Ginseng Populations Research Articles

Variation in ginsenoside content and composition within the American ginseng population.

The contents of 57 ginsenosides were measured in American ginseng roots collected from a single population grown in Jilin. Ginsenoside contents and compositions varied significantly among the population. The most abundant ginsenoside within the American ginseng root and by population was either Rg1 or Re, followed by Rb1, mal-Rb1, Rd, Rc, pseudoginsenoside F11, Ro, and gypenoside XVII. According to the relative contents of Rg1 and Re, the population was grouped into two chemotypes, and there were significant differences in ginsenoside profiles between them. The contents of protopanaxadiol-type ginsenosides were relatively stable. The rate-limiting enzymes of malonyl ginsenosides biosynthesis were located in front of their corresponding neutral ginsenosides. The regulatory sites of the oleanolic acid-type ginsenosides biosynthetic pathway were in front of the β-amyrin.

Genetic population structure of the Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) detected by microsatellite analysis.

Panax vietnamensis Ha et Grushv. is a precious medicinal species native to the tropical forests of Vietnam. Due to habitat loss and over-harvesting, this species is endangered in Vietnam. To conserve the species, we investigated genetic variability and population structure using nine microsatellites for 148 individuals from seven populations across the current distribution range of P. vietnamensis in Vietnam. We determined a moderate genetic diversity within populations (HO = 0.367, HE = 0.437) and relatively low population differentiation (the Weir and Cockerham index of 0.172 and the Hedrick index of 0.254) and showed significant differentiation (P < 0.05), which suggested fragmented habitats, over-utilization and over-harvesting of P. vietnamensis. Different clustering methods revealed that individuals were grouped into two major clusters, which were associated with gene flow across the geographical range of P. vietnamensis. This study also detected that ginseng populations can have undergone a recent bottleneck. We recommend measures in future P. vietnamensis conservation and breeding programs.

Linalool as a novel natural factor enhancing ginsenoside production in hairy root cultures of American ginseng

Ginsenosides are triterpenoid saponins, accumulated in root of Panax qiunquefolius. These secondary metabolites have numerous pharmacological properties such as: antimicrobial, antioxidant, anti-inflammation, anticancer. They have been found to regulate the functioning of the nervous and endocrine systems, thus maintaining homeostasis. Root harvesting for ginsenoside extraction for pharmaceutical industry destroys the entire plant, limiting its natural occurrence and impacts on wild populations of ginseng. The present study showed that hairy root cultures of P. quinquefolius, after using linalool as elicitor, can increase ginsenoside yield without the use of field-grown plants and independently of the vegetative season. The content of seven ginsenosides (Rb1, Rb2, Rb3, Rc, Rd, Rg1, Re) was determined. We found linalool to stimulate most studied saponin accumulation regardless of exposure time (24 and 72 h). Shorter time of elicitation and 0.1 µM linalool in medium proved to be optimum conditions to obtain the highest total saponin content (29% higher level than that of untreated roots) and Rg-group metabolites (2.28 fold higher amount than untreated roots). Ginsenosides, belonging to protopanaxadiol derivatives, were found to have different dynamics of their content changes depending on linalool concentration. The highest increase in untreated roots was noted for compound Rd. Therefore, elicitation with linalool can be an effective method of enhancing ginsenoside production in P. quinquefolium hairy root cultures cultivated in shake flasks.

Evaluation of genetic diversity of ginseng fruit color germplasm resources: based on SSR analysis

Illumina Xten was employed for shallow sequencing of Panax ginseng(ginseng) samples, MISA for screening of SSR loci, and Primer 3 for primer design. Polymorphic primers were screened from 180 primers. From the successfully amplified polymorphic primers, 15 primers which featured clear peak shape, good polymorphism, and ease of statistics were selected and used to evaluate the genetic diversity and germplasm resources of 36 ginseng accessions with different fruit colors from Jilin province. The results showed that red-fruit ginseng population had high genetic diversity with the average number of alleles(N_a) of 1.031 and haploid genetic diversity(h) of 0.172. The neighbor-joining cluster analysis demonstrated that the germplasms of red-fruit and yellow-fruit ginseng populations were obviously intermixed, and pick-fruit ginseng germplasms clustered into a single clade. The results of STRUCTURE analysis showed high proportion of single genotype in pick-fruit ginseng germplasm and abundant genotypes in red-fruit and yellow-fruit ginseng germplasms with obvious germplasm mixing. AMOVA revealed that genetic variation occurred mainly within populations(62.00%, P&lt;0.001), and rarely among populations(39%, P&lt;0.001), but homogenization was obvious among different populations. In summary, pink-fruit ginseng population may contain rare genotypes, which is the basis for breeding of high-quality high-yield, and multi-resistance varieties, genetic improvement of varieties, and sustainable development and utilization of ginseng germplasm resources.

