| Chrysanthemum morifolium of the genus Chrysanthemum in the composite familyis a perennial root flower, the head of the four most important cut flowers, originatedin China. Flower color is one of the most important ornamental traits of chrysanthemum.Now seven color varieties except blue have been cultivated using flower colorVariation, many extraordinary variety such as two color, there color, multi-color werealso appeared. The Chrysanthemum color chimera with varieties of morphologicalcharacters, possessing high ornament value, is an important species resource in breeding,as well as an ideal material for studying the color inheritance and mechanism offormation and regulation.A yellow-purple bi-color chimera was studied as experiment material, named’su-07’. Conventional cutting propagation can maintain chimeric traits, but the formationmechanism and genetic mechanism of chimeric design and color is not clear. In thisstudy, a chrysanthemum color chimera mentioned above was studied as experimentmaterial, preliminary studying about the biological characteristics and the maincomponents of the petal flower pigments,then cloning the sequence of cDNA thatrelated with the color formation, and analyzing the flower color differential expressionof genes related to flower color by real-time quantitative PCR technique. The resultswere obtained are as follows.1.Through investigation and analysis of the biological characteristics inchrysanthemum color chimera, it revealed that the experiment material is thelight-sensitive plant and the flower bud differentiation began in early August andcompleted in around60days; Chimeric plants can be divided into four kinds of formsand chimeric color can be divided into five kinds of basic forms.2.According to investigating and analyzing of genetic stability of the test materialsapplying morphology, cultivation science and tissue culture techniques, it turned out that separation of chimeric plants in2012was48.27%, while in2013was46.55%, theratio decreased slightly, shows that the chimeric color traits genetic instability. The pureyellow strain had been studied out owning to the successful separating and fixingchimeric characters by tissue culture of petals, and the pure purple lines weresuccessfully isolated by the cutting propagation technique purepurple.3. the content of total flavonoids was determination by visible and ultravioletSpectrophotometry in Chrysanthemum, the t test found that significant differences inflavonoid contentin purple petals and yellow petals (P<0.01).With the extracts ofdifferent color petals as experiment materials, the ultra-visible spectroscopic and highperformance liquid chromatography (HPLC) analysis that the pigment of the yellowpetal was chalcone substances and the purple one was chalcone substances andanthocyanins, the HPLC showde that purple petals at520nm containing at least9kindsof substances, and yellow petals under the wavelength without any characteristicabsorption peak.4. The full-length cDNA sequences of ANS and GAPDH were cloned applyingmolecular biology technology in the chrysanthemum petals. The coding regionsequence of ANS was1068bp, coded355amino acid residues and a termination codon;Homologous analysis showed that the homology with other8species of compositaeplants is81-98%.GAPDH gene coding region sequence is1026bp, coded341aminoacid residues and a termination codon, homology analysis showed that GAPDH genesequences rather conservative. ANS and GAPDH gene cloning in order to further studythe formation mechanism of the Mosaic color laid the foundation experiment.5. The pigment metabolism genes（CHS、CHI、F3H、DFR and ANS）of thechrysanthemum Chimaera flower has been analyzed using Real time quantitative PCRtechnique, five genes were highest expressed in Purple petals, the second was inChimeric petals. The expression quantity of DFR and ANS genes were significantlyhigher than in yellow petals in which the quantity was very little. |
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