Identification of the vascular plants of Churchill, Manitoba, using a DNA barcode library

Annals of Occupational and Environmental Medicine

Maria L Kuzmina¹  Karen L Johnson³  Hannah R Barron²  Paul DN Hebert¹ 

[1] Biodiversity Institute of Ontario, University of Guelph, Guelph, ON, Canada

[2] Trent University, Environmental and Life Sciences, Peterborough, ON, Canada

[3] The Manitoba Museum, Botany Department, Manitoba, Canada

关键词: Biomonitoring;    Climate change;    Species resolution;    ITS2;    matK;    rbcL;    DNA barcoding;    Arctic;   

Others  :  1085781
DOI  :  10.1186/1472-6785-12-25

 received in 2012-08-20, accepted in 2012-11-26,  发布年份 2012

【 摘 要 】

Background

Because arctic plant communities are highly vulnerable to climate change, shifts in their composition require rapid, accurate identifications, often for specimens that lack diagnostic floral characters. The present study examines the role that DNA barcoding can play in aiding floristic evaluations in the arctic by testing the effectiveness of the core plant barcode regions (rbcL, matK) and a supplemental ribosomal DNA (ITS2) marker for a well-studied flora near Churchill, Manitoba.

Results

This investigation examined 900 specimens representing 312 of the 354 species of vascular plants known from Churchill. Sequencing success was high for rbcL: 95% for fresh specimens and 85% for herbarium samples (mean age 20 years). ITS2 worked equally well for the fresh and herbarium material (89% and 88%). However, sequencing success was lower for matK, despite two rounds of PCR amplification, which reflected less effective primer binding and sensitivity to the DNA degradation (76% of fresh, 45% of herbaria samples). A species was considered as taxonomically resolved if its members showed at least one diagnostic difference from any other taxon in the study and formed a monophyletic clade. The highest species resolution (69%) was obtained by combining information from all three genes. The joint sequence information for rbcL and matK distinguished 54% of 286 species, while rbcL and ITS2 distinguished 63% of 285 species. Discrimination of species within Salix, which constituted 8% of the flora, was particularly problematic. Despite incomplete resolution, the barcode results revealed 22 misidentified herbarium specimens, and enabled the identification of field specimens which were otherwise too immature to identify. Although seven cases of ITS2 paralogy were noted in the families Cyperaceae, Juncaceae and Juncaginaceae, this intergenic spacer played an important role in resolving congeneric plant species at Churchill.

Conclusions

Our results provided fast and cost-effective solution to create a comprehensive, effective DNA barcode reference library for a local flora.

【 授权许可】

   
2012 Kuzmina et al.; licensee BioMed Central Ltd.

【 图 表 】

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