1 Introduction

Anoectochilus roxburghii, also known as Jinxianlian, is widely recognized in Asia as an important medicinal and edible plant and is also valued as an ornamental species. Phytochemical studies indicate that it contains polysaccharides, flavonoids, glycosides, alkaloids, organic acids, steroids, triterpenes, volatile compounds, and the characteristic active constituent kinsenoside, which together underpin its broad medicinal value (Hou et al., 2025). Pharmacological and clinical-use reports show that this species has been used for diabetes, hyperlipidemia, liver disease, inflammatory disorders, cancers, chronic hepatitis B, hyperuricemia, and related conditions, with reported activities including antidiabetic, hepatoprotective, antioxidant, anti-inflammatory, immunomodulatory, renal-protective, and antineoplastic effects (Wang et al., 2022). In addition to medicinal use, A. roxburghii has entered food, nutraceutical, cosmetic, and functional health-product markets, and has been developed into soups, beverages, jelly, face masks, soap, and other products, reflecting a continuing expansion from traditional herbal use toward diversified industrial applications (Wang et al., 2018). Patent mining further shows that current technological development is concentrated in pharmaceutical preparations and food products, especially downstream applications such as anti-inflammatory, anti-infective, analgesic, and antipyretic products, while companies remain the main research and development actors in this field (Shi et al., 2023). At the same time, industrial development still faces important bottlenecks, including slow variety breeding, insufficient quality-control systems, weak product innovation, limited brand recognition, and the absence of unified evaluation standards across regions; notably, A. roxburghii has not yet been included in the 2020 Chinese Pharmacopoeia, which constrains standardization, safety evaluation, and high-quality industrial development (Hong et al., 2016; Zou et al., 2025).

Despite this high value, wild A. roxburghii resources have undergone persistent depletion for more than two decades. Early resource surveys already emphasized the need for rational preservation and exploitation, while later field investigations in Xishuangbanna found that intensive collection had left the species only sporadically distributed in relatively inaccessible places, making artificial reproduction a practical necessity. More recent studies consistently attribute population decline to overexploitation, habitat destruction, ecological deterioration, slow growth, low seed germination, low natural propagation rates, and strict habitat requirements (Wang et al., 2018; Hou et al., 2025). Surveys in central Vietnam similarly recorded only 123 individuals across 9 of 20 survey lines in Bach Ma National Park, illustrating the scarcity and uneven distribution of extant populations even in protected landscapes (Ho et al., 2025). This conservation pressure is now formally recognized: A. roxburghii has been described as vulnerable, near-threatened, or endangered in international and national conservation frameworks, and has been listed as a National Level II Protected Plant in China (Wang et al., 2022; Hou et al., 2025). Its vulnerability is intensified by ecological specialization. The species depends strongly on precise light conditions for growth, photosynthesis, and flavonoid accumulation, and its sensitivity to abiotic stress suggests limited buffering capacity under environmental change. Climate modeling therefore indicates that, alongside direct human disturbance, future shifts in temperature and precipitation may further alter suitable habitats, making in situ conservation, germplasm rescue from contracting populations, and regionally guided artificial cultivation increasingly urgent.

This study will explore the key technical pathways for the coordinated development of Anoectochilus roxburghii resource conservation and artificial replacement cultivation. Against the background of continuous wild resource decline and increasing market demand, artificial replacement cultivation has become a core approach to relieving resource pressure, ensuring the supply of high-quality raw materials, and promoting sustainable industrial development. In recent years, tissue culture and in vitro rapid propagation technologies have enabled efficient multiplication, genetic consistency, and disease-free seedling production, while large-scale seedling systems and protocorm-like body (PLB) propagation have significantly improved propagation efficiency and transplant survival, providing important technical support for germplasm conservation and industrial production. Meanwhile, technologies such as light-environment regulation, wild-imitated cultivation, endophyte regulation, and under-forest ecological cultivation can further promote active-component accumulation, improve medicinal quality, and strengthen the connection between cultivated materials and wild-type quality. Based on this, this paper systematically analyzes the significance of wild resource conservation of A. roxburghii, discusses key technologies including seedling propagation, cultivation model optimization, precise environmental regulation, medicinal quality formation, and whole-process quality control, summarizes pathways for standardized production, coordinated industrial development, and high-value utilization, and proposes optimization strategies for current problems in artificial replacement cultivation, aiming to provide theoretical and practical references for resource conservation, cultivation technology optimization, and high-quality industrial development of A. roxburghii.

