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. 2025 Jun 18;20(6):e0326237. doi: 10.1371/journal.pone.0326237

Post-harvest treatment of wild-simulated ginseng under climate-smart environmental conditions

Solhee Kim 1, Taegon Kim 1,2, Jungyeon Kim 3,4,*, Kyo Suh 4,5,*
Editor: Chun-Hua Wang6
PMCID: PMC12176193  PMID: 40532029

Abstract

Wild-simulated ginseng (WSG; Panax ginseng C.A. Meyer) is prized for its unique ginsenoside profile and medicinal properties; however, traditional cultivation faces challenges, such as declining survival rates and prolonged growth periods. We aimed to develop an enhanced post-harvest treatment using climate-smart conditions to improve survival rates, boost root mass, and preserve the characteristic ginsenoside profile of WSG. Three-year-old WSG roots were transplanted and cultivated in a controlled smart facility in regulated light and irrigation conditions for five months using premium ginseng soil, a certified organic ginseng soil medium. Growth performance was monitored and ginsenoside profiles were analysed via ultra-performance liquid chromatography. Post-treatment, WSG exhibited a 2.5-fold increase in root weight and an overall survival rate of 97%. Total ginsenoside content reached 10.458 mg/g dried ginseng, with notably high levels of Re (7.716 mg/g) and the presence of rare compounds, such as Compound K and Rg3. The root-to-shoot ratio exceeded 1.23, indicating efficient resource allocation. These results demonstrate that climate-smart post-harvest treatment effectively enhances root development and maintains the medicinal quality of WSG, offering a promising strategy to overcome the limitations of conventional cultivation and improve its commercial viability.

Introduction

Wild-simulated ginseng (WSG; Panax ginseng C.A. Meyer) is a perennial herb belonging to the family Araliaceae, naturally growing in East Asian mountain regions [1]. WSG cultivation requires natural mountainous areas without artificial facilities, such as shade structures, using direct seeding or transplanting of nursery plants without pesticides or chemical fertilizers [2]. Although genetically identical to conventional ginseng—which requires a six-year growth period and increased agricultural chemical inputs after the third year—WSG grows naturally for 7 to over 20 years [3]. This fundamental difference in cultivation methods, especially in growth duration and chemical use, distinguishes WSG from field-cultivated ginseng (FCG).

WSG exhibits unique ginsenoside profiles and age-dependent concentration increases, along with distinctive compounds not detected in FCG [4,5]. Several studies have demonstrated that WSG synthesizes a wider array of ginsenosides than that by FCG, primarily due to environmental stresses in its natural habitat [2]. These stresses activate secondary metabolism, producing a range of bioactive compounds with health benefits [6]. This broader spectrum of ginsenosides enhances its therapeutic properties, including anti-cancer [7–9], anti-inflammatory [10,11], and immunomodulatory effects [12].

Recent societal changes, including aging population in Korea and increased health consciousness following the COVID-19 pandemic, have driven growing demand for WSG. By 2023, WSG was cultivated on 13,858 hectares by 3,792 forestry households [13]. Annual production of 254,111 kg yields a market value of 62.9 billion won, with prices up to 250,000 won per kg. These figures position WSG as one of the most valuable forest products of Korea, reflecting its premium market status.

Although WSG can be certified for sale after three years of growth, its intermediate root size at this stage affects market acceptance, with trade favouring roots aged at least seven years [14]. However, Korea Forest Service statistics show that WSG survival rates drastically decline with cultivation duration (Korea Forest Service, 2024). The average survival rate is 77.8% for one-year-old roots, decreasing to 55.2% at three years and 38.8% at five years. For seven-year-old roots, which are in high demand, the rate declines to about 25.3%. This trend reflects an annual loss rate of 15–20%, with the largest decrease (16.5%) occurring from four to five years.

Here, we aimed to develop an enhanced post-harvest treatment using climate-smart conditions to improve survival rates, boost root mass, and preserve the characteristic ginsenoside profile of WSG. We employed a controlled system that manages temperature, humidity, and light for WSG harvested during dormancy (3–4 years), applying a short-term (up to six months) post-harvest treatment in a controlled soil environment.

