摘要
植物NRT2基因家族(高亲和硝酸盐转运蛋白基因)在植物吸收和转运硝酸盐过程中发挥重要作用。本研究根据NRT2基因家族的保守序列设计兼并引物,扩增出一条821bp序列,以此序列为信息探针,通过电子克隆和RT-PCR的方法从烟草中克隆获得一个包含1593bp开放阅读框、编码530个氨基酸的c DNA序列,命名为Nt NRT2.4。生物信息学分析表明,该基因编码的蛋白质具有NRT家族的共有结构特征,与烟草已发现的三个本家族基因同源性达到77.54%,与拟南芥NRT2家族基因同源性达到81.63%,其启动子中包含多个与硝酸盐、光照及根系特异表达相关的元件;组织特异表达分析结果显示,烟草Nt NRT2.4基因的组织表达差异较大,在根部的表达量较高,在叶片和茎部的表达量低;不同氮素形态、光照和蔗糖处理下的基因表达分析结果表明,硝态氮、光照和蔗糖处理诱导Nt NRT2.4表达,而铵态氮抑制其表达。
NRT2 gene plays a critical role in nitrate absorption and transport in plants. 821 bp sequence of NRT2 gene was screened from c DNA library. Based on the sequence, Nt NRT2.4 was isolated from tobacco（Nicotiana tabacum L.） by in silico cloning and RT-PCR. Nt NRT2.4 includes an open reading frame（ORF） of 1593 bp and encode 530 deduced amino acid residues（AAR）. Phylogenetic analysis suggested that Nt NRT2.4 was high homologous to its paralogous genes from tobacco（77.54%） and orthologous genes from Arabidopsis（81.63%）, which were also induced by nitrate stresses. Promoter analysis suggested Nt NRT2.4 had large amount of motif involved in N and light metabolism. Reverse transcription-polymerase chain reaction（RT-PCR） and quantitative real-time PCR（q RT-PCR） were used to determine expression patterns of Nt NRT2.4 in tobacco. Results revealed that Nt NRT2.4 expressed more in tobacco roots and less in leaves and stems. Expression patterns under light and sugar responses suggested that Nt NRT2.4 was involved in response to light and sugar treatment, with significant different responsive profiles.
出处
《中国烟草学报》
EI
CAS
CSCD
北大核心
2016年第1期84-91,共8页
Acta Tabacaria Sinica
基金
贵州省烟草公司毕节市公司科技攻关项目(毕节合2014-59)
河南省高等学校重点科研项目(15A210029)
