青花菜抗坏血酸过氧化物酶基因boapx2的克隆与表达-植物病理学报.pdfVIP

植物病理学报 ACTA PHYTOPATHOLOGICA SINICA　 42(4):374-380(2012) 青花菜抗坏血酸过氧化物酶基因 BoAPX2 的克隆与表达分析 1* 2 1 蒋 明 , 张志仙 , 袁菱婧 1 2 ( 台州学院生命科学学院, 临海317000; 台州市农业科学研究院, 临海317000) 摘要:抗坏血酸过氧化物酶(Ascorbate peroxidase,APX)是H O 清除剂,在植物抗逆反应中起着重要作用。 根据NCBI发 2 2 布的过氧化物体抗坏血酸过氧化物酶(Peroxisomal APX,pAPX)基因序列设计简并引物,分别以青花菜(Brassica oleracea var. italica)叶片DNA和cDNA为材料进行PCR扩增,克隆到一个APX基因,命名为BoAPX2,序列提交GenBank,登录号 为JN172929。 BoAPX2 的基因组序列长为2 013 bp,具有8个内含子,编码区全长864 bp,编码287个氨基酸。 序列分析结 果表明,BoAPX2与已知的过氧化物体 APX 有较高的相似性,氨基酸的C 端具 1个跨膜结构域。 RT-PCR 结果表明, BoAPX2 的表达受霜霉病菌(Hyaloperonosporaparasitica)、H O 、水杨酸和NaCl诱导。 2 2 关键词:青花菜;霜霉病;抗坏血酸过氧化物酶;克隆;表达 Cloning and expression analysis of an ascorbate peroxidase gene BoAPX2 from Brassica oleraceavar. italica 1 2 1 1 　 JIANGMing ,ZHANGZhi-xian ,YUANLing-jing 　 ( CollegeofLife 2 Science,Taizhou University,Linhai317000,China; Taizhou Academy of Agricultural Research,Linhai317000,China) Abstract: Ascorbate peroxidase (APX),a scavenger of hydrogen peroxide,playsan important role in plant stressresistance. DegenerateprimersweredesignedaccordingtoperoxisomalAPX sequencespublishedinNC- BI. APX gene,designated BoAPX2,wasisolated from DNA and cDNA of leaves of Brassica oleracea var. italica,respectively. Gene sequence was submitted to GenBank with an accession number of JN172929. The full length genomicDNAofBoAPX2 was2013bpinlengthwith8 introns,andthecompletecoding sequence was864 bp in length encoding287 amino acids. Sequence analysisresultsindicated that BoAPX2 sharedhigh homology to known peroxisomalAPXsand wasp