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以红掌佛焰苞S5(盛花期:红色)、S8(衰老期:绿色)时期为试验材料进行数字基因表达谱(digital gene expression profiling,DGE)测序,分别得到12 766 374条(1.28 G)、18 583 390条(1.86 G)可分析序列(clean reads),被成功注释有差异表达的基因序列2 316条。为探究红掌佛焰苞颜色变化过程中关键基因的表达水平,将差异基因定位在花青素生物合成途径11条表达量变化的序列上,并通过实时荧光定量PCR对其中的4条序列进行检验,验证了DGE测序结果的准确性。同时研究这4个序列在红掌佛焰苞生长过程中的表达水平,结果表明:在佛焰苞发育初期,3条序列相对表达量逐渐增高,1条在S2时期降低,但均在S3时期达到最高或较高值;在S4、S5、S6、S7、S8时期,随着佛焰苞的成熟,4条序列表达量大体上均逐渐下降,在S8衰老期表达量最低。研究结果证明了这4条基因序列是红掌佛焰苞花色形成中的关键基因,S3时期是花青素生物合成的高峰期。
A total of 12 766 374 (1.28 G) fragments were obtained by digital gene expression profiling (DGE) sequencing using S5 (full bloom: red) and S8 (aging: green) , And 2 316 segments of 18 583 390 (1.86 G) clean reads successfully annotated with differentially expressed genes. In order to explore the expression level of key genes in the process of the color change of the spathe of Anthurium andraeanum, the differential genes were mapped on the sequences of the 11 expression levels of anthocyanin biosynthesis pathway, and 4 of them were sequenced by real-time fluorescence quantitative PCR Test, verified the accuracy of DGE sequencing results. The results showed that in the early stage of spathe development, the relative expression of three sequences gradually increased, while one decreased in S2, but both in S3 Reaching the highest or the highest value. In the S4, S5, S6, S7 and S8 periods, the expression of four sequences decreased gradually with the maturation of spathe and reached the lowest in the senescence stage of S8. The results show that these four gene sequences are the key genes in the flower bud formation of Anthurium, and S3 is the peak of anthocyanin biosynthesis.