YANG Hong-xia,QU Bai-hong,LIU Zhen-kun,et al.Molecular Cloning and Expression Analysis on PyDFR Gene of Pingguoli in Yanbian[J].Northern Horticulture,2015,39(04):99-103.[doi:10.11937/bfyy.201504023]
延边苹果梨PyDFR基因的克隆及表达分析
- Title:
- Molecular Cloning and Expression Analysis on PyDFR Gene of Pingguoli in Yanbian
- Keywords:
- Pingguoli; Dihyciroflavonol 4-reductase; gene cloning; sequence analysis; expression patterns
- 文献标志码:
- A
- 摘要:
- 以延边苹果梨果实为试材，采用同源克隆及实时荧光定量PCR的试验方法，研究PyDFR基因在果实着色过程中的表达特性及其与花青苷积累的关系。结果表明：PyDFR基因cDNA全长1 039 bp，推断其含有1个843个核苷酸的开放阅读框（ORF），编码281个氨基酸，推导的蛋白分子量为32 kDa，理论等电点pI为6.32。同源性分析表明，PyDFR基因与沙梨的（JQ749637.1）DFR基因的同源性高达99%；实时荧光定量PCR分析表明，PyDFR基因受到光诱导表达量迅速上升，去袋后1 d即达到最大值，随后迅速下降，最终稳定在同一表达水平，与果实着色和花色苷含量测定的结果相对应，即PyDFR基因对果实花色素苷积累有一定的促进作用。
- Abstract:
- Taking the Pingguoli in Yanbian as material，the relationship between the expression and the accumulation of anthocyanin during the pigmentation of Pingguoli was analyzed by the homology cloning technology,and the real-time fluorescent quantitative PCR technique.The results showed that,the full-length cDNA of PyDFR included 1 039 basepair and opened reading frame encompassed 843 bp encoding a polypeptide of 281 amino acids residues with a calculated molecular weight(MV) of 32 kDa and isoelectric point theory(pI) of 6.32.The homology analysis revealed that this cDNA and Pyrus pyrifolia （JQ749637.1) was 99%.Real-time fluorescent quantitative RT-PCR results indicated the expression level of PyDFR were markedly enhanced in the sunlit side，and reached a maximum value at 1 day after debagging，then started decline rapidly，but finally expressed stably in the same level.The color code and anthocyanins content produced results essentially in agreement with these figures.In addition that had a certain role in promoting the accumulation of fruit anthocyanin.
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备注/Memo
第一作者简介：杨宏霞（1989-），女，硕士研究生，研究方向为果树生理学。E-mail：hongxiayangss@163.com.责任作者：曲柏宏（1963-），男，博士，教授，博士生导师，现主要从事园艺植物栽培与生理等研究工作。E-mail：bhqu@ybu.edu.cn.基金项目：国家自然科学基金资助项目（31160389）。