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以转rolABC基因枳橙B、D、E系及野生植株为材料,制备标准品,采用SYBR Green I染料,构建rol基因和β-actin基因的双标准曲线,校正引物扩增效率后,用2-△△Ct法对rol基因在嫩茎、嫩叶、功能叶、树皮和根中的mRNA水平进行相对定量,建立适合于rolA、rolB、rolC基因实时荧光定量RT-PCR分析方法.初步认为rolC基因在嫩茎、嫩叶、功能叶和树皮中表达量最高;其次是rolA基因,主要在嫩茎中表达;rolB基因表达量最低,主要在根系中表达.
Using the SYBR Green I dye, the double standard curve of rol gene and β-actin gene was constructed by using rolABC gene Citrange orange B, D, E lines and wild plants as materials. After correcting the amplification efficiency of primers, - △△ Ct method for rol gene roly, tender leaves, functional leaves, bark and root mRNA levels relative quantitative determination of the rolA, rolB, rolC gene real-time quantitative RT-PCR analysis method rolC gene was highest in young stem, young leaves, functional leaves and bark; followed by rolA gene, which was mainly expressed in young stems; rolB gene was the lowest, mainly expressed in roots.