Maturation, conversion and metabolic profile of somatic embryos derived from Klussia odoratissimaMozaff as affected by light spectra ​

Maturation, conversion and metabolic profile of somatic embryos derived from Klussia odoratissimaMozaff as affected by light spectra ​ | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Maturation, conversion and metabolic profile of somatic embryos derived from Klussia odoratissimaMozaff as affected by light spectra ​ Farnoosh Khosravi, Fariba Rafiei, Morteza Ebrahimi, Mohammad Rabiei This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4938119/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 04 Nov, 2024 Read the published version in Plant Cell, Tissue and Organ Culture (PCTOC) → Version 1 posted 4 You are reading this latest preprint version Abstract The present study investigated potential application of somatic embryogenesis (SE) for conservation of Klussia odoratissima Mozaff. As the efficiency of SE greatly varies depending on the culture conditions including light, effects of five light spectrums including a fluorescent light and four types of light - emitting diode (LED) (red, blue, 3red-1blue, and3blue:1red) for an exposure time of a month on the maturation of K.odoratissima were also studied. To better understand the effect of light treatment on conversion of K.odoratissima cultures, two months after treatment, fresh and dry weight and metabolite contents were evaluated. Samples of the same age in situ plant and air-dry plant were also considered for relative comparison. Study of developmental stage of K.odoratissima under in vitro conditions revealed that a combination of 3red:1blue LED provided the highest number of cotyledonary (mature) embryos per callus. Hence, this light treatment was identified as the best treatment for the conversion of somatic embryos in K.odoratissima. Consequently, the highest fresh and dried weights were recorded in the cultures grown under 3blue:1red and blue LED treatment. Phthalide content of in situ plant was significantly greater than other treatments. Interestingly, the phthalide content was significantly higher in tissue culture samples compared to air-dry plants, which suggests tissue culture as an alternative technique for the production of bioactive compound in K.odoratissima. Cultures were incubated in 3blue:1red LED followed by blue LED possessed higher contents of phthalides. Application of LEDs is promising approach in micopropagation of K.odoratissima. Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Iranian wild celery, known as mountain celery (MC) or “Kelus” ( Klussia odoratissima Mozaff), is a perennial plant belonging to the family of Apiaceae. The plant has enormous economic and industrial applications, albeit being considered an endangered species. K. odoratissima grows in central Zagros Mountain ranges of Iran. It possesses a wide range of medicinal properties including sedative, antimicrobial, hypotensive and antioxidative activities (Ahmadipour et al. 2015 and references therein). The health benefits of K. odoratissima are mainly conferred by phthalides. Phthalides, including Butylidene phthalides and Ligustilides, are a group of secondary metabolites, which are recognized as the major flavoring component in K. odoratissima . Unlike most endemic plants that are vulnerable to herbivores’ browsing (Cubas et al. 2019 ), K. odoratissima is threatened by excessive and illegal harvest at early growth stage (Seifipour Naghneh et al. 2022 ). The production of secondary metabolites in plants is naturally influenced by some regional and environmental factors, which limit the commercialization of K. odoratissima production. The extraction of secondary metabolites from K. odoratissima in nature poses a risk of extinction. Plant tissue culture techniques have been recently highlighted as the means of in vitro production of secondary metabolites (Hashim et al. 2021 ). Somatic embryogenesis (SE) is a tissue culture technique in which somatic cells gain the capacity to produce embryos that finally develop into a whole plant (Aguilar-Hernández and Loyola-Vargas 2018 ; Garcia et al. 2019 ). This technique plays a vital role in plant regeneration and rapid reproduction (Tomiczak et al. 2019 ). The SE process includes several distinct stages: induction, embryo proliferation, maturation, and germination/conversion of embryos into plantlets (Ramírez-Mosqueda, 2022). In dicotyledonous plants, somatic embryos follow the same successive morphological developmental steps as described for zygotic embryogenesis: globular, heart-shaped, torpedo and cotyledonary (Ramírez-Mosqueda, 2022). According to recent studies, somatic embryos in suspension culture are useful tools for the generation of secondary metabolites (Murthy et al. 2023 ). This method has been successfully used to produce biomass and various secondary metabolites in several medicinal plant species such as Salvia officinalis (Kintzios et al.1999), Gentiana straminea (Cai et al. 2009 ), Rauwolfia serpentine (Harisaranraj et al. 2009), and Eurycoma longifolia Jack (Iriawati et al. 2014). These studies have demonstrated that the production of plant bioactive metabolites is affected by different biotic and abiotic elicitors. Application of elicitors appears to be a viable method for increasing secondary metabolites in medicinal plants. In this respect, light has emerged as one of the unique and efficient elicitors for enhancing the in vitro production of pharmacologically important secondary metabolites (Hashim et al. 2021 ). Various types of light (UV, fluorescent, and LEDs) have been reported as stimulators for the production of plant secondary metabolites (Hashim et al. 2021 ). In recent years, the application of light emitting diodes (LEDs) has attracted a significant attention as an alternative artificial light source for controlled environmental agriculture as well as plant tissue culture experiments (Almeida et al. 2019 ). LEDs have several advantages relative to traditional light sources including: small size, durability, long operating lifetime, and relatively cool emitting surfaces, which make them an ideal light source for indoor environments. An outstanding influence of LED lighting on plant regeneration and secondary metabolite production has been reported by numerous studies (Fan et al. 2013 ; Cio´c, et al. 2018; Almeida et al. 2019 ; Hashim et al. 2021 ). However, the response of different plants to LEDs has been inconsistent and there was no specific pattern among various species (Bula et al. 1999; Gupta and Jatothu 2013). In 1995, Munoz estimated that between18000-25000 wild plant species were at risk of extinction, and in vitro culture techniques were proposed as the sole solution for satisfactory conservation of the majority of endangered species. As for K. odoratissima , attempts to conserve the plant had little progress due to poor seed setting and germination as well as long-time seed dormancy (Askari-Khorasgani et al. 2013 ). Hence, tissue culture seems to be an effective method for conservation of K. odoratissima . Few reports have studies in vitro culture in K. odoratissima . Most researches have focused on traditional methods such as adjusting the basic medium and growth regulators, and its artificial light source was the traditional fluorescent lamp. Therefore, the objective of the present study was to investigate in vitro tissue culture in tandem with different LED spectra on maturation and development of somatic embryogenesis in K. odoratissima . Since areal parts of K. odoratissima are rich sources of phthalides, we have also compared the concentration of phthalide across samples grown under different light conditions. This study provides an effective regeneration method for conservation of K. odoratissima and could pave the way for the proliferation of this endangered plant in bioreactor systems. Material and Methods Plant Material and Seed Germination Seeds of K.odoratissima were collected from its natural habitat in Iran's “Chaharmahal and Bakhtiari” province. Seeds were first treated with 70% ethanol for 1 min, and then washed with sterile distilled water. Subsequently, seeds were dipped in freshly prepared hypochlorite sodium solution (1% w/v with a few droplets of tween-20) for 15 min. Seeds were then rinsed at least three times with sterile distilled water and cultured in a hormone-free (HF) MS basal medium (Murashige and Skoog) (pH 5.8) containing 3.0% (w/v) sucrose before incubating in a dark condition for 2 months at 4°C. Induction and Proliferation of Embryogenic Callus For the induction of embryogenic callus, we used three different explants including; cotyledonary leaves, hypocotyls (both derived from in vitro germinated seeds), and zygotic embryos. The explants were cultured in MS medium supplemented with 1 mg/L 2,4-dichlorophenoxyaceticacid (2,4-D) and 0.25 mg/L Kinetin as described by Ebrahimi et al.( 2018). The induction medium was supplemented with 30 g/L of sucrose and 7 g/L of agar and dispensed into petri dishes. All treatments had three replicates, each consisted of 10 explants. The cotyledonary leaves were cut in half and cultured with the adaxial side in the medium (Ebrahimi et al. 2018 ). The cultures were incubated at 25 ± 2°C in a dark culture room. After a month, the SE induction rate (% of explants containing SE/total explants) and the mean number of somatic embryos per explant were recorded. The embryo-forming capacity (EFC) index was also calculated using the following equations according to Raomai et al. ( 2014 ): EFC = SE induction rate × mean number of SEs per explant/100 Besides, embryogenic calli were sub-cultured in three cycles on MS basal medium containing 1mg/L of 2,4-D for embryogenic callus proliferation. To avoid genetic variation, we used single-embryogenic callus for the proliferation and the provision of samples for the light treatment experiments. The cultures were kept in the same condition as mentioned above. Light treatments In order to investigate the effects of different light spectra on maturation of somatic embryos derived from K. odoratissima , five light treatments were applied including a fluorescent lamp, red LED (660-675nm), blue LED (460-475nm), 3red:1blue LED and 3blue:1red LED (Fig. 1 ). Somatic embryos were incubated for a month on MS medium (pH 5.8) enriched with 30g/L sucrose and 6.2 g/L agar under diurnal temperature of 23 ± 1°C, the relative humidity of 60%, a 16L:8D photoperiod, and a photosynthetic photon flux density of 50 µmol m − 2 s − 1 . The experiment was conducted in a completely randomized design (CRD) with three replications containing 200mg somatic embryogenesis mass (approximately 80–110 embryos). Within each LED environment, petri dishes were rotationally switched their locations on a daily basis to assure of the homogeneity in the light intensity for all samples. After a month, the number of somatic embryos at each developmental stage (globular, heart, torpedo, and cotyledonary) was recorded. In addition, the number of regenerated embryos at each LED treatment was recorded. The samples were transferred on fresh MS medium and regenerated embryos were grown under the same LED conditions for another additional two months. Following this period, different characteristics including fresh weight (FW), dry weight (DW), leaf area index (mm 2 ) and chlorophyll content (mg/g FW) of samples were measured. Leaf area index was calculated using ImageJ software. The total chlorophyll content (Chl) and the content of carotenoids (Car) were quantified spectrophotometrically using the Lichtenthaler equations and represented as mg/g FW (Lichtenthaler 1978). Secondary metabolites To understand the effect of LEDs on metabolite profile of K. odoratissima , 2.5g fresh leaves from in vitro cultured plantlets as well as in situ plants were harvested for the GC-MS analysis. These plants had the same age (two months). Likewise, a sample of air-dry of K. odoratissima was also used to measure the concentration and composition of bioactive compounds. It is noteworthy that dry powder of K. odoratissima is available in the market year around. The headspace (HS) sampling technique was implemented to ensure preserving the volatile constituents and avoiding any loss. The procedures for GC-MS analysis of K. odoratissima , were previously described by Seifipour Naghneh et al. ( 2022 ). The relative percentage of metabolic constituents was evaluated from the total peak area (TIC) by apparatus software. Metabolic compounds were identified by comparing their mass spectra with those stored in Wiley NBS75K.L and NIST/EPA/NIH (2002 version, National Institute of Standards and Technology, Gaithersburg, MD, USA) mass spectral libraries using various search engines (PBM, Nist02). Given the area under the peak is a function of the compound's concentration in the sample, the GC-MS data were presented as the area under the peak (Trivedi et al. 2017 ; Khazaeie et al., 2023). Statistical analysis Data analysis was performed using the SAS software (v.9.3), and the averages with the standard errors were compared by one-way ANOVA. The Duncan’s multiple range test was used to separate the treatment means (P < 0.05). Results and Discussion Induction and Proliferation of Embryogenic Callus Somatic embryogenesis (SE) is a pivotal process in plant biotechnology that enables the regeneration of plants from vegetative or non-reproductive tissues. The efficiency of SE greatly varies depending on the plant species, type of explant, the culture conditions, and the growth regulators involved. In our study, the callus induction was started 12–14 days following the culture of explants and the percentage of SE after a month was evaluated. Our finding revealed a significant difference (P ≤ 0.05) in the induction of somatic embryos on explants derived from germinated seeds and zygotic embryos under laboratory conditions (Fig. 2a). Although callus formation occurred more rapidly in hypocotyl explants compared to others, none of resulting calli exhibited an embryogenic potential. Therefore, this explant was excluded from further analysis. Similarly, Ebrahimi et al. ( 2018 ) reported that K. odoratissima explants were unable to induce somatic embryos irrespective of callus formation on the hypocotyls. Based on our results, the percentage of explants containing embryos exceeded 60 and 40% in zygotic embryos and cotyledonary leaves, respectively. Moreover, the average number of embryos per cotyledonary leaf reached 16, whereas this number was eight for zygotic embryos (Fig. 2b). Although the EFC index was higher in cotyledonary leaves compared to zygotic embryos but the change was insignificant (Fig. 2c). Following the formation of embryogenic calli, somatic embryos were formed on lateral edges and adaxial surface of cotyledonary leaves in the fourth week. In contrast, the formation of embryo on zygotic embryos was induced after three weeks albeit without callus formation (Fig. 3 ). In the study reported by Hu et al. (2016), zygotic embryos produced callus in indirect SE, but they had a lower frequency of callus formation compared to other explants such as leaves or immature embryos. Alternatively, the callus formed from zygotic embryos may have a lower capacity to develop into somatic embryos compared to callus from other explants. While there is no strong evidence to support this hypothesis, this may be due to the lack of dedifferentiation ability of zygotic embryos (von Arnold et al. 2002 ). In particular, mature and well-developed embryos are less inclined to deviate towards callus formation. The ability of an explant to undergo direct or indirect embryogenesis was historically thought to be determined by the age of the explant. The further the explant is from the zygotic embryo stage, the more reprogramming (callus formation) needed to convert the explant into a somatic embryo (Merckle et al. 1995). On the other hand, young tissues of cotyledonary leaves contain higher levels of totipotency, which efficiently respond to laboratory culture conditions. Therefore, such tissues become less differentiated compared to zygotic embryos, making them more favorable for SE (Bhojwani and Razdan 1996 ; Thorpe 2007 ). Investigation into in vitro propagation of Quassia amara L. (Simaroubaceae) using the vegetative embryogenesis revealed that leaf and internode explants from older plants were ineffective due to severe phenolic exudation (Martin and Madassery 2005 ). On the other hand, cotyledon explants were superior for the induction of vegetative embryos. Embryos were developed from both axial side and cutting point of the embryonic axis, with more embryos forming at the proximal end compared to the middle and distal regions of the cotyledons. The effect of LED on developmental stages and conversion of somatic embryos In tissue culturing, environmental factors play a central role in plant morphogenesis (Gupta and Jatothu 2013). Developmental stages of somatic embryos have been shown to be influenced by light spectrum (Almeida et al. 2019 ). In the present research, the influence of light wavelength on development of somatic embryos of K. odoratissima formed per callus (200 mg of initial FW) was evaluated after a month (Fig. 4 ). Our morphological studies indicated that light treatment significantly affected the development and regeneration of somatic embryos in the K. odoratissima (Table 1 ). In this way, the number of cotyledonary (mature) embryos per callus under 