The knotty politics of ginseng conservation and management in Appalachia

American ginseng (Panax quinquefolius) is a long-lived understory species in Appalachian forests and the most valuable medicinal plant in North America. Indeed, "digging" for ginseng roots is an important livelihood strategy throughout Appalachia. Increasingly, however, concern for ginseng populations is escalating, and state and federal policies have introduced new harvesting restrictions, as well as new law enforcement efforts that target ginseng diggers. Here I am interested in troubling the high-profile narrative that ginseng populations are crashing due to the unscrupulous practices of Appalachian diggers. I draw on ecological research, historical documents, and my own ethnographic fieldwork to argue that we need a fuller understanding of both ginseng population demographics and the potential causes for ginseng decline before we embrace a narrative that disenfranchises those who depend on and, in many cases, have helped steward this enigmatic plant. This research speaks to growing tensions between rural livelihoods and conservation efforts worldwide.

Effects of altered climates on American ginseng population dynamics

AbstractAs climates change, species with locally adapted populations may be particularly vulnerable as specialization narrows the range of conditions under which populations can persist. Populations adapted to local climate as well as other site‐specific characteristics like soils present challenges for inferring how changing climates affect fitness, as climatic and nonclimatic variables that constitute local conditions decouple. We conducted two transplant experiments involving American ginseng to test how climatic conditions affect performance while controlling for effects of other site characteristics. We first out‐planted populations from differing elevations to gardens arrayed along an elevation/climate gradient. We also grew maternal plants under temperatures corresponding to home‐site and future conditions (16.4–22.4°C), transplanting resultant progeny to two home‐sites at different elevations (400 m, 800 m). Source populations responded idiosyncratically to elevation reflecting how nonclimatic site characteristics strongly affected plant fitness. Germination rates declined for seeds from maternal plants exposed to warmer temperatures, which compounded with diminished seed production of maternal plants, suggested that population growth may decline rapidly as warm years become hotter and more frequent. Controlling for maternal temperature effects provided evidence that plants are adapted to home‐site conditions, both climatic and nonclimatic, with population growth rates for out‐planted populations ranging from below population replacement levels (λ = 0.58) to well above (λ = 1.33). Evidence of local adaptation to climatic and nonclimatic environmental components, in combination with negative fitness impacts of warming climates on offspring via maternal effects, suggests that changing climate may imperil ginseng and other similar understory species.

Variety origin authentication of Panax ginseng C.A. Mey. and industrial ginseng products using SNP-based allele-specific PCR method

Accurate identification of variety origin of Panax ginseng is quite necessary not only for variety conservation but also for the quality control of ginseng intermediate and terminal products. However, DNA degradation during the manufacturing processes and the low intra-specific polymorphism within P. ginseng lead to botanical origin authentication of industrial ginseng products at the variety level quite difficult. In this study, a multiple allele-specific PCR system was established based on two ‘Huangguo’ variety-specific SNPs that were respectively exploited from chloroplast ccsA and rpoC2 genes. The multiple allele-specific PCR method produced DNA amplicons of longer than 200 bp from seriously degraded samples and could detect 1 % of intentional adulteration of ‘Hongguo’ variety down to 0.01 ng level of genomic DNA. Furthermore, the developed real time allele-specific PCR enabled high-throughput genotyping of ‘Huangguo’ from local ginseng populations. This study provided a uniform method for variety origin authenticity control through the P. ginseng industrial chain and therefore can be used to maintain the high quality of industrial P. ginseng products.

