Background/Objectives: For over two millennia, Ganoderma lucidum has served as a traditional remedy, yet its translation into evidence-based therapy remains stymied by a persistent obstacle. Potent in vitro activities consistently fail to translate in vivo, a shortfall rooted in the poor systemic bioavailability of its signature triterpenoids and polysaccharides. We contend that the defining hurdle is no longer compound discovery but whether emerging formulation technologies genuinely overcome these barriers or merely sidestep them in ways that obscure the underlying physiology. Methods: We construct a structure–activity–formulation (SAF) prioritization framework. We critically survey a broad spectrum of delivery platforms—including lipid-based, polymeric, micellar, tellurium nanorod, and G. lucidum-derived vesicular systems—against their demonstrated capacity to improve oral exposure. Clinical assessments and regulatory considerations are integrated to anchor the analysis in translational reality. Results: Bioavailability has been systematically treated as a post hoc variable, even though the very structural features conferring bioactivity impose the steepest systemic barriers. To date, no G. lucidum nanoformulation has advanced to human trials, and the regulatory terrain for these complex natural-product nanomedicines remains uncharted. We distill three experimentally tractable propositions—testing lipid-mediated absorption, triple-helix conformational dependency, and co-delivery synergy—that offer discrete entry points for rigorous investigation. Conclusions: We argue that the field must pivot decisively from descriptive cataloging to hypothesis-driven, comparative investigation, anchored by reference-material standardization and proactive regulatory dialogue. Confronting the bioavailability bottleneck as a primary design parameter—rather than an ancillary nuisance—represents the most credible route from historical remedy to evidence-based therapeutic.

Santalum album L. is an economically important species that can produce highly valued essential oil (EO) consisting mainly of santalols in its aromatic heartwood. Elicitors stimulate the production of EO, but the underlying mechanism remains unknown. In this study, the regulatory role and ability of a cytokinin (6-benzyladenine, BA) to promote the accumulation of secondary metabolites, was investigated. The levels of EO and total flavonoids increased 2.3- and 1.5-fold, respectively one year after BA treatment. The top significantly enriched Kyoto Encyclopedia of Genes and Genomes pathways of upregulated differentially expressed genes (DEGs) and differentially accumulated metabolites contained ‘terpenoid biosynthesis’, ‘phenylpropanoid biosynthesis’, and ‘flavonoids biosynthesis’, displaying similar profiles to those detected in natural heartwood from 10-year-old trees. Prominently, the content of santalols was increased by approximately 90-fold by BA relative to the control and was associated with the contents of flavonoids and phenolics. BA also enhanced the contents of cytokinins, an auxin (IAA), gibberellins, jasmonic acid and an ethylene precursor while DEGs associated with the biosynthesis and signaling of these hormones were significantly upregulated. Dual-LUC assays confirmed that identified SaMYC2a-2c and three SaERFs activated SaSSY and SaCYP736A167 expression. Moreover, exogenous BA regulated EO biosynthesis in sandalwood suspension cells but could not directly activate the expression of these two genes in these cells. We infer from these findings that exogenously applied BA positively participated in the biosynthesis of sandalwood EO by interacting with other hormones. These findings also reveal details of the regulatory mechanism underlying the cytokinin-induced production of secondary metabolites, which determine the fragrance and color of S. album heartwood.