3red:1blue treatment was significantly (P ≤ 0.05) greater than that under other light treatments (Table 2 ). Additionally, the somatic embryos subjected to 3red:1blue treatment, were able to undergo conversion phase (Table 2 ). Table 1 Analysis of variance for effect of different light treatments on embryogenic cell maturation and regeneration of K. odoratissima Source of variation df Mean of square Globular Heart Torpedo Cotyledonary Regenerated Light treatment 4 126.67 ns 17.17 ** 13.43 ** 417.57 ** 12.4 * Error 10 313.3 0.47 0.67 17.07 2.53 ns, *, ** means non-significant, significant at p < 0.05 and significant in p < 0.01, respectively. Table 2 Mean comparisons for effect of different light treatments on development and regeneration of somatic embryos in K. odoratissima Light treatment Mean number of somatic embryos Globular Heart Torpedo Cotyledonary Regenerated Flourescent 60 a ± 10.01 3.67 b ± 0.33 3.67 cd ±0.67 22.3 b ± 2.96 9.33 ab ± 0.67 Red LED 70 a ±10.4 3.33 b ± 0.34 5.33 b ±0.33 20.4 b ± 0.66 6.67 bc ± 1.67 Blue LED 60 a ±10.05 1.67 c ± 0.67 4.67 b ±0.66 12.7 c ± 2.67 5.67 bc ± 0.66 3Red:1Blue LED 66.7 a ±12.02 7.64 a ± 0.31 8 a ± 0.001 41.7 a ± 3.35 10.7 a ± 1.61 3Blue:1Red LED 53.3 a ±8.83 2 c ± 0.001 2.33 d ± 0.33 13 c ±1.01 7.33 c ± 0.33 Data are expressed as mean ± SE; Different letters indicate significant differences using the Duncan’s Multiple Range Test (p < 0.05). The minimum number of cotyledonary embryos was observed under blue LED and 3blue:1red LED (Table 2 ). In Fritillaria cirrhosa , the maximum number of SEs with cotyledonary leaves was observed in white light followed by a combination of 8red:1blue LED (Chen et al. 2020 ). On the other hand, in sugarcane, the highest number of somatic embryos reaching to maturation phase was reported under LED lights with the medium blue wavelength (Heringer et al. 2017 ). Therefore, depending on plant species, different types of growth chambers equipped with LED lights can be considered to achieve the best performance. LED lamps have become a common approach for optimizing plant growth conditions, and this approach can successfully be used for SE of several species such as Dimocarpus longan (Li et al. 2018 ), Saccharum spp. (Heringer et al. 2017 ) and Vitis vinifera (Tittmann et al. 2015 ). A review of literature showed that light is selectively perceived by plant photoreceptors including cryptochromes and phototropins. Although cryptochromes react with blue and UV-A lights, phytochromes are reactive to red and far-red light (Huang et al. 2014 ). The plant photoreceptors perceive different wavelengths and trigger complex signal transduction to modulate gene expression profile and subsequently induce developmental responses such as induction, maturation and conversion in somatic embryos (Pedmale et al. 2016 ; Li et al. 2018 ; Gupta and Jatothu 2013). The present findings showed that 3red:1blue LED played an important role in maturation of SE in K. odoratissima so that application of this light combination in SE could coordinate photoreceptors with proteins, ions, hormones and other factors to regulate gene expression patterns with accompanying changes in SE phases. Interestingly, we noticed that light treatment could affect synchronization of somatic embryos. In this regard, the embryos grown under red LED and 3red:1blue LED had more uniformity compared to other light conditions (Fig. 4 ). Synchronization of somatic embryo cultures could greatly increase the efficiency of SE for in vitro propagation, which makes the technique ideal to produce artificial seeds (Nadel et al. 1990 ; Alwael et al. 2017 ). Thus, if all resultant cultures could be synchronized, this would have shortened the duration of culture with concomitant increase in propagation efficiency and uniformity (Nadel et al. 1990 ). A review of literature indicated that researchers prefer to use hormonal treatments to increase synchronization in SE cultures. To the best of our knowledge, this report is the first to note that light treatment could also influence the uniformity of SE in plant species. Therefore, we suggest to consider light as an underlying factor in synchronization of SE in plant tissue culture technique. To better understand the effect of light treatment on conversion of K. odoratissima cultures, fresh and dry weight, leaf area index with chlorophyll and carotenoid contents were evaluated (Fig. 5 ). Our findings showed a significant variation in the fresh and dry weights for light treatments following additional two months of the experiment (Table 3 ). Table 3 Analysis of variance for effect of different light treatments on conversion of somatic embryos derived from K. odoratissima Source of variation df Mean of square FW (mg) DW (mg) LAI (mm 2 ) Chl (mg/g FW) Car(mg/g FW) Light treatment 4 9473.33 ** 93.45 ** 83.85 * 0.017 * 0.000037 ** Error 10 1044.33 8.83 20.51 0.004 0.000004 The symbols *, ** means significant at p < 0.05 and significant in p < 0.01, respectively. The highest fresh weight was recorded in the cultures grown under combination of 3blue:1red and blue LEDs followed by Fluorescent lamp. In contrast, the lowest fresh weight was obtained under the red LED (Table 4 ). The dry weights were significantly enhanced in the cultures treated under 3blue:1red LED and blue LED (p ≤ 0.05) whereas no significant difference was observed in the dry weight of the samples incubated under the red LED and 3red:1blue LED treatments (Table 4 ). Likewise, the same trend was recorded for leaf area index in the samples grown under the combination of 3blue:1red and blue LED lights (Table 4 ). Table 4 Mean comparisons for effect of different light treatments on growth performance of converted cultures in K. odoratissima Light treatment Mean number of somatic embryos FW (mg) DW (mg) LAI (mm 2 ) Chl (mg/g FW) Car (mg/g FW) Flourescent 587.01 a ± 19.43 52.51 b ± 1.61 22.16 ab ± 1.75 0.84 ab ±0.051 0.02 c ± 0.0011 Red LED 503.33 b ± 11.47 45.77 c ± 1.33 16.67 b ± 0.88 0.73 b ± 0.023 0.015 c ± 0.0006 Blue LED 587.04 a ± 33.12 54.75 a ± 2.89 25.88 a ± 4.27 0.85 a ± 0.029 0.021 ab ± 0.0011 3Red:1Blue LED 513.67 b ±7.84 46.63 c ± 0.88 17.02 b ± 0.58 0.75 b ± 0.023 0.018 bc ± 0.0003 3Blue:1Red LED 627.01 a ± 8.54 59.67 a ± 2.78 28.56 a ± 3.42 0.92 a ± 0.052 0.023 a ± 0.002 Data are expressed as mean ± SE; Different letters indicate significant differences using the Duncan’s Multiple Range Test (p < 0.05). The chlorophyll content is a measure of well-being of a plant under different conditions. Our finding showed that light quality significantly affected total chlorophyll and carotenoid contents in the leaves of K. odoratissima plantlets (Table 3 ). The chlorophyll content was correlated with the culture growth in the present study. Consequently, cultures exposed to the 3blue:1red LED and blue LED exhibited the highest content of chlorophyll and carotenoids and the cultures grown under red LED showed the lowest content of the pigments (Table 4 ). Our results are consistent with those reporting plants grown under blue light had a higher chlorophyll content in comparison to plants grown under red light (Fan et al. 2013 ; Hung et al. 2016 ). Blue light seems to play a crucial role in the activity of photosystem I and II as well as the photosynthetic electron transport capacity (Miao et al. 2016 ). In plant species such as Gerbera jamesonii and Myrtus communis , the accumulation of photosynthetic pigments was reduced when plants were subjected to red LED (Pawłowska et al. 2018 ; Cio´c et al. 2018). In line with our findings, the accumulation of carotenoids in Sinningia speciosa (Simlat et al. 2016 ) and Stevia rebaudiana (Zheng and van Labeke 2017 ) cultures was stimulated by blue light spectrum. Findings from the present study, corroborate earlier literature that reported light quantity, quality and duration were involved in controlling morphogenesis, growth and differentiation of plant cells, tissues and organ cultures (Moshe and Dalia 2007 ). Different sources of LED provide wavelengths that are matched with plant photoreceptors, which leads to optimal production and metabolome (Bourget 2008 ; Massa et al. 2008 ; Morrow 2008 ). Plants apparently sense changes in light quality through photoreceptors, which in turn regulate growth and development through modulating signaling pathways (Rechenmacher et al. 2010 ). Light treatments and secondary metabolites In plants, environmental factors have a significant role in the biosynthesis of secondary metabolites (Khajali and Rafiei 2024 ). Several studies have identified the influence of light related factors such as light intensity, light quality, and light period on metabolite accumulation of different medicinal plants (Khazaeie et al. 2023 and the references therein). Despite a great deal of information on the application of LEDs on plant secondary metabolites, limited information is available on optimizing LEDs to improve secondary metabolites in endangered plant species. We demonstrated the effect of different light treatments on the metabolic compounds in in vitro grown aerial parts of the K. oddoratisma. Chemical composition of K. oddoratisma among different conditions including in situ , dried powder sample from in situ plant, florescent, red, blue, 3red:1blue and 3blue:1red indicated 14, 6, 9, 5, 8, 8 and 9 bioactive constituents, respectively (Table 5 , 6 ). A more diverse variation of bioactive compounds was recorded for two-old-month fresh plant grows in situ (Table 6 ). The composition of plant secondary metabolites is affected by different factors including environmental condition, interaction between genotype and environment, method of distillation, storage condition, physiological stage, time of harvest, and season (Khajali and Rafiei 2024 ). Table 5 The composition and concentration (AUP/FW) of bioactive compounds in K. odoratissima cultures grown under different light treatment and two-month-old dried sample RT Compound’s name KI Cal Flourescent Red Blue 3Red:Blue 3Blue:Red Dried sample 14.209 3,9-Epoxy-p-mentha-1,8(10)-diene 1199 247181 ± 8421 19.895 Neophytadiene 1817 280265 ± 10023 18.8 (Z-Butylidenephthalide) 1675 3880705 ± 32421 1238428 ± 17400 3617628 ± 40221 2839095 ± 18664 4374798 ± 18728 1192851 ± 13235 19.447 ((E)-Ligustilide) 1830 6294434 ± 29671 1694471 ± 23551 6819459 ± 33265 4956396 ± 50210 6364303 ± 37425 1020396 ± 29840 16.955 isoledene 1373 - - 442749 ± 2655 14.978 (α-Copaene) 1377 320392 ± 9885 - 986654 ± 7677 481636 ± 8965 325730 ± 10245 336192 ± 5922 16.94 (β-Isocomene) 1412 - - - 170230 ± 8545 15.679 (β-Copaene) 1430 2621460 ± 4744 357727 ± 11250 7446177 ± 12565 4300660 ± 6700 3090324 ± 12024 15.553 .beta.-ylangene (β-Ylangene) 1423 1225541 ± 5860 289332 ± 8600 4901094 ± 12433 2773455 ± 14254 2599152 ± 7855 15.158 (Germacrene D) 1484 374215 ± 13442 894846 ± 14000 431077 ± 1977 444565 ± 2166 16.58 6.alpha.-Cadina-4,9-diene, (-)- - - 2122920 ± 12922 1239421 ± 8500 1572100 ± 23412 16.585 .gamma.-Muurolene (γ-Muurolene) 1485 1197285 ± 11971 - - - - 15.927 trans-.beta.-Farnesene ((E)-β-Farnesene) 1473 6052262 ± 26329 20644815 ± 21459 12431334 ± 54200 9108300 ± 32950 15.942 cis-.beta.-Farnesene ((Z)-β-Farnesene) 1476 - 1341066 ± 8710 - - - 16.955 (-)-.alpha.-Panasinsen (α-Panasinsanene) 1527 945045 ± 10200 - - Others 3468376 ± 75980 3238994 ± 47684 3238994 ± 67234 2456886 ± 33865 693234 ± 20124 2594982 ± 21000 Data are expressed as mean ± SE Table 6 The composition and concentration (AUP/FW) of bioactive compounds in two-month-old in situ K. odoratissima RT Compound’s name KI Cal Two-month-old 13.371 D-Limonene 1046 65608 ± 446 23.239 β-Cubebene 1390 74028 ± 4235 24.437 Elixene 1453 1988650 ± 24004 24.812 (E)-β-Farnesene 1473 1311422 ± 14221 25.377 Germacrene D 1484 440701 ± 6412 25.274 γ-Cadinene 1497 49992 ± 2810 25.820 δ-Cadinene 1526 28151 ± 108 28.322 Z-Butylidene phthalide 1675 1234885 ± 28760 29.398 (E)-3-Butylidene phthalide 1744 194856 ± 4650 29.715 Z-Ligustilide 1765 24982212 ± 78700 30.625 E-Ligustilide 1830 702727 ± 9695 30.907 Caffeine 1582 8972991 ± 38439 6.876 (E)-2-Hexenal 866 1669101 ± 19010 15.951 Nonanal 1105 786261 ± 8218 Data are expressed as mean ± SE The natural habitat of K. oddoratisma is recognized by an altitude of > 2500m and the temperature of barely above 20°C during the vegetative stage (Seifipour Naghneh et al. 2022 ), which are totally different from a controlled condition. It is presumed that such unique growth environment could differently affect emission and composition of secondary metabolite in K. oddoratisma grown in situ compare to those obtained from tissue culture technique. In the present study, the composition and the relative concentration of bioactive compounds were significantly changed across different light conditions (Table 5 ). Phthalides constituted a large amount of secondary metabolites in K. odoratissima . These compounds are known to have a wide range of health benefits including antibacterial, antifungal, insecticidal, cytotoxic, anti-inflammatory properties (Seifipour Naghneh et al. 2022 and the references therein). In Drosera rotundifolia , the amount of secondary metabolites was significantly higher, in the field grown samples compared to in vitro plantlets (Tienaho et al. 2021 ). Wilken et al. ( 2005 ) used different tissue culture techniques in three different plant species including Hypericum perforatum , Cymbopogon citratus and Fabiana imbricate. They reported that the concentration of secondary metabolites produced in tissue culture technique was significantly lower when compared to the field grown plants. Interestingly, phthalide content was significantly (p ≤ 0.05) higher in tissue culture samples compared to dried sample derived from in situ plant (Table 5 ). In our study, under in vitro condition, 3blue:1red LED followed by blue LED and fluorescent light were the best treatments for production of phthalides. However, the concentration of phthalides was minimal in cultures grown under red LED condition (Table 5 ). Secondary metabolites are compounds that play an important role in the interaction of plants with their environment (Oksman-Caldentey and Inzé 2005; Khajali and Rafiei 2024 ). Therefore, it is perceived that their biosynthesis in plant tissues is modified by changing environmental conditions including light (Tomaszewiczet al., 2022 ). In C. delgadii plants, blue light was the most efficient for the accumulation of flavonoid glycosides, such as astragalin and rutin. A stimulatory effect of this light quality on the accumulation of secondary metabolites was also noticed for other species, e.g., Rhodiola imbricate (Kapoor et al. 2018 ) and Capsicum annuum (Yap et al. 2021 ). It has been reported that blue light stimulates the biosynthesis of secondary metabolites in plants and in the meantime, enhances the synthesis of compounds such as antioxidants. Such increase is attributed to the specific wavelength of blue light that affects plant metabolic pathways (Larsen et al. 2022 ). Under in vitro condition, Manivannan et al. ( 2015 ) demonstrated that blue LED treatment significantly increased the total phenol and flavonoid contents in the aerial parts of Rehmannia glutinosa . Moreover, Lian et al. ( 2019 ) reported that blue LED was associated with increased antioxidant activity and enhanced contents of total flavonoid and phenolic compounds in Gynura procumbens , with a notable increase in cyanidin-monoglucosides under blue light. A recent study indicated that high-intensity blue light supplementation for 4 hours during the night could induce the expression of MYBs, CRY2/3, SPAs, and HY5, thereby enhancing anthocyanin and catechin accumulation in tea leaves (Zheng et al. 2019 ). Furthermore, Wang et al. ( 2020 ) reported that blue light extensively regulates multiple physiological processes and secondary metabolism in young tea shoots, significantly affecting gene transcription primarily in the pathways of photosynthesis, lipid metabolism, and flavonoid synthesis under high-intensity blue light. Recent studies shows blue light has a significant impact on the biosynthesis of terpenes and cannabionoids in plants by enhancing the activity of key biosynthetic pathways and enzymes (Morello 2022; Desaulniers Brousseau 2021). Although application of LEDs in plant regeneration is promising, but any given plant species needs ascertaining wavelengths in order to maximize the yield and quality (Khazaeie et al. 2023). Based on analysis of GC–MS data, we found that 3blue:1red LED followed by blue LED and fluorescent light were the best environments for the production of phthalide, the most outstanding secondary compound in K. odoratissima. It seems that these light treatments can favorably affect light-responsive genes and such changes corresponded to biosynthesis of phthalides in K. odoratissima . Conclusion In general, it was noticed that the cotyledonary leaves were the most competent explant for somatic embryogenesis in K. odoratissima under the conditions of this study. The developmental stage of K. odoratissima under in vitro conditions requires a specific lighting treatment. In this regard, a combination of 3red:1blue LED provided the highest number of cotyledonary (mature) embryos per callus. This light treatment was the best treatments for conversion of somatic embryos in K. odoratissima . Consequently, the highest fresh and dried weights were recorded in the cultures grown under 3blue:1red and blue LED treatment. Interestingly, phthalide content was significantly (p ≤ 