Genetic characteristics of &lt;i&gt;Panax vietnamensis &lt;/i&gt;Ha &amp; Grushv. populations based on ssr

The results of genetic analysis of two Panax vietnamensis populations, collected in Tac-ngo ginseng garden, Tra Linh commune, Quang Nam Province and Mang ri ginseng garden, Kon Tum Province, showed that the population of ngoc linh ginseng in Quang Nam has higher alen frequency compared to the Ngoc Linh ginseng population in Kon Tum (AR = 2.27 &gt; AR = 2.05). The population of ngoc linh ginseng in Quang Nam has the expected heterozygous frequency (He) higher than the expected heterozygous frequency of the ginseng population in Kon Tum (0.42 &gt; 0.4). The observed heterozygous frequency (Ho) of the ngoc linh population in Quang Nam was also higher than the observed heterozygous frequency compared to the ngoc linh ginseng population in Kon Tum (0.49 &gt; 0.43). These results demonstrated that both populations of ngoc linh ginseng in Quang Nam and Kon Tum are maintaining high heterozygous frequencies; self-pollination has not affected to the genetic structure of these both populations yet. However, the population of ngoc linh ginseng in Quang Nam showed a higher level of diversity and heterozygous frequency than the ngoc linh ginseng population in Kon Tum.

Partial-root Harvest of American Ginseng (Panax quinquefolius L.): A Non-Destructive Method for Harvesting Root Tissue for Ginsenoside Analysis

American ginseng (Panax quinquefolius) roots have long been harvested for use in herbal medicine. Overharvesting has threatened long-term viability of wild American ginseng populations. Research has been ongoing to determine factors affecting the variation of ginsenosides in roots. Given the conservation concerns regarding wild American ginseng, we began experimenting with a partial-root harvest method in 2014 for extracting tissue for ginsenoside analysis without killing individual plants or causing long-term declines in wild populations. We took partial-root harvest samples from 57 plants in four wild populations throughout western North Carolina and monitored morphological attributes of these and 56 paired, unharvested plants of similar size for four years after harvest. Partial-root samples were taken from an additional 162 plants from 16 new populations in 2015 and 2016. Morphological attributes of these plants were monitored annually or biannually. In the paired plant study, annual reemergence did not differ between harvested and unharvested plants in any year after harvest. Leaf area was significantly lower in harvested plants than unharvested controls in the first year postharvest, but these differences did not persist after the first year. In the unpaired study, preharvest-postharvest comparisons were more variable, likely due to different harvest years and interannual variation in weather. Our results demonstrate that partial-root harvest could be an effective way for ginsenoside researchers to reduce their impact on wild and cultivated American ginseng populations and it may represent a non-destructive harvest protocol for root tissue phytochemical analysis.

Simultaneous Determination and Analysis of Major Ginsenosides in Wild American Ginseng Grown in Tennessee.

Ginsenosides are the major constituent that is responsible for the health effects of American ginseng. The ginsenoside profile of wild American ginseng is ultimately the result of germplasm, climate, geography, vegetation species, water, and soil conditions. This is the first report to address the ginsenoside profile of wild American ginseng grown in Tennessee (TN), the third leading state for production of wild American ginseng. In the present study, ten major ginsenosides in wild American ginseng roots grown in TN, including Rb1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg1, Rg2, and Rg3, were determined simultaneously. The chemotypic differences among TN wild ginseng, cultivated American ginseng, and Asian ginseng were assessed based on the widely used markers of ginsenoside profiling, including the top three ginsenosides, ratios of PPD/PPT, Rg1/Rb1, Rg1/Re, and Rb2/Rc. Our findings showed marked variation in ginsenoside profile for TN wild ginseng populations. Nevertheless, TN wild ginseng has significant higher ginsenoside content and more ginsenoside diversity than the cultivated ginseng. The total ginsenoside content in TN wild ginseng, as well as ginsenosides Rg1 and Re, increases with the age of the roots. Marked chemotypic differences between TN wild ginseng and cultivated American ginseng were observed based on the chemotypic markers. Surprisingly, we found that TN wild ginseng is close to Asian ginseng with regard to these characteristics in chemical composition. This study verified an accessible method to scientifically elucidate the difference in chemical constituents to distinguish wild from the cultivated American ginseng. This work is critical for the ecological and biological assessments of wild American ginseng so as to facilitate long-term sustainability of the wild population.