Callicarpa peichieniana is an important traditional Chinese medicinal plant with pharmacological benefits for digestive system diseases and wounds, as well as high ornamental value. The goal of this study is to establish an effective in vitro regeneration system in order to satisfy the expanding market demand. Extracts from algae can enhance the proliferation and rooting effect of adventitious buds and can improve the survival rate of transplantation. This study developed an in vitro regeneration system using apical bud explants of C. peichieniana associated with Chlorella sorokiniana (an alga species). Inter simple sequence repeat (ISSR) molecular markers confirmed the genetic fidelity of the regenerated plantlets. The highest number of adventitious buds (5.00 buds) was induced from the apical buds with 0.5 mg/L 6-BA in a Murashige and Skoog (MS) medium, and the highest proliferation coefficient (5.83) was achieved with 2.0 mg/L 6-BA. A rooting rate of 100% was achieved by using 0.1 mg/L NAA, MS with 50% macroelements, and 20 g/L sucrose, averaging 6.36 roots per explant and a root length of 1.32 cm. In all micropropagation stages, C. sorokiniana coexisted and proliferated alongside C. peichieniana materials. ISSR showed that the genetic fidelity of C. peichieniana regenerated plants was 95.45%. Coconut coir/perlite = 1∶1 (v/v) was identified as the optimal transplantation substrate, achieving a 100% survival rate. The “C. peichieniana–C. sorokiniana association” in vitro regeneration system established in this study not only enables the mass production of high-quality regenerated plantlets but provides new ideas and demonstrations for culturing multiple species in the same in vitro system.

Paphiopedilum orchids have a high ornamental value, and flower abundance is a key horticultural trait. Most Paphiopedilum plants exhibit weak tillering ability, with their tiller buds often entering a dormant state post-formation. Tiller production plays a crucial role in enhancing flower abundance and is potentially regulated by plant hormones. However, the effect of hormones on tillering in Paphiopedilum plants is still unclear. In this study, we investigated the promotion of tillering in P. callosum through exogenous root irrigation of benzylaminopurine (BAP). We observed a dose-dependent promotion of tiller production by BAP, with the strongest effect observed at a concentration of 400 mg/L. By comparing the expression of key genes in P. ‘SCBG Yingchun’ (with strong tiller ability) and P callosum (with weak tiller ability), we found that BAP promotes tillering by interacting with abscisic acid (ABA). This interaction involves down-regulation of the ABA degradation gene PcCYP707A, leading to a reduction in ABA content, and the subsequent down-regulation of dormancy-associated genes (PcDRMH1, PcSVP) to release bud dormancy. Additionally, BAP promotes sustained outgrowth of tiller buds by increasing the level of indole-3-acetic acid (IAA) through up-regulation of the IAA synthesis gene PcYUC2 and the transport gene PcPINIC. Our results indicated that the application of BAP promotes lateral bud outgrowth and increases tiller production in P. callosum. Through transcriptome analysis, we found that the BAP-promotion of tillering involves not only changes in endogenous IAA, ABA, and CTKs content but is also associated with the regulation of metabolism-related genes and dormancy-associated genes. This study presents the first comprehensive report of BAP-promoted tillering in P. callosum, providing a foundational basis for further mechanistic studies on tiller development in Paphiopedilum species and other non-model plants.

Gynura divaricata (L.) DC., a herb with economic, ecological, and medical importance, is widely cultivated in tropical and subtropical areas of Southeast Asia. G. divaricata can bloom and set seeds, but only a small number of seeds form and very few are viable, so seedlings are rarely employed for propagation purposes. In this study, callus and adventitious shoots were induced from leaves. Adventitious shoot propagation is reported for the first time for this species. The optimal medium for direct induction of adventitious shoots from leaf explants was Murashige and Skoog (MS) medium containing 4.0 mg L− 1 thidiazuron (TDZ), forming 7.78 adventitious shoot buds per explant. When 2.0 mg L− 1 TDZ and 0.5 mg L− 1 α-naphthaleneacetic acid (NAA) or 2.0 mg L− 1 6-benzyladenine (BA) and 0.5 mg L− 1 NAA were added to MS medium, callus was induced initially, followed by 9.78 or 8.87 adventitious shoot buds (per explant), which developed from callus. Adventitious shoots could not be induced by 2,4-dichlorophenoxyacetic acid, NAA or kinetin. The addition of L-ascorbic acid (AsA) to MS medium reduced the level of hyperhydricity of shoots and plantlets. The optimal medium for shoot proliferation was MS medium with 4.0 mg L− 1 BA and 50 mg L− 1 AsA. Optimal rooting of G. divaricata was achieved on MS medium with 0.5 mg L− 1 indole-3-butyric acid, with 97.33