2 Significance of Wild Resource Conservation

2.1 Genetic value

Wild germplasm resources of Anoectochilus roxburghii have irreplaceable genetic value because they preserve the full range of natural variation formed under long-term habitat selection (Priyanka et al., 2021; Yadav et al., 2024). As an endangered medicinal orchid with slow growth, low natural propagation, and narrow habitat requirements, the loss of wild populations would directly reduce the species’ adaptive and evolutionary potential. Climate contraction is especially important for peripheral populations, because populations at distribution margins can contain unique adaptive variation and are at greatest risk of disappearance, making targeted germplasm rescue necessary (Hou et al., 2025). More broadly, work on wild plant germplasm shows that natural populations are valuable gene pools for future breeding, stress resistance, and long-term improvement, a principle that is highly relevant to A. roxburghii conservation.

Recent molecular studies also show why wild germplasm should be treated as a strategic breeding resource rather than only a protected object. Transcriptome assembly in A. roxburghii generated 138 385 unigenes and identified 44 045 SSRs, providing marker resources for assessing diversity across accessions and supporting more effective breeding and conservation (Zhang et al., 2024). DNA barcode work further indicates that matK and ITS can support reliable species identification, with ITS offering better phylogenetic resolution, which is useful for germplasm authentication and genetic resource management (Nhàn, 2025). At the trait level, tetraploid materials produce more major secondary metabolites than diploids, showing that conserving genetically distinctive materials can directly expand medicinal and breeding value (Zhang et al., 2025).

2.2 Ecosystems and population restoration

Protecting the native ecosystem is fundamental because A. roxburghii depends on highly specific forest microhabitats rather than broad environmental tolerance. Field and distribution studies show that the species occurs mainly under evergreen or semi-deciduous broadleaf forest with high humidity, large canopy cover, and limited disturbance, while over-collection, deforestation, and forest conversion have sharply reduced wild occurrence (Ho et al., 2025). Its growth, photosynthesis, and flavonoid accumulation are strongly affected by light conditions, and its narrow ecological amplitude indicates limited buffering capacity under environmental change. Because stable future refugia are predicted mainly in Fujian, Guangdong, Guangxi, Yunnan, and Guizhou, these areas should remain priorities for in situ protection and reserve-gap assessment (Hou et al., 2025).

Population restoration should therefore combine habitat protection with scientifically guided reinforcement. Surveys in Bach Ma National Park found only 123 individuals across 9 of 20 survey lines, usually in small clusters of 4-15 plants, illustrating both rarity and fragmentation (Ho et al., 2025). Earlier regional studies likewise proposed ecological study, tissue-culture multiplication, and wild tending to increase population size and provide a basis for recovery. Restoration also needs to account for stress biology and disease control: drought severely threatens the species, candidate drought-resistance genes have now been identified for molecular breeding, and soft rot can reduce cultivated yield by 70%~80%, which means healthy reintroduction materials require both abiotic and biotic resilience (Xing et al., 2022; Jiang et al., 2025).

2.3 Conservation systems and sustainable utilization

A sustainable conservation system for A. roxburghii should integrate in situ protection, ex situ germplasm preservation, dynamic monitoring, improved-variety propagation, and regulated industrial utilization. Industry analyses identify weak wild-resource protection, slow variety breeding, and insufficient quality-system research as key constraints, and recommend stronger breeding-resource protection, dynamic monitoring, and improved propagation systems (Hong et al., 2016). General germplasm research likewise shows that effective conservation requires collection, storage, analysis, documentation, and exchange, using complementary methods such as slow-growth culture, cryopreservation, DNA banks, botanical gardens, and genetic reserves (Priyanka et al., 2021). For rare plants, in situ conservation is generally preferred, but ex situ systems are essential to prevent genetic loss and to maintain material for future recovery and breeding (Yadav et al., 2024).

Sustainable utilization depends on replacing destructive wild harvesting with high-quality artificial cultivation that still maintains medicinal characteristics. Large-scale micropropagation protocols now achieve shoot induction of 91.67%, rooting of 93.33%, and transplant survival of 90.2%, while tetraploid propagation systems also support high rooting and acclimatization rates, showing that ex situ multiplication can relieve harvest pressure on wild populations (Zhang et al., 2025). Tissue culture is already regarded as the main seedling-raising method because it enables rapid, genetically consistent, disease-free propagation and supports germplasm protection (Li and Li, 2025). At the same time, under-forest and wild-imitated cultivation are important because they better align production with ecological requirements, and wild-imitated systems can shift endophytic communities toward wild-like states while increasing kinsenoside-associated microbial functions, offering a stronger path toward coordinated conservation and utilization.