0.05) greater in tissue culture samples compared to dried powder of two-month-old in situ plants, which suggests tissue culture as an alternative technique for the production of bioactive compound in K. odoratissima . Cultures were incubated in 3blue:1red LED followed by blue LED and florescent light possessed higher contents of phthalides. Based on this research, it was shown that LEDs can be an effective alternative to florescent lamps in micropropagation of K. odoratissima . Declarations Acknowledgment Authors acknowledge Shahrekord University and Agricultural Biotechnology Research Institute of Iran-Central Branch (ABRII-CB) for spiritual and financial supporting. This research was part of a project in ABRII-CB entitled “Optimization of shoot culture and production of Kelus ( Kelussia odoratissima Mozaff.) in bioreactor- ID: 4-05-05-010-010551”. Funding This research was part of a project in ABRII-CB entitled “Optimization of shoot culture and production of Kelus ( Kelussia odoratissima Mozaff.) in bioreactor- ID: 4-05-05-010-010551”. Morteza Ebrahimi has received research support from “Agricultural Biotechnology Research Institute of Iran-Central Branch (ABRII-CB) “ Competing Interests The authors have no relevant financial or non-financial interests to disclose. Author Contributions All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Morteza Ebrahimi, Fariba Rafiei, Farnoosh Khosravi and Mohammad Rabiei. The first draft of the manuscript was written by Fariba Rafiei and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. References Aguilar-Hernández V, Loyola-Vargas VM (2018) Advanced proteomic approaches to elucidate somatic embryogenesis. Front Plant Sci 9:1658. https://doi.org/10.3389/fpls.2018.01658 Ahmadipour B, Hassanpour H, Asadi E, et al (2015) Kelussia odoratissima Mozzaf - a promising medicinal herb to prevent pulmonary hypertension in broiler chickens reared at high altitude. J Ethnopharmacol 159:49–54. https://doi.org/10.1016/j.jep.2014.10.043 Almeida FA, Vale EM, Reis RS, et al (2019) LED lamps enhance somatic embryo maturation in association with the differential accumulation of proteins in the Carica papaya L. “Golden” embryogenic callus. Plant Physiol Biochem 143:109–118. https://doi.org/10.1016/j.plaphy.2019.08.029 Alwael HA, Naik PM, Al-Khayri JM (2017) Synchronization of Somatic Embryogenesis in Date Palm Suspension Culture Using Abscisic Acid. In: Al-Khayri JM, Jain SM, Johnson DV (eds) Date Palm Biotechnology Protocols Volume I: Tissue Culture Applications. Springer New York, New York, NY, pp 215–226. https://doi.org/10.1007/978-1-4939-7156-5_18 Askari-Khorasgani O, Mortazaeinezhad F, Otroshy M, Golparvar AR (2013) Breaking seed dormancy of endangered medicinal plant Kelussia odoratissima using zygotic embryo culture technique. ISSN 2051:0853 Bhoite HA, Palshikar GS (2014) Plant Tissue Culture: A Review. World J Pharm Sci 565–572 Bhojwani SS, Razdan MK (1996) Plant tissue culture: Theory and practice: Theory and practice. Elsevier Science & Technology Bourget CM (2008) An introduction to light-emitting diodes. HortScience 43:1944-1946 Bowes BG (1999) A Colour Atlas of Plant Propagation and Conservation. Manson Publishing, London, England Bula RJ, Morrow RC, Tibbitts TW, et al (1991) Light-emitting diodes as a radiation source for plants. HortScience 26:203–205. https://doi.org/10.21273/hortsci.26.2.203 Cai Y, Liu Y, Liu Z, et al (2009) High-frequency embryogenesis and regeneration of plants with high content of gentiopicroside from the Chinese medicinal plant Gentiana straminea Maxim. In Vitro Cellular & Developmental Biology - Plant 45:730–739. https://doi.org/10.1007/s11627-009-9225-7 Chen C-C, Lee M-R, Wu C-R, et al (2020) LED Lights Affecting Morphogenesis and Isosteroidal Alkaloid Contents in Fritillaria cirrhosa D. Don-An Important Chinese Medicinal Herb. Plants 9:. https://doi.org/10.3390/plants9101351 Cioć M, Szewczyk A, Żupnik M, et al (2018) LED lighting affects plant growth, morphogenesis and phytochemical contents of Myrtus communis L. in vitro. Plant Cell Tissue Organ Cult 132:433–447. https://doi.org/10.1007/s11240-017-1340-2 Cubas J, Irl SDH, Villafuerte R, et al (2019) Endemic plant species are more palatable to introduced herbivores than non-endemics. Proc Biol Sci 286:20190136. https://doi.org/10.1098/rspb.2019.0136 Desaulniers Brousseau V, Wu B-S, MacPherson S, et al (2021) Cannabinoids and terpenes: How production of photo-protectants can be manipulated to enhance cannabis sativa L. phytochemistry. Front Plant Sci 12:620021. https://doi.org/10.3389/fpls.2021.620021 Dutta Gupta S, Jatothu B (2013) Fundamentals and applications of light-emitting diodes (LEDs) in in vitro plant growth and morphogenesis. Plant Biotechnol Rep 7:211–220. https://doi.org/10.1007/s11816-013-0277-0 Ebrahimi M, Mokhtari A, Amirian R (2018) A highly efficient method for somatic embryogenesis of Kelussia odorotissima Mozaff., an endangered medicinal plant. Plant Cell Tissue Organ Cult 132:99–110. https://doi.org/10.1007/s11240-017-1314-4 Ebrahimie E, Habashy AA, Mohammadie-Dehcheshmeh M, et al (2006) Direct shoot regeneration from mature embryo as a rapid and genotype-independent pathway in tissue culture of heterogeneous diverse sets of cumin ( Cuminum cyminum L.) genotypes. In Vitro Cellular & Developmental Biology - Plant 42:455–460. https://doi.org/10.1079/IVP2006789 Fan X, Zang J, Xu Z, et al (2013) Effects of different light quality on growth, chlorophyll concentration and chlorophyll biosynthesis precursors of non-heading Chinese cabbage ( Brassica campestris L.). Acta Physiol Plant 35:2721–2726. https://doi.org/10.1007/s11738-013-1304-z Garcia C, Furtado de Almeida A-A, Costa M, et al (2019) Abnormalities in somatic embryogenesis caused by 2,4-D: an overview. Plant Cell Tiss Organ Cult 137:193–212. https://doi.org/10.1007/s11240-019-01569-8 Guppta, S.D., Jatothu, B., 2013. Fundamentals and applications of light-emitting diodes (LEDs) in in vitro plant growth and morphogenesis. Plant Biotechnol Rep 7: 211–220. Harisaranraj R, Suresh K. (2009). Evaluation of the Chemical Composition Rauwolfia serpentina and Ephedra vulgaris . ABR 3:174-178. Hashim M, Ahmad B, Drouet S, et al (2021) Comparative Effects of Different Light Sources on the Production of Key Secondary Metabolites in Plants In Vitro Cultures. Plants 10:. https://doi.org/10.3390/plants10081521 Heringer AS, Reis RS, Passamani LZ, et al (2017) Comparative proteomics analysis of the effect of combined red and blue lights on sugarcane somatic embryogenesis. Acta Physiol Plant 39:52. https://doi.org/10.1007/s11738-017-2349-1 Hu R, Sun Y, Wu B, et al (2017) Somatic Embryogenesis of Immature Cunninghamia lanceolata (Lamb.) Hook Zygotic Embryos. Sci Rep 7:56. https://doi.org/10.1038/s41598-017-00156-1 Huang X, Ouyang X, Deng XW (2014) Beyond repression of photomorphogenesis: role switching of COP/DET/FUS in light signaling. Curr Opin Plant Biol 21:96–103. https://doi.org/10.1016/j.pbi.2014.07.003 Hung CD, Hong C-H, Kim S-K, et al (2016) LED light for in vitro and ex vitro efficient growth of economically important highbush blueberry ( Vaccinium corymbosum L.). Acta Physiol Plant 38:1–9. https://doi.org/10.1007/s11738-016-2164-0 Iriawati, Rahmawati A, Esyanti RR (2014) Analysis of secondary metabolite production in somatic embryo of pasak bumi ( Eurycoma longifolia Jack.). Procedia Chem 13:112–118. https://doi.org/10.1016/j.proche.2014.12.014 Kapoor S, Raghuvanshi R, Bhardwaj P, et al (2018) Influence of light quality on growth, secondary metabolites production and antioxidant activity in callus culture of Rhodiola imbricata Edgew. J Photochem Photobiol B 183:258–265. https://doi.org/10.1016/j.jphotobiol.2018.04.018 Khajali F and Rafiei F (2024) A review of plant anti-nutritional factors in animal health and production: the classification, biological properties, and the passivation strategy. JAF Res 18:101290. https://doi.org/10.1016/j.jafr.2024.101290 Khazaei M, Rafiei F, Sabzalian MR, Houshmand S (2023) Potential application of light emitting diodes to improve molecular, physiological, and growth responses in peppermint plant. JSFA Rep 3:663–673. https://doi.org/10.1002/jsf2.177 Kintzios S, Nikolaou A, Skoula M (1999) Somatic embryogenesis and in vitro rosmarinic acid accumulation in Salvia officinalis and S. fruticosa leaf callus cultures. Plant Cell Reports 18:462–466. https://doi.org/10.1007/s002990050604 Kumar Trivedi M (2017) Chromatographic and spectroscopic characterization of the consciousness energy healing treated Withania somnifera (ashwagandha) root extract. Eur J Biophys 5:38. https://doi.org/10.11648/j.ejb.20170502.12 Larsen DH, Li H, Shrestha S, et al (2022) Lack of blue light regulation of antioxidants and chilling tolerance in basil. Front Plant Sci 13:852654. https://doi.org/10.3389/fpls.2022.852654 Li H, Lin Y, Chen X, Bai Y, Wang C, Xu X, et al. (2018) Effects of blue light on flavonoid accumulation linked to the expression of miR393, miR394 and miR395 in longan embryogenic calli. PLoS One 13:e0191444. https://doi.org/10.1371/ journal.pone.0191444 Lian TT, Moe MM, Kim YJ, Bang KS (2019) Effects of different colored LEDs on the enhancement of biologically active ingredients in callus cultures of Gynura procumbens (lour.) Merr. Molecules 24:4336. https://doi.org/10.3390/molecules24234336 Lichtenthaler HK (1987) Chlorophylls and Carotenoids: Pigments of photosynthetic biomembranes. In: Methods in Enzymology. Elsevier, pp 350–382 Manivannan A, Soundararajan P, Halimah N, et al (2015) Blue LED light enhances growth, phytochemical contents, and antioxidant enzyme activities of Rehmannia glutinosa cultured in vitro. Hortic Environ Biotechnol 56:105–113. https://doi.org/10.1007/s13580-015-0114-1 Martin KP, Madassery J (2005) Direct and indirect somatic embryogenesis on cotyledon explants of Quassia amara L., an antileukaemic drug plant. In Vitro Cell Dev Biol Plant 4:54–57. https://doi.org/10.1079/IVP2004588 Massa GD, Kim HH, Wheeler R, Mitchel AC (2008) Plant Productivity in Response to LED Lighting. HortScience 43:1951-1956. Merkle SA, Parrott WA, Flinn BS (1995) Morphogenic Aspects of Somatic Embryogenesis. In: Thorpe, T.A., Ed., In Vitro Embryogenesis in Plants, Kluwer Academic Publishers, Dondrecht, 155-203. https://doi.org/10.1007/978-94-011-0485-2_5 Miao Y-X, Wang X-Z, Gao L-H, et al (2016) Blue light is more essential than red light for maintaining the activities of photosystem II and I and photosynthetic electron transport capacity in cucumber leaves. Journal of Integrative Agriculture 15:87–100. https://doi.org/10.1016/S2095-3119(15)61202-3 Moshe R and Dalia E (2007) On the effect of light on shoot regeneration in petunia. Plant Cell Tissue Organ Cult. 89: 49-54. Morello V, Brousseau VD, Wu N, et al (2022) Light quality impacts vertical growth rate, phytochemical yield and cannabinoid production efficiency in cannabis sativa. Plants 11:2982. https://doi.org/10.3390/plants11212982 Morrow RC (2008) LED lighting in horticulture. HortScience 43:1947–1950. Munoz MC (1995) Micropropagation of endangered plants. Ecologia Medi 21:291-297. Murthy HN, Joseph KS, Hahn J-E, et al (2023) Suspension culture of somatic embryos for the production of high-value secondary metabolites. Physiol Mol Biol Plants 29:1153–1177. https://doi.org/10.1007/s12298-023-01365-x Nadel BL, Altman A, Ziv M (1990) Regulation of somatic embryogenesis in celery cell suspensions. Plant Cell Tissue Organ Cult 20:119–124. https://doi.org/10.1007/BF00114709 Nhut DT, Huy NP, Tai NT, et al (2015) Light-emitting diodes and their potential in callus growth, plantlet development and saponin accumulation during somatic embryogenesis of Panax vietnamensis Ha et Grushv. Biotechnol Biotechnol Equip 29:299–308. https://doi.org/10.1080/13102818.2014.1000210 Oksman-Caldentey K-M, Inzé D (2004) Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. Trends Plant Sci 9:433–440. https://doi.org/10.1016/j.tplants.2004.07.006 Pawłowska B, Żupnik M, Szewczyk-Taranek B, Cioć M (2018) Impact of LED light sources on morphogenesis and levels of photosynthetic pigments in Gerbera jamesonii grown in vitro. Hortic Environ Biotechnol 59:115–123. https://doi.org/10.1007/s13580-018-0012-4 Pedmale UV, Huang S-SC, Zander M, et al (2016) Cryptochromes interact directly with PIFs to control plant growth in limiting blue light. Cell 164:233–245. https://doi.org/10.1016/j.cell.2015.12.018 Ramírez-Mosqueda MA (2022a) Overview of Somatic Embryogenesis. In: Ramírez-Mosqueda MA (ed) Somatic Embryogenesis: Methods and Protocols. Springer US, New York, NY, pp 1–8 Ramírez-Mosqueda MA (ed) (2022b) Somatic embryogenesis: Methods and protocols, 1st edn. Springer, New York, NY Raomai S, Kumaria S, Tandon P (2014) Plant regeneration through direct somatic embryogenesis from immature zygotic embryos of the medicinal plant, Paris polyphylla Sm. Plant Cell Tissue Organ Cult 118:445–455. https://doi.org/10.1007/s11240-014-0496-2 Rechenmacher C, Schmitt JL, Droste A (2010) Spore germination and gametophyte development of Cyathea atrovirens (Langsd. & Fisch.) Domin (Cyatheaceae) under different pH conditions. Braz J Biol 70:1155–1160. https://doi.org/10.1590/s1519-69842010000600004 Seifipour Naghneh N, Rafiei F, Shahbazi E, Gheisari MM (2022) Chemical bioactive profile of mountain celery as affected by vegetative stages. Nat Prod Res 36:5643–5646. https://doi.org/10.1080/14786419.2021.2022667 Simlat M, Ślęzak P, Moś M, et al (2016) The effect of light quality on seed germination, seedling growth and selected biochemical properties of Stevia rebaudiana Bertoni. Sci Hortic (Amsterdam) 211:295–304. https://doi.org/10.1016/j.scienta.2016.09.009 Suresh K, Babu SS (2009) Production of Reserpine in Somatic Embryos of Rauwolfia serpentina Cultured in Bioreactors by the Induction of Elicitor (Methyl Jasmonate) Thorpe TA (2007) History of plant tissue culture. Mol Biotechnol 37:169–180. https://doi.org/10.1007/s12033-007-0031-3 Tienaho J, Reshamwala D, Karonen M, et al (2021) Field-Grown and In Vitro Propagated Round-Leaved Sundew ( Drosera rotundifolia L.) Show Differences in Metabolic Profiles and Biological Activities. Molecules 26:. https://doi.org/10.3390/molecules26123581 Tittmann S, Borner M, Harst M, Bleser E (2015) Influence of led-illumination to the regeneration potential of somatic embryos of Vitis vinifera 'Chardonnay' - preliminary studies. Acta Hortic. (Wagening.) 1083:445–453. https://doi.org/10.17660/ ActaHortic.2015.1083.57. Tomaszewicz W, Cioć M, Dos Santos Szewczyk K, et al (2022) Enhancing In Vitro Production of the Tree Fern Cyathea delgadii and Modifying Secondary Metabolite Profiles by LED Lighting. Cells 11:. https://doi.org/10.3390/cells11030486 Tomiczak K, Mikuła A, Niedziela A, et al (2019) Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application. Front Plant Sci 10:762. https://doi.org/10.3389/fpls.2019.00762 Trivedi MK, Branton A, Trivedi D, et al . (2017) Chromatographic and spectroscopic characterization of the consciousness energy healing treated Withania Somnifera (ashwagandha) root extract. Euro J Biophysic 5:38-47. https:// doi.org/ 10.11648/j.ejb.20170502.12. Wang P, Chen S, Gu M, et al (2020) Exploration of the effects of different blue LED light intensities on flavonoid and lipid metabolism in tea plants via transcriptomics and metabolomics. Int J Mol Sci 21:4606. https://doi.org/10.3390/ijms21134606 Wilken D, González EJ, Hohe A, et al (2005) Comparison of secondary plant metabolite production in cell suspension, callus culture and temporary immersion system. In: liquid culture systems for in vitro plant propagation. Springer, Netherlands, pp. 525–537. von Arnold S, Sabala I, Bozhkov P, et al (2002) Developmental pathways of somatic embryogenesis. Plant Cell Tissue Organ Cult 69:233–249. https://doi.org/10.1023/A:1015673200621 Xu Y, Yang M, Cheng F, et al (2020) Effects of LED photoperiods and light qualities on in vitro growth and chlorophyll fluorescence of Cunninghamia lanceolata . BMC Plant Biol 20:269. https://doi.org/10.1186/s12870-020-02480-7 Yan J, Peng P, Duan G, et al (2021) Multiple analyses of various factors affecting the plantlet regeneration of Picea mongolica (H. Q. Wu) W.D. Xu from somatic embryos. Sci Rep 11:6694. https://doi.org/10.1038/s41598-021-83948-w Yap ESP, Uthairatanakij A, Laohakunjit N, et al (2021) Plant growth and metabolic changes in “Super Hot” chili fruit ( Capsicum annuum ) exposed to supplemental LED lights. Plant Sci 305:110826. https://doi.org/10.1016/j.plantsci.2021.110826 Zheng C, Ma J-Q, Ma C-L, et al (2019) Regulation of growth and flavonoid formation of tea plants ( Camellia sinensis ) by blue and green light. J Agric Food Chem 67:2408–2419. https://doi.org/10.1021/acs.jafc.8b07050 Zheng L, Van Labeke M-C (2017) Long-term effects of red- and blue-light emitting diodes on leaf anatomy and photosynthetic efficiency of three ornamental pot plants. 