Utilization of RAPD Markers to Assess Genetic Diversity of Wild Populations of North American Ginseng (Panax quinquefolium)

Abstract

The Catskill Mountains of New York State are an important source of wild-collected American ginseng (Panax quinquefolium) and, increasingly, of woods-cultivated ginseng. The objective of this study was to assess genetic diversity among 9 different wild ginseng populations in and adjacent to the Catskill Mountain region of New York State and to compare these to wild populations from other states including Kentucky, Tennessee, North Carolina, Pennsylvania, and Virginia, and one cultivated population from Wisconsin. Randomly amplified polymorphic DNA (RAPD) markers were used to estimate the genetic distance among samples from the 15 populations. Pooled DNA from 10 plants of each of 8 New York populations was initially screened with 64 random primers; subsequently, the 15 primers that exhibited the greatest number of reproducible polymorphic markers were selected for further experimentation. Gel electrophoresis with the selected 15 primers produced 124 highly reproducible polymorphic bands. The ratio of discordant bands to total bands scored was used to estimate the genetic distance within and among populations. Multidimensional scaling (MDS) of the relation matrix showed distinctly separate clusters between New York and non-New York populations, indicating separation between these two groupings. The MDS analysis was confirmed using pooled chi-square tests for fragment homogeneity. This study shows that RAPD markers can be used as population-specific markers for Panax quinquefolium, and may eventually be utilized as markers for ginsenoside assessment.

Key words

North America - population - Panax quinquefolium - Araliaceae - genetic diversity - Catskill Mountains - ginsenoside - RAPD - HPLC

References

• 1 Bai D, Brandle J, Reeleder R. Genetic diversity in North American ginseng (Panax quinquefolius L.) grown in Ontario detected by RAPD analysis.  Genome. 1997;  40 111-5
• 2 Boehm C L, Harrison H C, Jung G, Nienhuis J. Organization of American and Asian ginseng germplasm using randomly amplified polymorphic DNA (RAPD) markers.  J Am Soc Hortic Sci. 1999;  124 252-6
• 3 Schluter C, Punja Z K. Genetic diversity among natural and cultivated field populations and seed lots of American ginseng (Panax quinquefolius L.) in Canada.  Int J Plant Sci. 2002;  163 427-39
• 4 Lim Y P, Shin C S, Lee S J, Youn Y N, Jo J S. Survey of proper primers and genetic analysis of Korean ginseng (Panax ginseng C.A. Meyer) variants using the RAPD technique.  Korean J Ginseng Sci. 1993;  17 153-8
• 5 Um J Y, Chung H S, Kim M S, Na H J, Kwon H J, Kim J J. et al . Molecular authentication of Panax ginseng species by RAPD analysis and PCR-RFLP.  Biol Pharm Bull. 2001;  24 872-5
• 6 Kim J H, Yuk J A, Cha S K, Kim H H, Seong B J, Kim S I. et al . Genetic variation in pure lines of Panax ginseng based on by RAPD analysis.  Korean J Med Crop Sci. 2003;  11 102-8
• 7 Ha W Y, Yau F CF, But P PH, Wang J, Shaw P C. Direct amplification of length polymorphism analysis differentiates Panax ginseng from P. quinquefolius .  Planta Med. 2001;  67 587-9
• 8 Ha W Y, Shaw P C, Liu J, Yau F CF, Wang J. Authentication of Panax ginseng and Panax quinquefolius using amplified fragment length polymorphism(AFLP) and directed amplification of minisatellite region DNA(DAMD).  J Agric Food Chem. 2002;  50 1871-5
• 9 Ngan F, Shaw P, But P, Wang J. Molecular authentication of Panax species.  Phytochemistry. 1999;  50 787-91
• 10 Wang J, Ha W Y, Ngan F N, But P PH, Shaw P C. Application of sequence characterized amplified region (SCAR) analysis to authenticate Panax Species and their adulterants.  Planta Med. 2001;  67 781-3
• 11 Persons W S. American ginseng farming in its native woodland habitat. Proceedings of the International Ginseng Conference Vancouver; 1994
• 12 Grubbs H J, Case M AC. Allozyme variation in American ginseng (Panax quinquefolius L.): Variation, breeding system, and implications for current conservation practice.  Conserv Genet. 2004;  5 13-24
• 13 Proctor J TA, Bailey W G. Ginseng: industry, botany, and culture. In: Janick J, editor Horticultural reviews. New York; Van Nostrand Reinhold Company; 1987: 187-236
• 14 Sun M, Wong K C. Genetic structure of three orchid species with contrasting breeding systems using rapd and allozyme markers.  J Am Soc Hortic Sci. 2001;  88 2180-8
• 15 Morgan D JW, Reitz S R, Atkinson P W, Trumble J T. The resolution of Californian populations of Liriomyza huidobrensis and Liriomyza trifolii (Diptera: Agromyzidae) using PCR.  Heredity. 2000;  85 53-61
• 16 Bernatzky R, Tanksley S D. Toward a saturated linkage map in tomato based on isozymes and random cDNA sequences.  Genetics. 1986;  112 887-98
• 17 Nienhuis J, Tivang J, Skroch P. Genetic relationships among cultivars and landraces of Lima bean (Phaseolus lunatus L.) as measured by RAPD markers.  J Am Soc Hortic Sci. 1995;  120 300-6
• 18 Tivang J, Skroch P W, Nienhuis J. Randomly amplified polymorphic DNA (RAPD) variation among and within Artichoke (Cynara Scolymus L.) cultivars and breeding populations.  J Am Soc Hortic Sci. 1996;  121 783-8
• 19 Lim W, Mudge K W, Vermeylen F. Effects of population, age, and cultivation methods on ginsenoside content of American ginseng (Panax quinquefolium).  J Agric Food Chem. 2005;  53 8498-505
• 20 Charron D, Gagnon D. The demography of northern populations of Panax quinquefolium (American ginseng).  J Ecol. 1991;  79 431-45
• 21 Gagnon D, Nantel P. Prediction of population extinction of ginseng and wild leek under harvesting pressure.  Bull Ecol Soc America. 1992;  73 182
• 22 McGraw J B. Evidence for decline in stature of American ginseng plants from herbarium specimens.  Biol Conserv. 2001;  98 25-32
• 23 Cruse-Sanders J M, Hamrick J L. Genetic diversity in harvested and protected populations of wild American ginseng, Panax quinquefolius. (Araliaceae).  Am J Bot. 2004;  91 540-8