3 Artificial Cultivation Technical Pathways

3.1 Seedling propagation technology of Anoectochilus roxburghii

Seedling propagation is the primary technical foundation for artificial replacement cultivation of Anoectochilus roxburghii, because conventional propagation is constrained by low natural regeneration and unstable seedling supply (Wang et al., 2022). Tissue culture has therefore become the main seedling-raising route, especially for rapid multiplication, germplasm conservation, and standardized industrial production (Li and Li, 2025). Large-scale nodal-segment culture systems have already achieved 91.67% shoot induction on 1/2 MS with 1.5 mg/L BA, a proliferation coefficient of 4.33 on 1/2 MS with BA, Kn, and NAA, and 93.33% rooting on medium containing NAA, IBA, and banana mash. After acclimatization, these regenerated plantlets reached 90.2% survival in a sand:peat substrate covered with live moss, showing that tissue-cultured seedlings can enter ex vitro production at high efficiency.

Recent studies further show that protocorm and protocorm-like body systems can markedly increase propagation efficiency and broaden the usable germplasm base. An optimized PLB induction, proliferation, regeneration, and rooting protocol produced an 89% PLB induction rate, a 400% secondary PLB proliferation rate, 10.5 shoots per PLB mass, and 98% rooting, while all plantlets survived acclimation (Wang et al., 2022). A newer floral-bud/protocorm route achieved 83.0% protocorm regeneration, 50.33 PLBs per explant, 35 shoots per PLB cluster, and 85% greenhouse survival after 12 months, while kinsenoside content continued to increase during greenhouse cultivation (Luan et al., 2026). For tetraploid materials, B5 medium plus 1 mg/L BA and 0.05 mg/L NAA improved shoot proliferation, red light raised shoot induction to 92%, and MS with 1.0 mg/L IBA plus 1.0 mg/L NAA produced 92% rooting, followed by 80% survival after pot transfer (Zhang et al., 2025).

3.2 Optimization of artificial cultivation models for Anoectochilus roxburghii

Beyond seedling production, the success of artificial replacement cultivation depends on selecting cultivation models that balance survival, biomass accumulation, medicinal quality, and ecological similarity to wild habitats. Early practice showed that under-forest cultivation was proposed specifically to address resource exhaustion by using shaded forest environments to simulate native ecological conditions. Substrate-based field domestication studies on Dinghushan Mountain found that peat: perlite at 4:1 or peat:sand:sawdust at 3:1:1 supported 94.3%-97.6% seedling survival and good growth under natural conditions. Quality-oriented cultivation analysis likewise suggested that high-quality production was associated with MS medium and a humus soil:river sand:perlite substrate of 600:200:200, although that conclusion still requires broader validation (Xu et al., 2017). These results show that matrix design is not a minor management variable, but a core determinant of establishment success and product quality.

Current model optimization is moving from simple soil cultivation toward wild-imitated, microbe-assisted, and hydroponic or soilless systems. Wild-imitated cultivation changed the endophytic community within three months so that it converged toward wild-tending plants, and bacterial diversity was positively associated with kinsenoside content, especially through Burkholderia-Caballeronia-Paraburkholderia. Arbuscular mycorrhizal inoculation with Glomus intraradices improved root architecture, biomass, polysaccharides, flavonoids, and stalk-rot resistance, supporting a model in which beneficial fungi are integrated into production systems rather than treated as incidental (Gu et al., 2025). Hydroponic culture systems also show promise: in vitro Erlenmeyer-flask culture with cotton-layer substrate and SH medium improved shoot growth, while ex vitro hydroponics with cotton-layer substrate and 1/2 SH solution gave the best biomass and kinsenoside accumulation (Luan et al., 2025). More broadly, soilless cultivation offers a pathway for precise rhizosphere regulation, higher nutrient-use efficiency, and microbiome engineering, though cost and substrate sustainability remain practical constraints (Tuxun et al., 2025).

3.3 Precise environmental regulation technology for Anoectochilus roxburghii cultivation

Because A. roxburghii has narrow ecological amplitude, precise environmental regulation is essential for replacing wild collection with stable artificial production. Light regulation is one of the clearest examples. Orthogonal light experiments showed that light intensity, red:blue ratio, and photoperiod all significantly affected growth and metabolite accumulation, with 60 μmol·m⁻²·s⁻¹, red:blue=2:1, and a 10 h photoperiod favoring growth and polysaccharides, while 100 μmol·m⁻²·s⁻¹, red:blue=1:1, and a 14 h photoperiod favored flavonoids (Chen et al., 2021). Supplemental blue light also promoted leaf number, stem diameter, biomass, chlorophyll a, flavonoids, and polyphenols, whereas yellow light increased soluble sugar and polysaccharides (Wang et al., 2018). During in vitro propagation, light effects are stage-specific: darkness or weak light favors PLB proliferation, but light is required for PLB differentiation, and red light can enhance tetraploid shoot proliferation (Figure 1) (Wang et al., 2022; Zhang et al., 2025).

