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Genetic Analysis of Cypripedium Reginae

AJAS · 2020 Plant Sciences (inferred)

Alisa Halchenko, Garima Rastogi · and

Overview

Showy lady’s slippers orchids (Cypripedium reginae) are critically endangered in New Hampshire, existing in less than 5 places in the wild. The intent of this experiment was to develop an approach for using PCR and DNA sequencing of microsatellite regions to assess genetic diversity of wild Cyp. reginae populations. A lack of genetic diversity can be a contributing factor for a population’s decline especially when facing environmental challenges such as global climate change. We performed PCR on DNA samples extracted from plants in an artificial fen located in Lyme, NH. These plants were grown in axenic seed culture from seeds collected from the Eshqua Bog in Vermont. We utilized PCR primers that had been previously used to target variable microsatellite DNA regions in another species of Cypripedium. We targeted these specific microsatellite regions because they are likely to change between two distinct Cypripedium reginae populations. Our experiments identified a set of primers that could be used for our Cyp. reginae samples and yielded PCR products for one microsatellite region from 16 plants that were submitted for DNA sequencing. This produced sequencing results for the very short microsatellite segments of DNA. The obtained sequences were analyzed for quality and aligned to determine the amount of differences among individuals. There were few differences, with the sequences generally aligning. This suggests our analytic approach works for Cyp. reginae and can be used to analyze the genetic diversity of this orchid species. Based on these results, the plants within the Lyme fen were not substantially genetically different from one another, but comparisons to populations in the wild are needed to fully confirm this suspicion. We would like to analyze the genetic diversity of plants from wild fens to determine if they have robust genetic diversity within and among different fens.

Competition history

  • AJAS 2020 Category not listed

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Source: AAAS Annual Meeting (Confex) / American Junior Academy of Science

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