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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (11): 1949-1955.doi: 10.3724/SP.J.1006.2011.01949


Cloning of nia Gene and Its Differential Expression Induced by Different Nitrogen Forms in Sugar Beet (Beta vulgaris L.)

DING Guang-Zhou1,2, HOU Jing2,CHEN Li1,MA Feng-Ming2,*,CHEN Lian-Jiang1,*   

  1. 1 Sugar Beet Institute of Chinese Academy of Agricultural Sciences, Harbin 150080, China; 2 College of Agriculture, Northeast Agricultural University, Harbin 150030, China
  • Received:2011-04-02 Revised:2011-07-15 Online:2011-11-12 Published:2011-09-06
  • Contact: 马凤鸣, E-mail: fengming_ma@sohu.com, Tel: 0451-55191947; 陈连江, E-mail: chen4004@163.com, Tel: 0451-86609481

Abstract: From diploid species Ty7, a cDNA clone related to nitrate reduction was isolated by Homology-based cloning. According to its sequences information, a novel full-length cDNA termed nia (accession number: EU163265) was obtained by using rapid amplification of cDNA ends (RACE). Nia was 3 247 bp long containing a 2 718 bp ORF, encoded 905 amino acids with a theoretical molecular weight of 102 kD and an isoelectric point of 6.12. Southern bloting proved that nia gene of Ty7 existed in the form of low copies. It was also proved that this clone was the style of NADH-NR. To reveal the effect of nitrate and ammonium nitrogen on nia expression, we determined NR activity in sugar beet under the treatment of nitrate and ammonium. Gene transcripts of nia were detected by semi-quantitative PCR. Efficiency of mRNA synthesis from each sample was estimated by quantitative PCR of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The results showed that when nia gene was induced by ammonium, the gene expression was promoted with low concentration of ammonium ion, while inhibited by high concentration of ammonium ion. When nia gene was induced by nitrate, being positively correlated with the concentration, the gene expression was enhanced with the increase of the concentration. By 30 mmol L–1 nitrate induction for four hours, nia gene expression reached the highest value, and at about six hours of induction, the expression decreased significantly.

Key words: Nia gene cloning, Nitrogen form and processing, Gene differential expression

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