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Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4938119","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":349130260,"identity":"acce6bbe-f41f-4338-a7c4-95a02fa2140f","order_by":0,"name":"Farnoosh Khosravi","email":"","orcid":"","institution":"Shahrekord University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Farnoosh","middleName":"","lastName":"Khosravi","suffix":""},{"id":349130261,"identity":"724e141d-635f-4155-92b9-ed2dd70cc5de","order_by":1,"name":"Fariba Rafiei","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAUlEQVRIiWNgGAWjYBAC9gYGhsNgFvPBBoaEChCDuQGvFp4DMC1siUAtZ0BaGAlrYYZoSWBgYGwDsQhpYT/78HBBDUM+fxtz44OH82qj+duBWn5UbMOthSfd4PCMYwyWM44xNhskbjueO+MwYwNjz5nbOLXYM6QxHOZhYzBguN/YJpG47VhuA1ALM2Mbbi08/M+AWv4xGMgfY2z/kTjnWO58glokgLbwtjEYGBwD+j2xoSZ3A2EtQFtm9kkYGAL9IpFw7EDuRqCWg/j8wsOfxvy54JuNgdwx9ocff9TU5c47f/jggx8VuLVAgQSMAYnWA4TUI4M6UhSPglEwCkbBCAEARE5afUGYR8IAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-7695-1588","institution":"Shahrekord University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Fariba","middleName":"","lastName":"Rafiei","suffix":""},{"id":349130262,"identity":"36c594f5-e3a2-48ab-b123-3cf3e6cc0dde","order_by":2,"name":"Morteza Ebrahimi","email":"","orcid":"","institution":"ABRII: Agricultural Biotechnology Research Institute of Iran","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Morteza","middleName":"","lastName":"Ebrahimi","suffix":""},{"id":349130263,"identity":"f4e2b742-c090-4cf2-935f-909b6dbcf8ea","order_by":3,"name":"Mohammad Rabiei","email":"","orcid":"","institution":"Shahrekord University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Rabiei","suffix":""}],"badges":[],"createdAt":"2024-08-19 10:59:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4938119/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4938119/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11240-024-02903-5","type":"published","date":"2024-11-04T15:57:57+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":65894035,"identity":"b96036ab-1382-43db-91d1-ce956d7d1436","added_by":"auto","created_at":"2024-10-04 06:07:24","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":488620,"visible":true,"origin":"","legend":"\u003cp\u003eApplication of different light treatments; from left to right: Fluorescent, Red LED, Blue LED, 3Red:1Blue LED and 3Blue:1Red LED on maturation and development of aerial part of somatic embryos in \u003cem\u003eK. odoratissima\u003c/em\u003e.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/c4fdfc3ea0037eec7c48a8bd.png"},{"id":65892212,"identity":"07a8b68a-7aa3-4e84-b772-cdc9b85b253f","added_by":"auto","created_at":"2024-10-04 05:35:24","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":8014,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of explant (zygotic embryo and cotyledonary leave) on SE induction rate (a), the mean number of somatic embryos/explant (b) and the embryo-forming capacity (EFC) index (c). Values are mean ± SE.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/9cf5a9585bdc879ce8bea242.png"},{"id":65893666,"identity":"30994575-078d-464c-ae7e-20ab8498ccde","added_by":"auto","created_at":"2024-10-04 05:59:24","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":371774,"visible":true,"origin":"","legend":"\u003cp\u003eFormation of embryo on zygotic embryos after three weeks (a); Formation of embryo on cotyledonary leaf after four weeks (b)\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/4b460aff5601970fb6759952.png"},{"id":65892207,"identity":"39f5a5ca-885a-4e58-8b5b-579683ca9ca7","added_by":"auto","created_at":"2024-10-04 05:35:24","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":101583,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of LED on maturation of somatic embryos after 30 days, Fluorescent (a), Red LED (b), Blue LED (c), 3Red:1Blue LED (d) and 3Blue:1Red LED (e)\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/37dded0904c600f695b0560e.png"},{"id":65892213,"identity":"9bdfc62e-3b83-4627-a7d4-6858bfed1d12","added_by":"auto","created_at":"2024-10-04 05:35:25","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":144795,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of light treatments on growth and developmental stages of regenerated samples after one month \u0026nbsp;(1M) and two months (2M) respectively; \u0026nbsp;Fluorescent light (F), Red LED (R), Blue LED (B), 3Red:1Blue LED (3R:1B) and 3Blue:1Red LED (3B:1R)\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/1555cb80ce6d0f61f87345c1.png"},{"id":68749972,"identity":"dec0caa5-8343-45cb-9300-6a34c579f892","added_by":"auto","created_at":"2024-11-11 16:08:10","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2529567,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/230b6aed-ca9c-4b7a-be61-21869b9f5a30.pdf"},{"id":65892211,"identity":"2c2f3f7e-7d9e-46e8-8130-bddcdcf68eb6","added_by":"auto","created_at":"2024-10-04 05:35:24","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":172226,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarydata.docx","url":"https://assets-eu.researchsquare.com/files/rs-4938119/v1/e14a85a8aae885daeb49a68f.docx"}],"financialInterests":"","formattedTitle":"Maturation, conversion and metabolic profile of somatic embryos derived from Klussia odoratissimaMozaff as affected by light spectra ​","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIranian wild celery, known as mountain celery (MC) or \u0026ldquo;Kelus\u0026rdquo; (\u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003eKlussia odoratissima\u003c/span\u003e Mozaff), is a perennial plant belonging to the family of Apiaceae. The plant has enormous economic and industrial applications, albeit being considered an endangered species. \u003cem\u003eK. odoratissima\u003c/em\u003e grows in central Zagros Mountain ranges of Iran. It possesses a wide range of medicinal properties including sedative, antimicrobial, hypotensive and antioxidative activities (Ahmadipour et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e and references therein). The health benefits of \u003cem\u003eK. odoratissima\u003c/em\u003e are mainly conferred by phthalides. Phthalides, including Butylidene phthalides and Ligustilides, are a group of secondary metabolites, which are recognized as the major flavoring component in \u003cem\u003eK. odoratissima\u003c/em\u003e. Unlike most endemic plants that are vulnerable to herbivores\u0026rsquo; browsing (Cubas et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), K. \u003cem\u003eodoratissima\u003c/em\u003e is threatened by excessive and illegal harvest at early growth stage (Seifipour Naghneh et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The production of secondary metabolites in plants is naturally influenced by some regional and environmental factors, which limit the commercialization of \u003cem\u003eK. odoratissima\u003c/em\u003e production. The extraction of secondary metabolites from \u003cem\u003eK. odoratissima\u003c/em\u003e in nature poses a risk of extinction. Plant tissue culture techniques have been recently highlighted as the means of \u003cem\u003ein vitro\u003c/em\u003e production of secondary metabolites (Hashim et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Somatic embryogenesis (SE) is a tissue culture technique in which somatic cells gain the capacity to produce embryos that finally develop into a whole plant (Aguilar-Hern\u0026aacute;ndez and Loyola-Vargas \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Garcia et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). This technique plays a vital role in plant regeneration and rapid reproduction (Tomiczak et al. \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The SE process includes several distinct stages: induction, embryo proliferation, maturation, and germination/conversion of embryos into plantlets (Ram\u0026iacute;rez-Mosqueda, 2022). In dicotyledonous plants, somatic embryos follow the same successive morphological developmental steps as described for zygotic embryogenesis: globular, heart-shaped, torpedo and cotyledonary (Ram\u0026iacute;rez-Mosqueda, 2022). According to recent studies, somatic embryos in suspension culture are useful tools for the generation of secondary metabolites (Murthy et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). This method has been successfully used to produce biomass and various secondary metabolites in several medicinal plant species such as \u003cem\u003eSalvia officinalis\u003c/em\u003e (Kintzios et al.1999), \u003cem\u003eGentiana straminea\u003c/em\u003e (Cai et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), \u003cem\u003eRauwolfia serpentine\u003c/em\u003e (Harisaranraj et al. 2009), and \u003cem\u003eEurycoma longifolia\u003c/em\u003e Jack (Iriawati et al. 2014). These studies have demonstrated that the production of plant bioactive metabolites is affected by different biotic and abiotic elicitors. Application of elicitors appears to be a viable method for increasing secondary metabolites in medicinal plants. In this respect, light has emerged as one of the unique and efficient elicitors for enhancing the \u003cem\u003ein vitro\u003c/em\u003e production of pharmacologically important secondary metabolites (Hashim et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eVarious types of light (UV, fluorescent, and LEDs) have been reported as stimulators for the production of plant secondary metabolites (Hashim et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In recent years, the application of light emitting diodes (LEDs) has attracted a significant attention as an alternative artificial light source for controlled environmental agriculture as well as plant tissue culture experiments (Almeida et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). LEDs have several advantages relative to traditional light sources including: small size, durability, long operating lifetime, and relatively cool emitting surfaces, which make them an ideal light source for indoor environments. An outstanding influence of LED lighting on plant regeneration and secondary metabolite production has been reported by numerous studies (Fan et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Cio\u0026acute;c, et al. 2018; Almeida et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Hashim et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). However, the response of different plants to LEDs has been inconsistent and there was no specific pattern among various species (Bula et al. 1999; Gupta and Jatothu 2013).\u003c/p\u003e \u003cp\u003eIn 1995, Munoz estimated that between18000-25000 wild plant species were at risk of extinction, and \u003cem\u003ein vitro\u003c/em\u003e culture techniques were proposed as the sole solution for satisfactory conservation of the majority of endangered species. As for \u003cem\u003eK. odoratissima\u003c/em\u003e, attempts to conserve the plant had little progress due to poor seed setting and germination as well as long-time seed dormancy (Askari-Khorasgani et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Hence, tissue culture seems to be an effective method for conservation of \u003cem\u003eK. odoratissima\u003c/em\u003e. Few reports have studies \u003cem\u003ein vitro\u003c/em\u003e culture in \u003cem\u003eK. odoratissima\u003c/em\u003e. Most researches have focused on traditional methods such as adjusting the basic medium and growth regulators, and its artificial light source was the traditional fluorescent lamp. Therefore, the objective of the present study was to investigate \u003cem\u003ein vitro\u003c/em\u003e tissue culture in tandem with different LED spectra on maturation and development of somatic embryogenesis in \u003cem\u003eK. odoratissima\u003c/em\u003e. Since areal parts of \u003cem\u003eK. odoratissima\u003c/em\u003e are rich sources of phthalides, we have also compared the concentration of phthalide across samples grown under different light conditions. This study provides an effective regeneration method for conservation of \u003cem\u003eK. odoratissima\u003c/em\u003e and could pave the way for the proliferation of this endangered plant in bioreactor systems.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant Material and Seed Germination\u003c/h2\u003e \u003cp\u003eSeeds of \u003cem\u003eK.odoratissima\u003c/em\u003e were collected from its natural habitat in Iran's \u0026ldquo;Chaharmahal and Bakhtiari\u0026rdquo; province. Seeds were first treated with 70% ethanol for 1 min, and then washed with sterile distilled water. Subsequently, seeds were dipped in freshly prepared hypochlorite sodium solution (1% w/v with a few droplets of tween-20) for 15 min. Seeds were then rinsed at least three times with sterile distilled water and cultured in a hormone-free (HF) MS basal medium (Murashige and Skoog) (pH 5.8) containing 3.0% (w/v) sucrose before incubating in a dark condition for 2 months at 4\u0026deg;C.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eInduction and Proliferation of Embryogenic Callus\u003c/h2\u003e \u003cp\u003eFor the induction of embryogenic callus, we used three different explants including; cotyledonary leaves, hypocotyls (both derived from \u003cem\u003ein vitro\u003c/em\u003e germinated seeds), and zygotic embryos. The explants were cultured in MS medium supplemented with 1 mg/L 2,4-dichlorophenoxyaceticacid (2,4-D) and 0.25 mg/L Kinetin as described by Ebrahimi et al.( 2018). The induction medium was supplemented with 30 g/L of sucrose and 7 g/L of agar and dispensed into petri dishes. All treatments had three replicates, each consisted of 10 explants. The cotyledonary leaves were cut in half and cultured with the adaxial side in the medium (Ebrahimi et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The cultures were incubated at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C in a dark culture room. After a month, the SE induction rate (% of explants containing SE/total explants) and the mean number of somatic embryos per explant were recorded. The embryo-forming capacity (EFC) index was also calculated using the following equations according to Raomai et al. (\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2014\u003c/span\u003e):\u003c/p\u003e \u003cp\u003eEFC\u0026thinsp;=\u0026thinsp;SE induction rate \u0026times; mean number of SEs per explant/100\u003c/p\u003e \u003cp\u003eBesides, embryogenic calli were sub-cultured in three cycles on MS basal medium containing 1mg/L of 2,4-D for embryogenic callus proliferation. To avoid genetic variation, we used single-embryogenic callus for the proliferation and the provision of samples for the light treatment experiments. The cultures were kept in the same condition as mentioned above.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eLight treatments\u003c/h2\u003e \u003cp\u003eIn order to investigate the effects of different light spectra on maturation of somatic embryos derived from \u003cem\u003eK. odoratissima\u003c/em\u003e, five light treatments were applied including a fluorescent lamp, red LED (660-675nm), blue LED (460-475nm), 3red:1blue LED and 3blue:1red LED (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Somatic embryos were incubated for a month on MS medium (pH 5.8) enriched with 30g/L sucrose and 6.2 g/L agar under diurnal temperature of 23\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, the relative humidity of 60%, a 16L:8D photoperiod, and a photosynthetic photon flux density of 50 \u0026micro;mol m\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003es\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The experiment was conducted in a completely randomized design (CRD) with three replications containing 200mg somatic embryogenesis mass (approximately 80\u0026ndash;110 embryos). Within each LED environment, petri dishes were rotationally switched their locations on a daily basis to assure of the homogeneity in the light intensity for all samples. After a month, the number of somatic embryos at each developmental stage (globular, heart, torpedo, and cotyledonary) was recorded. In addition, the number of regenerated embryos at each LED treatment was recorded. The samples were transferred on fresh MS medium and regenerated embryos were grown under the same LED conditions for another additional two months. Following this period, different characteristics including fresh weight (FW), dry weight (DW), leaf area index (mm\u003csup\u003e2\u003c/sup\u003e) and chlorophyll content (mg/g FW) of samples were measured. Leaf area index was calculated using ImageJ software. The total chlorophyll content (Chl) and the content of carotenoids (Car) were quantified spectrophotometrically using the Lichtenthaler equations and represented as mg/g FW (Lichtenthaler 1978).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eSecondary metabolites\u003c/h2\u003e \u003cp\u003eTo understand the effect of LEDs on metabolite profile of \u003cem\u003eK. odoratissima\u003c/em\u003e, 2.5g fresh leaves from \u003cem\u003ein vitro\u003c/em\u003e cultured plantlets as well as \u003cem\u003ein situ\u003c/em\u003e plants were harvested for the GC-MS analysis. These plants had the same age (two months). Likewise, a sample of air-dry of \u003cem\u003eK. odoratissima\u003c/em\u003e was also used to measure the concentration and composition of bioactive compounds. It is noteworthy that dry powder of \u003cem\u003eK. odoratissima\u003c/em\u003e is available in the market year around. The headspace (HS) sampling technique was implemented to ensure preserving the volatile constituents and avoiding any loss. The procedures for GC-MS analysis of \u003cem\u003eK. odoratissima\u003c/em\u003e, were previously described by Seifipour Naghneh et al. (\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The relative percentage of metabolic constituents was evaluated from the total peak area (TIC) by apparatus software. Metabolic compounds were identified by comparing their mass spectra with those stored in Wiley NBS75K.L and NIST/EPA/NIH (2002 version, National Institute of Standards and Technology, Gaithersburg, MD, USA) mass spectral libraries using various search engines (PBM, Nist02). Given the area under the peak is a function of the compound's concentration in the sample, the GC-MS data were presented as the area under the peak (Trivedi et al. \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Khazaeie et al., 2023).