Temperature, humidity, shading, and planting density also require fine control across different cultivation stages. Under Dinghushan conditions, optimal growth was observed at 23~28 ℃, pH 4.75, and about 90% relative humidity, with shade-net use and avoidance of excessive substrate moisture. Shading studies in micropropagated Anoectochilus plantlets found that mild to medium shade generally supported biomass production and faster growth, while species responses still differed (Chen et al., 2017). Newer work on shade-tolerant responses in A. roxburghii showed that moderate dense planting improved stem diameter, leaf area, SOD activity, and stem lignin, while increasing far-red light reduced stalk-rot incidence and strengthened defense-related enzyme activity rather than triggering a typical shade-avoidance response (Guo et al., 2025). Precise regulation also extends to the transplant stage, where hormone balance, osmotic adjustment, antioxidant activation, substrate improvement, and later-stage light management jointly determine root-system recovery and seedling establishment after tissue culture (Li and Li, 2025).

4 Quality Formation and Quality Control

4.1 Accumulation patterns and influencing factors of active components in Anoectochilus roxburghii

The medicinal quality of artificially cultivated Anoectochilus roxburghii is determined by the accumulation of multiple active components, chiefly kinsenoside, flavonoids, polysaccharides, phenolic acids, and nucleosides, rather than by any single compound (Zou et al., 2025). Kinsenoside is especially important because A. roxburghii is its exclusive medicinal source, but its content varies substantially among varieties, production areas, and cultivation periods (Zou et al., 2025). Variety effects are clear: among nine varieties, Hongxiadaye had the highest kinsenoside content, while Xiaoyuanye uniquely contained both kinsenoside and its isomer (Figure 2) (Wang et al., 2025). Regional heterogeneity is equally strong, with targeted and untargeted metabolomics showing large differences in flavonoid profiles among Fujian habitats, and Youxi samples having higher average levels of eight quantified flavonoids despite Yongchun showing the richest unique-metabolite diversity (Lyu et al., 2024).

Active-component accumulation is also shaped by developmental stage and culture system. In greenhouse-acclimatized plants from floral-bud-derived propagules, whole-plant kinsenoside rose with cultivation time and peaked at 2.591% after 12 months (Zou et al., 2025). In hydroponic systems, ex vitro cotton-layer and cocopeat systems supplied with 1/2 SH nutrient solution produced higher whole-plant kinsenoside accumulation of 1.24%~1.32% dry weight (Luan et al., 2025). Rhizome bioreactor culture showed that biomass, kinsenoside, and polysaccharide accumulation peaked at 30 days, reaching 2980.5 mg/L and 5 672.9 mg/L for kinsenoside and polysaccharides, respectively, under optimized inoculum, aeration, and light conditions (Wei et al., 2020). At the mechanistic level, transcriptomic analysis linked higher kinsenoside accumulation to elevated expression of ACAA, Hbd, CoAT, PaaH, and UGT genes, while light-regulation studies further showed that red-blue light activates phenylpropanoid, flavonoid, and kinsenoside biosynthetic pathways (Wang et al., 2025; Luo et al., 2025).

4.2 Regulatory effects of cultivation environment on the medicinal quality of Anoectochilus roxburghii

Among environmental factors, light quality is the most consistently validated regulator of medicinal quality in cultivated A. roxburghii. Blue film increased fresh weight, leaf area, chlorophyll, antioxidant enzyme activities, polysaccharides, and flavones, whereas red film promoted plant height and phenolic accumulation. Supplemental blue light likewise increased biomass, total flavonoids, and total polyphenols, while yellow light preferentially increased soluble sugars and polysaccharides (Wang et al., 2018). More precise orthogonal optimization showed that light intensity, red:blue ratio, and photoperiod all significantly affect growth and metabolite accumulation, with 60 μmol·m⁻²·s⁻¹, 2:1 red:blue, and a 10 h photoperiod favoring growth and polysaccharides, while 100 μmol·m⁻²·s⁻¹, 1:1 red:blue, and a 14 h photoperiod favored flavonoids (Chen et al., 2021). Recent metabolomic work further showed that an R2B1 light regime substantially increased amino acids, polyphenols, and kinsenoside, with kinsenoside rising by 81.25% in leaves and 72.24% in stems relative to white light (Wu et al., 2023; Luo et al., 2025).