Materials and methods

Cultivation in smart facility for post-treated WSG

We obtained certified three-year-old WSG plants from the Pyeongchang Wild Ginseng Association (Gangwon-do, Republic of Korea). We transplanted the plants on April 20, 2022 and cultivated them for five months in the smart facility with environmental controls for light, irrigation, and temperature regulation. Environmental conditions were based on known physiological thresholds for WSG; the temperature was maintained below 21°C and light intensity under 200 PPFD (≈5,000 lux) to prevent photoinhibition and stress-induced growth inhibition [15,16]. Humidity was not actively regulated but remained stable under shaded indoor conditions. We used premium ginseng soil (Shinsung Mineral Co., Ltd., Cheongan-myeon, Republic of Korea)—the first certified organic ginseng cultivation medium in Korea (Certification No. 1-2-109)—as the post-harvest treatment medium. Further details on cultivation techniques, environmental management, and container design are provided in S1 Appendix.

Ginsenoside analysis

We analysed nine major ginsenosides (Rb1, Rg1, Rg3, Rd, Re, Rf, Mc1, F2, and Compound K) using the Waters ACQUITY® ultra-performance liquid chromatography (UPLC) system (Waters Corp., Milford, MA, USA). We operated the instrument at a gas temperature of 300 °C and a flow rate of 5 L/min. The nebulizer pressure was maintained at 45 psi, while the sheath gas temperature and flow rate were set at 250 °C and 11 L/min, respectively. Both positive and negative modes were employed, with a capillary voltage of 3,500 V and a nozzle voltage of 500 V. To ensure consistency with previous studies and eliminate moisture-related variations, we used dried post-treated WSG samples for all analyses, allowing direct comparison with existing data on both WSG and FCG.

Impact of post-treatment protocol on WSG growth and survival characteristics

To evaluate post-treated WSG growth patterns, we monitored root weight changes over five months. Specimens were initially subjected to visual screening for preliminary separation, then weighed to assign them into two groups based on a 1.0 g fresh weight threshold: Group A (> 1.0 g; mean: 1.58 ± 0.21 g) and Group B (≤ 1.0 g; mean: 0.74 ± 0.18 g). This grouping criterion was informed by a bimodal weight distribution observed across 220 specimens (Fig 1A). The final harvest at five months revealed substantial growth in both groups: Group A achieved an average root weight of 3.85 g (a 2.44-fold increase), whereas Group B reached an average of 1.53 g (a 2.07-fold increase) (Fig 1B).

Fig 1. Post-harvest treatment of wild-simulated ginseng (WSG).

Fig 1

(A) Smart facility for post-treated WSG at Pyeongchang campus in Seoul National University. (B) Changes in average root weight of post-treated WSG over the treatment period. Group A (initial n = 220) and Group B (initial n = 220) represent larger and smaller roots, respectively. Samples (n = 40) were excluded at 2 months to measure the ratio of underground to shoot fresh weight. The survival rate was 96.8% for Group A and 97.3% for Group B. *** represents p value < 0.0001 (t test; FDR adjusted).

To evaluate the effect of post-treatment on the ginsenoside profiles of WSG, UPLC analysis was performed. The total ginsenoside content of post-treated WSG was 10.458 mg/g of dried wild-simulated ginseng (DG) weight and considering an average root weight of 4.0 g, each root contained approximately 41.83 mg of total ginsenosides (Table 1). Of the analysed ginsenosides, Re was the most abundant (7.716 mg/g DG), followed by Rb1 (1.293 mg/g DG) and Rg1 (1.195 mg/g DG). Ginsenoside compounds K and Rg3, which are known to be present in WSG, however, absent in FCG, were detected in post-treated WSG [5].

Table 1. Analysis of ginsenoside content using UPLC in post-treated WSG.

Ginsenoside Post-treated WSG
(Unit: mg/g, DG)
PPD Rb1 1.293 ± 0.024
Rg3 0.017 ± 0.001
Rd 0.218 ± 0.017
Mc1 0.002 ± 0.000
F2 0.011 ± 0.001
CK 0.006 ± 0.001
PPT Rg1 1.195 ± 0.121
Re 7.716 ± 0.257
Rf 0.001 ± 0.000
Total 10.458 ± 0.421

Data represent mean values ± standard error (mg/g). DG: weight of dried wild-simulated ginseng.

