Molecular cloning and gene expression differences of the anthocyanin biosynthesis-related genes in the red/green skin color mutant of pear (Pyrus communis L.)
Molecular cloning and gene expression differences of the anthocyanin biosynthesis-related genes in the red/green skin color mutant of pear (Pyrus communis L.)
复制标题
DOI：
10.1007/s11295-013-0644-6
复制
发表时间：
2013-06
影响因子：
2.4
通讯作者：
Ya-nan Yang;Guang-Wu Zhao;Wen-quan Yue;Shaoling Zhang;C. Gu;Jun Wu
中科院分区：
文献类型：
--
作者：
Ya-nan Yang;Guang-Wu Zhao;Wen-quan Yue;Shaoling Zhang;C. Gu;Jun Wu
为了揭示梨品种"早红Doyenne du Comice"及其绿色变异株系红/绿色突变的分子机制，克隆了两个品种7个花色苷合成基因（PAL、CHS、CHI、DFR、F3 H、ANS、UFGT）的全长cDNA。已向美国国家生物技术信息中心（NCBI）提交的登录号分别为KC 460392、KC 460393、KC 460394、KC 460395、KC 460396、KC 460397和KC 460398。然而，在颜色突变体之间没有序列差异，这意味着皮肤颜色的变化不是由这些基因中的任何一个突变引起的。同时，采用实时荧光定量PCR（qRT-PCR）检测这7个基因的表达水平。结果表明，多数结构基因在红皮品种果实发育过程中表达上调，但CHI1和UFGT基因仅在果实发育早期表达。qRT-PCR检测了MYB 10、bHLH和WD 40的表达水平，发现MYB 10基因在红色突变体中的表达量在早期显著高于绿色突变体，而bHL和WD 40在后期的表达量较高。这些数据表明，花青素合成途径中的结构基因的表达差异导致突变体的肤色变化。然而，MYB 10、bHLH和WD 40似乎不是调控花色苷生物合成和决定红/绿色突变体的关键转录因子。
To reveal the molecular mechanisms that led to the red/green color mutation of pear between the cultivar ‘Early red Doyenne du Comice’ and its green variant strain, the full-length cDNA of the seven anthocyanin biosynthesis genes (PAL,CHS,CHI,DFR,F3H,ANS,UFGT) was cloned in both cultivars. The accession number has been submitted to National Center for Biotechnology Information (NCBI) as KC460392, KC460393, KC460394, KC460395, KC460396, KC460397, and KC460398, respectively. However, there was no sequence difference between the color mutants, which means that the skin color change was not caused by mutation of any of these genes. Meanwhile, the expression levels of these seven genes were examined by quantitative real-time PCR (qRT-PCR). Results showed that most of the structural genes were up-regulated in the red-skinned cultivar during fruit development, but theCHIandUFGTgenes were highly expressed only at an early stage. The expression levels of the transcription factorsMYB10,bHLH, andWD40were also investigated by qRT-PCR, and theMYB10gene was found to be expressed at significantly higher levels in the red variety than in the green mutant at the early stage, while the expression levels ofbHLHandWD40were higher at a later stage. These data indicate that the expression difference of structural genes in the anthocyanin biosynthesis pathway led to the skin color change of the mutant. However,MYB10,bHLH, andWD40do not appear to be the key transcription factors that regulate the biosynthesis of anthocyanin and determine the red/green color mutant.