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All that is gold does not glitter? Herbarium samples and DNA phylogeny.

Plantains (Plantago L.) are widely distributed herbs and shrubs from the asterid family Plantaginaeae. There are more than 200 species of Plantago (Rahn, 1996). Plantains exhibit the transition from the insect-pollinated to wind-pollinated flowers (Preston et al., 2011) and unusually fast evolution (Ronsted et al., 2002; Cho et al., 2004): for 5.5 million years, this group produce hundreds of species and colonized of all continents including Antarctic. Unfortunately, the most detailed phylogeny of genus includes only 23 species (Hoggard et al., 2003). We decided to sequence as many species of genus as possible. Since the fresh material is often unavailable, we use herbarium samples from U.S. and international herbarium collections. With the collaboration of International Barcode of Life Project (iBoL), we checked several phylogenetic markers (coi5, rbcL, trnlF, ITS) and found that nuclear ITS2 pseudogene allows to make trees with the best resolution on the species level. ITS2 has the relatively short length (~330 bp) and therefore is suitable for amplification from the partially degraded DNA extracted from herbarium samples. Using two different protocols, we extracted DNA from more than 200 herbarium samples, and have now the comprehensive dataset which elucidates relations between age and weight of tissue sample, and concentration and quality of extracted DNA. One of our plots (dependence between the age of sample and DNA purity) is given below:

One can see that the estimated (LOESS method) curve does not constitute any real dependence, the slope is almost zero. In all, the "old DNA" may be as good as DNA extracted from freshly collected samples, even XIX century herbarium collections will work for our purposes.

Supported by Department of Biology of Minot State University. We are grateful to many people, including herbarium curators of different U. S. and Russia herbarium collections.

Hye Ji Lee (1) *, Alexey Shipunov (1)

(1) Department of Biology, Minot State University, Minot, North Dakota, U.S.

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Author:Lee, Hye Ji; Shipunov, Alexey
Publication:Proceedings of the North Dakota Academy of Science
Geographic Code:1U4ND
Date:Jan 1, 2014
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