The post-treatment protocol successfully addresses a key challenge in WSG cultivation—maintaining ginsenoside profiles while achieving greater root mass and survival rate. Excluding the 40 specimens harvested at two months from Group A, only seven specimens in Group A and six in Group B were lost during the treatment period, resulting in an estimated survival rate of 97%. This high survival rate, combined with the substantial weight gains observed in both groups, suggests that the post-treatment protocol effectively supported root development while maintaining plant viability. These results indicate that post-treatment conditions promoted proportional growth regardless of initial root size, as both groups exhibited similar relative weight increases over the five months. The enhanced root development observed in our study (a 2.5-fold increase over five months) considerably exceeds the growth rates reported in traditional WSG cultivation, where annual root weight increases typically range from 30 to 50% [17]. The post-treated WSG also contained notable amounts of ginsenosides, such as CK and Rg3 [5], which are significantly more abundant in WSG than in FCG (Table 1). This suggests that post-treatment enhanced size and maintained high survival while preserving the distinctive characteristics of WSG.

Pre-treatment ginsenoside profiles were not analyzed in this study due to the study design, which prioritized monitoring of post-treatment growth and biochemical outcomes. As a result, absolute changes in ginsenoside content before and after treatment could not be quantified. Nevertheless, the detection of rare ginsenosides such as Compound K and Rg3—compounds typically absent in three-year-old FCG—may indicate a potential induction of secondary metabolite biosynthesis under climate-smart post-harvest conditions. This interpretation remains speculative in the absence of baseline values, and future studies should include paired pre- and post-treatment ginsenoside analyses to validate the observed biochemical trends.

In addition, the long-term reusability of the certified organic soil used in this study was not evaluated. Future investigations should assess its structural integrity, nutrient retention, and microbial stability across multiple planting cycles to determine its applicability for repeated use in commercial WSG production systems.

Conclusions

This study demonstrates that post-harvest treatment of WSG under controlled environmental conditions enhances root development while preserving its distinctive medicinal properties through systematic management of environmental factors. Key findings include a 2.5-fold weight increase over five months, survival rates exceeding 96%, and retention of the characteristic ginsenoside profiles.

The post-treatment protocol addresses several crucial challenges in traditional WSG cultivation. Offering controlled post-harvest conditions that preserve the unique properties of WSG may resolve the issues of the industry with unstable production and variable quality. The preservation of rare ginsenosides, such as Compound K and Rg3, confirms that this approach maintains the medicinal value characteristic of WSG while improving production efficiency. The post-treated WSG evaluated in this study occupies a distinct position between conventional field-cultivated and wild-simulated ginseng. It shows improved root growth and preservation of pharmacologically relevant ginsenosides under a shorter cultivation window. While the production cost is expected to be higher than that of FCG, it remains significantly lower than that of long-term WSG cultivation. This balance of cost and efficacy may offer a practical and scalable alternative for premium ginseng markets.

Achieving significant growth within five months, high survival rates, and desirable pharmacological properties suggests potential economic benefits. This method could improve the commercial value of previously unmarketable WSG while ensuring consistent quality standards.

Future research should investigate treatment conditions for different genetic stocks and seasonal effects. This research represents a step toward overcoming current limitations in WSG cultivation while preserving its valuable medicinal properties.

Supporting information

S1 Appendix. Cultivation techniques, environmental management, and container design.

(DOCX)

pone.0326237.s001.docx (32.1KB, docx)
S1 Fig. Changes in chlorophyll fluorescence (Fv/Fm) of WSG leaves under different light and irrigation conditions over a 15-day period.

Light/H2O: natural light with water, Light/HOCl: natural light with HOCl, Dark/H2O: shaded with water, Dark/HOCl: shaded with HOCl. Measurements were taken at 1, 5, 8, 12, and 15 days after treatment. Vertical bars indicate standard errors.

(DOCX)

pone.0326237.s002.docx (60.3KB, docx)
S2 Fig. Comparison of shoot development under different light and irrigation conditions.

(A) Three petioles from plants grown under light and dark conditions. (B) Representative leaf morphology of middle leaflets under different light and irrigation treatments (Light/H2O, Light/HOCl, Dark/H2O, Dark/HOCl). Scale bar = 5 cm.