Santalum album is an economically important plant in the craft, spices and medicine industries. The main chemical constituents found in sandalwood essential oils are sesquiterpenes. 3-Hydroxy-3-methylglutaryl monoacyl-coenzyme A reductase (HMGR) is one of the rate-limiting enzymes required for the synthesis of sandal sesquiterpenes, but there are no studies on the HMGR gene in S. album. In this study, the full-length ORFs of the upper rate-limiting enzyme genes SaHMGR1 and SaHMGR2, which lie upstream of the MVA metabolic pathway of sandal sesquiterpenes, were cloned for the first time. Bioinformatics and phylogenetic analyses were conducted. The results showed that SaHMGR1 and SaHMGR2 had typical domains of HMGR class I enzymes in the HMGR superfamily, including four catalytic sites, six NADPH-binding sites, five substrate binding regions, four inhibitor binding sites, and several dimer interface regions. A phylogenetic analysis showed that SaHMGR1 and SaHMGR2 were highly conserved relative to corresponding genes in other plants. An analysis of subcellular localization showed that these SaHMGR genes were located in the endoplasmic reticulum. SaHMGR1 and SaHMGR2 were detected by real-time PCR in roots, sapwood, heartwood, young leaves, mature leaves and twigs. Highest expression was in roots. SaHMGR1 expression was higher in mature leaves than in heartwood while SaHMGR2 expression was lower in mature leaves than in heartwood. Expression in Escherichia coli strain DH5α with plasmid pET-32a (+) was also used to verify the functionality of both HMGR proteins, which catalyzed the formation of MVA from HMG-CoA. In E. coli, the enzymatic activity of SaHMGR1 was higher than that of SaHMGR2. These findings provide a basis for further studies on the function of SaHMGR genes and the regulation of sesquiterpene biosynthesis in S. album.

Turfgrasses growing on tropical island reefs have developed morphological, structural, physiological and ecological strategies that confer them with tolerance to salt, high temperatures and drought. They are thus able to adaptively survive in extreme environments with high salt concentrations, high temperatures, strongly alkaline soil, and high light intensities. Research on tropical island turfgrasses advance knowledge on island ecological restoration, salt tolerance, and resource development. This study focused on five turfgrass species that are used as lawn turfgrass in southern China: Lepturus repens (G. Forst.) R. Br., Thuarea involuta (Forst.) R. Br, Zoysia matrella (L.) Merr, Cynodon dactylon (L.) Persoon and Axonopus compressus (Sw.) P. Beauv. For all species, using an in vitro tissue culture system based on Murashige and Skoog (MS) basal medium for shoot proliferation and growth, the physiological responses to different NaCl concentrations were assessed, and NaCl tolerance was evaluated. Based on physiological or biochemical parameters, salt tolerance of the species was A. compressus (0.06) > Z. matrella (0.09) > C. dactylon (0.14) > L. repens (0.42) > T. involuta (0.67) at 50 mM NaCl; Z. matrella (0.04) > A. compressus (0.25) > C. dactylon (0.35) > L. repens (0.46) > T. involuta (0.67) at 100 mM; C. dactylon (0.30) > L. repens (0.37) > A. compressus (0.48) > Z. matrella (0.60) > T. involuta (0.95) at 200 mM NaCl; C. dactylon (0.33) > L. repens (0.46) > Z. matrella (0.72) > A. compressus (0.83) = T. involuta (0.83) at 400 mM NaCl The correlation between physiological and biochemical parameters of these five turfgrasses was also assessed, with a strong correlation with MDA content in T. involuta, with proline content and POD, SOD activity in L. repens and Z. matrella, with proline content and CAT, SOD activity in C. dactylon, and with proline content and POD activity in A. compressus, suggesting their different roles in the protective mechanism underlying different concentrations of salt stress tolerance.

