虎杖基因PcMYB1真核表达载体构建及遗传转化
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摘要
在虎杖(Polygonum cuspidatum Sieb.et Zucc.)叶片中获得了1个R2R3-MYB转录因子基因,命名为PcMYB1。推测其在苯丙烷代谢中起负调控作用。为了探索PcMYB1的的功能,构建了PcMYB1基因的真核过量表达载体,通过农杆菌介导,利用花絮浸染法转化拟南芥,转化效率为0.11%。经潮霉素筛选和PCR鉴定,结果表明PcMYB1成功转入拟南芥中。
One R2R3-MYB transcription factor named PcMYB1 was isolated from the leaf of Polygonum cuspidatum.It was speculated that it played a negative regulatory role in phenylpropanoid metabolism.In order to evaluate the function of PcMYB1,the plant expression vector pCAMBIA-35 SPcMYB1 was constructed and introduced into Arabidopsis thalianathrough Agrobacterium tumefaciens by means of the floral dip method.The transformants were screened on the medium containing hygromycin and the integration of PcMYB1 gene into the transgenic plants was verified via the PCR.The results showed that the transformation efficiency was 0.11% and PcMYB1 was successfully transferred into Arabidopsis thaliana.
One R2R3-MYB transcription factor named PcMYB1 was isolated from the leaf of Polygonum cuspidatum.It was speculated that it played a negative regulatory role in phenylpropanoid metabolism.In order to evaluate the function of PcMYB1,the plant expression vector pCAMBIA-35 SPcMYB1 was constructed and introduced into Arabidopsis thalianathrough Agrobacterium tumefaciens by means of the floral dip method.The transformants were screened on the medium containing hygromycin and the integration of PcMYB1 gene into the transgenic plants was verified via the PCR.The results showed that the transformation efficiency was 0.11% and PcMYB1 was successfully transferred into Arabidopsis thaliana.
引文
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[12]邢文,金晓玲.调控植物类黄酮生物合成的MYB转录因子研究进展[J].分子植物育种,2015,13(3):689~696.
[13]李针针,刘兆磊,陈发棣,等.荷花R2R3-MYB转录因子NnMYB4对拟南芥木质素合成的影响[J].南京农业大学学报,2016,39(6):932~938.
[14]Fornale S,Shi X H,Chai C,et al.ZmMYB31directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux[J].Plant J,2010,64:633~644.
[2]时圣明,潘明佳,王文倩,等.虎杖的化学成分及药理作用研究进展[J].药物评价研究,2016,39(2):317~321.
[3]柳忠玉,雷健,李晓筱,等.虎杖转录因子PcMYB1基因的克隆及原核表达[J].河南农业科学,2018,47(1):96~102.
[4]Cho S,NamKoong K,Shin M,et al.Cardiovascular protective effects and clinical applications of resveratrol[J].J Med Food,2017,20:323~334.
[5]王延谦,张波,陈文杰,等.小麦R2R3-MYB转录因子TaMYB3-4D的克隆及功能分析[J].西北植物学报,2015,35(4):646~652.
[6]Xu F,Ning Y,Zhang W,et al.An R2R3~MYB transcription facter as a negative regulator of the flavonoid biosynthesis pathway in Cinkgo biloba[J].Functional Integrative Genomics,2014,14:177~189.
[7]Legay S,Lacombe E,Goicoechea M,et al,Molecular characterization of EgMYB1,aputative transcriptional repressor of the lignin biosynthetic pathway[J].Plant Sci,2007,173:542~549.
[8]Li C,Lu S.Genome-wide characterization and comparative analysis of R2R3-MYB transcription factors shows the complexity of MYBassociated regulatory networks in Salvia miltiorrhiza[J].BMC Genomics,2014,15:277~289.
[9]Legay S,Sivadon P,Blervacq A S,et al.EgMYB1,an R2R3MYB transcriptional factor from eucalyptus negatively regulates secondary cell wall formation in Arabidopsis and poplar[J].New Phytol,2010,188:774~786.
[10]Fornale S,Shi X H,Chai C,et al.ZmMYB31directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux[J].Plant J,2010,64:633~644.
[11]张新,崔广艳,陈翠娜,等.农杆菌介导DR1172基因转化拟南芥[J].基因组学与应用生物学,2012,(6):582~586.
[12]邢文,金晓玲.调控植物类黄酮生物合成的MYB转录因子研究进展[J].分子植物育种,2015,13(3):689~696.
[13]李针针,刘兆磊,陈发棣,等.荷花R2R3-MYB转录因子NnMYB4对拟南芥木质素合成的影响[J].南京农业大学学报,2016,39(6):932~938.
[14]Fornale S,Shi X H,Chai C,et al.ZmMYB31directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux[J].Plant J,2010,64:633~644.