(DOCX)

pone.0326237.s003.docx (270.6KB, docx)
S3 Fig. The average weight of WSG roots under different light and irrigation conditions after two months of treatment.

The initial weight of transplanted roots was 1.70 ± 0.40 g (mean ± SD, n = 39). Bars represent mean values ± standard errors (n.s.: not significant, ***: p < 0.001, ****: p < 0.0001).

(DOCX)

pone.0326237.s004.docx (42.7KB, docx)

Acknowledgments

We sincerely appreciate the invaluable assistance of the Green Bio Research Facility Center at the Institutes of Green Bio Science & Technology with the ginsenoside analysis.

Abbrevations

WSG

wild-simulated ginseng

FCG

field-cultivated ginseng

DG

dried ginseng

PPT

protopanaxatriol

CK

Compound K.

Data Availability

All relevant data are within the manuscript and its Supporting information files.

Funding Statement

The author(s) received no specific funding for this work.

References

  • 1.Nah SY. Ginseng; recent advances and trends. J Ginseng Res. 1997;21:1–12. [Google Scholar]
  • 2.Kim Y-J, Jeon J-N, Jang M-G, Oh JY, Kwon W-S, Jung S-K, et al. Ginsenoside profiles and related gene expression during foliation in Panax ginseng Meyer. J Ginseng Res. 2014;38(1):66–72. doi: 10.1016/j.jgr.2013.11.001 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Kim K, Um Y, Eo H-J, Park HW, Jeon KS, Kim H-J. Study on the correlation between the ginsenoside contents and growth characteristics of wild-simulated ginseng with different year-roots (Panax ginseng C.A. Meyer). Korean J Plant Res. 2020;33:255–62. doi: 10.7732/kjpr.2020.33.4.255 [DOI] [Google Scholar]
  • 4.Kim K, Huh JH, Um Y, Jeon KS, Kim HJ. The comparative of growth characteristics and ginsenoside contents in wild-simulated ginseng (Panax ginseng C.A. Meyer) on different years by soil properties of cultivation regions. Korean J Plant Res. 2020;33:651–8. doi: 10.7732/kjpr.2020.33.6.651 [DOI] [Google Scholar]
  • 5.Ma R, Yang P, Jing C, Fu B, Teng X, Zhao D, et al. Comparison of the metabolomic and proteomic profiles associated with triterpene and phytosterol accumulation between wild and cultivated ginseng. Plant Physiol Biochem. 2023;195:288–99. doi: 10.1016/j.plaphy.2023.01.020 [DOI] [PubMed] [Google Scholar]
  • 6.Hyun SH, Ahn H-Y, Kim H-J, Kim SW, So S-H, In G, et al. Immuno-enhancement effects of Korean Red Ginseng in healthy adults: a randomized, double-blind, placebo-controlled trial. J Ginseng Res. 2021;45(1):191–8. doi: 10.1016/j.jgr.2020.08.003 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Choi K. Botanical characteristics, pharmacological effects and medicinal components of Korean Panax ginseng C A Meyer. Acta Pharmacol Sin. 2008;29(9):1109–18. doi: 10.1111/j.1745-7254.2008.00869.x [DOI] [PubMed] [Google Scholar]
  • 8.Helms S. Cancer prevention and therapeutics: Panax ginseng. Altern Med Rev. 2004;9(3):259–74. [PubMed] [Google Scholar]
  • 9.Hong H, Baatar D, Hwang SG. Anticancer activities of ginsenosides, the main active components of Ginseng. Evid Based Complement Alternat Med. 2021;2021:8858006. doi: 10.1155/2021/8858006 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Lin C-H, Lin Y-A, Chen S-L, Hsu M-C, Hsu C-C. American Ginseng attenuates eccentric exercise-induced muscle damage via the modulation of lipid peroxidation and inflammatory adaptation in males. Nutrients. 2021;14(1):78. doi: 10.3390/nu14010078 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Park HJ, Jo S-M, Seo SH, Lee M, Lee Y, Kang I. Anti-inflammatory potential of cultured ginseng roots extract in lipopolysaccharide-stimulated mouse macrophages and adipocytes. Int J Environ Res Public Health. 2020;17(13):4716. doi: 10.3390/ijerph17134716 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Saba E, Lee YY, Kim M, Kim S-H, Hong S-B, Rhee MH. A comparative study on immune-stimulatory and antioxidant activities of various types of ginseng extracts in murine and rodent models. J Ginseng Res. 2018;42(4):577–84. doi: 10.1016/j.jgr.2018.07.004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Korea Forest Service. 2023 Wild-Simulated Ginseng Statistical Report. 2024. [Google Scholar]
  • 14.Sheban KC, Woodbury DJ, Duguid MC. Importance of environmental factors on plantings of wild-simulated American Ginseng. Agroforest Syst. 2021;96(1):147–60. doi: 10.1007/s10457-021-00705-8 [DOI] [Google Scholar]
  • 15.Aro EM, Virgin I, Andersson B. Photoinhibition of Photosystem II. Inactivation, protein damage and turnover. Biochim Biophys Acta. 1993;1143(2):113–34. doi: 10.1016/0005-2728(93)90134-2 [DOI] [PubMed] [Google Scholar]
  • 16.Song J, Yang J, Jeong BR. Growth and photosynthetic responses to increased LED light intensity in Korean Ginseng (Panax ginseng C.A. Meyer) Sprouts. Agronomy. 2023;13(9):2375. doi: 10.3390/agronomy13092375 [DOI] [Google Scholar]
  • 17.Jeong B-G, Jung G-R, Kim MS, Moon H-G, Park S-J, Chun J. Ginsenoside contents and antioxidant activities of cultivated mountain ginseng (Panax ginseng C.A. Meyer) with different ages. Korean J Food Preserv. 2019;26(1):90–100. doi: 10.11002/kjfp.2019.26.1.90 [DOI] [Google Scholar]