Santalum album is highly valued for its fragrant essential oil from heartwood. Volatile terpenoids involved in aroma formation in plants can be emitted in response to a variety of environmental stresses. However, the regulatory mechanisms underpinning the response of S. album to external stresses are not yet known. In this study, the regulatory mechanism of S. album leaves was investigated after the application of methyl jasmonate (MeJA), a stressor. Eight classes of volatile organic compounds were identified in S. album leaves, including terpenes, aldehydes, alcohols, ketones, esters, benzenoids, alkanes and heterocyclic compounds. In total, 15 terpenoids, such as ocimene, linalool, nerolidol and α-farnesene, were considerably induced 6 h after MeJA treatment, amounting to 38.01 % of all volatiles. Notably, transcript levels of the genes in the MVA pathway were enhanced by 2- to 8-fold after 6 h of MeJA treatment compared to the control. Exogenously applied MeJA resulted in the significant upregulation of ten terpene synthase (SaTPS) genes. In vitro enzyme activity assays confirmed that four SaTPS recombinant proteins converted substrates into (E)-nerolidol and linalool. Overexpression of the four SaTPS genes produced (E)-nerolidol, (Z)-nerolidol and linalool in sandalwood callus. These results suggest that the four SaTPS genes have the ability to synthesize nerolidol and linalool in S. album leaves in response to MeJA treatment. This study provides insight into the regulatory mechanism of the biosynthesis of terpenoids in S. album in response to an environmental stress.

Soil salinity, a major environmental stress, restricts agricultural production worldwide. Gynura divaricata (L.) DC. is widely cultivated on tropical islands in China and has both edible and medicinal value. NaCl stress and growth indicators, antioxidant enzyme activity, as well as MDA, proline, and soluble sugar content, were determined. Based on the transcriptomic data of tissue-cultured G. divaricata plantlets grown in control (0 mM NaCl) and salt stress (50 and 200 mM NaCl) conditions, gene expression patterns were examined. KEGG enrichment analysis of differentially expressed genes indicated significant enrichment of plant hormone signaling, MAPK signaling, and starch and sucrose metabolism pathways. These findings allowed key biological pathways and salt stress-responsive genes to be identified, thus providing a molecular basis for breeding salt-tolerant G. divaricata varieties. This transcriptomic analysis revealed a complex tolerance mechanism of G. divaricata in response to NaCl, laying a foundation for screening and cloning key genes related to NaCl tolerance and studying their interactions. These findings would allow the molecular basis of NaCl tolerance to be explored, with the aim of breeding NaCl-tolerant varieties.

Platycerium wallichii Hook is a rare and endangered tropical epiphytic fern with ornamental and medicinal value. It is usually propagated by spore germination to produce sporophytes. This study established an in vitro protocols for spore germination, leading to filament formation and development of prothallia, and finally development of sporophytes following fertilization. The prothallia could be proliferated by 8.7 time within 30 days. The development of archegonia and antheridia on prothallia was asynchronous and prothallia were mainly dioecious (over 90

