Determination of plant phylogeny involving introns from the RuBisCo small subunit encoding gene

摘要

A process for the determination of the phylogeny of plant genetic material comprises the characterisation of at least one intron from the ribulose 1,5-bisphosphate carboxylase (RuBisCo) encoding gene therefrom, and comparison of said intron(s) with at least one previously-characterised intron from a RuBisCo small subunit encoding gene. The comparison may involve introns having substantially identical flanking sequences, the sequences of the introns, or the length of one or more of the introns (optionally including intervening sequences). The genetic material may be genomic DNA. The process may be used to determine the phylogeny of processed plant genetic material, or the genus, species or variety of plant genetic material. A preferred process comprises measurement of the length of the introns in a double-stranded nucleic acid by: (i) separation of the two strands of the nucleic acid, thereby to yield single-stranded molecules; performing, at least once, the following: (ii) annealing, to the single-stranded molecules (the 'template strands'), oligonucleotide primers specific to conserved sequences adjacent to the intron(s), whose length is to be measured, of the RuBisCo small subunit encoding gene; (iii) extending the bound oligonucleotide primers to yield complementary copies of the template strands; (iv) separation of the resultant double-stranded products to yield single-stranded molecules; wherein the phylogeny of the plant genetic material is determined by measurement of the lengths of the extended oligonucleotide primers, and comparison of the lengths of said extended primers with the length of extended oligonucleotide primers previously measured from at least one intron of a RuBisCo small subunit encoding gene. Two oligonucleotide primers may be used, one specific to a conserved sequence 5' to the first intron, and the other specific to a conserved sequence 3' to the last intron, of a RuBisCo small subunit encoding gene.