Decision Letter 0

Chun-Hua Wang

Dear Dr. Kim,

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Reviewers' comments:

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Reviewer #2: Yes

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Reviewer #1: This manuscript presents a valuable study that proposes a post-cultivation treatment strategy for wild-simulated ginseng (WSG) under climate-smart conditions. The approach aims to enhance the added value of WSG by improving survival rates, increasing root biomass, and maintaining a characteristic ginsenoside profile. While the study is significant, the following revisions are recommended to enhance clarity and scientific rigor:

1. Clarification of Grouping Criteria: The basis for dividing WSG into two groups requires further explanation. The use of subjective descriptors such as "large" and "small" is ambiguous and should be replaced with clearly defined, quantifiable parameters (e.g., weight range, root diameter).

2. Terminology Concerning Plant Age: Referring to three-year-old WSG as "seedlings" appears to be inappropriate. It is suggested that the authors use terminology that accurately reflects the developmental stage of the plants based on established ginseng cultivation standards.

3. Inclusion of Pre-treatment Ginsenoside Data: Table 1 presents only the ginsenoside content following post-treatment. To allow for meaningful interpretation of the treatment’s effect, it is essential to also include the ginsenoside profile prior to treatment.

4. Accuracy of Initial Root Data in Figure S4: The description of the initial WSG root weight as "more than 1 g" in Figure S4 lacks precision. Providing exact weight measurements or a statistical summary (e.g., mean ± SD) would improve the credibility and reproducibility of the data.

Reviewer #2: The manuscript cultivates an artificial intelligence climate management system for WSG, which improves survival rate and shortens growth period while ensuring effective ingredient content. I hope the following information can be added to the article:

How are the optimal temperatures and humidity for WSG growth determined?

How many times can organic nutrient soil be planted?

Is there any difference in morphology between artificially cultivated and wild WSGs? How much is the cost difference?

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PLoS One. 2025 Jun 18;20(6):e0326237. doi: 10.1371/journal.pone.0326237.r003

Author response to Decision Letter 1


26 May 2025

Responses to Reviewers

Many thanks to the editors and reviewers for their constructive comments that will help improve our manuscript. Each comment was revised and the contents were highlighted with red colour. The responses to individual comments are in Italics below:

Reviewer #1:

This manuscript presents a valuable study that proposes a post-cultivation treatment strategy for wild-simulated ginseng (WSG) under climate-smart conditions. The approach aims to enhance the added value of WSG by improving survival rates, increasing root biomass, and maintaining a characteristic ginsenoside profile. While the study is significant, the following revisions are recommended to enhance clarity and scientific rigor:

1. Clarification of Grouping Criteria: The basis for dividing WSG into two groups requires further explanation. The use of subjective descriptors such as "large" and "small" is ambiguous and should be replaced with clearly defined, quantifiable parameters (e.g., weight range, root diameter).

