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Journal of Plant Research

, Volume 115, Issue 1, pp 0065–0070 | Cite as

Development of primer sets for PCR amplification of the PgiC gene in ferns

  • H. Ishikawa
  • Y. Watano
  • K. Kano
  • M. Ito
  • S. Kurita
TECHNICAL NOTE

Abstract

Polymerase chain reaction (PCR)-based nuclear DNA markers were developed for fern species. We first determined the partial nucleotide sequence of cDNA of the pgiC gene encoding cytosolic phosphoglucose isomerase from Dryopteris caudipinna, and then PCR primers for exon-primed, intron-crossing (EPIC) amplifications were designed. The EPIC primers are universally applicable to the most derived indusiate fern families such as Dryopteridaceae, Thelypteridaceae, and Woodsiaceae. The PCR products of primers 14F/16R containing two introns are moderate in size (534 bp–ca.1000 bp) and are possibly of value in phylogenetic reconstruction at specific and generic levels. Codominant nuclear DNA markers applicable to the estimation of mating systems and other population genetic studies were also developed by a combination of single-strand conformation polymorphism (SSCP) and EPIC amplification using primers 14F/15R and 15F/16R. In order to provide a case study using these markers, allelic variation of PCR products using 15F/16R was examined in populations of Arachniodes standishii (Dryopteridaceae).

Key words Codominant nuclear DNA marker Homosporous ferns PCR primers PgiC Phosphoglucose isomerase SSCP 

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Copyright information

© The Botanical Society of Japan and Springer-Verlag Tokyo 2002

Authors and Affiliations

  • H. Ishikawa
    • 1
  • Y. Watano
    • 2
  • K. Kano
    • 2
  • M. Ito
    • 1
  • S. Kurita
    • 1
  1. 1.Department of Biology, Chiba University, Inage, Chiba, JapanJP
  2. 2.Department of Biology, Kanazawa University, Kakuma, Kanazawa, JapanJP

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