蒙古沙冬青AmMYB4-like基因的克隆与表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2017.05.016

摘要/Abstract

摘要： 基于已有研究工作基础,从蒙古沙冬青[Ammopiptanthus mongolicus（Maxim）Cheng. f]中分离到1个编码MYB类转录因子基因,命名为AmMYB4-like。该序列长度为1 145 bp,含有一个由960 bp 组成的开放阅读框,编码319个氨基酸,具有R2和R3保守结构域,属于R2R3-MYB转录因子。荧光定量PCR分析结果表明,AmMYB4-like 在叶片中参与低温和干旱胁迫应答,在根中主要参与干旱胁迫应答。构建了pPZP212-AmMYB4-like 植物表达载体,旨为进一步研究AmMYB4-like 的功能奠定基础。

关键词: 蒙古沙冬青, AmMYB4-like, 基因克隆, 表达分析, 植物表达载体构建

Abstract: Based on our previous work,a gene encoding MYB transcription factor was cloned from Ammopiptanthus mongolicus,and it was designated as AmMYB4-like. The full length of the AmMYB4-like cDNA was 1,145 bp,including a 960 bp opening reading frame（ORF）which encoded a 319-amino acid peptide. AmMYB4-like contained R2 and R3 conserved domains and belonged to the R2R3-MYB subfamily. Quantitative real-time PCR（qRT-PCR）analyses revealed that AmMYB4-like was up-regulated in leaves under low temperature and drought stresses,but it was mainly induced by drought stress in roots. Plant expression vector pPZP212- AmMYB4-like was constructed for further research of AmMYB4-like.

Key words: Ammopiptanthus mongolicus, AmMYB4-lik, gene cloning, expression analysis, construction of plant expression vector

肖自华,高飞,孙会改,周宜君. 蒙古沙冬青AmMYB4-like基因的克隆与表达分析[J]. 生物技术通报, 2017, 33(5): 108-116.

XIAO Zi-hua GAO Fei SUN Hui-gai ZHOU Yi-jun. Cloning and Expression Analysis of AmMYB4-like in Ammopiptanthus mongolicus[J]. Biotechnology Bulletin, 2017, 33(5): 108-116.