� We thank the reviewer for pointing out the need to clarify the grouping criteria for WSG specimens. In response, we have revised the manuscript to remove subjective descriptors such as “large” and “small,” and instead provide an objective, quantifiable grouping based on root weight.

� Specifically, specimens were initially screened visually to facilitate preliminary sorting, then weighed and divided using a 1.0 g fresh weight threshold. This value was chosen based on the bimodal weight distribution observed in the 220 initial samples, which revealed a natural separation near 1.0 g. Group A included specimens >1.0 g (mean: 1.58 ± 0.21 g), and Group B included those ≤1.0 g (mean: 0.74 ± 0.18 g). These changes have been incorporated into the revised manuscript as follows: Revised text in Page 4, Lines 89-92:

“Specimens were initially subjected to visual screening for preliminary separation, then weighed to assign them into two groups based on a 1.0 g fresh weight threshold: Group A (> 1.0 g; mean: 1.58 ± 0.21 g) and Group B (≤ 1.0 g; mean: 0.74 ± 0.18 g). This grouping criterion was informed by a bimodal weight distribution observed across 220 specimens (Fig. 1A). …”

2. Terminology Concerning Plant Age: Referring to three-year-old WSG as "seedlings" appears to be inappropriate. It is suggested that the authors use terminology that accurately reflects the developmental stage of the plants based on established ginseng cultivation standards.

� Referring to three-year-old WSG as “seedlings” was inaccurate and has been corrected in the revised manuscript. Based on established cultivation standards, this developmental stage exceeds the seedling phase. We have therefore replaced the term with “three-year-old wild-simulated ginseng plants”, which more accurately describes the biological maturity of the material used in this study.

“WSG cultivation requires natural mountainous areas without artificial facilities, …, using direct seeding or transplanting of nursery plants without pesticides or chemical fertilizers [2]. ” : Revised text in Page 3, Lines 36.

“We obtained certified three-year-old WSG plants from the Pyeongchang Wild Ginseng Association (Gangwon-do, Republic of Korea). We transplanted the plants on April 20, …” : Revised text in Page 4, Lines 67-68.

3. Inclusion of Pre-treatment Ginsenoside Data: Table 1 presents only the ginsenoside content following post-treatment. To allow for meaningful interpretation of the treatment’s effect, it is essential to also include the ginsenoside profile prior to treatment.

� We appreciate the reviewer’s observation regarding the need for pre-treatment ginsenoside data to enable direct evaluation of the treatment’s biochemical effects. In this study, however, ginsenoside profiling was conducted only after treatment, and baseline ginsenoside levels were not measured. This decision reflected the study design, which prioritized post-treatment root growth tracking and chemical composition assessment rather than pre-post biochemical comparison.

� To address this limitation, we have added a clarifying statement in the Discussion section that explains the absence of pre-treatment data, and we acknowledge that this limits our ability to quantify biochemical changes induced by the treatment. However, we also note the detection of rare ginsenosides such as Compound K and Rg3—compounds typically absent in three-year-old field-cultivated ginseng (FCG)—which may suggest a positive effect of the treatment on secondary metabolite biosynthesis. Added text in Discussion (Page 6, Lines 118-125):

“Pre-treatment ginsenoside profiles were not analyzed in this study due to the study design, which prioritized monitoring of post-treatment growth and biochemical outcomes. As a result, absolute changes in ginsenoside content before and after treatment could not be quantified. Nevertheless, the detection of rare ginsenosides such as Compound K and Rg3—compounds typically absent in three-year-old field-cultivated ginseng (FCG)—may indicate a potential induction of secondary metabolite biosynthesis under climate-smart post-harvest conditions. This interpretation remains speculative in the absence of baseline values, and future studies should include paired pre- and post-treatment ginsenoside analyses to validate the observed biochemical trends.”