In order to investigate the function and regulation mechanism of NDH dehydrogenase gene in Santalum album,the technique of RACE was used to amplify the full-length sequence of SaNDH6 with heartwood as material.The technique of quantitative real-time fluorescence PCR(RT-qPCR)was employed to analyze its expression in different tissues and after hormone induction.The subcellular location was determined by Arabidopsis thaliana protoplast transient expression.2 kb cis-acting element upstream of start codon ATG was analyzed by PlantCARE online service,and the transcription factors which could bind the cis-acting elements was predicted by PlantRegMap software.The results were as follows:(1)SaNDH6 encoded 303 amino acids.It was a hydrophobin and located in chloroplast.(2)The phylogenetic tree analysis indicated that SaNDH6 had a more closely evolutionary relationship with NDH6 from woody plants.(3)Plant care analysis showed that the promoter sequence of SaNDH6 contained a large number of light responsive cis-acting elements such as ACE,AE-box,Box 4,G-Box and GT1-motif.It also contained abscisic acid(ABA)responsive element ABRE,jasmonic acid methyl ester(MeJA)responsive elements CGTCA-motif and TGACG-motif,gibberellin(GA3)responsive elements P-box,ARE cis-acting regulatory element essential for the anaerobic induction,and TC-rich repeats element involved in defense and stress responsiveness.(4)The results of plantRegMap analysis showed that there were 76 transcription factors that could bind to the SaNDH6 promoter,and among which,ERF transcription factor was the most(up to 40 TFs).(5)SaNDH6 can be expressed in the tissues of roots,heartwoods,calluses and leaves,but had a higher expression level in the tissue of leaves;under 1×104 mol·L-1 MeJA and GA3 treatments,the expression level of SaNDH6 were significantly elevated after 3 h when compared with 0 h,respectively.In conclusion,SaNDH6 was a nucleus gene encoding protein,its expression was induced by light and some hormones,and it might be involved in against some defense and stress processes in S.album.

Tetrastigma hemsleyanum is a perennial evergreen vine of the Vitaceae. The entire herb is used in traditional Chinese medicine as a broad-spectrum plant-based antibiotic, so it has high economic and social value. Wild T. hemsleyanum resources are scarce, so it has been declared an endangered and rare medicinal plant. Seed yield is low and vegetative propagation by cuttings results in limited plant production, so development of the T. hemsleyanum industry requires optimized propagation protocols and the development of new biotechnologies to proliferate this plant in commercial quantities. In this study, shoot organogenesis was successfully induced from leaves and petioles. Two plant growth regulators, 6-benzyladenine (BA) and thidiazuron, induced callus and adventitious shoots, but the ideal adventitious shoot induction medium was Murashige and Skoog (MS) medium containing 1.0 mg L−1 BA and 0.1 mg L−1 α-naphthaleneacetic acid (NAA). This resulted in a shoot proliferation coefficient (SPC) of 6.73 within 30 d at a light intensity of 100 µmol m−2 s−1. When light intensity was increased from 50 to 200 µmol m−2 s−1, SPC (7.35), chlorophyll a (Chl a), Chl b, and total Chl (a + b) content increased. On MS medium containing 0.1–2.0 mg L−1 NAA or indole-3-butyric acid, 100

Background The extract from Metasequoia glyptostroboides Hu et Cheng, a rare and endangered species native to China, exhibits numerous biological and pharmacological activities. The species is recalcitrant to rooting during micropropagation, a challenge that has yet to be resolved. In this study, transcriptomic and hormonal analyses were conducted to appreciate the molecular mechanism of adventitious root (AR) formation in optimized rooting conditions. Results The use of 2/5-strength Woody Plant Medium (WPM) significantly promoted AR formation of M. glyptostroboides shoots while the content of endogenous auxin, cytokinins and gibberellins (GAs) varied at different stages of AR formation. Transcriptomic analysis showed the significant up- or down-regulation of differentially expressed genes (DEGs) associated with plant hormone signal transduction and the phenylpropanoid biosynthesis pathway in response to 2/5-strength WPM. DEGs related to the biosynthesis of indole-3-acetic acid, cytokinins and GAs were identified. Transcript factors involved in 13 families were also revealed. A weighted gene co-expression network analysis indicated a strong correlation between hormones and genes involved in plant hormone signal transduction and the phenylpropanoid biosynthetic pathway. Conclusions These results indicate that the AR-promoting potential of 2/5-strength WPM in M. glyptostroboides was due to complex interactions between hormones and the expression of genes related to plant hormone signal transduction and the phenylpropanoid biosynthetic pathway.

