Two cDNAs representing allelles of the nitrate reductase gene of potato (Solanum tuberosum L. cv. Desiree): sequence analysis, genomic organization and expression

John Langford Wray, N Harris, JM Foster, A Kumar, other 2

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)

Abstract

Two, different, full-length cDNAs, StNR2 and StNR3, of 3049 and 3066 nucleotides, respectively, were isolated from a Solanum tuberosum L. cv. Desiree leaf cDNA library by an RT-PCR approach. Conceptual translation of the longest open reading frame of each cDNA showed that they could encode a protein of 911 amino acids in each case with an M-r of 102.6 and 102.5 kDa, respectively, and with 99.7% identity with each other. The cDNAs have a high degree of sequence similarity with all higher plant nitrate reductases (NRs) and possess structural characteristics expected of NADH-NR proteins, consistent with enzyme activity data, Southern analysis of genomic DNA suggested the presence of a single NR gene in the potato genome whilst studies using the mapping population F1840, and the full-length stNR2 cDNA as hybridization probe, identified a single NR locus within the potato genome that is located on chromosome XI. The two cDNAs identified here are probably derived from two transcribing alleles of this single gene. Distribution of total NR transcript and of NADH-NR activity, in different organs of compost-grown plants, depended on the level of nitrate supplied: at low nitrate level transcript and activity were detected only in leaf and stem tissue whilst at high nitrate level they could also be detected in root and stolen. An RT-PCR approach was used to differentiate between the transcripts derived from the two identified alleles and to show that the pattern of transcription of the two identified alleles of the potato nia gene, in the organs studied, is the same.

Original languageEnglish
Pages (from-to)1017-1026
Number of pages10
JournalJournal of Experimental Botany
Volume51
Publication statusPublished - Jun 2000

Keywords

  • gene expression
  • genomic organization
  • nitrate reductase
  • potato
  • Solanum tuberosum L. cv. Desiree
  • REGULATORY PHOSPHORYLATION SITE
  • HIGHER-PLANTS
  • POSTTRANSLATIONAL REGULATION
  • HINGE-1 REGION
  • MESSENGER-RNA
  • ENCODING GENE
  • NADH-BINDING
  • PETIOLE SAP
  • NITROGEN
  • DOMAIN

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