参考文献

[1] Paz-Ares J, Ghosal D, Wienand U, et al. The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators[J]. EMBO J, 1987, 6（12）：3553-3558.
[2] Chen YH, Yang XY, He K, et al. The MYB transcription factor superfamily of Arabidopsis：expression analysis and phylogenetic comparison with the rice MYB family[J]. Plant Mol Biol, 2006, 60（1）：107-124.
[3] Cao ZH, Zhang SZ, Wang RK, et al. Genome wide analysis of the apple MYB transcription factor family allows the identification of MdoMYB121 gene conferring abiotic stress tolerance in plants[J]. PLoS One, 2013, 8（7）：e69955.
[4] Hou XJ, Li SB, Liu SR et al. Genome-wide classification and evolutionary and expression analyses of citrus MYB transcription factor families in sweet orange[J]. PLoS One, 2014, 9：e112375.
[5] Muthamilarasan M, Khandelwal R, Yadav CB, et al. Identification and molecular characterization of MYB transcription factor superfamily in C4 model plant foxtail millet（Setaria italica L.）[J]. PLoS One, 2014, 9：e109920.
[6] Du H, Zhang L, Liu L, et al. Biochemical and molecular characterization of plant MYB transcription factor family[J]. Biochemistry（Mosc）, 2009, 74（1）：5-16.
[7] Ambawat S, Sharma P, Yadav NR, et al. MYB transcription factor genes as regulators for plants：an overview[J]. Physiol Mol Biol Plants, 2013, 19（3）：307-321.
[8] Lepiniec L, Debeaujon I, Routaboul JM, et al. Genetics and biochemistry of seed flavonoids[J]. Annu Rev Plant Biol, 2006, 57（1）：405-430.
[9] Gonzalez A, Zhao M, Leavitt JM, et al. Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings[J]. Plant J, 2008, 53（5）：814-827.
[10] 杜海, 冉凤, 马珊珊, 等. GmMYB042 基因对类黄酮生物合成的调控作用[J]. 作物学报, 2016, 42（1）：1-10.
[11] Zhang JJ, Zhang SS, Li H, et al. Identification of transcription factors ZmMYB111 and ZmMYB148 involved in phenylpropanoid metabolism[J]. Front Plant Sci, 2016, 7：131-134.
[12] 周宜君, 刘春兰, 冯金朝, 等. 沙冬青抗旱、抗寒机理的研究进展[J]. 中国沙漠, 2001, 21（3）：312-315.
[13] 郭生祥, 刘志银, 郝昕. 沙冬青的研究进展[J]. 甘肃林业科技, 2005, 30（4）：5-8.
[14] Zhou YJ, Gao F, Liu R, et al. De novo sequencing and analysis of root transcriptome using 454 pyrosequencing to discover putative genes associated with drought tolerance in Ammopiptanthus mongolicus[J]. BMC Genomics, 2012, 13, 266.
[15] Gao F, Wang J, Wei SJ, et al. Transcriptomic analysis of drought stress responses in Ammopiptanthus mongolicus leaves using the RNA-Seq technique[J]. PLoS One, 2015, 10, e0124382.
[16] Wu YQ, Wei W, Pang X, et al. Comparative transcriptome profiling of a desert evergreen shrub, Ammopiptanthus mongolicus, in response to drought and cold stresses[J]. BMC Genomics, 2014, 15：671.
[17] Pang T, Ye CY, Xia X, et al. De novo sequencing and transcriptome analysis of the desert shrub, Ammopiptanthus mongolicus, during cold acclimation using Illumina/Solexa[J]. BMC Genomics, 2013, 14, 488.
[18] Chen JH, Sun Y, Sun F, et al. Tobacco plants ectopically expressing the Ammopiptanthus mongolicus AmCBL1 gene display enhanced tolerance to multiple abiotic stresses[J]. Plant Growth Regul, 2011, 6（3）：259-269.
[19] 李章磊, 高飞, 曹玉震, 等. 蒙古沙冬青AmDREB2. 1 基因的克隆及表达分析[J]. 生物技术通报, 2015, 31（3）：108-114.
[20] Gao F, Wang N, Li HY, et al. Identification of drought-responsive microRNAs and their targets in Ammopiptanthus mongolicus by using high-throughput sequencing[J]. Sci Rep, 2016, 6：34601.
[21] Shi J, Liu M, Shi J, et al. Reference gene selection for qPCR in Ammopiptanthus mongolicus under abiotic stresses and expression analysis of seven ROS-scavenging enzyme genes[J]. Plant Cell Rep, 2012, 31（7）：1245-54.
[22] Du H, Yang SS, Liang Z, et al. Genome-wide analysis of the MYB transcription factor superfamily in soybean[J]. BMC Plant Biol, 2012, 12：106.
[23] He Q, Jones DC, Li W, et al. Genome-Wide Identification of R2R3-MYB genes and expression analyses during abiotic stress in Gossypium raimondii[J]. Sci Rep, 2016, 6：22980.
[24] Wilkins O, Nahal H, Foong J, et al. Expansion and diversification of the Populus R2R3-MYB family of transcription factors[J]. Plant Physiol, 2009, 149（2）：981-993.
[25] Du H, Feng BR, Yang SS, et al. The R2R3-MYB transcription factor gene family in maize[J]. PLoS One, 2012, 7, e37463.
[26] Stracke R, Werber M, Weisshaar B. The R2R3-MYB gene family in Arabidopsis thaliana[J]. Curr Opin Plant Biol, 2001, 4（5）：447-456.
[27] Matus JT, Aquea F, Arce-Johnson P. Analysis of the grape MYB R2R3 subfamily reveals expanded wine quality-related clades and conserved gene structure organization across Vitis and Arabidopsis genomes[J]. BMC Plant Biol, 2008, 8（1）, 1-15.
[28] Katiyar A, Smita S, Lenka SK, et al. Genome-wide classification and expression analysis of MYB transcription factor families in rice and Arabidopsis[J]. BMC Genomics, 2012, 13, 544.
[29] 乔孟, 于延冲, 向凤宁. 拟南芥R2R3 -MYB类转录因子在环境胁迫中的作用[J]. 生命科学, 2009, 21（1）：145-150.