CAJ | 학술논문

以烟草根尖组织cDNA为模板，RT-PCR结合电子克隆得到NtWRKY-R1基因的完整ORF，序列分析显示，NtWRKY-R1具有WRKY转录因子家族典型的WRKYGQK保守结构域及C2 H2的锌指结构，属于WRKY转录因子家族第Ⅱ类。采用生物信息学方法对NtWRKY-R1蛋白的理化性质、进化关系和磷酸化位点进行预测和分析，结果表明，NtWRKY-R1与茄科植物保守结构域的同源性及系统进化关系最近；磷酸化位点分析显示该蛋白可能通过磷酸化作用修饰，进而对其相应的代谢活动进行调节。通过构建真核表达载体、建立瞬时表达分析体系，结果显示，NtWRKY-R1基因的过表达导致JA信号途径标记基因PDF1．2及烟碱合成关键酶基因PMT1表达量降低，推测NtWRKY-R1基因影响JA信号途径，进而调控烟碱合成。
이연초근첨조직cDNA위모판，RT-PCR결합전자극륭득도NtWRKY-R1기인적완정ORF，서렬분석현시，NtWRKY-R1구유WRKY전록인자가족전형적WRKYGQK보수결구역급C2 H2적자지결구，속우WRKY전록인자가족제Ⅱ류。채용생물신식학방법대NtWRKY-R1단백적이화성질、진화관계화린산화위점진행예측화분석，결과표명，NtWRKY-R1여가과식물보수결구역적동원성급계통진화관계최근；린산화위점분석현시해단백가능통과린산화작용수식，진이대기상응적대사활동진행조절。통과구건진핵표체재체、건립순시표체분석체계，결과현시，NtWRKY-R1기인적과표체도치JA신호도경표기기인PDF1．2급연감합성관건매기인PMT1표체량강저，추측NtWRKY-R1기인영향JA신호도경，진이조공연감합성。
The intact ORF of NtWRKY-R1 gene was isolated by RT-PCR combined with in silico clo-ning from tobacco root tip tissues .Sequence analysis showed that NtWRKY -R1 had conserved WRKYGOK sequence and a C2H2 -type zinc finger motif , and belonged to group Ⅱ WRKY transcription factor family . Bioinformatics methods were used to predict and analyze the physical and chemical characters , evolutionary re-lationships and phosphorylation sites of NtWRKY -R1 protein.The results showed NtWRKY -R1 had a clos-er affinity with Solanaceae in homology and evolutionary relationships .Phosphorylation site analysis showed that NtWRKY-R1 might regulate corresponding metabolic activity through phosphorylation modification .By constructing eukaryotic expression vector and establishing transient expression analysis system , the over-ex-pression of NtWRKY-R1 gene resulted in down-regulated expression of nicotine biosynthesis key gene PMT1 and JA signaling pathway marker gene PDF1.2.It was thought that NtWRKY-R1 gene affected JA signaling pathway and then regulated nicotine biosynthesis .