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eData analysis was performed using the SAS software (v.9.3), and the averages with the standard errors were compared by one-way ANOVA. The Duncan\u0026rsquo;s multiple range test was used to separate the treatment means (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eInduction and Proliferation of Embryogenic Callus\u003c/h2\u003e \u003cp\u003eSomatic embryogenesis (SE) is a pivotal process in plant biotechnology that enables the regeneration of plants from vegetative or non-reproductive tissues. The efficiency of SE greatly varies depending on the plant species, type of explant, the culture conditions, and the growth regulators involved.\u003c/p\u003e \u003cp\u003eIn our study, the callus induction was started 12\u0026ndash;14 days following the culture of explants and the percentage of SE after a month was evaluated. Our finding revealed a significant difference (P\u0026thinsp;\u0026le;\u0026thinsp;0.05) in the induction of somatic embryos on explants derived from germinated seeds and zygotic embryos under laboratory conditions (Fig.\u0026nbsp;2a). Although callus formation occurred more rapidly in hypocotyl explants compared to others, none of resulting calli exhibited an embryogenic potential. Therefore, this explant was excluded from further analysis. Similarly, Ebrahimi et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported that \u003cem\u003eK. odoratissima\u003c/em\u003e explants were unable to induce somatic embryos irrespective of callus formation on the hypocotyls.\u003c/p\u003e \u003cp\u003eBased on our results, the percentage of explants containing embryos exceeded 60 and 40% in zygotic embryos and cotyledonary leaves, respectively. Moreover, the average number of embryos per cotyledonary leaf reached 16, whereas this number was eight for zygotic embryos (Fig.\u0026nbsp;2b). Although the EFC index was higher in cotyledonary leaves compared to zygotic embryos but the change was insignificant (Fig.\u0026nbsp;2c).\u003c/p\u003e \u003cp\u003eFollowing the formation of embryogenic calli, somatic embryos were formed on lateral edges and adaxial surface of cotyledonary leaves in the fourth week. In contrast, the formation of embryo on zygotic embryos was induced after three weeks albeit without callus formation (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the study reported by Hu et al. (2016), zygotic embryos produced callus in indirect SE, but they had a lower frequency of callus formation compared to other explants such as leaves or immature embryos. Alternatively, the callus formed from zygotic embryos may have a lower capacity to develop into somatic embryos compared to callus from other explants. While there is no strong evidence to support this hypothesis, this may be due to the lack of dedifferentiation ability of zygotic embryos (von Arnold et al. \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). In particular, mature and well-developed embryos are less inclined to deviate towards callus formation. The ability of an explant to undergo direct or indirect embryogenesis was historically thought to be determined by the age of the explant. The further the explant is from the zygotic embryo stage, the more reprogramming (callus formation) needed to convert the explant into a somatic embryo (Merckle et al. 1995). On the other hand, young tissues of cotyledonary leaves contain higher levels of totipotency, which efficiently respond to laboratory culture conditions. Therefore, such tissues become less differentiated compared to zygotic embryos, making them more favorable for SE (Bhojwani and Razdan \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Thorpe \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eInvestigation into \u003cem\u003ein vitro\u003c/em\u003e propagation of \u003cem\u003eQuassia amara\u003c/em\u003e L. (Simaroubaceae) using the vegetative embryogenesis revealed that leaf and internode explants from older plants were ineffective due to severe phenolic exudation (Martin and Madassery \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). On the other hand, cotyledon explants were superior for the induction of vegetative embryos. Embryos were developed from both axial side and cutting point of the embryonic axis, with more embryos forming at the proximal end compared to the middle and distal regions of the cotyledons.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eThe effect of LED on developmental stages and conversion of somatic embryos\u003c/h2\u003e \u003cp\u003eIn tissue culturing, environmental factors play a central role in plant morphogenesis (Gupta and Jatothu 2013). Developmental stages of somatic embryos have been shown to be influenced by light spectrum (Almeida et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In the present research, the influence of light wavelength on development of somatic embryos of \u003cem\u003eK. odoratissima\u003c/em\u003e formed per callus (200 mg of initial FW) was evaluated after a month (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Our morphological studies indicated that light treatment significantly affected the development and regeneration of somatic embryos in the \u003cem\u003eK.\u003c/em\u003e \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003eodoratissima\u003c/span\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In this way, the number of cotyledonary (mature) embryos per callus under 3red:1blue treatment was significantly (P\u0026thinsp;\u0026le;\u0026thinsp;0.05) greater than that under other light treatments (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Additionally, the somatic embryos subjected to 3red:1blue treatment, were able to undergo conversion phase (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAnalysis of variance for effect of different light treatments on embryogenic cell maturation and regeneration of \u003cem\u003eK. odoratissima\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSource of variation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c7\" namest=\"c3\"\u003e \u003cp\u003eMean of square\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGlobular\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHeart\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTorpedo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCotyledonary\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRegenerated\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLight treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e126.67\u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.17\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13.43\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e417.57\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e12.4\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eError\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e313.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003ens, *, ** means non-significant, significant at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 and significant in p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, respectively.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean comparisons for effect of different light treatments on development and regeneration of somatic embryos in \u003cem\u003eK. odoratissima\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLight treatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eMean number of somatic embryos\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGlobular\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHeart\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTorpedo\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCotyledonary\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eRegenerated\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlourescent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e60\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;10.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.67\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.67 \u003csup\u003ecd\u003c/sup\u003e \u0026plusmn;0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.3\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.33\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRed LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e70 \u003csup\u003ea\u003c/sup\u003e \u0026plusmn;10.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.33\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.33 \u003csup\u003eb\u003c/sup\u003e \u0026plusmn;0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.4\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.67\u003csup\u003ebc\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBlue LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e60 \u003csup\u003ea\u003c/sup\u003e \u0026plusmn;10.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.67\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.67 \u003csup\u003eb\u003c/sup\u003e \u0026plusmn;0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.7\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.67\u003csup\u003ebc\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3Red:1Blue LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e66.7 \u003csup\u003ea\u003c/sup\u003e \u0026plusmn;12.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.64\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.7\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;3.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.7\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3Blue:1Red LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e53.3 \u003csup\u003ea\u003c/sup\u003e \u0026plusmn;8.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.33\u003csup\u003ed\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13 \u003csup\u003ec\u003c/sup\u003e\u0026plusmn;1.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.33\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003eData are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE; Different letters indicate significant differences using the Duncan\u0026rsquo;s Multiple Range Test (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe minimum number of cotyledonary embryos was observed under blue LED and 3blue:1red LED (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In \u003cem\u003eFritillaria cirrhosa\u003c/em\u003e, the maximum number of SEs with cotyledonary leaves was observed in white light followed by a combination of 8red:1blue LED (Chen et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). On the other hand, in sugarcane, the highest number of somatic embryos reaching to maturation phase was reported under LED lights with the medium blue wavelength (Heringer et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Therefore, depending on plant species, different types of growth chambers equipped with LED lights can be considered to achieve the best performance.\u003c/p\u003e \u003cp\u003eLED lamps have become a common approach for optimizing plant growth conditions, and this approach can successfully be used for SE of several species such as \u003cem\u003eDimocarpus longan\u003c/em\u003e (Li et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), \u003cem\u003eSaccharum\u003c/em\u003e spp. (Heringer et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and \u003cem\u003eVitis vinifera\u003c/em\u003e (Tittmann et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). A review of literature showed that light is selectively perceived by plant photoreceptors including cryptochromes and phototropins. Although cryptochromes react with blue and UV-A lights, phytochromes are reactive to red and far-red light (Huang et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The plant photoreceptors perceive different wavelengths and trigger complex signal transduction to modulate gene expression profile and subsequently induce developmental responses such as induction, maturation and conversion in somatic embryos (Pedmale et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Gupta and Jatothu 2013). The present findings showed that 3red:1blue LED played an important role in maturation of SE in \u003cem\u003eK. odoratissima\u003c/em\u003e so that application of this light combination in SE could coordinate photoreceptors with proteins, ions, hormones and other factors to regulate gene expression patterns with accompanying changes in SE phases.\u003c/p\u003e \u003cp\u003eInterestingly, we noticed that light treatment could affect synchronization of somatic embryos. In this regard, the embryos grown under red LED and 3red:1blue LED had more uniformity compared to other light conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Synchronization of somatic embryo cultures could greatly increase the efficiency of SE for \u003cem\u003ein vitro\u003c/em\u003e propagation, which makes the technique ideal to produce artificial seeds (Nadel et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Alwael et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Thus, if all resultant cultures could be synchronized, this would have shortened the duration of culture with concomitant increase in propagation efficiency and uniformity (Nadel et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). A review of literature indicated that researchers prefer to use hormonal treatments to increase synchronization in SE cultures. To the best of our knowledge, this report is the first to note that light treatment could also influence the uniformity of SE in plant species. Therefore, we suggest to consider light as an underlying factor in synchronization of SE in plant tissue culture technique.\u003c/p\u003e \u003cp\u003eTo better understand the effect of light treatment on conversion of \u003cem\u003eK.\u003c/em\u003e \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003eodoratissima\u003c/span\u003e cultures, fresh and dry weight, leaf area index with chlorophyll and carotenoid contents were evaluated (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Our findings showed a significant variation in the fresh and dry weights for light treatments following additional two months of the experiment (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAnalysis of variance for effect of different light treatments on conversion of somatic embryos derived from \u003cem\u003eK. odoratissima\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSource of variation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c7\" namest=\"c3\"\u003e \u003cp\u003eMean of square\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFW (mg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDW (mg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLAI (mm\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eChl (mg/g FW)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCar(mg/g FW)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLight treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9473.33\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e93.45\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e83.85\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.017\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.000037\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eError\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1044.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.000004\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eThe symbols *, ** means significant at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 and significant in p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, respectively.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe highest fresh weight was recorded in the cultures grown under combination of 3blue:1red and blue LEDs followed by Fluorescent lamp. In contrast, the lowest fresh weight was obtained under the red LED (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The dry weights were significantly enhanced in the cultures treated under 3blue:1red LED and blue LED (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) whereas no significant difference was observed in the dry weight of the samples incubated under the red LED and 3red:1blue LED treatments (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Likewise, the same trend was recorded for leaf area index in the samples grown under the combination of 3blue:1red and blue LED lights (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean comparisons for effect of different light treatments on growth performance of converted cultures in \u003cem\u003eK. odoratissima\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLight treatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eMean number of somatic embryos\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFW (mg)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDW (mg)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLAI (mm\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eChl (mg/g FW)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCar (mg/g FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlourescent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e587.01\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;19.