摘要
采用RACE和RT-PCR方法,克隆白三叶拉丁诺(Trifolium repens cv.‘Ladino’)的cDNA基因序列,命名为TrFQR1,对该序列进行生物信息学分析,利用实时荧光定量PCR检测TrFQR1的表达模式。结果表明:(1)TrFQR1序列长为1 003bp,开放阅读框为612bp,编码203个氨基酸,该蛋白的理论分子质量21.88kD,等电点为5.96,属于亲水性稳定蛋白,无信号肽,无跨膜结构域,在进化上高度保守,N端11~15位氨基酸和C端112~165位氨基酸是FMN结合位点。(2)TrFQR1表达模式分析显示,白三叶TrFQR1基因对于8种处理均有响应,但不同处理响应不同,其中:用25μmol/L SNP、10mmol/L H_2O_2、5mmol/L CaCl_2处理1.5h以及15%PEG处理3h时,白三叶TrFQR1基因相对表达量显著增加;在200mmol/L NaCl和600μmol/L CdSO4处理下,TrFQR1基因相对表达量随处理时间的增加而增加;4℃处理6h和0.02mmol/L NaHS处理1.5h时,TrFQR1基因相对表达量显著减少。
In order to study the biological function of TrFQR1 in Trifolium repens cv.‘Ladino’,we cloned cDNA sequence of TrFQR1 by RACE-PCR and RT-PCR and analyzed with bioinformatics software.The expression pattern of TrFQR1 was detected by real-time fluorescence quantitative PCR(RT-qPCR).The results showed that:(1)TrFQR1 was obtained at a length of 1 003 bp,which open reading frame contained 612 bases and encoded 203 amino acid.The molecular weight was 21.88 kD and the isoelectric point was 5.96.TrFQR1 was a hydrophilic protein that highly conserved in evolution without signal peptide and transmembrane domains.The N terminal 11-15 aa and the C terminal 112-165 aa were the FMN binding sites.(2)RT-qPCR results showed that TrFQR1 could respond to the all eight treatments.However,TrFQR1 presented different expression trends to those treatments.At 1.5 h after the treatment of 25μmol/L SNP,10 mmol/L H 2O 2 or 5 mmol/L CaCl 2 and 3h after the treatment of 15%PEG,the relative expression levels of TrFQR1 were upregulated significantly.The relative expression levels of TrFQR1 increased with the increase of treatment time under 200 mmol/L NaCl or 600μmol/L CdSO 4.The relative expression of TrFQR1 gene were significantly decreased at 6 h after the treatment of 4℃and 1.5 h after the treatment of 0.02 mmol/L NaHS.
作者
吴星
张艳
雍斌
李州
彭燕
WU Xing;ZHANG Yan;YONG Bin;LI Zhou;PENG Yan(College of Animal Science and Technology,Sichuan Agricultural University,Chengdu 611130,China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2018年第3期431-438,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(NSFC31372371)
