Nuclear DNA content of Hydrastis canadensis L. and genome size stability of in vitro regenerated plantlets
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Knowing the genome size is an important step towards deciding and planning for genome sequencing of a given species. Using flow cytometry, nuclear DNA content of Hydrastis canadensis was estimated, and genome size stability of its in vitro regenerated plantlets were assessed. The nuclear DNA content of H. canadensis was estimated to be 2.62 ± 0.020 pg/2C. This is the first report to estimate nuclear DNA content of H. canadensis; therefore this study provides valuable information that will facilitate genome sequencing and subsequent molecular studies of this economically important medicinal plant. Comparison of genome size between in vitro regenerated (explant source was from wild plants) and wild plants of H. canadensis did not reveal any significant difference (P ≤ 0.05) in nuclear DNA content. This suggests that micropropagation of H. canadensis, even after numerous sub-culturing and long-term culture periods produced in vitro plantlets with a stable genome size. These results provide further evidence that micropropagation techniques have the potential to be used as a source of planting stock, along with seeds, for restoring locally threatened H. canadensis wild populations and for commercial cultivation to supply the growing herbal market.
KeywordsGermplasm conservation Medicinal plant Goldenseal Yellow root Flow cytometry Tissue culture
Arumuganathan and Earle’s buffer
We wish to thank Dr. Jaroslav Doležel of the Institute of Experimental Botany, Olomouc, Czech Republic for kindly providing P. sativum cv. Ctirad seeds used as internal reference standard, and Dr. Kathy Brundage of the West Virginia University Flow Cytometry Core Facility for her technical help with Flow Cytometry. The Flow Cytometry Core facility is supported by National Institutes of Health grant # RR016440, whereas this project was supported by the National Research Initiative Competitive Grant no. 2007-35318-18455 from the USDA National Institute of Food and Agriculture.
- AHPA (2007) Tonnage survey of select North American wild-harvested plants, 2004–2005. American Herbal Products Association, Silver Spring, Maryland. Available via DIALOG. http://www.ahpa.org/Default.aspx?tabid=68#2004tonnagesurvey. Accessed on 18 Nov 2009
- Bannerman J (1997) Goldenseal in world trade: pressures and potentials. HerbalGram 41:51–52Google Scholar
- Bedir E, Lata H, Schaneberg B, Moraes RM (2004) In vitro plant regeneration from leaf-derived callus in H. canadensis (Hydrastis candensis). Acta Hort 629:277–281Google Scholar
- Bennett MD, Leitch IJ (2005) Plant DNA C-values database release 4.0. Royal Botanic Gardens, Kew. Availabbe via DIALOG. http://data.kew.org/cvalues. Accessed on 21 Oct 2009
- Bennett MD, Leitch IJ, Price HJ, Johnston JS (2003) Comparisons with Caenorhabditis (~100 Mb) and Drosophila (~175 Mb) using flow cytometry showgenome size in Arabidopsis to be ~157 Mb and thus ~25% larger than the Arabidopsis genome initiative estimate of ~125 MB. Ann Bot 91:547–557CrossRefPubMedGoogle Scholar
- Blumenthal M (1999) Herb market levels after five years of bloom: 1999 sales in mainstream market up only 11% in first half of 1999 after 55% increase in 1998. HerbalGram 47:64–65Google Scholar
- CITES (1997) Proposal for the inclusion of Hydrastis Canadensis. Appendix II, CITESGoogle Scholar
- Foster S, Duke J (2000) A field guide to medicinal plants and herbs of eastern and central North America, 2nd edn. Houghton Mifflin Company, New York, pp 57–58Google Scholar
- Hall KC, Camper ND (2002) Tissue culture of goldenseal (Hydrastis canadensis L.). In Vitro Cell Dev Biol-Plant 38:293–295Google Scholar
- SAS Institute, Inc (2003) SAS proprietary software version 9.1. SAS Inst, Cary, NCGoogle Scholar