4. Accuracy of Initial Root Data in Figure S4: The description of the initial WSG root weight as "more than 1 g" in Figure S4 lacks precision. Providing exact weight measurements or a statistical summary (e.g., mean ± SD) would improve the credibility and reproducibility of the data.

� We have revised the caption of Figure S4 to include the actual initial root weight statistics used in the short-term treatment experiment. Specifically, the initial weight of transplanted roots was 1.70 ± 0.40 g (mean ± SD, n = 39), replacing the previously vague “more than 1 g” description and caption. Revised SI text in Page 10:

“The experiment involved three-year-old or older wild-simulated ginseng roots with a mean initial weight of 1.70 ± 0.40 g (mean ± SD, n = 39). Measurements were taken two months after transplantation.”

“Figure S4. The average weight of WSG roots under different light and irrigation conditions after two months of treatment. The initial weight of transplanted roots was 1.70 ± 0.40 g (mean ± SD, n = 39). Bars represent mean values ± standard errors (n.s.: not significant, ***: p < 0.001, ****: p < 0.0001).”

Reviewer #2:

The manuscript cultivates an artificial intelligence climate management system for WSG, which improves survival rate and shortens growth period while ensuring effective ingredient content. I hope the following information can be added to the article:

1. How are the optimal temperatures and humidity for WSG growth determined?

� We thank the reviewer for raising this important point regarding the basis for environmental control during post-harvest treatment. In this study, temperature and humidity conditions were not set based on a single defined "optimal value" derived from empirical testing, but were instead informed by known physiological characteristics of WSG and agronomic standards reported in the literature.

� Specifically, our Supplementary Information (Section S2.3) describes the rationale for environmental control:

- WSG is highly sensitive to high temperatures, with light saturation levels dropping sharply above 30°C.

- Therefore, we maintained air and soil temperatures below 21°C, and light intensity below 200 PPFD (approx. 5,000 lux), using natural light and polyethylene shade netting.

- Soil moisture was closely monitored and managed through a combination of bottom-watering and mist irrigation, adjusted seasonally to maintain appropriate substrate conditions.

- While humidity was not mechanically controlled, the shaded, naturally cool indoor conditions of the treatment facility helped maintain stable relative humidity in the range suitable for ginseng post-harvest handling.

� This protocol was guided by previously reported environmental thresholds (e.g., Kim et al., 2019) and standard WSG practices recommended by Korean forestry and medicinal plant research institutes. The goal was to provide a low-stress environment favorable for root biomass retention and survival during the dormancy transition, rather than to experimentally define an optimal climate envelope.

“… Environmental conditions were based on known physiological thresholds for WSG; the temperature was maintained below 21℃ and light intensity under 200 PPFD (≈5,000 lux) to prevent photoinhibition and stress-induced growth inhibition [1-3]. Humidity was not actively regulated but remained stable under shaded indoor conditions. … Further details on cultivation techniques, environmental management, and container design are provided in Supplementary Sections S1 and S2.” Added text in Materials and Methods (Page 4, Lines 70-77).

“These conditions were selected based on the known physiological sensitivities of Panax ginseng. Elevated temperatures and high light intensity are known to impair photosynthetic efficiency and reduce biomass accumulation due to photoinhibition and increased respiration [4]. Prior studies have shown that ginseng exhibits optimal performance when light intensity is kept below 200 PPFD, particularly under cooler conditions [5, 6]. While humidity was not mechanically controlled, the shaded and enclosed indoor environment provided moderate and consistent humidity levels comparable to those in natural understory habitats.” Revised SI text in Page 7.

2. How many times can organic nutrient soil be planted?

� We acknowledge the reviewer’s interest in the long-term reusability of the certified organic nutrient soil used in this study. However, we did not evaluate repeated planting cycles using the same soil in this experiment, as our focus was limited to the short-term post-harvest treatment and physiological response of WSG.

� We agree that the reusability of the soil medium—particularly its physicochemical stability and nutrient retention over multiple planting cycles—is a critical topic for practical implementation. We have added a statement in the Discussion section identifying this aspect as a key area for future investigation. Added text in Discussion (Page 6-7, Lines 126-129):

“In addition, the long-term reusability of the certified organic soil used in this study was not evaluated. Future investigations should assess its structural integrity, nutrient retention, and microbial stability across multiple planting cycles to determine its applicability for repeated use in commercial WSG production systems.”

