摘要
旨在利用基因工程手段将AeSePAP1基因转入到小麦,为获得耐低磷胁迫的小麦品种奠定基础。以西尔斯山羊草的叶片为材料,根据小麦和二穗短柄草的PAP1基因设计特异引物,采用同源克隆的方法获得西尔斯山羊草紫色酸性磷酸酶PAP1基因的cDNA序列,并将其命名为AeSePAP1。该基因长1.03kb,编码335个氨基酸,分子质量为37.95ku,等电点为5.55。与小麦PAP1基因相比,西尔斯山羊草PAP1基因的cDNA的编码区有36处发生碱基替代,包括24处转换和12处颠换,有16处编码的氨基酸残基不同,其序列一致性达94.66%。对AeSePAP1基因编码的蛋白质进行生物信息学分析发现,该蛋白具有信号肽和跨膜结构,定位于质膜或分泌到胞外。系统进化树分析表明,西尔斯山羊草紫色酸性磷酸酶PAP1基因与普通小麦、短柄草、谷子的PAP1基因同源性较高。
In order to help for wheat breeding, we could get some low phosphorus tolerance germplasm resources via the methods of the genetic transformation of AeSePAP1 into wheat cultivars. In this study, PAP1 gene was cloned from Aegilops searsii with designed primers according to the sequence of PAP1 genes from wheat （Triticum aestivum ） and slender falsebrome （Brachypodium distachyon） based on homologous cloning method and named AeSePAP1 . The cloned AeSePAP1 gene was 1.03 kb in size and encoded 335 amino acids, with molecular mass of 37. 95 ku and isoelectric point 5.55. Sequence alignment of AeSePAP1 with PAP1 of wheat showed that there were 36 single nucleotide polymorphisms, 24 locus transitions and 12 locus transversion andAeSePAP1 had high hornology toPAPJ of wheat with the identities of 94.66%. Amino acid sequence alignment showed that 16 deduced amino acids of AeSePAP1 were different fromPAP1 of wheat. Bioinformatics analysis indicated that the N-terminal region of AeSePAP1 displayed signal sequences and trans-memberance region. The phylogenic tree results showed that PAP1 gene from Aegilops searsii shared high similarities to the previously reportedPAPI genes from other plants, such as Triticum aestivum, Brachypodium distachyon and Setaria italica.
出处
《西北农业学报》
CAS
CSCD
北大核心
2014年第10期105-111,共7页
Acta Agriculturae Boreali-occidentalis Sinica
基金
西北农林科技大学基本科研业务费项目(QN2011003)
唐仲英育种基金
关键词
西尔斯山羊草
紫色酸性磷酸酶
耐低磷基因
抗逆性
Aegilops searsii
Purple acid phosphotase
Low phosphorus tolerance gene
Resistance