The cultivation substrate, habitat-like microclimate, and microbial symbiosis also strongly regulate medicinal quality. Habitat metabolomics showed that flavonoid accumulation is significantly associated with temperature and humidity, while morphological performance was better in more humid habitats (Lyu et al., 2024). Practical cultivation studies identified suitable conditions of 23~28 ℃, about 90% relative humidity, pH 4.75, and shaded management, with peat-based mixed substrates supporting 94.3%~97.6% seedling survival (Xu et al., 2017). Wild-imitated cultivation increased kinsenoside-associated quality by shifting the endophytic community toward that of wild-tending plants, and bacterial diversity showed a significant positive correlation with kinsenoside content, especially for Burkholderia-Caballeronia-Paraburkholderia. Beneficial fungi and bacteria exert similar effects: endophytic fungal strains increased biomass and induced flavonoids, kinsenoside, and polysaccharides, while dual Bacillus velezensis inoculation increased fresh weight by 82.6% and 106.6%, raised kinsenoside by 9.33% and 21.65% per gram, and enhanced flavonoids and rhizosphere enzyme activities (Ye et al., 2020; Wei et al., 2020).

4.3 Construction of a whole-process quality control system for Anoectochilus roxburghii

A whole-process quality control system for A. roxburghii must begin at the source with authenticated germplasm and continue through cultivation, harvesting, processing, storage, and final product evaluation. Current reviews agree that quality is jointly shaped by germplasm, producing area, harvesting, processing, preparation, and storage. Germplasm authentication should combine morphological identification with molecular tools, because SSR markers and DNA barcoding have been proposed for classification, resource preservation, and varietal discrimination (Zou et al., 2025). Because content differs markedly among strains and origins, source control should include elite-variety selection, origin traceability, and chemotype classification rather than simple species confirmation (Wang et al., 2025). During cultivation, quality control should integrate standardized light, substrate, humidity, and microbial-management protocols, since each of these factors measurably alters active-compound accumulation (Xu et al., 2017; Wei et al., 2020; Chen et al., 2021).

Post-harvest control is equally necessary because active components decline during processing and storage. Different drying temperatures affect kinsenoside, total flavonoids, eight individual flavonoids, and six nucleosides (Zou et al., 2025). Among drying methods, vacuum drying at (60±2) ℃ best preserved appearance, aroma, and active ingredients overall, although freeze-drying better maintained natural appearance and hot-air or microwave drying preserved specific sensory or polysaccharide traits. Storage studies showed that polysaccharides, kinsenoside, and flavonoids all declined over eight months, vacuum packaging better preserved polysaccharides and flavonoids, and kinsenoside degradation was driven mainly by endogenous enzymes rather than packaging method (Wei et al., 2022). At the analytical end, single-index control is inadequate, so multi-component methods such as LC-MS/MS, metabolomics, and integrated phenotype-barcode-chemotype-transcriptome frameworks should be used to establish Q-markers and comprehensive specifications; this is particularly important because A. roxburghii still lacks a unified pharmacopoeial standard and consistent cross-provincial evaluation system (Zhang et al., 2022; Wang et al., 2025).

5 Standardized Production and Industrial Pathways

5.1 Construction of a standardized production system for Anoectochilus roxburghii

A standardized production system for Anoectochilus roxburghii should begin with stable seedling supply, because wild resources are endangered, conventional propagation is inefficient, and industrial cultivation depends on reproducible micropropagation rather than continued wild collection (Hong et al., 2016; Wang et al., 2022). Current evidence already provides workable technical benchmarks for standardized seedling production: nodal-segment culture achieved 91.67% shoot induction, a proliferation rate of 4.33, 93.33% rooting, and 90.2% transplant survival under optimized media and substrate conditions. PLB-based systems further improved efficiency, reaching 89% PLB induction, 400% secondary PLB proliferation, 98% rooting, and full survival after acclimation, which makes them especially suitable for large-scale factory seedling production. More recent protocols for tetraploid or floral-bud-derived materials show that standardized propagation can also be linked to elite-germplasm utilization, with high shoot induction, rooting, and long-term greenhouse survival (Zhang et al., 2025; Luan et al., 2026).