Genetic diversity among cultivated and wild Panax ginseng populations revealed by high-resolution microsatellite markers

BackgroundGinseng (Panax ginseng Meyer) is one of the world's most valuable medicinal plants with numerous pharmacological effects. Ginseng has been cultivated from wild mountain ginseng collections for a few hundred years. However, the genetic diversity of cultivated and wild ginseng populations is not fully understood. MethodsWe developed 92 polymorphic microsatellite markers based on whole-genome sequence data. We selected five markers that represent clear allele diversity for each of their corresponding loci to elucidate genetic diversity. These markers were applied to 147 individual plants, including cultivars, breeding lines, and wild populations in Korea and neighboring countries. ResultsMost of the 92 markers displayed multiple-band patterns, resulting from genome duplication, which causes confusion in interpretation of their target locus. The five high-resolution markers revealed 3 to 8 alleles from each single locus. The proportion of heterozygosity (He) ranged from 0.027 to 0.190, with an average of 0.132, which is notably lower than that of previous studies. Polymorphism information content of the markers ranged from 0.199 to 0.701, with an average of 0.454. There was no statistically significant difference in genetic diversity between cultivated and wild ginseng groups, and they showed intermingled positioning in the phylogenetic relationship. ConclusionGinseng has a relatively high level of genetic diversity, and cultivated and wild groups have similar levels of genetic diversity. Collectively, our data demonstrate that current breeding populations have abundant genetic diversity for breeding of elite ginseng cultivars.

The Case of the 30-Year Persistence of the Single Known Population of Panax quinquefolius L. (Araliaceae) in Louisiana

Abstract There were a total of 12 documented Panax quinquefolius (American Ginseng) in the only known population in Louisiana in 1986. Our reassessment of the population in 2016 found 16 plants. We suggest that the long-term persistence of this tiny American Ginseng population indicates that simple matrix-population models may be overestimating quasi-extinction numbers and minimum viable populations, perhaps as a result of failing to sufficiently consider influences of environmental conditions. We further suggest that this, the southernmost documented population of the species in the US, has endured beyond expectation but remains at acute risk of extirpation and that any population-specific genes are at risk of loss if no intervention is made to protect the population.

Genotypic and Organ Variation in Ginsenoside Contents from American Ginseng Populations

Variation in ginsenoside content was investigated as a function of population/genotype, plant organ, and age using four geographically isolated wild populations and one landrace population of american ginseng (Panax quinquefolius L.). The contents of individual and total ginsenosides were affected by the main and two-way interactions between population, organ, and age. Ginsenoside Re was not detected in roots of the wild population plants but was found in leaves and in both organs of the landrace population. A positive relationship between root age and total root ginsenosides was detected in two wild populations. Individual root ginsenosides were highly correlated with certain leaf ginsenosides in wild populations rather than in landrace populations. Therefore, the results suggest that certain leaf ginsenosides would be applied for potential biomarkers to estimate individual root ginsenosides. Principal component analysis (PCA) scores plot indicates that all wild populations were segregated from the single landrace population. However, cluster analysis indicates that differences existed between organs, and between the wild and landrace populations. Overall, the result suggests that the variation of individual and total ginsenoside contents would be influenced by a combination of population, plant organ, and root age.

Genetic structure of the American ginseng (Panax quinquefoliusL.) in Eastern Canada using reduced-representation high-throughput sequencing

The American ginseng (Panax quinquefolius L.) has been used for a wide range of medicinal purposes for more than 300 years, and is at risk in most of its range because of harvesting in natural populations, herbivory, and habitat loss. Its genetic structure is largely unknown in the previously glaciated areas of Eastern Canada, although such information could provide useful information for restoration strategies. We generated and analysed data from a reduced-representation high-throughput sequencing approach with a BAMOVA population model to partition the genetic variation within and among six natural populations of American ginseng in Eastern Canada. We found that an important and significant fraction of the genetic variation was structured among populations ([Formula: see text] = 42%; FST= 34%) at the geographical scale of the study (&lt;250 km). No clear evidence of isolation-by-distance was observed. This important genetic structure observed among American ginseng populations from a region that was covered by ice during the last glaciations is similar to what had been found in previous studies on southern populations or throughout the species range.

