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擎天凤梨‘Ostsra’是一种观赏红色苞片为主的高档盆栽花卉。一般认为植物苞片的呈色物质除叶绿素外多来源于花色素,而CHS、F3’H和DFR是在花色素的合成过程中的关键酶。本研究采用采用简并引物及同源克隆法获得了881 bp的CHS基因,编码276个氨基酸的序列,GenBank登录号为KX364270;采用cDNA全长文库构建、EST批量测序及目标单克隆序列测通法,获得了F3’H和DFR基因。F3’H基因长1 176 bp,可编码325个氨基酸序列,GenBank登录号为KX364271;DFR基因长1 209 bp,可编码167个氨基酸的序列,GenBank登录号为KX364272。采用实时定量PCR法检测CHS、F3’H和DFR在‘Ostsra’苞片呈色过程中的表达变化,结果表明,3个基因在苞片变色过程中(绿苞-半红苞-全红苞)都呈现出递增的趋势,即随着苞片颜色的变深,3基因的表达量逐渐增加,全红状态时,表达量最高,而作为对照的绿叶,表达水平都是最低的。
Chingti pineapple ’Ostsra’ is a kind of ornamental red bracts mainly of high-grade potted flowers. It is generally believed that the braches of plants are mostly derived from anthocyanidins except for chlorophyll, whereas CHS, F3’H and DFR are key enzymes in the synthesis of anthocyanidins. In this study, a 881 bp CHS gene was obtained by using degenerate primers and homology cloning method, which encodes a 276 amino acid sequence with GenBank accession number KX364270. Using full-length cDNA library, EST batch sequencing and target monoclonal sequencing Method, obtained F3’H and DFR genes. The F3’H gene is 1 176 bp long and encodes a protein of 325 amino acids with GenBank accession number KX364271. The DFR gene is 1 209 bp in length and encodes a 167 amino acid sequence with the GenBank accession number KX364272. The changes of the expression of CHS, F3’H and DFR in bracts of ’Ostsra’ were detected by real-time quantitative PCR. The results showed that during the discoloration of bracts, the three genes (green bud - half red bracts - all red bracts ) Showed an increasing trend, that is, as the bracts become darker in color, the expression level of the 3 gene gradually increases. In the red state, the expression level is the highest, while the green leaf as a control has the lowest expression level.