Hippeastrum, a highly diverse genus in the Amaryllidaceae family, is a valuable ornamental bulbous flowering plant. Somatic embryogenesis (SE) is an efficient method for mass production of Hippeastrum plantlets. Previous studies have been devoted to the in vitro propagation of Hippeastrum, but the SE and its regulatory networks are rarely reported. In this study, we established a direct SE method of Hippeastrum ‘Bangkok Rose’ using leaf bases as explants. MS supplemented with 1.00 mg · L-1 NAA + 1.00 mg · L-1 KT + 0.25 mg · L-1 TDZ was the optimal medium for SE. Histological observations showed that the bipolar somatic embryo originated from the epidermal cell layer and underwent initiation, globular, scutellar and coleoptile stages. During SE, endogenous hormones of IAA, CTK, ABA, and SA were highly accumulated. Transcriptomic analysis revealed the genes encoding auxin biosynthesis/metabolic enzymes and efflux carriers were induced, while the auxin receptor of TIR1 and ARF transcriptional repressor of Aux/IAA were down-regulated and up-regulated, respectively, leading to suppression of auxin signaling. In contrast, cytokine signaling was promoted at the early stage of SE, as biosynthesis, transport, and signaling components were up-regulated. Various stress-related genes were up-regulated at the early or late stages of SE. Chromatin remodeling could also be dynamically regulated via distinct expression enzymes that control histone methylation and acetylation during SE. Moreover, key SE regulators, including WOXs and SERKs were highly expressed along with SE. Overall, the present study provides insights into the SE regulatory mechanisms of the Hippeastrum.

Axonopus compressus (Swartz) Beauv., a perennial herb in the Poaceae that has been introduced to a number of tropical and subtropical countries and regions, can serve as a lawn ground cover while its leaves can be developed as a bioenergy resource. It also displays some resistance to heavy metals, allowing it to be planted in areas and urban green spaces polluted by heavy metals. A. compressus is also used as a traditional Chinese medicine. This study is the first report on tissue culture of A. compressus. Stem explants induced shoot clusters on 6-benzyladenine (BA)-containing Murashige and Skoog (MS) medium, with 2.0 mg L–1 BA inducing a shoot proliferation coefficient (SPC) of 10.89 within 30 d, while MS medium containing 2.0 mg L–1 BA and 0.1 mg L–1 α-naphthaleneacetic acid (NAA) amplified SPC to 12.88 within 30 d. SPC on MS medium supplemented with kinetin (KIN) or thidiazuron (TDZ) was not as high as on medium with BA and never exceeded 3.57 within 30 d. A. compressus formed adventitious roots easily, within 15 d, most efficiently on ½MS medium supplemented with 0.1 to 0.5 mg L–1 IAA, NAA, or IBA, or even on auxin-free medium. Resulting plantlets displayed a high survival rate (> 98

The primary constituents of the essential oil derived from Santalum album L. are (Z)-α-santalol, (Z)-β-santalol, (Z)-α-exo-bergamotol, and (Z)-epi-β- santalol. SaCYP736A167 plays a pivotal role in the biosynthesis of these sesquiterpene alcohols within S. album, but the mechanisms governing the expression of the SaCYP736A167 gene is far from being deciphered. In this research, a promoter sequence of the SaCYP736A167 gene, spanning 2808 base pairs, was isolated from S. album. A bioinformatics analysis of the 2384-bp SaCYP736A167 promoter (PSaCYP736A167) showed that abundant stress-inducible cis-acting elements were distributed in different regions of PSaCYP736A167. The histochemical β-glucuronidase (GUS) staining of T1 transgenic Nicotiana tabacum plants harboring PSaCYP736A167 demonstrated that the predominant GUS activity was exhibited in the parenchyma cells of the stem cortex and phloem, suggesting that PSaCYP736A167 is a tissue-specific expression promoter. GUS fluorometric assays of transiently transgenic N. benthamiana leaves revealed that seven distinct segments of PSaCYP736A167 exhibited notably varied levels of GUS activity. A 936-base pair sequence upstream of the transcription initiation codon ATG constitutes the core promoter section of PSaCYP736A167. Our findings shed light on the regulatory mechanisms controlling the transcription of the SaCYP736A167 gene, potentially serving as a novel tissue-specific promoter for applications in transgenic plant biotechnology.