A simple real-time polymerase chain reaction (PCR)-based assay for authentication of the Chinese Panax ginseng cultivar Damaya from a local ginseng population.

Panax ginseng is one of the most important medicinal plants in the Orient. Owing to its increasing demand in the world market, cultivated ginseng has become the main source of medicinal material. Among the Chinese ginseng cultivars, Damaya commands higher prices and is grown in significant proportions among the local ginseng population. Due to the lack of rapid and accurate authentication methods, Damaya is distributed among different cultivars in the local ginseng population in China. Here, we identified a unique, Damaya-specific single nucleotide polymorphism (SNP) site present in the second intron of mitochondrial cytochrome c oxidase subunit 2 (cox2). Based on this SNP, a Damaya cultivar-specific primer was designed and an allele-specific polymerase chain reaction (PCR) was optimized for the effective molecular authentication of Damaya. We designed a method by combining a simple DNA isolation method with real-time allele-specific PCR using SYBR Green I fluorescent dye, and proved its efficacy in clearly discriminated Damaya cultivar from other Chinese ginseng cultivars according to the allelic discrimination analysis. Hence, this study provides a simple and rapid assay for the differentiation and conservation of Damaya from the local Chinese ginseng population.

Genetic diversity and population structure of ginseng in China based on RAPD analysis

AbstractPopulation genetic diversity was estimated from forty-four individual ginseng (Panax ginsengC.A. Meyer) plants collected from seven geographical populations located in Heilongjiang, Liaoning, and Jilin Provinces of China as well as the People’s Republic of Korea by using randomly amplified polymorphic DNA (RAPD) markers. Overall, 41 polymorphic loci were amplified using ten primer pairs. The polymorphism percentage ranged from 50% to 100% among seven local populations of ginseng, indicating that there is plentiful genetic diversity in wild ginseng populations. The genetic diversity at the species level was higher than that at the population level. Variance analysis showed that there was a significant difference among populations in genetic diversity. The genetic differentiation coefficient (i.e., FST) indicates that 43% of the variation occurred among populations, which indicates that substantial genetic differentiation occurred among populations. At the same time, the measured value of gene flow (Nm) was 0.66 based on the observed genetic differentiation coefficient among populations, suggesting there was moderate gene flow among populations.

Analysis of wood thrush (Hylocichla mustelina) movement patterns to explain the spatial structure of American ginseng (Panax quinquefolius) populations

AbstractAmerican ginseng (Panax quinquefolius) is America's premier wild‐harvested, medicinal plant that inhabits the forest understory of eastern deciduous forests. Recent research revealed that birds, particularly wood thrushes (Hylocichla mustelina), disperse ginseng seeds by regurgitating viable seeds 15–37 min after consuming the berries. We carried out two studies to examine the potential effect of thrushes on spatial dispersion patterns of ginseng. First, to analyze how far wood thrushes could disperse seeds, two wood thrushes were outfitted with radio transmitters and tracked for multiple days. Second, for 28 natural populations of ginseng, we created a clustering index to quantify to what degree populations were structured into spatially separated units. To further detect spatial impacts of thrushes, we analyzed inter‐cluster distances and the overall spread of ginseng populations. Over the time period in which wood thrushes retain ginseng in their guts, the seeds would be dispersed a mean distance of 15.2–21.7 m. The observed distances ranged from 0–96.6 m. These distances were comparable to the overall spread metric for ginseng populations with wood thrushes, which had increased spacing in comparison to those without wood thrushes. The mean dispersion index differed for populations with and without wood thrushes. It is imperative to understand the interaction between wood thrushes and ginseng to facilitate conservation, as both species are experiencing population declines. Dispersal of seeds by wood thrushes could play an important role in allowing species such as ginseng to respond to climate change, deer browse, overharvesting, and other environmental stresses.

