New insights into allelic diversity of a phosphoenolpyruvate carboxylase in the conifer Picea abies (L.) Karst.

Abstract

Two genomic full-length alleles of a phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) were isolated and analysed in the gymnosperm Norway spruce [Picea abies (L.) Karst.]. Mendelian segregation analysis confirmed that the two alleles belong to the same DNA gene locus. Southern analysis was performed using exon and intron probes, separately. A small gene family was discovered when exon probes were used. The isolated gene locus was termed PEPC-1 and found to exhibit the largest PEPC genomic sequence characterised so far in the plant kingdom. This is due to comparatively long intron sequences, and to an additional intron in the 3'-untranslated region. Sequence homology of the two full-length PEPC-1 alleles is 99.8%. From exon variation, one single exchange of an amino acid was deduced. The introns harboured polymorphic regions that were analysed for variability among three different spruce individuals. Within-PEPC-1 genotyping revealed that all three individuals could be unambiguously distinguished from each other. The results are discussed with respect not only to exon/intron site evolution among alleles but also to particular genomic features of a gymnosperm PEPC in comparison to PEPC in angiosperms.

Characterisation of the phosphoenolpyruvate carboxylase gene family in sugarcane (Saccharum spp.)

“…STS genetic markers and genomic organisation of PEPC genes STS markers for the sugarcane PEPC genes were developed as already proposed for a Picea PEPC gene (Ipsen and Ziegenhagen 2001). Using our primers, we showed that STS PEPC markers can also be successfully used in related grass genera.…”

Molecular identification of individual oak and fir trees from maternal tissues of their fruits or seeds

“…The three primer pairs were taken from a previously characterised full-length sequence of PEPC-1 in Norway spruce (Ipsen and Ziegenhagen 2001) and selected for different sizes of the expected PCR products. The three primer pairs were taken from a previously characterised full-length sequence of PEPC-1 in Norway spruce (Ipsen and Ziegenhagen 2001) and selected for different sizes of the expected PCR products.…”

“…The endosperm haplotypes revealed that four of the five donor trees could be unambiguously distinguished from each other and that with one exception Fig. To prove that nuclear DNA can also, in principle, be analysed from single wings, we PCR-amplified different regions of the single copy nuclear gene PEPC-1 (Ipsen and Ziegenhagen 2001). A Agarose gel electrophoresis of total DNA extracted from the pericarps following the CTAB-based minipreparation protocol by Dumolin et al (1995).…”