發(fā)布時(shí)間：2018-06-05 23:41

  本文選題：報(bào)春苣苔 + 花色素苷��； 參考：《仲愷農(nóng)業(yè)工程學(xué)院》2017年碩士論文

【摘要】：報(bào)春苣苔屬(Primulina)植物花色艷麗且變異豐富,整株花期較長(zhǎng),極具觀賞性。本研究從遺傳和發(fā)育兩個(gè)角度分別對(duì)幾種不同顏色的報(bào)春苣苔屬植物及鐘冠報(bào)春苣苔不同發(fā)育階段的花色素苷成分和含量進(jìn)行測(cè)定,揭示了報(bào)春苣苔花色艷麗的生化基礎(chǔ),并且以懷集報(bào)春苣苔為試材,建立了該屬植物的轉(zhuǎn)基因體系,為今后了解該屬植物花色豐富變異的生理及遺傳機(jī)制奠定了基礎(chǔ),有助于開(kāi)展雜交育種及分子育種以進(jìn)行花色種質(zhì)創(chuàng)新。主要研究結(jié)果如下:(1)遺傳方面,以38種報(bào)春苣苔屬植物的新鮮花朵為試材,采用pH示差法測(cè)總花色素苷含量。結(jié)果顯示花色越深,花色素苷的總含量越多,按照含量差異,從中選出13個(gè)物種用于花色素苷成分的鑒定,其中鐘冠報(bào)春苣苔含有的花色素苷種類(lèi)最多,為17種,白色的馬壩報(bào)春苣苔和懷集報(bào)春苣苔及黃色的黃花牛耳朵不含有花色素苷,主要顯色成分為黃酮類(lèi)物質(zhì)。(2)發(fā)育方面,以顏色最豐富的=鐘冠報(bào)春苣苔為試材,通過(guò)花器官特異性研究發(fā)現(xiàn),花瓣部分呈紫色是由于該部位藍(lán)紫色的花色素(錦葵素、矮牽牛素及飛燕草素)的比例之和達(dá)到64.0%;而花筒部分的紅色,是由于該部位矢車(chē)菊素及芍藥花素分別占據(jù)了36.7%及20.0%。伴隨著花發(fā)育過(guò)程,紅粉色的花色素占據(jù)比例逐漸減小,而藍(lán)紫色花色素所占比例漸趨增加,這可能是花瓣/花筒的比例伴隨著花發(fā)育漸趨增大。開(kāi)花前總花色素苷含量緩慢上升并趨于穩(wěn)態(tài)(S2階段有一峰值);而花盛開(kāi)之后,含量急劇降低,這可能與花衰老進(jìn)程的花色素苷降解相關(guān)。(3)轉(zhuǎn)基因方面,以白色花的懷集報(bào)春苣苔為試材,利用農(nóng)桿菌介導(dǎo)法建立了報(bào)春苣苔屬植物高效快速轉(zhuǎn)化體系,為花色育種提供條件。本實(shí)驗(yàn)以葉片為外植體進(jìn)行植物組織培養(yǎng),其中愈傷組織誘導(dǎo)培養(yǎng)基成分為MS培養(yǎng)基包含0.1 mg·L~(-1) TDZ和0.1mg·L~(-1) NAA,芽誘導(dǎo)培養(yǎng)基為MS培養(yǎng)基包含0.5 mg·L~(-1) TDZ和0.1 mg·L~(-1) NAA,所有的培養(yǎng)基均包含6 g·L~(-1)瓊脂和30 g·L~(-1)蔗糖�；謴�(fù)培養(yǎng)基中抗生素起始濃度為頭孢噻肟500 mg·L~(-1),后期依次遞減。用20 mg·L~(-1)的潮霉素經(jīng)過(guò)4次篩選后進(jìn)行PCR檢測(cè),轉(zhuǎn)化效率為20%。
[Abstract]:The flowers of the genus Primulina were colorful and varied, and the whole plant had a long flowering period and was very ornamental. In this study, the anthocyanin composition and content of several different color plants of the genus Cesneria and Zhong Guan in different developmental stages were determined from the perspective of heredity and development respectively, and the biochemical basis of the brilliant color of the flower was revealed. Moreover, the transgenic system of this genus was established by using Cecilaria villosum as the material, which laid a foundation for understanding the physiological and genetic mechanism of the rich variation of flower color in this genus in the future. It is helpful to carry out cross breeding and molecular breeding to carry out flower color germplasm innovation. The main results were as follows: (1) in the aspect of heredity, the total anthocyanin content was measured by pH difference method with 38 fresh flowers of Cesneria. The results showed that the deeper the flower color, the more the total content of anthocyanin. According to the content difference, 13 species were selected for the identification of anthocyanin components, among which 17 species of anthocyanin were found in Zhong Guan. The ears of white and yellow cattle did not contain anthocyanin, and the main color component was flavonoids. The most colorful = Zhong Guan was used as the test material. Studies on flower organ specificity showed that the purple part of the petals was due to the sum of 64.0% of the anthocyanins (mallow, peachin and verdanin) in this part of the flower tube, while the red part of the flower tube was red. This is because cornwort and paeoniflorin occupy 36.7% and 20.0% respectively. With the development of flower, the proportion of red and pink anthocyanins decreased gradually, while the proportion of blue and purple anthocyanins gradually increased, which may be the proportion of petals / flower tube increased with the flower development. The content of total anthocyanin increased slowly before anthesis and tended to have a peak value in the stable phase of S 2, but decreased sharply after blooming, which may be related to the degradation of anthocyanin in the process of flower senescence. An efficient and rapid transformation system was established by means of Agrobacterium tumefaciens-mediated method. In this experiment, the leaves were used as explants for plant tissue culture. The callus induction medium consisted of MS medium containing 0.1 mg / L TDZ and 0.1mg L ~ (1) NAA, MS medium containing 0.5 mg / L ~ (-1) TDZ and 0.1 mg / L ~ (-1) NAA, and all medium containing 6 g / L ~ (-1) Agar and 30 g / L ~ (-1) sucrose. The initial concentration of antibiotics in the recovery medium was cefotaxime 500 mg / L ~ (-1), which decreased at the later stage. Hygromycin (20 mg / L) was used to detect PCR for 4 times, and the conversion efficiency was 20%.
【學(xué)位授予單位】：仲愷農(nóng)業(yè)工程學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S68

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