Microsatellite Analysis of &amp;lt;i&amp;gt;Panax ginseng&amp;lt;/i&amp;gt; Natural Populations in Russia

Panax ginseng C.A. Meyer is an endangered species in Russia. To restore this species, effective protective measures, including the reintroduction into favorable habitats, must be worked out considering the specificity of genetic structure of ginseng populations. One hundred and thirty-nine P. ginseng plants were collected from the forests of nine administrative areas of Primorsky Territory of Russia and transferred to a collection nursery for further investigation. Microsatellite markers were used to study the genetic diversity and the genetic structure of ginseng populations. For populations studied with SSR, the number of observed alleles was ranging from 15 to 25, allelic richness from 1.83 to 3.04, polymorphic loci from 62.5% to 87.7%, observed heterozygosity from 0.410 to 0.512 (an average of 0.453) and expected heterozygosity from 0.304 to 0.479, with an average of 0.393. The values of the inbreeding coefficient within populations (Fis) ranged from -0.447 to 0.056, and their average value was -0.296. Genetic differentiation among populations was significant (Fst = 0.115) but an isolation-by-distance pattern was not detected. UPGMA and MS-tree confirmed the presence of genetic structure within P. ginseng and visualized genetic relationships of populations with similar pattern. STRUCTURE analysis revealed the genetic admixture between different ginseng populations. It was established with SSR markers that P. ginseng still preserves substantial genetic resources although all populations are largely exhausted. Because ginseng populations are significantly differentiated all of them should be restored. Considering the admixture of ginseng populations it would be advisable to apply the individual assignment test to verify the content of indigenous populations and to identify the "true" population plants to serve as stock material for reintroduction.

American ginseng (Panax quinquefolius L.) floristic associations in Pennsylvania: guidance for identifying calcium-rich forest farming sites

In the eastern United States, there is interest among forest landowners in American ginseng husbandry, and particularly in cultivating this plant on forestlands using a wild-simulated forest farming approach. This study documented the flora and soil conditions associated with wild and wild-simulated ginseng populations throughout Pennsylvania (PA) to develop floristic “indicators” that can be used to identify supportive growing sites on forestlands. A total of 243 plant species were documented associates of ginseng across PA: 32 over-story trees, 37 shrubs and understory trees, 15 vines, 143 herbs, and 16 ferns. Statistical analysis revealed a largely shared floristic assemblage throughout the state although some associates did differ according to region and physiographic province. Previous studies have suggested that a soil calcium content, especially soils having at least 3,360 kg ha−1 (3,000 lbs ac), appear to be particularly conducive to wild and wild-simulated ginseng occurrence and/or vigor, and indicator species analysis in this study revealed that three of the top plant associates that can be used for determining sites that meet this calcium threshold in PA are white ash, Jack-in-the-pulpit, and rattlesnake fern. These results suggest that successful adoption of wild-simulated ginseng forest farming is likely to be improved in forested areas where these species are found collectively as a dominant component of local plant assemblages.

Sunfleck effects on physiology, growth, and local demography of American ginseng (Panax quinquefolius L.)

The light environment of the forest understory is heterogeneous, characterized by low levels of diffuse light punctuated by short periods of direct solar radiation known as sunflecks. Less well understood is the degree to which sunflecks are responsible for spatially varying performance of understory plants. We studied natural variation in the light environment, as well as physiology, growth, and microsite demographics of American ginseng (Panax quinquefolius L.) to test whether clustering of plants within a population was related to sunfleck variation. HOBO dataloggers were positioned in 10 clusters within a single ginseng population to measure the local understory light environment during mid-season (June 27 to August 9 2011). Photosynthesis and transpiration were measured with a Li-Cor 6400 portable photosynthesis system. Population census data were used to obtain measurements of the relative growth rate (2011–2012), seed production efficiency (2011) and seed germination (2009–2012) of plants in the same subpopulations. All measured dependent variables were regressed against the percentage of mean photosynthetic photon flux density contributed from sunflecks and mean length of the longest sunfleck. Light saturated photosynthetic rates of ginseng showed a positive linear relationship to both sunfleck characteristics, while transpiration was unaffected. The relative growth rate of established ginseng plants also had a positive linear relationship with the length of the longest sunfleck. However, germination rate was negatively affected by an increasing percentage of photosynthetic photon flux density from sunflecks. Seed production efficiency was not influenced by either sunfleck characteristic. Due to negative effects on germination, initial ginseng cluster formation is likely not associated with sunflecks, however photosynthetic and growth responses suggest that fitness effects are positive for established individuals. A spatially and temporally varying light environment is beneficial for ginseng population growth.