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Joe-Ann McCoy, Jeanine M. Davis, N. Dwight Camper, Ikhlas Khan and Avula Bharathi

Black cohosh [Actaea racemosa L. syn. Cimicifuga racemosa (L.) Nuttal] is a native North American medicinal plant traditionally harvested for its rhizomes and roots. Black cohosh products were listed in the top 10 selling herbal supplements from 2002 to 2005. As a result of increasing commercial demand, there is a need to develop propagation protocols suitable for production purposes to replace current methods of harvesting from wild populations. The objectives of this study were to 1) determine optimal rhizome propagule division size for successful regeneration, 2) analyze triterpene glycoside concentrations, 3) quantify survival rates after 3 years of production, and 4) evaluate net yield results. Experimental sites included a shade cloth structure in an agricultural research field, a shaded forest interior, and a shaded, disturbed forest edge. Plant emergence, growth, and survival were assessed at each site over a 3-year period. Optimal rhizome division size for propagation was a 10 to 30-g section originating from terminal rhizome portions. Rhizome survival averaged 97% among all treatments tested by year 3 at three sites. No differences in mean triterpene glycoside concentrations were detected between rhizome size classes or sites tested. Mean cimiracemoside concentrations ranged from 0.80 to 1.39 mg·g–1 d/w tissue, deoxyactein 0.47 to 0.92 mg·g–1, and actein 10.41 to 13.69 mg·g–1. No differences in triterpene levels were detected between flowering and nonflowering plants, nor were yields reduced. Net yields from a shade cloth production site were 9 and 17 times higher than a disturbed forest edge and forest site respectively. Black cohosh is a strong candidate for commercial propagation under adequate site selection.

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Kent E. Cushman, Rita M. Moraes, Patrick D. Gerard, Ebru Bedir, Bladimiro Silva and Ikhlas A. Khan

Leaves of American mayapple (Podophyllum peltatum) are being investigated as an alternative and renewable source of podophyllotoxin, a pharmaceutical compound used in the manufacture of several drugs. This study examined long-term performance of mayapple populations subjected to different harvest strategies. A naturally occurring population in shade was subjected to leaf removal treatments of frequency (every year, every 2nd or 3rd year) or timing (early or late season). Plots were 1.0 m2, established during Spring 2001, and treatments were applied from 2001 to 2004. Control plots not previously harvested were also included each year. Plants did not tolerate the severest of leaf removal treatments: early harvest time in combination with annual harvest frequency. Early annual harvests reduced total leaf dry mass and total leaf area in a quadratic manner. Late harvest conducted annually, and early harvest conducted every other year, also reduced leaf dry mass and area but not as much as early annual harvest. Plants harvested every year, early, or early every year produced fewer sexual shoots than other treatment combinations. Contents of α-peltatin, β-peltatin, and total lignans were higher for leaves harvested early than those harvested late during each year of the study, demonstrating that lignan contents were affected by leaf age and not treatment. In conclusion, our results for plants grown in shade show that leaves can be removed late in the growing season every 2nd or 3rd year or early every 3rd year without reducing long-term performance of the population. This is more restrictive than that reported for populations in full sun where plants tolerated late harvests every year.

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Kent E. Cushman, Muhammad Maqbool, Hemant Lata, Ebru Bedir, Ikhlas A. Khan and Rita M. Moraes

Four levels of shade (0%, 30%, 55%, and 80%) were used to determine their effect on growth and lignan content of american mayapple (Podophyllum peltatum L.). Mayapple rhizomes were harvested from the wild and transplanted into plant beds on 20 Dec. 2001 using a randomized complete block design with four blocks. Growth and lignan content were recorded during spring of 2002 and 2003. Leaf samples were analyzed for the following lignans: podophyllotoxin, alpha-peltatin, and beta-peltatin. Increasing levels of shade increased shoot longevity, leaf area per plant (cm2/plant), and shoot height. Shade did not affect shoot emergence, total leaf area (cm2·m-2), or leaf dry mass (g·m-2 or g/plant). Regardless of year, podophyllotoxin and total lignan contents at 0% shade were significantly greater than those at 80% shade, and the overall trend was for decreasing contents with increasing shade. Shade did not affect alpha-peltatin content. Content of beta-peltatin was greatest at 0% shade compared to the other three shade treatments. Year affected alpha-peltatin and beta-peltatin contents, with less content of each in 2003 than in 2002. There were large numerical decreases in podophyllotoxin yield (podophyllotoxin content per unit area, mg·m-2) as shade increased from 0% to 80%, but these differences were only marginally significant (P = 0.0897). In contrast, podophyllotoxin yield was significantly greater in 2003 than in 2002 as total leaf area and dry mass significantly increased. Increasing levels of shade slightly decreased air and soil temperatures. Our results indicate that american mayapple is not a shade-requiring species. Under full sun (0% shade) shoots did not persist as long as under shade and leaves were smaller and thicker, but total lignan content was significantly greater than under shade. It appears that growers of specialty crops serving the pharmaceutical industry can establish and cultivate american mayapple under full sun, thus providing leaf biomass with high podophyllotoxin content while avoiding the cost of expensive shade structures.

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Kent Cushman, Muhammad Maqbool, Ebru Bedir, Hemant Lata, Ikhlas Khan and Rita Moraes

Podophyllotoxin is a pharmaceutical compound extracted from rhizomes of Indian mayapple (Podophyllumemodi). Leaves of American mayapple (P. peltatum) also contain podophyllotoxin, and the species is being investigated as a domestic, renewable, and alternative source of the compound. The objective of this study was to explore strategies of leaf removal that would not adversely affect regrowth of American mayapple shoots in subsequent years. Plots were established in two locations among naturally occurring populations in the wild, one in full sun and one in partial shade. Plots were 1.0 m2 and leaves were removed from plants every spring, every other spring, or every third spring. In addition, leaves were removed in early spring, soon after shoots emerged and leaves had fully expanded, or in late spring, when leaves first showed evidence of yellowing and beginning to senesce. Sexual and asexual leaves were harvested separately. Leaf number, leaf area, and dry weight were recorded. Subsamples of leaf material were extracted to determine podophyllotoxin, α-peltatin, and β-peltatin contents. Results clearly showed that leaf removal every year, in combination with early harvest, was too severe and plants lost vigor over the 4-year period of this study. Plants subjected to this treatment combination produced significantly less leaf area and dry weight than any of the other treatment combinations. Results were similar for both sun and shade locations. Lignan content was not affected by treatment. Our results indicate that leaves can be removed from mayapple plants as often as every year provided harvests are not scheduled too soon after shoot emergence.