Polymerase chain reaction amplification of the RuBisCo small subunit genes and their novel application to plant tissue identification

Abstract

Degenerate polymerase chain reaction (PCR) primers and specific PCR conditions have been developed for the selective amplification of a polymorphic region of ribulose-1,5-bisphosphate carboxylase (RuBisCo) small subunit (ssu) genes. Reliable amplification has been achieved for genes from more than 20 plant species from a variety of taxonomic groups. Analysis of Nicotiana species provides strong evidence that the test loci are polymorphic at the interspecies level but show little polymorphic variation at the intraspecies level. This specific multilocus PCR approach provides a powerful counterpart to random amplification of polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) analysis for the identification of plant tissues.