏色變淺,苞片快速生長(zhǎng)。苞片Ⅲ期,苞片綠色已經(jīng)基本退去,略微帶淺黃色,苞片生長(zhǎng)速度稍微放緩。苞片Ⅳ期的苞片已經(jīng)到達(dá)成熟期,純白色,到達(dá)苞片的最大尺寸,生長(zhǎng)停止。2.珙桐苞片發(fā)育過(guò)程中主要內(nèi)源激素的變化規(guī)律使用高效液相色譜對(duì)珙桐苞片不同發(fā)育過(guò)程中的內(nèi)源激素含量的測(cè)定顯示:在苞片與葉片的比較中發(fā)現(xiàn),珙桐苞片中的內(nèi)源激素含量遠(yuǎn)遠(yuǎn)高于珙桐葉片中的含量。珙桐苞片發(fā)育的整個(gè)過(guò)程中主要內(nèi)源激素含量的變化總體上是不斷增加的;GA3與ABA的含量在珙桐苞片發(fā)育過(guò)程中是先上升,到最后階段稍微下降;而IAA與ZT的含量在珙桐苞片發(fā)育過(guò)程中呈上升趨勢(shì),在最后階段達(dá)到最大值。3.珙桐苞片ABA激素與DiASR表達(dá)量的變化規(guī)律從珙桐苞片轉(zhuǎn)錄組數(shù)據(jù)庫(kù)中挑選出在珙桐苞片中與ABA有關(guān)且高量表達(dá)的DiASR基因。通過(guò)對(duì)珙桐苞片發(fā)育過(guò)程中內(nèi)源激素含量的測(cè)定發(fā)現(xiàn):ABA激素含量隨著苞片生長(zhǎng)發(fā)育而穩(wěn)步增長(zhǎng)的同時(shí),當(dāng)?shù)竭_(dá)苞片Ⅲ期時(shí)達(dá)到最大值(ⅢⅡⅣⅠ),珙桐苞片發(fā)育Ⅲ期與其它幾期的ABA激素含量有顯著性差異。根據(jù)珙桐苞片各發(fā)育階段的熒光定量結(jié)果表明:苞片的DiASR基因表達(dá)量是花序的200倍、是葉片的110倍,DiASR基因在苞片Ⅲ期的表達(dá)量最高,DiASR在珙桐苞片發(fā)育Ⅱ期表達(dá)量劇增,到Ⅲ期達(dá)到最高值(ⅢⅣⅡⅠ)。可以看出ABA含量和調(diào)控ABA表達(dá)的DiASR基因的變化情況基本一致,都在苞片Ⅲ期達(dá)到最大值,在苞片Ⅰ期達(dá)到最小值。依據(jù)兩者的數(shù)據(jù)分析。發(fā)現(xiàn)兩者的表達(dá)水平具有顯著相關(guān)性。
[Abstract]:Dovetree (Davidia involucrata Baill.) is Nyssaceae relic plants, life on earth for nearly 60 million years, is the most valuable species, has high research value and application of Western Davidia involucrata in landscape are very high appraisal. Its most representative is white bracts round its head inflorescence and paired. Bracts is in a special class of plant organs appeared in the reproductive development, in broad sense, which is defined as with flower or inflorescence related leaf, bract development of Davidia involucrata is ideal material, not only a unique phenotype of bract of Davidia involucrata, and compared with bracts and leaves at the same period of growth. The growth rate is far more than the leaf bracts also germination. This study mainly developed in the process of physiological and biochemical analysis and morphological observation from bracts: samples of different developmental stages of acquisition bracts, record bract color, size of bract growth data and bracts The related physiological indexes during the growth of chlorophyll content, the growth stage of Davidia bract segment analysis, focuses on the analysis of changes of endogenous hormones in different developmental stages of Davidia bracts. The actual observation results and hormonal changes, combined with the experimental group have Davidia transcriptome database selection and bract of Davidia involucrata bract growth ASR gene related to the extraction of total RNA, gene cloning and expression of the specific protein with prokaryotic protein. Suitable for use in Davidia involucrate bract developed reference genes for quantitative analysis, according to the expression pattern of leaf and bract, bract and quantitative analysis of qRT-PCR in different developmental stages of sample, expression of genes related to verification by qRT-PCR. The experimental results are as follows: 1 Davidia bract morphology development regularity of Davidia bract growth stage can be divided into 4 stages: in stage I of bract, bract of Davidia involucrata In the early formation of a green bracts and leaves, with mixed buds, bud scales is not completely detached, very small and ordinary leaf shape difference, color difference is very small. Bracts II, bract and leaf color gradually Dovetree in color difference, color shallow, bracts bracts of rapid growth. Bracts green, has receded slightly pale yellow bracts slightly slower. The growth rate of bract bract stage has reached maturity, the pure white, reaching the maximum size of bracts, determination of endogenous hormone content of growth cessation changes of endogenous.2. in the process of development of bract of Davidia involucrata using high performance liquid chromatography of Davidia bracts in the process of development of different display: in the bracts and leaves, the content of endogenous hormones in the bract of Davidia involucrata is far higher than the content in the leaves. The main content of Dovetree endogenous hormones in the whole process of development in the bract of Davidia involucrata The overall change is increasing; the content of GA3 and ABA in the development process of Davidia bracts is increased to the final stage decreased slightly; while the content of IAA and ZT increased in Davidia bracts during development, reached the maximum value of.3. ABA DiASR hormone and bract of Davidia involucrata expression quantity pick out in Davidia bracts in ABA and the high expression of DiASR gene from bract of Davidia involucrata transcriptome in the last stage. Through the determination of the contents of endogenous hormones in the process of development of Davidia involucrate bract found: ABA with growth hormone content of bracts and steady growth at the same time, when the maximum arrival bracts of third stage (II III IV I), there was significant difference in development stage and bract of Davidia involucrata ABA hormone content in several other period. According to the quantitative fluorescence in different developmental stages of bract of Davidia involucrata results: DiASR gene expression is the bract inflorescence 200 times, 110 times the amount of leaves, DiASR gene expression was highest in bracts III, DiASR in Davidia bract developed phase II expression increase, to reach the highest value (stage III IV II I). See the changes in the content of ABA and the regulation of ABA expression of DiASR gene can be basically the same, are bracts in stage III reached the maximum in the bract stage and reached the minimum. Based on the analysis of the data found. Their expression level has a significant correlation.

【學(xué)位授予單位】：中南林業(yè)科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S792.99
，

本文編號(hào)：1691657