生菜AMT1基因克隆及其在TiO_2/ZnO纳米材料处理下的表达  

Molecular Cloning and Expression Analysis of LsAMT1 Gene From Lettuce Under TiO_2/ZnO Nanomaterials

作　　者：汪玉洁[1] 陈日远[1] 刘厚诚[1] 宋世威[1] 孙光闻[1] 

机构地区：[1]华南农业大学园艺学院,广东广州510642

出　　处：《核农学报》2018年第4期665-672,共8页Journal of Nuclear Agricultural Sciences

基　　金：广东省自然科学基金(2015A030313397);现代农业产业技术体系专项基金(CARS-25-C-04)

摘　　要：为检测生菜在TiO_2/ZnO纳米材料处理下LsAMT1基因的表达情况,以生菜为试验材料,采用RTPCR结合RACE技术,以及实时荧光定量PCR技术,克隆LsAMT1基因序列并对其进行表达分析。结果表明,LsAMT1基因的开放阅读框为1 533 bp,编码510个氨基酸,推测蛋白质分子量为54.36 k D,具有编码了一组植物的高亲和铵转运蛋白AMT1家族的保守特征序列:DFAGSGVVHMVGGIAGLWGALIEGPR,与菊科的刺苞菜蓟氨基酸序列同源性高达93%。LsAMT1仅在生菜根部表达,在生菜的全生长期,LsAMT1的表达量随着时间的推移呈现先上升后下降的趋势;在生长中后期,纳米材料处理的生菜根部中LsAMT1的表达量显著高于对照,说明该基因的表达与纳米材料的处理有一定的关联。本研究为进一步探究TiO_2/ZnO纳米材料影响生菜氮素吸收和利用的分子机理奠定了基础。In order to observe expressions of LsAMT1 in lettuce treated by Ti O2/Zn O nanomaterials,the full-length c DNA of AMT1 was cloned using reverse transcription polymerase chain reaction（RT-PCR） and rapid-amplification of c DNA ends（RACE）. The results showed that the open reading frame of LsAMT1 is 1 533 bp,which encoded a polypeptide of510 amino acids with an estimated molecular mass of 54. 36 k D. It has the highly conserved featured sequence of AMT1 family DFAGSGVVHMVGGIAGLWGALIEGPR showed as high as 93% homology to AMT1 in Cynara cardunculus var.scolymus. Real-time quantitative PCR showed that LsAMT1 was expressed only in the roots of lettuce,and the expression level of LsAMT1 in the whole growth period showed a tendency of increasing first and then decreasing. The expression of AMT1 in roots of lettuce treated with nanomaterials was significantly higher than that of control at the middle and late growth stage,indicating that the expression of this gene was related to nanomaterials. The present study might provide a significant foundation for further studies on molecular mechanism of nitrogen absorption and use of lettuce treated by Ti O2/Zn O nanomaterials.

关 键 词：生菜 LsAMT1 克隆 纳米材料 基因表达 

分 类 号：S636.2[农业科学—蔬菜学]