Genetic Diversity of Paris polyphylla var. yunnanensis, a Traditional Chinese Medicinal Herb, Detected by ISSR Markers

Abstract

Paris polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz. is an important Chinese medicinal herb. Because of overharvesting, the wild populations of this herb have greatly declined and become fragmentized. In this paper, ISSR markers were used to determine the genetic diversity and genetic structure of this variety represented by a total of 153 individuals from three natural populations and three cultivated populations. Fourteen primers produced a total of 251 bands, of which 227 were polymorphic (PPB = 90.44 %). For the natural populations, the results showed that genetic differentiation was mainly within populations (GST = 0.1952), with low genetic diversity at the population level. At the population level, genetic diversity of the cultivated populations was relatively higher than that of the natural populations (PPB = 57.24 % vs. 53.38 %, HE = 0.153 vs. 0.151, HO = 0.241 vs. 0.235). This pattern can be explained by the recent introduction and artificial selection of cultivars from comparatively wide areas of origin, and subsequent gene flow among populations in cultivation. Although the neighbour-joining cluster analysis seemed to suggest that there was conspicuous genetic differentiation between the natural and cultivated populations, the AMOVA showed that only 4.84 % of the total variance existed between groups of natural and cultivated populations, while 67.51 % of the variance occurred within populations. In the end, some suggestions for conservation of this important herb are proposed.

Key words

Trilliaceae - Paris polyphylla var. yunnanensis - ISSR - genetic diversity

References

• 1 Li H. The phylogeny of the genus Paris L.  Acta Bot Yunn. 1984;  6 351-62.
• 2 Matsuda H, Pongpiriyadacha Y, Morikawa T, Kishi A, Kataoka S, Yoshikawa M. Protective effects of steroid saponins from Paris polyphylla var. yunnanensis on ethanol- or indomethacin-induced gastric mucosal lesions in rats: structural requirement for activity and mode of action.  Bio Med Chem Lett. 2003;  13 1101-6.
• 3 Zhou L G, Yang C Z, Li J Q, Wang S L, Wu J Y. Heptasaccharide and octasaccharide isolated from Paris polyphylla var. yunnanensis and their plant growth-regulatory activity.  Plant Sci. 2003;  165 571-5.
• 4 Heywood V H, Iriondo J M. Plant conservation: old problems, new perspectives. In: Iriondo JM, Heywood VH, editors. Plant conservation biology: Emerging tools and strategies. Biological Conservation.  Amsterdam: Elsevier. Science;  2003 321-35.
• 5 Oostermeijer J GB, Luijten S H, den Nijs J CM. Integrating demographic and genetic approaches in plant conservation.  Biol Conserv. 2003;  113 389-98.
• 6 Barrett S CH, Kohn J R. Genetic and evolutionary consequences of small population size in plants: implications for conservation. In: Falk DA, Holsinger KE, editors. Genetics andconservation of rare plants.  New York: Oxford University. Press;  1991 3-30.
• 7 Doyle J. DNA protocols for plants - CTAB total DNA isolation. In: Hewitt GM, Johnston A, editors Molecular techniques in taxonomy. Berlin; Springer 1991: 283-93.
• 8 Williams J GK, Kubelik A R, Livak K J, Rafalski J A, Tingey S V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers.  Nucleic Acids Res. 1990;  18 6531-5.
• 9 Yeh F C, Yang R C, Boyle T. POPGENE, Microsoft Windows-based freeware for population genetic analysis. Release 1.31. Edmonton; University of Alberta 1999.
• 10 Miller M P. AMOVA-PREP 1.01: A Program for the preparation of AMOVA input files from dominant-markers raw data. Computer software distributed by author 1998.
• 11 Excoffier L, Smouse P E, Quattro J M. Analysis of molecular variance inferred from metric distances among DNA haplotypes: Applications to human mitochondrial DNA restriction data.  Genetics. 1992;  131 479-91.
• 12 Rohlf F J. NTSYS-pc: Numerical taxonomy and multivariate Analysisasystem,Version 2.1 New York; Exeter Software 2000.
• 13 McDermott J M, McDonald B A. Gene flow in plant pathosystems.  Annu Rev Phytol. 1993;  31 353-73.
• 14 Nybom H. Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants.  Mol Ecol. 2004;  13 1143-55.
• 15 Jacquemyn H, Brys R, Honnay O, Hermy M, Roldán-Ruiz I. Sexual reproduction, clonal diversity and genetic differentiation in patchily distributed populations of the temperate forest herb Paris quadrifolia (Trilliaceae).  Oecologia. 2006;  147 434-44.
• 16 Li Q, Xiao M, Guo L, Wang L, Tang L, Xu Y. et al . Genetic diversity and genetic structure of an endangered species, Trillium tschonoskii .  Biochem Genet. 2005;  43 445-58.
• 17 Tomimatsu H, Ohara M. Genetic diversity and local population structure of fragmented populations of Trillium camschatcense (Trilliaceae).  Biol Conserv. 2003;  109 249-58.
• 18 Zhang J Y, Yu H, Zhang S G, Ding C. RAPD variation within and among four populations of Paris polyphylla .  Biol Sci. 2004;  12 517-22.
• 19 Hamrick J L, Godt M JW. Allozyme diversity in plant species. In: Brown ADH, Clegg MT, Kahler AL, Weir BS, editors. Plant population genetics, breeding and genetics resources.  Sunderland:. Sinauer;  1990 43-63.
• 20 Li H. The genus Paris (Trilliaceae). Beijing; Science Press 1998
• 21 Sage T L, Griffin S , Pontieri V, Drobac P, Cole W W, Barrett S CH. Stigmatic self-incompatibility and mating patterns in Trillium grandiflorum and Trillium erectum (Melanthiaceae).  Ann Bot. 2001;  88 829-41.
• 22 Zhou L G, Wu J Y, Wang S L. Low-temperature stratification strategies and growth regulators for rapid induction of Paris polyphylla var. yunnanensis seed germination.  Plant Growth Regul. 2003;  41 179-83.
• 23 Hutchison D W, Templeton A R. Correlation of pairwise genetic and geographic distance measures: inferring the relative influences of gene flow and drift on the distribution of genetic variability.  Evolution. 1999;  53 1898-914.
• 24 Slatkin M. Gene flow and the geographic structure of natural populations.  Science. 1987;  236 787-92.
• 25 Wright S. The genetical structure of populations.  Ann Eugenics. 1951;  15 323-54.