Standardization must then extend from nursery protocols to cultivation management, environmental control, and quality criteria. Industry reviews identify the lag in variety breeding, insufficient quality systems, and weak product innovation as central bottlenecks, so production standards should define improved-variety propagation, substrate and greenhouse management, harmless pest control, and dynamic monitoring from seedling to harvest (Hong et al., 2016). Practical cultivation studies already outline the core variables that can be standardized, including ecological conditions, greenhouse location, seedbed construction, fertilizer and water management, and harvest management. Evidence also supports standardizing light and substrate regimes, because blue light improves biomass, flavonoids, and polysaccharides, while suitable mixed substrates and high humidity favor transplant survival and plant quality (Ye et al., 2020). In practice, a standardized production system should integrate elite germplasm, rapid propagation, controlled environment cultivation, and multi-index quality evaluation so that cultivated material is both consistent and traceable (Wang et al., 2022; Zhang et al., 2025).

5.2 Coordinated development of resource conservation and industry of Anoectochilus roxburghii

The coordinated development of conservation and industry is necessary because A. roxburghii has high medicinal and economic value, but its wild populations have been severely depleted by over-collection, habitat loss, slow growth, and low natural regeneration. Artificial cultivation is therefore not only an industrial strategy but also a conservation substitute that reduces harvesting pressure on wild populations (Wang et al., 2018; Wang et al., 2022). The conservation side should combine in situ protection of stable habitats with ex situ germplasm preservation, especially for threatened peripheral populations and elite local resources. Distribution modeling further suggests that future cultivation bases can be planned in newly suitable northern regions to meet demand while reducing pressure on vulnerable southern wild populations (Hou et al., 2025).

Industrial development should be coupled to conservation through germplasm protection, regional planning, and wild-imitated cultivation. Earlier resource surveys already argued that preservation and rational exploitation must be considered together, rather than as separate agendas. Wild-imitated cultivation is especially promising because it shifts the endophytic community toward that of wild-tending plants and is associated with higher kinsenoside content, which helps align cultivated material with wild medicinal quality. Beneficial microbial strategies support the same conservation-industry linkage: endophytic fungi and Bacillus velezensis strains increase biomass and active compounds, which can raise cultivated yield and quality without increasing collection pressure on wild plants (Ye et al., 2020; Hou et al., 2025). The most sustainable pathway is therefore a coordinated model in which protected germplasm resources feed standardized artificial cultivation, and cultivation success in turn lowers incentives for destructive wild harvesting (Hong et al., 2016).

5.3 Extension of the Anoectochilus roxburghii industrial chain and high-value utilization

The industrial chain of A. roxburghii can be extended because the species is not only a medicinal plant but also a functional food and ornamental resource with diverse bioactive constituents and broad pharmacological relevance. It is already used in foods, soups, teas, beverages, and several dosage forms, and clinical or quasi-clinical applications have been reported for diabetes, hepatitis B, hyperuricemia, cough-variant asthma, and other conditions (Ye et al., 2020; Gam et al., 2020). These features support downstream diversification into health foods, botanical beverages, standardized extracts, and pharmaceutical intermediates rather than reliance on crude fresh-herb sales alone (Wang et al., 2018; Hou et al., 2025). PLBs and rhizome cultures also offer alternative production units for kinsenoside and polysaccharides, which creates opportunities for ingredient-oriented processing rather than depending exclusively on whole-plant harvests (Wang et al., 2022; Luan et al., 2026).

High-value utilization also depends on strengthening the midstream processing segment and building stronger product innovation capacity. Patent analysis indicates that current research and development already focuses on pharmaceutical preparations, food uses, and downstream products such as anti-inflammatory and anti-infective applications, while also pointing to the need for extraction and purification equipment in the midstream segment. The same analysis suggests further expansion into medical, cosmetic, and oral-care products, including immune regulation, skin repair, whitening, and tooth-protection applications. Because current industry assessments still identify weak innovation capability, low brand competence, and limited market cognition, future industrial upgrading should combine standardized raw-material supply with deep processing, brand development, and diversified consumer products (Hong et al., 2016). For A. roxburghii, the most viable industrial development pathway is therefore to use artificial replacement cultivation as the upstream base, and then extend toward high-value extracts, functional foods, pharmaceutical products, and other differentiated applications that increase output value while supporting conservation.

