cDNA-AFLP analysis on 2 Osmanthus fragrans cultivars with different flower color and molecular characteristics of OfMYB1 gene
cDNA-AFLP analysis on 2 Osmanthus fragrans cultivars with different flower color and molecular characteristics of OfMYB1 gene
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2个不同花色桂花品种的cDNA-AFLP分析及OfMYB1基因分子特征
DOI：
10.1007/s00468-015-1175-6
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发表时间：
2015-06-01
影响因子：
2.3
通讯作者：
Shang, Fude
中科院分区：
文献类型：
--
作者：
Han, Yuanji;Chen, Weicai;Shang, Fude
关键信息本研究以2个Osman thus fragrans品种的花瓣为材料，进行了cDNA-AFLP分析，共得到102个cDNA片段。结果表明，桂花品种‘丹桂’和‘银桂’在花色和挥发性物质方面存在较大差异。我们对这两个品种的花瓣进行了cDNA-AFLP分析，以分离与花色素和芳香物质产生有关的基因。用256对引物组合检测到390个差异表达片段，得到102个cDNA片段。在差异表达的转录本中，45个在“银桂”中上调，57个在“丹桂”中上调。这些差异表达的基因涉及多种代谢和信号转导途径。根据cDNA-AFLP分析，一个72-bp的片段在'丹桂'中上调，差异表达的转录衍生片段的序列比较表明这是一个MYB转录因子。MyB 1基因cDNA全长749 bp，编码区585 bp，编码195个氨基酸。系统发育分析表明，MyB 1基因属于含有“nyWsv/mE/DD 1 WP/s”基序的亚群。原生质体转化实验表明，OfMYB 1蛋白主要定位于细胞核。酵母单杂交实验证实OfMYB 1是一个转录激活因子，调控OfPAL基因的表达。在20. 0. 001 ~ 30. 002 μ g/L的条件下，对OfMYB 1和花青素生物合成途径基因（OfCHS、OfCHI、OfDFR和OfANS）的差异表达模式进行了研究。结果表明，OfMYB 1可能不调控这些基因的表达。
Key message In this study, we carried out cDNA-AFLP analysis on petals of 2 Osman thus fragrans cultivars, yielding 102 cDNA fragments. Of MYB1 gene was cloned and the molecular characteristics were studied.The Ostnanthus fragrans cultivars 'Dangui' and 'Yingui' differ considerably in floral coloration and volatiles. We carried out cDNA-AFLP analysis on petals of the two cultivars to isolate genes involved in the production of floral pigments and aromatic chemicals. Using 256 primer combinations, we detected 390 differentially expressed fragments, yielding 102 cDNA fragments. Of the differentially expressed transcripts, 45 were upregulated in `Yingui' and 57 were upregulated in 'Dangui'. The differentially expressed genes were involved in several metabolic and signal transduction pathways. According to the cDNA-AFLP analysis, a 72-bp fragment was upregulated in `Dangui', sequence comparison of the differentially expressed transcript-derived fragment showed this to be a MYB transcription factor. The cDNA of Of MYB1 was 749 bp long and contained a coding sequence of 585 bp encoding a polypeptide of 195 amino acids. A phylogenetic analysis placed the Of MYB1 gene within the subgroup containing "nyWsv/mE/DD1WP/s" motif. Protoplast transformation experiment showed that OfMYB1 protein accumulated in cell nuclei. Yeast one hybrid assay confirmed that OfMYB1 acted as a transcriptional activator to regulate expression of OfPAL gene. The differential expression patterns of OfMYB1 and anthocyanin biosynthetic pathway genes (OfCHS, OfCHI, OfDFR and OfANS) in the 2 0. fragrans cultivars suggested that OfMYB1 may not regulate the expression of these genes.