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e52.51\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e22.16\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.84 \u003csup\u003eab\u003c/sup\u003e\u0026plusmn;0.051\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.02\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0011\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRed LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e503.33\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;11.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e45.77\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.67\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.73\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.015\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0006\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBlue LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e587.04\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;33.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e54.75\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25.88\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;4.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.85\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.021\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0011\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3Red:1Blue LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e513.67 \u003csup\u003eb\u003c/sup\u003e\u0026plusmn;7.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46.63\u003csup\u003ec\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.02\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.75\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.018\u003csup\u003ebc\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0003\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3Blue:1Red LED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e627.01\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;8.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e59.67\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e28.56\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;3.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.92\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.052\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.023\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003eData are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE; Different letters indicate significant differences using the Duncan\u0026rsquo;s Multiple Range Test (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe chlorophyll content is a measure of well-being of a plant under different conditions. Our finding showed that light quality significantly affected total chlorophyll and carotenoid contents in the leaves of \u003cem\u003eK.\u003c/em\u003e \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003eodoratissima\u003c/span\u003e plantlets (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The chlorophyll content was correlated with the culture growth in the present study. Consequently, cultures exposed to the 3blue:1red LED and blue LED exhibited the highest content of chlorophyll and carotenoids and the cultures grown under red LED showed the lowest content of the pigments (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Our results are consistent with those reporting plants grown under blue light had a higher chlorophyll content in comparison to plants grown under red light (Fan et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Hung et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Blue light seems to play a crucial role in the activity of photosystem I and II as well as the photosynthetic electron transport capacity (Miao et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In plant species such as \u003cem\u003eGerbera jamesonii\u003c/em\u003e and \u003cem\u003eMyrtus communis\u003c/em\u003e, the accumulation of photosynthetic pigments was reduced when plants were subjected to red LED (Pawłowska et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Cio\u0026acute;c et al. 2018). In line with our findings, the accumulation of carotenoids in \u003cem\u003eSinningia speciosa\u003c/em\u003e (Simlat et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and \u003cem\u003eStevia rebaudiana\u003c/em\u003e (Zheng and van Labeke \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) cultures was stimulated by blue light spectrum. Findings from the present study, corroborate earlier literature that reported light quantity, quality and duration were involved in controlling morphogenesis, growth and differentiation of plant cells, tissues and organ cultures (Moshe and Dalia \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Different sources of LED provide wavelengths that are matched with plant photoreceptors, which leads to optimal production and metabolome (Bourget \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Massa et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Morrow \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Plants apparently sense changes in light quality through photoreceptors, which in turn regulate growth and development through modulating signaling pathways (Rechenmacher et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eLight treatments and secondary metabolites\u003c/h2\u003e \u003cp\u003eIn plants, environmental factors have a significant role in the biosynthesis of secondary metabolites (Khajali and Rafiei \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Several studies have identified the influence of light related factors such as light intensity, light quality, and light period on metabolite accumulation of different medicinal plants (Khazaeie et al. 2023 and the references therein). Despite a great deal of information on the application of LEDs on plant secondary metabolites, limited information is available on optimizing LEDs to improve secondary metabolites in endangered plant species. We demonstrated the effect of different light treatments on the metabolic compounds in \u003cem\u003ein vitro\u003c/em\u003e grown aerial parts of the \u003cem\u003eK. oddoratisma.\u003c/em\u003e Chemical composition of \u003cem\u003eK. oddoratisma\u003c/em\u003e among different conditions including \u003cem\u003ein situ\u003c/em\u003e, dried powder sample from \u003cem\u003ein situ\u003c/em\u003e plant, florescent, red, blue, 3red:1blue and 3blue:1red indicated 14, 6, 9, 5, 8, 8 and 9 bioactive constituents, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). A more diverse variation of bioactive compounds was recorded for two-old-month fresh plant grows \u003cem\u003ein situ\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The composition of plant secondary metabolites is affected by different factors including environmental condition, interaction between genotype and environment, method of distillation, storage condition, physiological stage, time of harvest, and season (Khajali and Rafiei \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe composition and concentration (AUP/FW) of bioactive compounds in \u003cem\u003eK. odoratissima\u003c/em\u003e cultures grown under different light treatment and two-month-old dried sample\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCompound\u0026rsquo;s name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKI \u003csub\u003eCal\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFlourescent\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRed\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eBlue\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3Red:Blue\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3Blue:Red\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eDried sample\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14.209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3,9-Epoxy-p-mentha-1,8(10)-diene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1199\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e247181\u0026thinsp;\u0026plusmn;\u0026thinsp;8421\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19.895\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNeophytadiene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1817\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e280265\u0026thinsp;\u0026plusmn;\u0026thinsp;10023\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(Z-Butylidenephthalide)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1675\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3880705\u0026thinsp;\u0026plusmn;\u0026thinsp;32421\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1238428\u0026thinsp;\u0026plusmn;\u0026thinsp;17400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3617628\u0026thinsp;\u0026plusmn;\u0026thinsp;40221\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2839095\u0026thinsp;\u0026plusmn;\u0026thinsp;18664\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4374798\u0026thinsp;\u0026plusmn;\u0026thinsp;18728\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e1192851\u0026thinsp;\u0026plusmn;\u0026thinsp;13235\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19.447\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e((E)-Ligustilide)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1830\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6294434\u0026thinsp;\u0026plusmn;\u0026thinsp;29671\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1694471\u0026thinsp;\u0026plusmn;\u0026thinsp;23551\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6819459\u0026thinsp;\u0026plusmn;\u0026thinsp;33265\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4956396\u0026thinsp;\u0026plusmn;\u0026thinsp;50210\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6364303\u0026thinsp;\u0026plusmn;\u0026thinsp;37425\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e1020396\u0026thinsp;\u0026plusmn;\u0026thinsp;29840\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16.955\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eisoledene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1373\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e442749\u0026thinsp;\u0026plusmn;\u0026thinsp;2655\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14.978\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(α-Copaene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1377\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e320392\u0026thinsp;\u0026plusmn;\u0026thinsp;9885\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e986654\u0026thinsp;\u0026plusmn;\u0026thinsp;7677\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e481636\u0026thinsp;\u0026plusmn;\u0026thinsp;8965\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e325730\u0026thinsp;\u0026plusmn;\u0026thinsp;10245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e336192\u0026thinsp;\u0026plusmn;\u0026thinsp;5922\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(β-Isocomene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1412\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e170230\u0026thinsp;\u0026plusmn;\u0026thinsp;8545\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.679\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(β-Copaene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1430\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2621460\u0026thinsp;\u0026plusmn;\u0026thinsp;4744\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e357727\u0026thinsp;\u0026plusmn;\u0026thinsp;11250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7446177\u0026thinsp;\u0026plusmn;\u0026thinsp;12565\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4300660\u0026thinsp;\u0026plusmn;\u0026thinsp;6700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3090324\u0026thinsp;\u0026plusmn;\u0026thinsp;12024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.553\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e.beta.-ylangene (β-Ylangene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1423\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1225541\u0026thinsp;\u0026plusmn;\u0026thinsp;5860\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e289332\u0026thinsp;\u0026plusmn;\u0026thinsp;8600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4901094\u0026thinsp;\u0026plusmn;\u0026thinsp;12433\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2773455\u0026thinsp;\u0026plusmn;\u0026thinsp;14254\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2599152\u0026thinsp;\u0026plusmn;\u0026thinsp;7855\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.158\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(Germacrene D)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e374215\u0026thinsp;\u0026plusmn;\u0026thinsp;13442\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e894846\u0026thinsp;\u0026plusmn;\u0026thinsp;14000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e431077\u0026thinsp;\u0026plusmn;\u0026thinsp;1977\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e444565\u0026thinsp;\u0026plusmn;\u0026thinsp;2166\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.alpha.-Cadina-4,9-diene, (-)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2122920\u0026thinsp;\u0026plusmn;\u0026thinsp;12922\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1239421\u0026thinsp;\u0026plusmn;\u0026thinsp;8500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1572100\u0026thinsp;\u0026plusmn;\u0026thinsp;23412\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16.585\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e.gamma.-Muurolene (γ-Muurolene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1485\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1197285\u0026thinsp;\u0026plusmn;\u0026thinsp;11971\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.927\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003etrans-.beta.-Farnesene ((E)-β-Farnesene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1473\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6052262\u0026thinsp;\u0026plusmn;\u0026thinsp;26329\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20644815\u0026thinsp;\u0026plusmn;\u0026thinsp;21459\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e12431334\u0026thinsp;\u0026plusmn;\u0026thinsp;54200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9108300\u0026thinsp;\u0026plusmn;\u0026thinsp;32950\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.942\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecis-.beta.-Farnesene ((Z)-β-Farnesene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1476\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1341066\u0026thinsp;\u0026plusmn;\u0026thinsp;8710\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16.955\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(-)-.alpha.