6 Problems and Optimization Strategies

6.1 Utilization efficiency of superior germplasm resources of Anoectochilus roxburghii still needs improvement

Current artificial replacement cultivation has reduced dependence on wild resources, but the efficient use of elite germplasm remains limited by slow variety breeding, insufficient protection of breeding resources, and weak integration between germplasm evaluation and industrial propagation (Wei et al., 2022). Existing evidence shows that polyploid breeding is a promising route to superior germplasm utilization, because tetraploid plants generally show higher major secondary metabolite production, stronger photosynthetic performance, larger roots and stomata, and better adaptation to unstable environments than diploid materials (Su et al., 2017; Huang et al., 2022; Zhang et al., 2025). Artificial induction methods have already provided a practical basis for this direction: mutagenic tetraploid breeding reported frequencies above 75% and improved medicinal yield, while optimized in vitro propagation protocols for tetraploid materials achieved high shoot induction, rooting, and transplant survival, making elite-line expansion technically feasible (Zhang et al., 2025). At the same time, utilization efficiency is still constrained by incomplete molecular characterization of germplasm, although the recent chromosome-level autotetraploid genome now offers a reference for functional genomics, marker development, and precise molecular breeding (Fang et al., 2025).

Optimization should therefore focus on building a coordinated “collection-evaluation-breeding-propagation” system for elite germplasm. First, wild, local, and cultivated resources should be systematically collected and conserved, while superior chemotypes and stress-tolerant lines are identified through combined phenotypic, phytochemical, and molecular screening (Ye et al., 2020). Second, molecular breeding should be accelerated by using genomic resources and drought-resistance candidate genes such as the ArWRKY57-ArWRKY70-ArLEA5 module to develop elite cultivars with both stable quality and stronger environmental resilience (Jiang et al., 2025; Fang et al., 2025). Third, rapid propagation systems should be matched to elite germplasm deployment, including PLB-based regeneration, nodal culture, and tetraploid micropropagation, so that breeding gains can be translated quickly into industrial seedling supply (Zhang et al., 2025). In short, the key problem is not the absence of superior germplasm, but the low efficiency with which conserved, identified, bred, and propagated elite materials are converted into standardized cultivation resources (Su et al., 2017; Huang et al., 2022).

6.2 Quality stability of artificially cultivated Anoectochilus roxburghii still needs enhancement

The main quality problem in cultivated A. roxburghii is that active constituents vary substantially with cultivation mode, light regime, microbial association, habitat conditions, harvest stage, and post-harvest handling, so cultivated material often lacks the consistency required for medicinal use (Luo et al., 2025). Environmental regulation is especially important: blue light improves growth, flavonoids, and polysaccharides, red-blue combinations can markedly increase phenolic acids, flavonoids, and kinsenoside, and optimized orthogonal light protocols differentially favor polysaccharides or flavonoids depending on light intensity, red:blue ratio, and photoperiod (Chen et al., 2021; Luo et al., 2025). Biological regulation is similarly influential, because wild-imitated cultivation shifts the endophytic community toward wild-type structure and is associated with higher kinsenoside, while endophytic fungi, mycorrhizal fungi, AM fungi, and beneficial bacteria all improve biomass and promote accumulation of flavonoids, polysaccharides, or kinsenoside (Ye et al., 2020; Zhang et al., 2020; Gu et al., 2025). Even after harvest, active compounds continue to change: polysaccharides, kinsenoside, and flavonoids all decline during storage, vacuum packaging better preserves polysaccharides and flavonoids, and kinsenoside degradation is influenced more by endogenous enzymes than packaging method (Wei et al., 2022).

Improving quality stability requires shifting from simple yield-oriented cultivation to process-oriented quality regulation. Cultivation standards should define target light formulas, substrate composition, humidity, temperature, and culture duration for different production objectives, since both bioreactor rhizome culture and greenhouse systems show clear optimum windows for metabolite accumulation (Jin et al., 2017; Chen et al., 2021). Microbial management should become part of routine cultivation, with prioritized use of beneficial endophytes, mycorrhizae, and AM fungi that simultaneously improve growth, quality, and disease resistance (Ye et al., 2020; Zhang et al., 2020; Gu et al., 2025). Harvest timing and elicitation strategies should also be optimized, because kinsenoside accumulation peaks at defined developmental stages and can be significantly enhanced by yeast extract, salicylic acid, or methyl jasmonate under controlled rhizome culture conditions (Jin et al., 2018; Luo et al., 2018). Finally, post-harvest drying, packaging, and storage must be standardized together with multi-component chemical monitoring, so that quality stability is controlled across the full chain rather than judged only at harvest (Wei et al., 2022).

