RESEARCH PROPOSAL
A COMPARATIVE STUDY ON PRIMULA VULGARIS HUDS. IN TERMS OF
MORPHOLOGICAL AND MOLECULAR PROPERTIES AT SUBSPECIFIC LEVEL
Zeynep ÇOLAK
14.03.2014
A Comparative Study on Primula vulgaris Huds. in Terms of Morphological and
Molecular Properties at Subspecific Level
Abstract
Primula vulgaris Huds has a systematic problem at subspecific level in that it has an
inadequate identification key in Flora of Turkey. Primula vulgaris has represented by two
subspecies in the Turkey. These taxa naturally grow together in the Northern of Blacksea.
Besides, these two subspecies have a similar morphology and flower colors. These makes it
difficult to distinguish them. Thus, we need new criteria except color and altitude. In this study,
18 populations of two subspecies of Primula vulgaris from different altitude in Trabzon will
be investigated based on nrDNA ITS and phenetic properties. At the end of the study, we can
find new criteria and prepare an identification key. Besides, similarity and dissimilarity of ITS
regions could be found. Eventually, we could be able to say hese taxa are “subspecies” or
“varieties” of Primula vulgaris. Thus, it can be contribute to the solution of the existing
systematic problem of species.
KeyWords: PrimulavulgarisHuds.,ITS, Phenetic
Introduction
Primula vulgaris Huds.is an important member of the Primula L. which is the largest
genus of Primulaceae family (Richards, 2003). Primroses (Primula L., Primulaceae) are
remarkable plants with features making them appropriate candidates for a wide range of scientific
research, especially in population biology, evolution of sex, plant-animal interactions, and
geographic diversity (Valentine 1947, 1948, 1955; Smith and Fletcher 1948; Whale 1984; Mast et
al. 2001; Richards 2003; Kalman et al. 2004, 2007).
This species has been represented by two subspecies in Turkey. These are P. vulgaris
Huds. subsp. vulgaris and P. vulgaris Huds. subsp. sibthorpii(Hoffmanns)W.W.Sm. & Forrest
(Lamond, 1978).
The common primrose has several color morphs. Two of them, traditionally treated as
subspecies vulgariswith white and yellow flowers and polymorphic sibthorpii with the
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predominance of dark (purple, pink) flowers, often occur at the same places along the eastern
Black Sea coast. There are no reliable morphological differences between color forms.
These two taxa are distributed in the same habitat in the North of our country although
called “subspecies” and they have a quite similar morphology so it’s quite difficult to distinguish
between these two taxa according to altitude of distribution and morphological character like
flower color.subsp. vulgaris has distribution in the range of 500-2100m altitude in Northern and
Southern but subsp. sibthorpiihas distribution in the range of 0-850m altitude only in the Northern
part of Turkey (Lemond, 1978). This shows that two subspecies naturally grow together in the
range of 500-850 m altitude. So, it makes difficult to distinguish them. Because of these reasons
we need to find new criteria for distinguishing these two taxa.
In this study, it’s aimed to study morphological and molecular profiles of large population
of Primula vulgaris in a comparative way.Thus, this is the first study in terms of these properties
about subspecies of P. vulgaris and new criteria can be determined for distinguishing these taxa.
Research Questions
1. What are the new criteria which can be used to distinguish these two taxa?
2. What are the important morphological characters can be used to distinguish them?
3. Is there a correlation between altitude and population distribution and color forms of
these taxa?
4. Are there any differences of ITS regions of these taxa?
5. Is there a correlation between morphological and molecular properties of these taxa?
Material and Methods
Forthe present study, plant material will be collected from different altitudes (02200m)in Trabzon-Turkey. Plant samples will be dried according to the herbarium techniques
and used for further morphological examination and measurements. The row data matrix of
morphological measurements will be used for CA and PCA analysis.
For molecular study, total genomic DNAs will be extracted from silica-dried leaves
following the modified CTAB extraction procedure of Doyle and Doyle (1987). After DNA
extraction the entire ITS regions (ITS1, 5.8S, and ITS2) will be amplified by PCR using
universal primers ITS4 an ITS5. After that, the nucleotide sequences will be analyzed by using
proper phylogenetic programs. Then, ITSpolymorphism can be determined.
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Result and Discussion
As a result of this study, an identification key can be prepared for P. vulgaris at
subspecific level. The most important traits in distinguishing the subspecies can be found. nrDNA
ITS profiles for 18 populations belong to subspecies can be analyzed to represent phylogenetic
relationship at intra and interspecific level among the examined populations. A dendogram can be
prepared according to morphological characters. And it can be found whether there is a
correlation between these morphological and molecular criteria.
References
Doyle, J., J. and Doyle, J., L., 1987. A Rapid DNA Isolation Procedure for Small
Quantities of Fresh Leaf Tissue, Phytochemical Bulletin, 19, 11 - 15.
Kalman, K., Medvegy, A. andMihalik, E., 2004. Pattern of the Floral Variation in the
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Kalman, K., Medvegy, A., Penzes,Zs. andMihalik, E., 2007. Morph-specificvariation of
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Lamond, J., 1978. In: Davis, P. H., Ed., Flora of Turkey and The East Aegean Islands,
Edinburgh University Press, Edinburgh, 6, 112 - 120.
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