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...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.展开更多
Transgenic citrange lines with rol ABC genes behave rosette branching and extreme dwarfing.To explore the regulatory mechanism of plant hormones in axillary shoot growth of transgenic citrange,a full length cD NA of C...Transgenic citrange lines with rol ABC genes behave rosette branching and extreme dwarfing.To explore the regulatory mechanism of plant hormones in axillary shoot growth of transgenic citrange,a full length cD NA of CPMAX2 was cloned from citrange [Citrus sinensis(L.) Osb × Poncirus trifoliate(L.) Raf.] by RT-PCR in this study.The expression of CPMAX2 was detected in axillary tender leaves of 3 transgenic citrange lines and the wild type.Additionally,we constructed an over-expression vector CPMAX2-p CAMBIA1301 for further study.The results showed that the c DNA sequence and its putative peptide sequence shared 99.66% and 99.14% of identity with its Citrus sinensis ortholog MAX2.The deduced amino acid sequence contained putatively one F-box and two LRR repeat domains that are highly conserved in MAX2 genes.CPMAX2 was obviously down-expressed in tender leaves of 3 transgenic citrange lines compared with the wild type.The results suggest CPMAX2 play an important role in regulating the rosette shoot growth in transgenic citrange with rol ABC genes.展开更多
文摘以转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.
基金financially supported by the National Natural Science Foundation of China(31301750)the National High Technology Research and Development Program of China(863 Program)(2011AA100205)
文摘Transgenic citrange lines with rol ABC genes behave rosette branching and extreme dwarfing.To explore the regulatory mechanism of plant hormones in axillary shoot growth of transgenic citrange,a full length cD NA of CPMAX2 was cloned from citrange [Citrus sinensis(L.) Osb × Poncirus trifoliate(L.) Raf.] by RT-PCR in this study.The expression of CPMAX2 was detected in axillary tender leaves of 3 transgenic citrange lines and the wild type.Additionally,we constructed an over-expression vector CPMAX2-p CAMBIA1301 for further study.The results showed that the c DNA sequence and its putative peptide sequence shared 99.66% and 99.14% of identity with its Citrus sinensis ortholog MAX2.The deduced amino acid sequence contained putatively one F-box and two LRR repeat domains that are highly conserved in MAX2 genes.CPMAX2 was obviously down-expressed in tender leaves of 3 transgenic citrange lines compared with the wild type.The results suggest CPMAX2 play an important role in regulating the rosette shoot growth in transgenic citrange with rol ABC genes.