-Panasinsen (α-Panasinsanene)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1527\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e945045\u0026thinsp;\u0026plusmn;\u0026thinsp;10200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOthers\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3468376\u0026thinsp;\u0026plusmn;\u0026thinsp;75980\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3238994\u0026thinsp;\u0026plusmn;\u0026thinsp;47684\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3238994\u0026thinsp;\u0026plusmn;\u0026thinsp;67234\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2456886\u0026thinsp;\u0026plusmn;\u0026thinsp;33865\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e693234\u0026thinsp;\u0026plusmn;\u0026thinsp;20124\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c10\"\u003e \u003cp\u003e2594982\u0026thinsp;\u0026plusmn;\u0026thinsp;21000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"10\"\u003eData are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe composition and concentration (AUP/FW) of bioactive compounds in two-month-old \u003cem\u003ein situ K. odoratissima\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCompound\u0026rsquo;s name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKI \u003csub\u003eCal\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTwo-month-old\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13.371\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-Limonene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1046\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e65608\u0026thinsp;\u0026plusmn;\u0026thinsp;446\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23.239\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eβ-Cubebene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1390\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e74028\u0026thinsp;\u0026plusmn;\u0026thinsp;4235\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24.437\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eElixene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1453\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1988650\u0026thinsp;\u0026plusmn;\u0026thinsp;24004\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24.812\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(E)-β-Farnesene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1473\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1311422\u0026thinsp;\u0026plusmn;\u0026thinsp;14221\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25.377\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGermacrene D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e440701\u0026thinsp;\u0026plusmn;\u0026thinsp;6412\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25.274\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eγ-Cadinene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1497\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e49992\u0026thinsp;\u0026plusmn;\u0026thinsp;2810\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25.820\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eδ-Cadinene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1526\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e28151\u0026thinsp;\u0026plusmn;\u0026thinsp;108\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28.322\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eZ-Butylidene phthalide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1675\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1234885\u0026thinsp;\u0026plusmn;\u0026thinsp;28760\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29.398\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(E)-3-Butylidene phthalide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1744\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e194856\u0026thinsp;\u0026plusmn;\u0026thinsp;4650\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29.715\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eZ-Ligustilide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1765\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e24982212\u0026thinsp;\u0026plusmn;\u0026thinsp;78700\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30.625\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eE-Ligustilide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1830\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e702727\u0026thinsp;\u0026plusmn;\u0026thinsp;9695\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30.907\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCaffeine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1582\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e8972991\u0026thinsp;\u0026plusmn;\u0026thinsp;38439\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6.876\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(E)-2-Hexenal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e866\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1669101\u0026thinsp;\u0026plusmn;\u0026thinsp;19010\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15.951\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNonanal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e786261\u0026thinsp;\u0026plusmn;\u0026thinsp;8218\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eData are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe natural habitat of \u003cem\u003eK. oddoratisma\u003c/em\u003e is recognized by an altitude of \u0026gt;\u0026thinsp;2500m and the temperature of barely above 20\u0026deg;C during the vegetative stage (Seifipour Naghneh et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), which are totally different from a controlled condition. It is presumed that such unique growth environment could differently affect emission and composition of secondary metabolite in \u003cem\u003eK. oddoratisma\u003c/em\u003e grown \u003cem\u003ein situ\u003c/em\u003e compare to those obtained from tissue culture technique.\u003c/p\u003e \u003cp\u003eIn the present study, the composition and the relative concentration of bioactive compounds were significantly changed across different light conditions (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Phthalides constituted a large amount of secondary metabolites in \u003cem\u003eK. odoratissima\u003c/em\u003e. These compounds are known to have a wide range of health benefits including antibacterial, antifungal, insecticidal, cytotoxic, anti-inflammatory properties (Seifipour Naghneh et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2022\u003c/span\u003e and the references therein). In \u003cem\u003eDrosera rotundifolia\u003c/em\u003e, the amount of secondary metabolites was significantly higher, in the field grown samples compared to \u003cem\u003ein vitro\u003c/em\u003e plantlets (Tienaho et al. \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Wilken et al. (\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) used different tissue culture techniques in three different plant species including \u003cem\u003eHypericum perforatum\u003c/em\u003e, \u003cem\u003eCymbopogon citratus\u003c/em\u003e and \u003cem\u003eFabiana imbricate.\u003c/em\u003e They reported that the concentration of secondary metabolites produced in tissue culture technique was significantly lower when compared to the field grown plants.\u003c/p\u003e \u003cp\u003eInterestingly, phthalide content was significantly (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) higher in tissue culture samples compared to dried sample derived from \u003cem\u003ein situ\u003c/em\u003e plant (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn our study, under \u003cem\u003ein vitro\u003c/em\u003e condition, 3blue:1red LED followed by blue LED and fluorescent light were the best treatments for production of phthalides. However, the concentration of phthalides was minimal in cultures grown under red LED condition (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSecondary metabolites are compounds that play an important role in the interaction of plants with their environment (Oksman-Caldentey and Inz\u0026eacute; 2005; Khajali and Rafiei \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Therefore, it is perceived that their biosynthesis in plant tissues is modified by changing environmental conditions including light (Tomaszewiczet al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In \u003cem\u003eC. delgadii\u003c/em\u003e plants, blue light was the most efficient for the accumulation of flavonoid glycosides, such as astragalin and rutin. A stimulatory effect of this light quality on the accumulation of secondary metabolites was also noticed for other species, e.g., \u003cem\u003eRhodiola imbricate\u003c/em\u003e (Kapoor et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) and \u003cem\u003eCapsicum annuum\u003c/em\u003e (Yap et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIt has been reported that blue light stimulates the biosynthesis of secondary metabolites in plants and in the meantime, enhances the synthesis of compounds such as antioxidants. Such increase is attributed to the specific wavelength of blue light that affects plant metabolic pathways (Larsen et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eUnder \u003cem\u003ein vitro\u003c/em\u003e condition, Manivannan et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) demonstrated that blue LED treatment significantly increased the total phenol and flavonoid contents in the aerial parts of \u003cem\u003eRehmannia glutinosa\u003c/em\u003e. Moreover, Lian et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) reported that blue LED was associated with increased antioxidant activity and enhanced contents of total flavonoid and phenolic compounds in \u003cem\u003eGynura procumbens\u003c/em\u003e, with a notable increase in cyanidin-monoglucosides under blue light.\u003c/p\u003e \u003cp\u003eA recent study indicated that high-intensity blue light supplementation for 4 hours during the night could induce the expression of MYBs, CRY2/3, SPAs, and HY5, thereby enhancing anthocyanin and catechin accumulation in tea leaves (Zheng et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Furthermore, Wang et al. (\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) reported that blue light extensively regulates multiple physiological processes and secondary metabolism in young tea shoots, significantly affecting gene transcription primarily in the pathways of photosynthesis, lipid metabolism, and flavonoid synthesis under high-intensity blue light. Recent studies shows blue light has a significant impact on the biosynthesis of terpenes and cannabionoids in plants by enhancing the activity of key biosynthetic pathways and enzymes (Morello 2022; Desaulniers Brousseau 2021).\u003c/p\u003e \u003cp\u003eAlthough application of LEDs in plant regeneration is promising, but any given plant species needs ascertaining wavelengths in order to maximize the yield and quality (Khazaeie et al. 2023). Based on analysis of GC\u0026ndash;MS data, we found that 3blue:1red LED followed by blue LED and fluorescent light were the best environments for the production of phthalide, the most outstanding secondary compound in \u003cem\u003eK. odoratissima.\u003c/em\u003e It seems that these light treatments can favorably affect light-responsive genes and such changes corresponded to biosynthesis of phthalides in \u003cem\u003eK. odoratissima\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn general, it was noticed that the cotyledonary leaves were the most competent explant for somatic embryogenesis in \u003cem\u003eK. odoratissima\u003c/em\u003e under the conditions of this study. The developmental stage of \u003cem\u003eK. odoratissima\u003c/em\u003e under \u003cem\u003ein vitro\u003c/em\u003e conditions requires a specific lighting treatment. In this regard, a combination of 3red:1blue LED provided the highest number of cotyledonary (mature) embryos per callus. This light treatment was the best treatments for conversion of somatic embryos in \u003cem\u003eK. odoratissima\u003c/em\u003e. Consequently, the highest fresh and dried weights were recorded in the cultures grown under 3blue:1red and blue LED treatment. Interestingly, phthalide content was significantly (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) greater in tissue culture samples compared to dried powder of two-month-old \u003cem\u003ein situ\u003c/em\u003e plants, which suggests tissue culture as an alternative technique for the production of bioactive compound in \u003cem\u003eK. odoratissima\u003c/em\u003e. Cultures were incubated in 3blue:1red LED followed by blue LED and florescent light possessed higher contents of phthalides. Based on this research, it was shown that LEDs can be an effective alternative to florescent lamps in micropropagation of \u003cem\u003eK. odoratissima\u003c/em\u003e.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors acknowledge Shahrekord University and Agricultural Biotechnology Research Institute of Iran-Central Branch (ABRII-CB) for spiritual and financial supporting. This research was part of a project in ABRII-CB entitled \u0026ldquo;Optimization of shoot culture and production of Kelus (\u003cem\u003eKelussia odoratissima\u003c/em\u003e Mozaff.) in bioreactor- ID: 4-05-05-010-010551\u0026rdquo;.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was part of a project in ABRII-CB entitled \u0026ldquo;Optimization of shoot culture and production of Kelus (\u003cem\u003eKelussia odoratissima\u003c/em\u003e Mozaff.) in bioreactor- ID: 4-05-05-010-010551\u0026rdquo;. Morteza Ebrahimi\u0026nbsp;has received research support\u0026nbsp;from \u0026ldquo;Agricultural Biotechnology Research Institute of Iran-Central Branch (ABRII-CB) \u0026ldquo;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAll authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Morteza Ebrahimi, Fariba Rafiei, Farnoosh Khosravi and Mohammad Rabiei. The first draft of the manuscript was written by Fariba Rafiei and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/em\u003e\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAguilar-Hern\u0026aacute;ndez V, Loyola-Vargas VM (2018) Advanced proteomic approaches to elucidate somatic embryogenesis. Front Plant Sci 9:1658. https://doi.org/10.3389/fpls.2018.01658\u003c/li\u003e\n\u003cli\u003eAhmadipour B, Hassanpour H, Asadi E, et al (2015) \u003cem\u003eKelussia odoratissima\u003c/em\u003e Mozzaf - a promising medicinal herb to prevent pulmonary hypertension in broiler chickens reared at high altitude. J Ethnopharmacol 159:49\u0026ndash;54. https://doi.org/10.1016/j.jep.2014.10.043\u003c/li\u003e\n\u003cli\u003eAlmeida FA, Vale EM, Reis RS, et al (2019) LED lamps enhance somatic embryo maturation in association with the differential accumulation of proteins in the \u003cem\u003eCarica papaya\u003c/em\u003e L. \u0026ldquo;Golden\u0026rdquo; embryogenic callus. Plant Physiol Biochem 143:109\u0026ndash;118. https://doi.org/10.1016/j.plaphy.2019.08.029\u003c/li\u003e\n\u003cli\u003eAlwael HA, Naik PM, Al-Khayri JM (2017) Synchronization of Somatic Embryogenesis in Date Palm Suspension Culture Using Abscisic Acid. In: Al-Khayri JM, Jain SM, Johnson DV (eds) Date Palm Biotechnology Protocols Volume I: Tissue Culture Applications. Springer New York, New York, NY, pp 215\u0026ndash;226. https://doi.org/10.1007/978-1-4939-7156-5_18\u003c/li\u003e\n\u003cli\u003eAskari-Khorasgani O, Mortazaeinezhad F, Otroshy M, Golparvar AR (2013) Breaking seed dormancy of endangered medicinal plant Kelussia odoratissima using zygotic embryo culture technique. ISSN 2051:0853\u003c/li\u003e\n\u003cli\u003eBhoite HA, Palshikar GS (2014) Plant Tissue Culture: A Review. World J Pharm Sci 565\u0026ndash;572\u003c/li\u003e\n\u003cli\u003eBhojwani SS, Razdan MK (1996) Plant tissue culture: Theory and practice: Theory and practice. Elsevier Science \u0026amp; Technology\u003c/li\u003e\n\u003cli\u003eBourget CM (2008) An introduction to light-emitting diodes. HortScience 43:1944-1946\u003c/li\u003e\n\u003cli\u003eBowes BG (1999) A Colour Atlas of Plant Propagation and Conservation. Manson Publishing, London, England\u003c/li\u003e\n\u003cli\u003eBula RJ, Morrow RC, Tibbitts TW, et al (1991) Light-emitting diodes as a radiation source for plants. HortScience 26:203\u0026ndash;205. https://doi.org/10.21273/hortsci.26.2.203\u003c/li\u003e\n\u003cli\u003eCai Y, Liu Y, Liu Z, et al (2009) High-frequency embryogenesis and regeneration of plants with high content of gentiopicroside from the Chinese medicinal plant \u003cem\u003eGentiana straminea\u003c/em\u003e Maxim. In Vitro Cellular \u0026amp; Developmental Biology - Plant 45:730\u0026ndash;739. https://doi.org/10.1007/s11627-009-9225-7\u003c/li\u003e\n\u003cli\u003eChen C-C, Lee M-R, Wu C-R, et al (2020) LED Lights Affecting Morphogenesis and Isosteroidal Alkaloid Contents in \u003cem\u003eFritillaria cirrhosa\u003c/em\u003e D. Don-An Important Chinese Medicinal Herb. Plants 9:. https://doi.org/10.3390/plants9101351\u003c/li\u003e\n\u003cli\u003eCioć M, Szewczyk A, Żupnik M, et al (2018) LED lighting affects plant growth, morphogenesis and phytochemical contents of \u003cem\u003eMyrtus communis\u003c/em\u003e L. in vitro. Plant Cell Tissue Organ Cult 132:433\u0026ndash;447. https://doi.org/10.1007/s11240-017-1340-2\u003c/li\u003e\n\u003cli\u003eCubas J, Irl SDH, Villafuerte R, et al (2019) Endemic plant species are more palatable to introduced herbivores than non-endemics. Proc Biol Sci 286:20190136. https://doi.org/10.1098/rspb.2019.0136\u003c/li\u003e\n\u003cli\u003eDesaulniers Brousseau V, Wu B-S, MacPherson S, et al (2021) Cannabinoids and terpenes: How production of photo-protectants can be manipulated to enhance cannabis sativa L. phytochemistry. Front Plant Sci 12:620021. https://doi.org/10.3389/fpls.2021.620021\u003c/li\u003e\n\u003cli\u003eDutta Gupta S, Jatothu B (2013) Fundamentals and applications of light-emitting diodes (LEDs) in in vitro plant growth and morphogenesis. Plant Biotechnol Rep 7:211\u0026ndash;220. https://doi.org/10.1007/s11816-013-0277-0\u003c/li\u003e\n\u003cli\u003eEbrahimi M, Mokhtari A, Amirian R (2018) A highly efficient method for somatic embryogenesis of Kelussia odorotissima Mozaff., an endangered medicinal plant. Plant Cell Tissue Organ Cult 132:99\u0026ndash;110. https://doi.org/10.1007/s11240-017-1314-4\u003c/li\u003e\n\u003cli\u003eEbrahimie E, Habashy AA, Mohammadie-Dehcheshmeh M, et al (2006) Direct shoot regeneration from mature embryo as a rapid and genotype-independent pathway in tissue culture of heterogeneous diverse sets of cumin (\u003cem\u003eCuminum cyminum\u003c/em\u003e L.) genotypes. In Vitro Cellular \u0026amp; Developmental Biology - Plant 42:455\u0026ndash;460. https://doi.org/10.1079/IVP2006789\u003c/li\u003e\n\u003cli\u003eFan X, Zang J, Xu Z, et al (2013) Effects of different light quality on growth, chlorophyll concentration and chlorophyll biosynthesis precursors of non-heading Chinese cabbage (\u003cem\u003eBrassica campestris\u003c/em\u003e L.). Acta Physiol Plant 35:2721\u0026ndash;2726. https://doi.org/10.1007/s11738-013-1304-z\u003c/li\u003e\n\u003cli\u003eGarcia C, Furtado de Almeida A-A, Costa M, et al (2019) Abnormalities in somatic embryogenesis caused by 2,4-D: an overview. Plant Cell Tiss Organ Cult 137:193\u0026ndash;212. https://doi.org/10.1007/s11240-019-01569-8\u003c/li\u003e\n\u003cli\u003eGuppta, S.D., Jatothu, B., 2013. Fundamentals and applications of light-emitting diodes (LEDs) in in vitro plant growth and morphogenesis. Plant Biotechnol Rep 7: 211\u0026ndash;220.\u003c/li\u003e\n\u003cli\u003eHarisaranraj R, Suresh K. (2009). Evaluation of the Chemical Composition \u003cem\u003eRauwolfia serpentina\u003c/em\u003e and \u003cem\u003eEphedra vulgaris\u003c/em\u003e. ABR 3:174-178.