3. Is there any difference in morphology between artificially cultivated and wild WSGs? How much is the cost difference?

� Thank you for raising this important question regarding the morphological and cost-related differences between artificially cultivated and wild-simulated ginseng (WSG).

� Regarding morphology, this study did not include a direct experimental comparison between wild-grown and artificially cultivated WSG. All samples were sourced from the same origin and treated under post-harvest conditions to assess survival rate, root mass, and ginsenoside profile. While morphological variations were not quantitatively evaluated, it is generally observed in field practice that wild WSG tends to exhibit more irregular and asymmetrical root structures, denser epidermal layers, and slower growth due to prolonged exposure to natural stressors. In contrast, roots cultivated under controlled environments are typically more uniform in shape, smoother in appearance, and larger in biomass. These tendencies were not formally assessed in this study and are mentioned here only as qualitative background.

� Regarding cost, no formal economic analysis was performed. However, based on current market data and cultivation practices, field-cultivated ginseng (FCG; 4–6 years) is typically priced at 30,000–70,000 KRW/kg, while wild-simulated ginseng aged seven years or more is valued at 200,000–250,000 KRW/kg. The post-treated WSG presented in this study is expected to fall between these two categories in terms of cost, requiring more resources than FCG but significantly less than wild cultivation. At the same time, it retains key pharmacological compounds—such as Re, Compound K, and Rg3—that are usually associated with higher-value ginseng products. To reflect this context, we have added the following statement to the Conclusion section (Page 6, Lines 139-147):

“… The post-treated WSG evaluated in this study occupies a distinct position between conventional field-cultivated and wild-simulated ginseng. It shows improved root growth and preservation of pharmacologically relevant ginsenosides under a shorter cultivation window. While the production cost is expected to be higher than that of FCG, it remains significantly lower than that of long-term WSG cultivation. This balance of cost and efficacy may offer a practical and scalable alternative for premium ginseng markets.

Achieving significant growth within five months, high survival rates, and desirable pharmacological properties suggests potential economic benefits. This method could improve the commercial value of previously unmarketable WSG while ensuring consistent quality standards.”

Attachment

Submitted filename: Responses to Reviewers.docx

pone.0326237.s006.docx (26.4KB, docx)

Decision Letter 1

Chun-Hua Wang

<p>Post-harvest Treatment of Wild-simulated Ginseng under Climate-smart Environmental Conditions

PONE-D-25-14014R1

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Acceptance letter

Chun-Hua Wang

PONE-D-25-14014R1

PLOS ONE

Dear Dr. Kim,

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Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 Appendix. Cultivation techniques, environmental management, and container design.

    (DOCX)

    pone.0326237.s001.docx (32.1KB, docx)
    S1 Fig. Changes in chlorophyll fluorescence (Fv/Fm) of WSG leaves under different light and irrigation conditions over a 15-day period.

    Light/H2O: natural light with water, Light/HOCl: natural light with HOCl, Dark/H2O: shaded with water, Dark/HOCl: shaded with HOCl. Measurements were taken at 1, 5, 8, 12, and 15 days after treatment. Vertical bars indicate standard errors.

    (DOCX)

    pone.0326237.s002.docx (60.3KB, docx)
    S2 Fig. Comparison of shoot development under different light and irrigation conditions.

    (A) Three petioles from plants grown under light and dark conditions. (B) Representative leaf morphology of middle leaflets under different light and irrigation treatments (Light/H2O, Light/HOCl, Dark/H2O, Dark/HOCl). Scale bar = 5 cm.

    (DOCX)

    pone.0326237.s003.docx (270.6KB, docx)
    S3 Fig. The average weight of WSG roots under different light and irrigation conditions after two months of treatment.

    The initial weight of transplanted roots was 1.70 ± 0.40 g (mean ± SD, n = 39). Bars represent mean values ± standard errors (n.s.: not significant, ***: p < 0.001, ****: p < 0.0001).

    (DOCX)

    pone.0326237.s004.docx (42.7KB, docx)
    Attachment

    Submitted filename: Responses to Reviewers.docx

    pone.0326237.s006.docx (26.4KB, docx)

    Data Availability Statement

    All relevant data are within the manuscript and its Supporting information files.


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