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ACTA HORTICULTURAE SINICA ›› 2011, Vol. 38 ›› Issue (12): 2309-2316.

• Vegetables • Previous Articles     Next Articles

Cloning and Expression Pattern Analysis of NRT2 Gene in Non-heading Chinese Cabbage

KONG Min,YANG Xue-dong,HOU Xi-lin*,LIU Tong-kun,and REN Jun   

  1. (State Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing Agricultural University,Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China,Ministry of Agriculture,Nanjing 210095,China)
  • Received:2011-09-20 Revised:2011-11-25 Online:2011-12-25 Published:2011-12-25
  • Contact: HOU Xi-lin

Abstract: In this study,a full-length of cDNA sequence of a high-affinity nitrate transporter gene BcNRT2 was cloned from non-heading Chinese cabbage(Brassica campestris ssp. chinensis Makino)cultivar‘Suzhouqing’using reverse transcript PCR(RT-PCR). Sequence analysis showed that the length of nucleotide sequence of this gene is 1 593 bp,containing a complete open reading frame to encode 530 amino acids. Nucleotide and amino acid sequence comparison indicates that BcNRT2 has a certain high similarity with the orthologous gene BnNRT2 in Brassica napus(98%,99%)and AtNRT2.1 in Arabidopsis thaliana(90%,95%),respectively. Conclusion was made that NRT2 gene is highly conserved among several plant species. Quantitative real-time PCR analysis showed that the expression of BcNRT2 has the highest level in root cells,and expression pattern of this gene belongs to induced system. After 0.5 h treatment of low concentrations of NO3-(0.2 mmol · L-1 KNO3),the expression of BcNRT2 was up-regulated rapidly in root and shoot,suggested that BcNRT2 may acts as a NO3- sensor or signal transducer. After treatment of high concentrations of NO3-(20 mmol · L-1 KNO3),the expression of BcNRT2 was highly up-regulated and lasted longer in root and shoot,which may be resulted from a high-level response of the regulation of a low-affinity nitrate transporter NRT1.1. BcNRT2 protein was located at the plasma membrane supported by subcellular localization assays experiment.

Key words: non-heading Chinese cabbage, nitrate, NRT2, real-time RT-PCR, subcellular localization