摘要
以叶用莴苣品种耐抽薹意大利生菜为试材,采用RT-PCR结合RACE技术,克隆磷转运蛋白Ls PHT1基因序列;并利用半定量PCR结合实时荧光定量PCR技术检测叶用莴苣在纳米材料处理下Ls PHT1基因的表达情况。结果表明:Ls PHT1基因的开放阅读框为1 605 bp,编码534个氨基酸,推测蛋白质分子量为58.5 k D,具有PHT1家族的保守特征序列GGDYPLSATIx SE,与菊花PHT氨基酸序列同源性高达88%。Ls PHT1仅在叶用莴苣根部表达,而在叶片中几乎不表达。在叶用莴苣全生长期内,Ls PHT1的表达量呈先上升后下降的变化趋势。在生长中后期,纳米材料处理的叶用莴苣根部Ls PHT1表达量显著高于对照,说明Ls PHT1的表达受纳米材料的诱导上调。
Taking lettuce(Lactuca sativa L.)from‘Naichoutaiyidalishengcai'as experimental material,adopting RACE combined with RT-PCR,this experiment cloned phosphate transporting protein Ls PHT1 gene sequence and tested the expression of Ls PHT1 gene of lettuce under nano materials treatment by semi-quantitative PCR combined with real-time fluorescent quantitative PCR.The results showed that the open reading frame of Ls PHT1 gene was 1 605 bp,encoding a polypeptide of 534 amino acids with an estimated protein molecular mass of 58.5 k D.It had the highly conserved PHT1 family signatures,GGDYPLSATIx SE.The similarity of amino acids between PHT with the homologous gene in Chrysanthemum was 88%.The Ls PHT1 was expressed only in lettuce roots,and almost not in lettuce leaves.The expression level of Ls PHT1 in the whole growth period showed a tendency of increasing first and then decreasing.The expression quantity of Ls PHT1 in roots of lettuce treated by nano materials was significantly higher than that of the contrast during the mid and late growth period,indicating that the expression of Ls PHT1 was up-regulated by the induction of nano materials.
出处
《中国蔬菜》
北大核心
2018年第2期27-33,共7页
China Vegetables
基金
广东省自然科学基金项目(2015A030313397)
现代农业产业技术体系专项(CARS-25-C-04)
