摘要
参照油梨炭疽病菌(Colletotrichum gloeosporioides)的环境pH应答转录因子基因pac1设计1对特异性引物,以芒果炭疽病菌(C.gloeosporioides)基因组DNA为模板,扩增获得大小为2 625 bp的目的基因片段。序列分析结果表明,目的片断包含完整的阅读框,与油梨炭疽病菌C.gloeosporioides pac1基因相应片段长度相同,二者碱基序列同源性达97%。用RT-PCR技术获得pac1基因cDNA片段,证实目的基因片段中存在3个大小分别为59,50,71 bp的内含子。拼接的全长cDNA长1 755 bp,推测编码的氨基酸序列长584个氨基酸残基,与推测的油梨炭疽病菌pac1基因所编码的蛋白质同样大小,二者氨基酸序列相似性达99%,只有1个氨基酸的差异,可以肯定克隆的目的片段为C.gloeosporioides pac1基因,为进一步研究C.gloeosporioides侵染芒果时的pH调控奠定了基础。
According to nucleotide sequence of pac1 gene of avocado anthracnose causal agent, a pair of primers specific-for pH-Responsive Transcription Regulator gene pac 1 were designed and used to amplify a fragment homologous to pac 1 gene from a strain A2 of Colletotrichum gloeosporioides isolated from mango with anthracnose symptom. The amplified DNA fragment was 2 625 bp in size and covered a whole 1 755-bp open reading frame like pacl gene, containing three putative introns （59, 50 and 71 bp in size, respectively）, which were verified after a 960-bp cDNA fragment covering these three introns was amplified and sequenced. Blast result showed a 98% similarity between this 2 625-bp fragment and pacl gene. Conceptual translation of putative cDNA indicates that this open reading frame encodes a polypeptide of 584 amino acids, which has almost the same amino acid sequence as the pac 1 gene with difference of one amino acid. Therefore, this 2 625-bp fragment can be confirmed to be pacl gene of C. gloeosporioides, which will be used in further study of pH regulation of pathogenesis of C. gloeosporioides on mango.
出处
《热带作物学报》
CSCD
2008年第4期501-505,共5页
Chinese Journal of Tropical Crops
基金
国家科技支撑计划项目(2007BAD48B05)
公益性行业(农业)科研专项经费项目(nyhyzx07-032-2)资助
关键词
芒果
胶孢炭疽菌
环境pH应答转录调控因子基因
mango Colletotrichum gloeosporioides pH-responsive transcription regulator gene