There are typographical errors with the pictures and tables in the original text, and these should be corrected in this corrigendum. The authors state that these corrections do not affect the methods, results, or conclusions of the original text.

Euryodendron excelsum H. T. Chang, a rare and endangered evergreen tree that is endemic to China. The micropropagation system of this species has been established, but some challenges associated with in vitro rooting remained to be improved. In this study, the in vitro rooting of E. excelsum plantlets were optimized by dark exposure, and the network of gene expression and endogenous hormones levels during dark-induced adventitious root (AR) formation were revealed. AR formation of E. excelsum plantlets were significantly promoted by dark exposure, especially by dark exposure for 15 d. In the stems of E. excelsum plantlets under the treatment of dark exposure for 15 d, lower level of abscisic acid (ABA), gibberellic acid 1 (GA 1 ), isopentenyladenine (IP), isopentenyladenosine (IPA) and zeatin (ZT), as well as higher level of GA 7 , jasmonic acid (JA) and salicylic acid (SA), promoted the whole course of AR formation. The higher level of trans-zeatin riboside (TZR) and T-zeatin (TZT) promoted the elongation of dark-induced AR, while higher level of indole-3-acetic acid (IAA) stimulated the process of AR primordia formation. Differentially expressed genes (DEGs) involved in hormone biosynthesis, plant hormone signal transduction and phenylpropanoid biosynthesis participated in the regulation of darkinduced AR development. The weighted gene co-expression network (WGCNA) analysis identified five modules that had highly correlation with phytohormone contents, and numerous hub genes associated with carotenoid biosynthesis, tryptophan metabolism, zeatin biosynthesis, alpha-Linolenic acid metabolism, phenylalanine metabolism, plant hormone signal transduction and phenylpropanoid biosynthesis were revealed. Those result will provide technical reference for in vitro rooting of woody species, and promote biological conservation and genetic engineering of rare and endangered species.

Sugarcane (Saccharum spp. hybrid) cultivar Yuetang 03-373 is a hybrid with superior agronomic characteristics that was obtained from the cross between Yuetang 92-1287 (♀) and Yuetang 93–159 (♂). In this study, transversely cut discs of immature leaves were used to induce adventitious shoots for the first time. Embryonic callus was induced from leaf explants on Murashige and Skoog (MS) medium with 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1 mg/l α-naphthaleneacetic acid (NAA). On MS medium containing 0.75 mg/l 6-benzyladenine and 0.1 mg/l NAA, 73.1

Thuarea involuta, a perennial herb that often grows on sandy beaches, has documented salt and drought tolerance, making it a useful plant species for the stabilization of island reefs. However, there are no protocols for its large-scale propagation, so effective in vitro protocols can assist to mass propagate healthy plants for ecorestoration. In this study, T. involuta stem segments were used as explants to establish a highly efficient in vitro shoot proliferation and regeneration system for the first time. The shoot proliferation coefficient (SPC) was highest when thidiazuron was used alone (SPC = 8.78) relative to the use of kinetin (KIN) or 6-benzyladenine (SPC = 6.53 and 8.30, respectively). Multiple shoots were proliferated on Murashige and Skoog (MS) medium supplemented with 1.0 mg L−1 BA and 0.1 mg L−1 α-naphthaleneacetic acid, leading to a SPC of 9.64 within 30 days. After individual shoots were transferred to MS medium with or without indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA), all formed roots within 15 days. In vitro plantlets transplanted to a peat and vermiculite (1:1) substrate displayed 96.67 An efficient in vitro shoot proliferation, callus induction, differentiation, plant regeneration and transplantation system in a salt-tolerant plant Thuarea involuta was established for the first time.