\u003c/li\u003e\n\u003cli\u003eHashim M, Ahmad B, Drouet S, et al (2021) Comparative Effects of Different Light Sources on the Production of Key Secondary Metabolites in Plants In Vitro Cultures. Plants 10:. https://doi.org/10.3390/plants10081521\u003c/li\u003e\n\u003cli\u003eHeringer AS, Reis RS, Passamani LZ, et al (2017) Comparative proteomics analysis of the effect of combined red and blue lights on sugarcane somatic embryogenesis. Acta Physiol Plant 39:52. https://doi.org/10.1007/s11738-017-2349-1\u003c/li\u003e\n\u003cli\u003eHu R, Sun Y, Wu B, et al (2017) Somatic Embryogenesis of Immature \u003cem\u003eCunninghamia lanceolata\u003c/em\u003e (Lamb.) Hook Zygotic Embryos. Sci Rep 7:56. https://doi.org/10.1038/s41598-017-00156-1\u003c/li\u003e\n\u003cli\u003eHuang X, Ouyang X, Deng XW (2014) Beyond repression of photomorphogenesis: role switching of COP/DET/FUS in light signaling. Curr Opin Plant Biol 21:96\u0026ndash;103. https://doi.org/10.1016/j.pbi.2014.07.003\u003c/li\u003e\n\u003cli\u003eHung CD, Hong C-H, Kim S-K, et al (2016) LED light for in vitro and ex vitro efficient growth of economically important highbush blueberry (\u003cem\u003eVaccinium corymbosum\u003c/em\u003e L.). Acta Physiol Plant 38:1\u0026ndash;9. https://doi.org/10.1007/s11738-016-2164-0\u003c/li\u003e\n\u003cli\u003eIriawati, Rahmawati A, Esyanti RR (2014) Analysis of secondary metabolite production in somatic embryo of pasak bumi (\u003cem\u003eEurycoma longifolia\u003c/em\u003e Jack.). Procedia Chem 13:112\u0026ndash;118. https://doi.org/10.1016/j.proche.2014.12.014\u003c/li\u003e\n\u003cli\u003eKapoor S, Raghuvanshi R, Bhardwaj P, et al (2018) Influence of light quality on growth, secondary metabolites production and antioxidant activity in callus culture of \u003cem\u003eRhodiola imbricata\u003c/em\u003e Edgew. J Photochem Photobiol B 183:258\u0026ndash;265. https://doi.org/10.1016/j.jphotobiol.2018.04.018\u003c/li\u003e\n\u003cli\u003eKhajali F and Rafiei F (2024) A review of plant anti-nutritional factors in animal health and production: the classification, biological properties, and the passivation strategy. JAF Res 18:101290. https://doi.org/10.1016/j.jafr.2024.101290\u003c/li\u003e\n\u003cli\u003eKhazaei M, Rafiei F, Sabzalian MR, Houshmand S (2023) Potential application of light emitting diodes to improve molecular, physiological, and growth responses in peppermint plant. JSFA Rep 3:663\u0026ndash;673. https://doi.org/10.1002/jsf2.177\u003c/li\u003e\n\u003cli\u003eKintzios S, Nikolaou A, Skoula M (1999) Somatic embryogenesis and in vitro rosmarinic acid accumulation in Salvia officinalis and S. fruticosa leaf callus cultures. Plant Cell Reports 18:462\u0026ndash;466. https://doi.org/10.1007/s002990050604\u003c/li\u003e\n\u003cli\u003eKumar Trivedi M (2017) Chromatographic and spectroscopic characterization of the consciousness energy healing treated \u003cem\u003eWithania somnifera\u003c/em\u003e (ashwagandha) root extract. Eur J Biophys 5:38. https://doi.org/10.11648/j.ejb.20170502.12\u003c/li\u003e\n\u003cli\u003eLarsen DH, Li H, Shrestha S, et al (2022) Lack of blue light regulation of antioxidants and chilling tolerance in basil. Front Plant Sci 13:852654. https://doi.org/10.3389/fpls.2022.852654\u003c/li\u003e\n\u003cli\u003eLi H, Lin Y, Chen X, Bai Y, Wang C, Xu X, et al. (2018) Effects of blue light on flavonoid accumulation linked to the expression of miR393, miR394 and miR395 in longan embryogenic calli. PLoS One 13:e0191444. https://doi.org/10.1371/ journal.pone.0191444\u003c/li\u003e\n\u003cli\u003eLian TT, Moe MM, Kim YJ, Bang KS (2019) Effects of different colored LEDs on the enhancement of biologically active ingredients in callus cultures of \u003cem\u003eGynura procumbens\u003c/em\u003e (lour.) Merr. Molecules 24:4336. https://doi.org/10.3390/molecules24234336\u003c/li\u003e\n\u003cli\u003eLichtenthaler HK (1987) Chlorophylls and Carotenoids: Pigments of photosynthetic biomembranes. In: Methods in Enzymology. Elsevier, pp 350\u0026ndash;382\u003c/li\u003e\n\u003cli\u003eManivannan A, Soundararajan P, Halimah N, et al (2015) Blue LED light enhances growth, phytochemical contents, and antioxidant enzyme activities of \u003cem\u003eRehmannia glutinosa\u003c/em\u003e cultured in vitro. Hortic Environ Biotechnol 56:105\u0026ndash;113. https://doi.org/10.1007/s13580-015-0114-1\u003c/li\u003e\n\u003cli\u003eMartin KP, Madassery J (2005) Direct and indirect somatic embryogenesis on cotyledon explants of \u003cem\u003eQuassia amara\u003c/em\u003e L., an antileukaemic drug plant. In Vitro Cell Dev Biol Plant 4:54\u0026ndash;57. https://doi.org/10.1079/IVP2004588\u003c/li\u003e\n\u003cli\u003eMassa GD, Kim HH, Wheeler R, Mitchel AC (2008) Plant Productivity in Response to LED Lighting. HortScience 43:1951-1956.\u003c/li\u003e\n\u003cli\u003eMerkle SA, Parrott WA, Flinn BS (1995) Morphogenic Aspects of Somatic Embryogenesis. In: Thorpe, T.A., Ed., In Vitro Embryogenesis in Plants, Kluwer Academic Publishers, Dondrecht, 155-203. https://doi.org/10.1007/978-94-011-0485-2_5\u003c/li\u003e\n\u003cli\u003eMiao Y-X, Wang X-Z, Gao L-H, et al (2016) Blue light is more essential than red light for maintaining the activities of photosystem II and I and photosynthetic electron transport capacity in cucumber leaves. Journal of Integrative Agriculture 15:87\u0026ndash;100. https://doi.org/10.1016/S2095-3119(15)61202-3\u003c/li\u003e\n\u003cli\u003eMoshe R and Dalia E (2007) On the effect of light on shoot regeneration in petunia. Plant Cell Tissue Organ Cult. 89: 49-54.\u003c/li\u003e\n\u003cli\u003eMorello V, Brousseau VD, Wu N, et al (2022) Light quality impacts vertical growth rate, phytochemical yield and cannabinoid production efficiency in cannabis sativa. Plants 11:2982. https://doi.org/10.3390/plants11212982\u003c/li\u003e\n\u003cli\u003eMorrow RC (2008) LED lighting in horticulture. HortScience 43:1947\u0026ndash;1950.\u003c/li\u003e\n\u003cli\u003eMunoz MC (1995) Micropropagation of endangered plants. Ecologia Medi 21:291-297.\u003c/li\u003e\n\u003cli\u003eMurthy HN, Joseph KS, Hahn J-E, et al (2023) Suspension culture of somatic embryos for the production of high-value secondary metabolites. Physiol Mol Biol Plants 29:1153\u0026ndash;1177. https://doi.org/10.1007/s12298-023-01365-x\u003c/li\u003e\n\u003cli\u003eNadel BL, Altman A, Ziv M (1990) Regulation of somatic embryogenesis in celery cell suspensions. Plant Cell Tissue Organ Cult 20:119\u0026ndash;124. https://doi.org/10.1007/BF00114709\u003c/li\u003e\n\u003cli\u003eNhut DT, Huy NP, Tai NT, et al (2015) Light-emitting diodes and their potential in callus growth, plantlet development and saponin accumulation during somatic embryogenesis of \u003cem\u003ePanax vietnamensis\u003c/em\u003e Ha \u003cem\u003eet\u003c/em\u003e Grushv. Biotechnol Biotechnol Equip 29:299\u0026ndash;308. https://doi.org/10.1080/13102818.2014.1000210\u003c/li\u003e\n\u003cli\u003eOksman-Caldentey K-M, Inz\u0026eacute; D (2004) Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. Trends Plant Sci 9:433\u0026ndash;440. https://doi.org/10.1016/j.tplants.2004.07.006\u003c/li\u003e\n\u003cli\u003ePawłowska B, Żupnik M, Szewczyk-Taranek B, Cioć M (2018) Impact of LED light sources on morphogenesis and levels of photosynthetic pigments in \u003cem\u003eGerbera jamesonii\u003c/em\u003e grown in vitro. Hortic Environ Biotechnol 59:115\u0026ndash;123. https://doi.org/10.1007/s13580-018-0012-4\u003c/li\u003e\n\u003cli\u003ePedmale UV, Huang S-SC, Zander M, et al (2016) Cryptochromes interact directly with PIFs to control plant growth in limiting blue light. Cell 164:233\u0026ndash;245. https://doi.org/10.1016/j.cell.2015.12.018\u003c/li\u003e\n\u003cli\u003eRam\u0026iacute;rez-Mosqueda MA (2022a) Overview of Somatic Embryogenesis. In: Ram\u0026iacute;rez-Mosqueda MA (ed) Somatic Embryogenesis: Methods and Protocols. Springer US, New York, NY, pp 1\u0026ndash;8\u003c/li\u003e\n\u003cli\u003eRam\u0026iacute;rez-Mosqueda MA (ed) (2022b) Somatic embryogenesis: Methods and protocols, 1st edn. Springer, New York, NY\u003c/li\u003e\n\u003cli\u003eRaomai S, Kumaria S, Tandon P (2014) Plant regeneration through direct somatic embryogenesis from immature zygotic embryos of the medicinal plant, \u003cem\u003eParis polyphylla\u003c/em\u003e Sm. Plant Cell Tissue Organ Cult 118:445\u0026ndash;455. https://doi.org/10.1007/s11240-014-0496-2\u003c/li\u003e\n\u003cli\u003eRechenmacher C, Schmitt JL, Droste A (2010) Spore germination and gametophyte development of Cyathea atrovirens (Langsd. \u0026amp; Fisch.) Domin (Cyatheaceae) under different pH conditions. Braz J Biol 70:1155\u0026ndash;1160. https://doi.org/10.1590/s1519-69842010000600004\u003c/li\u003e\n\u003cli\u003eSeifipour Naghneh N, Rafiei F, Shahbazi E, Gheisari MM (2022) Chemical bioactive profile of mountain celery as affected by vegetative stages. Nat Prod Res 36:5643\u0026ndash;5646. https://doi.org/10.1080/14786419.2021.2022667\u003c/li\u003e\n\u003cli\u003eSimlat M, Ślęzak P, Moś M, et al (2016) The effect of light quality on seed germination, seedling growth and selected biochemical properties of \u003cem\u003eStevia rebaudiana\u003c/em\u003e Bertoni. Sci Hortic (Amsterdam) 211:295\u0026ndash;304. https://doi.org/10.1016/j.scienta.2016.09.009\u003c/li\u003e\n\u003cli\u003eSuresh K, Babu SS (2009) Production of Reserpine in Somatic Embryos of Rauwolfia serpentina Cultured in Bioreactors by the Induction of Elicitor (Methyl Jasmonate)\u003c/li\u003e\n\u003cli\u003eThorpe TA (2007) History of plant tissue culture. Mol Biotechnol 37:169\u0026ndash;180. https://doi.org/10.1007/s12033-007-0031-3\u003c/li\u003e\n\u003cli\u003eTienaho J, Reshamwala D, Karonen M, et al (2021) Field-Grown and In Vitro Propagated Round-Leaved Sundew (\u003cem\u003eDrosera rotundifolia\u003c/em\u003e L.) Show Differences in Metabolic Profiles and Biological Activities. Molecules 26:. https://doi.org/10.3390/molecules26123581\u003c/li\u003e\n\u003cli\u003eTittmann S, Borner M, Harst M, Bleser E (2015) Influence of led-illumination to the regeneration potential of somatic embryos of Vitis vinifera \u0026apos;Chardonnay\u0026apos; - preliminary studies. Acta Hortic. (Wagening.) 1083:445\u0026ndash;453. https://doi.org/10.17660/ ActaHortic.2015.1083.57.\u003c/li\u003e\n\u003cli\u003eTomaszewicz W, Cioć M, Dos Santos Szewczyk K, et al (2022) Enhancing In Vitro Production of the Tree Fern Cyathea delgadii and Modifying Secondary Metabolite Profiles by LED Lighting. Cells 11:. https://doi.org/10.3390/cells11030486\u003c/li\u003e\n\u003cli\u003eTomiczak K, Mikuła A, Niedziela A, et al (2019) Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application. Front Plant Sci 10:762. https://doi.org/10.3389/fpls.2019.00762\u003c/li\u003e\n\u003cli\u003eTrivedi MK, Branton A, Trivedi D, et al\u003cem\u003e.\u003c/em\u003e (2017) Chromatographic and spectroscopic characterization of the consciousness energy healing treated \u003cem\u003eWithania Somnifera\u003c/em\u003e (ashwagandha) root extract. Euro J Biophysic 5:38-47. https://\u003cem\u003e doi.org/\u003c/em\u003e10.11648/j.ejb.20170502.12.\u003c/li\u003e\n\u003cli\u003eWang P, Chen S, Gu M, et al (2020) Exploration of the effects of different blue LED light intensities on flavonoid and lipid metabolism in tea plants via transcriptomics and metabolomics. Int J Mol Sci 21:4606. https://doi.org/10.3390/ijms21134606\u003c/li\u003e\n\u003cli\u003eWilken D, Gonz\u0026aacute;lez EJ, Hohe A, et al (2005) Comparison of secondary plant metabolite production in cell suspension, callus culture and temporary immersion system. In: liquid culture systems for in vitro plant propagation. Springer, Netherlands, pp. 525\u0026ndash;537.\u003c/li\u003e\n\u003cli\u003evon Arnold S, Sabala I, Bozhkov P, et al (2002) Developmental pathways of somatic embryogenesis. Plant Cell Tissue Organ Cult 69:233\u0026ndash;249. https://doi.org/10.1023/A:1015673200621\u003c/li\u003e\n\u003cli\u003eXu Y, Yang M, Cheng F, et al (2020) Effects of LED photoperiods and light qualities on in vitro growth and chlorophyll fluorescence of \u003cem\u003eCunninghamia lanceolata\u003c/em\u003e. BMC Plant Biol 20:269. https://doi.org/10.1186/s12870-020-02480-7\u003c/li\u003e\n\u003cli\u003eYan J, Peng P, Duan G, et al (2021) Multiple analyses of various factors affecting the plantlet regeneration of \u003cem\u003ePicea mongolica\u003c/em\u003e (H. Q. Wu) W.D. Xu from somatic embryos. Sci Rep 11:6694. https://doi.org/10.1038/s41598-021-83948-w\u003c/li\u003e\n\u003cli\u003eYap ESP, Uthairatanakij A, Laohakunjit N, et al (2021) Plant growth and metabolic changes in \u0026ldquo;Super Hot\u0026rdquo; chili fruit (\u003cem\u003eCapsicum annuum\u003c/em\u003e) exposed to supplemental LED lights. Plant Sci 305:110826. https://doi.org/10.1016/j.plantsci.2021.110826\u003c/li\u003e\n\u003cli\u003eZheng C, Ma J-Q, Ma C-L, et al (2019) Regulation of growth and flavonoid formation of tea plants ( \u003cem\u003eCamellia sinensis\u003c/em\u003e) by blue and green light. J Agric Food Chem 67:2408\u0026ndash;2419. https://doi.org/10.1021/acs.jafc.8b07050\u003c/li\u003e\n\u003cli\u003eZheng L, Van Labeke M-C (2017) Long-term effects of red- and blue-light emitting diodes on leaf anatomy and photosynthetic efficiency of three ornamental pot plants. Front Plant Sci 8:917. https://doi.org/10.3389/fpls.2017.00917\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"plant-cell-tissue-and-organ-culture-pctoc","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pcto","sideBox":"Learn more about [Plant Cell, Tissue and Organ Culture (PCTOC)](https://www.springer.com/journal/11240)","snPcode":"11240","submissionUrl":"https://submission.nature.com/new-submission/11240/3","title":"Plant Cell, Tissue and Organ Culture (PCTOC)","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-4938119/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4938119/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The present study investigated potential application of somatic embryogenesis (SE) for conservation of Klussia odoratissima Mozaff. As the efficiency of SE greatly varies depending on the culture conditions including light, effects of five light spectrums including a fluorescent light and four types of light - emitting diode (LED) (red, blue, 3red-1blue, and3blue:1red) for an exposure time of a month on the maturation of K.odoratissima were also studied. To better understand the effect of light treatment on conversion of K.odoratissima cultures, two months after treatment, fresh and dry weight and metabolite contents were evaluated. Samples of the same age in situ plant and air-dry plant were also considered for relative comparison. Study of developmental stage of K.odoratissima under in vitro conditions revealed that a combination of 3red:1blue LED provided the highest number of cotyledonary (mature) embryos per callus. Hence, this light treatment was identified as the best treatment for the conversion of somatic embryos in K.odoratissima. Consequently, the highest fresh and dried weights were recorded in the cultures grown under 3blue:1red and blue LED treatment. Phthalide content of in situ plant was significantly greater than other treatments. Interestingly, the phthalide content was significantly higher in tissue culture samples compared to air-dry plants, which suggests tissue culture as an alternative technique for the production of bioactive compound in K.odoratissima. Cultures were incubated in 3blue:1red LED followed by blue LED possessed higher contents of phthalides. Application of LEDs is promising approach in micopropagation of K.odoratissima.","manuscriptTitle":"Maturation, conversion and metabolic profile of somatic embryos derived from Klussia odoratissimaMozaff as affected by light spectra ​","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-04 05:35:19","doi":"10.21203/rs.3.rs-4938119/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2024-09-11T07:05:58+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-09-03T19:40:16+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-24T05:12:21+00:00","index":"","fulltext":""},{"type":"submitted","content":"Plant Cell, Tissue and Organ Culture (PCTOC)","date":"2024-08-20T23:18:34+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"plant-cell-tissue-and-organ-culture-pctoc","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pcto","sideBox":"Learn more about [Plant Cell, Tissue and Organ Culture (PCTOC)](https://www.springer.com/journal/11240)","snPcode":"11240","submissionUrl":"https://submission.nature.com/new-submission/11240/3","title":"Plant Cell, Tissue and Organ Culture (PCTOC)","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"096d8ad5-6c43-4b59-993b-01ac7f66da17","owner":[],"postedDate":"October 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-11-11T16:02:36+00:00","versionOfRecord":{"articleIdentity":"rs-4938119","link":"https://doi.org/10.1007/s11240-024-02903-5","journal":{"identity":"plant-cell-tissue-and-organ-culture-pctoc","isVorOnly":false,"title":"Plant Cell, Tissue and Organ Culture (PCTOC)"},"publishedOn":"2024-11-04 15:57:57","publishedOnDateReadable":"November 4th, 2024"},"versionCreatedAt":"2024-10-04 05:35:19","video":"","vorDoi":"10.1007/s11240-024-02903-5","vorDoiUrl":"https://doi.org/10.1007/s11240-024-02903-5","workflowStages":[]},"version":"v1","identity":"rs-4938119","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4938119","identity":"rs-4938119","version":["v1"]},"buildId":"CiT4i_kKBbxQbnFL0ufpk","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}