小麦WRKY转录因子VIGS基因沉默载体构建及验证
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摘要
WRKY转录因子家族广泛存在于各种植物中,有助于提高植物抗逆性。本研究用病毒介导的基因沉默方法沉默Brock中克隆到的TaWRKY基因,发现白粉菌成功侵染小麦叶片的比例上升,畸形胞比例下降。研究表明,TaWRKY基因在小麦抵抗白粉菌侵染过程中发挥着重要作用。
WRKY transcription factor family can help improve plant stress tolerance, which widely exist in various plants. After TaWRKY gene was silenced by VIGS method, it was found that the proportion of succeed Bgt inoculation increased, and the percentage of abnormal appressoria declined, such as papilla. The results indicated that TaWRKY gene played an important role in wheat-Bgt interaction.
WRKY transcription factor family can help improve plant stress tolerance, which widely exist in various plants. After TaWRKY gene was silenced by VIGS method, it was found that the proportion of succeed Bgt inoculation increased, and the percentage of abnormal appressoria declined, such as papilla. The results indicated that TaWRKY gene played an important role in wheat-Bgt interaction.
引文
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[17]谢政文,王连军,陈锦洋,等.植物WRKY转录因子及其生物学功能研究进展[J].中国农业科技导报,2016,18(3):46-54.
[18]BURCH S T M,SCHIFF M,LIU Y,et al.Efficient virusinduced gene silencing in Arabidopsis[J].Plant physiology,2006,142(1):21-27.
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[20]李淼淼,南富波,刘伟,等.VIGS技术在禾本科植物中的应用研究进展[J].麦类作物学报,2013,33(2):401-406.
[21]CAMPBELL J,HUANG L.Silencing of multiple genes in wheat using barley stripe mosaic virus[J].Journal of biotech research,2010,2(1):12-20.
[2]RUSHTON P J,MACDONALD H,HUTTLY A K,et al.Members of a new family of DNA-binding proteins bind to a conserved cis-element in the promoters of alpha-Amy2 genes[J].Plant molecular biology,1995,29:691-702.
[3]MAO G H,MENG X Z,LIU Y D,et al.Phosphorylation of a WRKY transcription factor by two pathogen-responsive MAPKs drives phytoalexin biosynthesis in Arabidopsis[J].Plant cell,2011,23(4):1639-1653.
[4]BIRKENBIHL R P,DIEZEL C,SOMSSICH I E.Arabidopsis WRKY33 is a key transcriptional regulator of hormonal and metabolic responses toward Botrytis cinerea infection[J].Plant physiol,2012,159(1):266-285.
[5]YAN Y,CHI Y,WANG Z,et al.Functional analysis of structurally related soybean Gm WRKY58 and Gm WRKY76 in plant growth and development[J].Journal of experimental botany,2016,67(15):4727-4742.
[6]CHOI C,HWANG S H,FANG I R,et al.Molecular characterization of Oryza sativa WRKY6,which binds to Wbox-like element 1 of the Oryza sativa pathogenesis-related(PR)10a promoter and confers reduced susceptibility to pathogens[J].New phytologist,2015,208(3):846-859.
[7]QIN Y,TIAN Y,LIU X.A wheat salinity-induced WRKY transcription factor Ta WRKY93 confers multiple abiotic stress tolerance in Arabidopsis thaliana[J].Biochemical and biophysical research communications,2015,464(2):428-433.
[8]MANGELSEN E,KILIAN J,BERENDZEN K W,et al.Phylogenetic and comparative gene expression analysis of barley(Hordeum vulgare)WRKY transcription factor family reveals putatively retained functions between monocots and dicots[J].BMC genomics,2008,9(1):194.
[9]GLCKNER G,EICHINGER L,SZAFRANSKI K,et al.Sequence and analysis of chromosome 2 of Dictyostelium discoideum[J].Nature,2002,418(6893):79-85.
[10]SCHMUTZ J,CANNON S B,SCHLUETER J,et al.Genome sequence of the palaeopolyploid soybean[J].Nature,2010,463(7278):178-183.
[11]YU S,CHONG-RUI A I,JING S J,et al.Research progress on functional analysis of rice WRKY,genes[J].水稻科学(英文版),2010,17(1):60-72.
[12]赵丹,赵继荣,黄茜,等.利用BSMV-VIGS技术快速分析小麦TNBL1基因的抗黄矮病功能[J].作物学报,2011,37(11):2106-2110.
[13]李淼淼,南富波,刘伟,等.VIGS技术在禾本科植物中的应用研究进展[J].麦类作物学报,2016,33(2):401-406.
[14]肖莹,刘君,刘晓颖,等.栽培小麦Brock中转录因子基因WRKY的克隆与表达分析[J].天津农业科学,2015,21(2):1-4.
[15]LI C,LIU X,FAN B,et al.Characterization and functional analysis of differentially expressed genes in-inoculated wheat near-isogenic lines by c DNA-AFLP and VIGS[J].Crop science,2014,54(5):2214.
[16]ISHIGURO S,NAKAMURA K.Characterization of a c DNA encoding a novel DNA-binding protein,SPF1,that recognizes SP8 sequences in the 5′upstream regions of genes coding for sporamin and beta-amylase from sweet potato[J].Molecular general genetics,1994,244:563-571.
[17]谢政文,王连军,陈锦洋,等.植物WRKY转录因子及其生物学功能研究进展[J].中国农业科技导报,2016,18(3):46-54.
[18]BURCH S T M,SCHIFF M,LIU Y,et al.Efficient virusinduced gene silencing in Arabidopsis[J].Plant physiology,2006,142(1):21-27.
[19]HOLZBERG S,BROSIO P,GROSS C,et al.Barley stripe mosaic virus-induced gene silencing in a monocot plant[J].Plant journal,2002,30(3):315-327.
[20]李淼淼,南富波,刘伟,等.VIGS技术在禾本科植物中的应用研究进展[J].麦类作物学报,2013,33(2):401-406.
[21]CAMPBELL J,HUANG L.Silencing of multiple genes in wheat using barley stripe mosaic virus[J].Journal of biotech research,2010,2(1):12-20.