Selection of candidate coding DNA barcoding regions for use on land plants

An in silico screen of 41 of the 81 coding regions of the Nicotiana plastid genome generated a shortlist of 12 candidates as DNA barcoding loci for land plants. These loci were evaluated for amplification and sequence variation against a reference set of 98 land plant taxa. The deployment of multiple primers and a modified multiplexed tandem polymerase chain reaction yielded 85–94% amplification across taxa, and mean sequence differences between sister taxa of 6.1 from 156 bases of accD to 22 from 493 bases of matK. We conclude that loci should be combined for effective diagnosis, and recommend further investigation of the following six loci: matK, rpoB, rpoC1, ndhJ, ycf5 and accD. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 1–11.     

Phylogenetic relationships in the grass family (Poaceae) based on the nuclear single copy locus topoisomerase 6 compared with chloroplast DNA

Phylogenetic relationships within the grass family were studied using a newly obtained locus of the nuclear single copy gene topoisomerase 6 (Topo6) spanning the four exons 8–11 and the chloroplast matK gene. Data were evaluated using maximum parsimony, maximum likelihood and Bayesian methods. All analyses showed genera Streptochaeta and Anomochloa as early diverging, followed by Pharus as sister to the rest of the Poaceae, and monophyly of the subfamily Anomochlooideae was supported by the nuclear dataset. The remaining grasses formed a strongly supported and monophyletic group, which split into the major clades BEP and PACMAD in the Topo6 analyses. Monophyly of the BEP clade was strongly supported by the Topo6 data. The results showed clearly incongruity between the two sets of data, such as the different subfamilial relationships of Bambusoideae, Ehrhartoideae and Pooideae. Most of the analysed species are representatives of subfamily Pooideae, which was analysed in more detail by PCR fragment length differences of another Topo6 region spanning the exons 17–19. Monophyly of Pooideae was strongly supported by the matK data, whereas the nuclear data placed Brachyelytrum outside of the remaining Pooideae. Relationships within the early evolutionary lineages remained largely unresolved in the phylogenetic trees, but the ‘core’ Pooideae (Aveneae/Poeae tribe complex and Hordeeae) were highly supported in all analyses. The differences in amplification lengths illustrate the tribe and subtribe classification of Pooideae. The comparatively conserved structure of the newly studied Topo6 region makes it a promising marker from the nuclear genome that could be successfully PCR-amplified to study higher-level phylogenetic relationships within grasses and perhaps between families within the order Poales.     

A reassessment of tribal affinities of Cratystylis and Haegiela (Asteraceae) based on three chloroplast DNA sequences

 The tribal affinities of Cratystylis and Haegiela were assessed using three chloroplast DNA sequences, the trnL/F spacer, the trnL intron and the matK coding region. The outgroup was represented by two species of the subfamily Barnadesioideae, whereas one to seven genera (45 species including Cratystylis and Haegiela) of the tribes of the Asteroideae [Anthemideae (6 genera), Astereae (7), Calenduleae (2), Gnaphalieae (7), Heliantheae s.l. (5), Inuleae s.str. (3), Plucheeae (3), Senecioneae (4)] and Cichorioideae, [Arctotideae (1), Cardueae (2), Lactuceae (2), Liabeae (1), Mutisieae (1) and Vernonieae (1)] comprise the ingroup. Phylogenetic analysis indicates that Cratystylis has strong support as a member of the tribe Plucheeae, whereas Haegiela is a member of Gnaphalieae. At some point in their taxonomic history, both genera have been placed in these tribes and there are good morphological and chemical characters that justify these placements. The monotypic Haegiela was once included in Epaltes (Plucheeae) and this study has confirmed the need for the separation of the two genera. The genus Cratystylis appears to be monophyletic. Received August 26, 2002; accepted September 19, 2002 Published online: February 7, 2003