6.3 The standardized industrial system of Anoectochilus roxburghii still needs improvement

Although artificial cultivation has supported expanding production, the industry still lacks a unified system linking germplasm, seedling production, cultivation management, quality evaluation, processing, storage, and downstream product development (Su et al., 2017; Wei et al., 2022). One major weakness is the absence of consistent quality standards across regions and products, compounded by species confusion and adulteration in the market because morphologically similar Anoectochilus, Goodyera, and Ludisia materials are sometimes used as substitutes (Ye et al., 2020; Fang et al., 2025). Another weakness is that industrial production often emphasizes planting scale more than chain integration, even though research already points to the importance of greenhouse management, harmless pest control, controlled environmental regulation, and post-harvest quality preservation (Zhang et al., 2020; Chen et al., 2021). The gap is therefore systemic: production technologies exist, but they are not yet assembled into a unified industrial specification from upstream breeding to midstream processing and downstream high-value utilization (Ye et al., 2020).

The optimization pathway is to build a full-chain standardized industrial system centered on traceability, quality markers, and coordinated industrial upgrading. Upstream, authenticated elite germplasm should be linked to standardized propagation and protected breeding-resource management, using DNA barcoding, chemotype classification, and molecular markers to prevent varietal confusion and improve source control (Ye et al., 2020; Fang et al., 2025). Midstream, production should be standardized through facility cultivation protocols, microbial-assisted quality enhancement, optimized drying and storage methods, and process specifications for extraction and purification of active ingredients (Ye et al., 2020; Gu et al., 2025). Downstream, the industry should expand from crude medicinal material sales toward functional foods, oral liquids, cosmetics, and pharmaceutical preparations, while using multi-component analytical tools to define stable Q-markers and improve product consistency (Wang et al., 2018; Zhang et al., 2020). Overall, the standardized industrial system of A. roxburghii still needs improvement because the next stage of artificial replacement cultivation is not simply more planting, but tighter integration of breeding, cultivation, quality control, processing, and high-value product development (Wei et al., 2022).

7 Conclusion and Prospects

Conservation of wild Anoectochilus roxburghii resources is the foundation for sustainable industrial development because natural populations have been severely reduced by over-collection, habitat loss, slow growth, low natural propagation, and narrow ecological requirements. Its threatened status is reflected in formal protection designations, including listing as a protected plant in China and recognition as vulnerable, near-threatened, or endangered in international conservation frameworks. Conservation planning now has clearer spatial guidance, because climate-based distribution modeling identifies Fujian, Guangdong, Guangxi, Yunnan, and Guizhou as stable highly suitable habitats that should be prioritized for in situ protection, while contracting peripheral populations should be urgently targeted for ex situ germplasm collection to preserve adaptive diversity. Earlier field-resource surveys and recent metabolomic studies also show that conserving wild populations is important not only for species survival but also for maintaining ecological, chemical, and regional diversity that underpins future breeding, quality evaluation, and rational utilization.

Artificial replacement cultivation of A. roxburghii is the core pathway for relieving resource pressure because market supply can no longer rely on wild harvesting, and artificial cultivation already serves as the main source of propagated material. Rapid propagation systems now provide a practical basis for large-scale replacement, including nodal culture with 91.67% shoot formation, 93.33% rooting, and 90.2% transplant survival, PLB systems with 89% induction, 400% secondary proliferation, 98% rooting, and full acclimatization survival, and tetraploid propagation systems that support both germplasm conservation and commercial multiplication. Beyond simple yield replacement, wild-imitated cultivation, under-forest cultivation, and karst-forest imitation-wild systems provide additional routes for reducing pressure on wild resources while improving medicinal continuity and ecological compatibility. Climate projections further suggest that establishing future cultivation bases in newly suitable northern regions such as Hubei, Anhui, and southern Henan could simultaneously meet market demand and reduce harvesting pressure on vulnerable southern wild populations.

Scientific and technological innovation will promote A. roxburghii resource conservation and high-quality industrial development because the main remaining bottlenecks are low breeding efficiency, unstable cultivated quality, weak quality systems, and limited industrial innovation capacity. Innovation is already improving propagation efficiency and product quality through optimized media, light regulation, elite tetraploid propagation, and alternative production systems such as rhizome bioreactors and PLB-based kinsenoside production. Molecular and omics tools are expanding the breeding and management toolbox, including DNA barcode identification, transcriptome resources, a high-quality reference genome, drought-tolerance gene modules, and metabolome-transcriptome analyses showing that red-blue light can strongly promote functional metabolite biosynthesis. At the cultivation end, microbial engineering, endophyte management, and environmentally controlled production can improve biomass, disease resistance, and active-ingredient accumulation, while industrial upgrading should extend toward standardized extraction, purification, quality markers, and diversified products in medicine, health food, and cosmetics.

Conflict of Interest Disclosure

The author affirms that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.

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