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Light Wavelengths Regulate Growth and Active Components of Cordyceps militaris Fruit Bodies
Journal of Food Biochemistry
Abstract
Cordyceps militaris is widely cultivated for functional food in China and Southeast Asia. Daylight was commonly used for development of C. militaris fruit bodies. In this study, effects of light wavelengths (pink light = 1/3,450–460 nm + 2/3,620–630 nm; red light = 620–630 nm; blue light = 450–460 nm) on fruit body's production and contents of bioactive components were investigated. The results are: pink light increased dried matter contents and bioefficiency of fruit bodies to the highest (40.06/6.77%) compared with blue light (24.44/4.86%), daylight (22.69/4.55%) and red light (22.06/4.17%), respectively. Pink light significantly enhanced accumulation of carotenoids. Red light obtained the highest contents of adenosine. Pink light is the most optimal for cordycepin accumulation. In order to get the highest production of adenosine, cordycepin, carotenoids and fruit body at the same time, pink light rather than only traditional daylight should be used in the fruit body development, or light wavelengths were regulated according to target compounds of the fruit bodies.
This study indicated that different light wavelengths have significant influences on fruit body development and accumulation of the main active compounds as cordycepin, adenosine and carotenoids. Pink light (1/3,450–460 nm + 2/3,620–630 nm) was optimal for production of adenosine, cordycepin and carotenoids. While the traditionally used daylight proved not suitable for cultivation of Cordyceps militaris. These results provided technological basis for cultivation of C. militaris and a insight to light reaction study on light wavelengths regulating accumulation of secondary metabolites.
... Yi et al. [14] reported that blue LEDs are necessary to induce the CM primordial and increase the carotenoid levels in their fruiting bodies. Moreover, Dong et al. [29] reported different wavelengths of light, namely pink light (1/3, 450-460 nm with 2/3, 620-630 nm), red light (620-630 nm), and blue light (450-460 nm), were found to promote the weight increase in CM fruiting bodies significantly. It was hypothesized that the differences in the wavelength ranges used in the published results and this study were responsible for the slight differences. ...
... In contrast, this study used a single wavelength, and another possibility was that it resulted from the slight difference in the culture conditions and the CM species. Moreover, Dong et al. [29] reported that pink-, blue-, fluorescent-, and red-light all promoted bioactive substance contents in CM, whereas pink-light showed the most significant accumulation of carotenoids and cordycepin. Red light yielded the highest adenosine content, suggesting that light wavelengths and LED lights combination could be adjusted according to the demand for the targeted compounds to be the cultured duration and stage. ...
... It has been documented that exposure to blue light impedes the elongation and expansion of CM cells while concurrently promoting the biosynthesis of cordycepin. [29] The same authors postulated that this mechanism was associated with cordycepin's capacity to impede DNA replication, explicitly functioning as an anti-mutagenic protective agent under short-wavelength light, resulting in elevated cordycepin yield. [29] In addition to the aforementioned, adenosine and cordycepin have been reported to exhibit preventative properties against cellular reactive oxygen species damage, anti-inflammatory effects, broad-spectrum antibiotic activity, cancer cell proliferation inhibition, apoptotic induction, and antioxidant activity in CM. [33] ...
Cordyceps militaris (CM) is a critical fungus for promoting health, while the cordycepin has been recognized as an indicator compound with bioactive functions such as induction of apoptosis, anti-inflammation, inhibition of angiogenesis, and antioxidant properties. However, the cultivation of CM required appropriate light to induce the fruiting body into regular growth. Light-emitting diodes (LED), which combine different wavelengths and conserve energy, offer promising potential for applications in CM cultivation. Therefore, this study involved using low-power and low-illumination LED light sources of different wavelengths for intervention in the photoperiod of CM cultures to observe the effects on the growth pattern, bioactive substance content, and antioxidant activity. This study showed that the growth patterns of CM developed normally when irradiated with low-brightness LED light sources of different wavelengths. Moreover, the total phenolic content (TPC), cordycepin, and relative antioxidant activity levels in CM fruiting body water extract were significantly elevated upon exposure irradiated with a glare blue light wavelength range compared to other groups (p < .05). Hence, this study proved a high potential for LED irradiation to be applicable in CM industrial development and exploration of practical knowledge in industrial biotechnology with facilitated CM products that can be economically produced.
... In recent years, there have been advances in light-emitting diode (LED) technology development, and many studies on the effects of LED on the growth of plants ( Hogewoning et al., 2010;Hernández and Kubota, 2012;Olle and Viršil _ e, 2013) and algae for horticultural applications (Mitchell, 2015). To the best of our knowledge, there were few reports on the application of LED light in the cultivation of C. militaris ( Dong et al., 2013;Yi et al., 2014). Particularly, limited information about the effect of different LED light combinations on the culturing of C. militaris is available. ...
... Light is an important factor for the production of fruiting bodies and bioactive compounds in C. militaris (Dong et al., 2013;Yi et al., 2014;Wu et al., 2016;Yang et al., 2016). However, currently, it takes two months or more to culture the fruiting bodies of C. militaris. ...
... When grown under varying LED wavelengths, the highest cordycepin, mannitol, and adenosine contents in C. militaris fermented rice were seen under green light, red light, and blue light, respectively (Table 3). The results recorded in this study showed a similar pattern to those of Dong et al. (2013) and Yi et al. (2014), in which the optimal LED light for production was different for adenosine and cordycepin in C. militaris fruiting bodies. In contrast to these results, Dong et al. (2012) showed that blue light could enhance cordycepin content in the liquid fermentation of C. militaris. ...
... In additional to the regulatory roles on the growth, development and reproduction, light can also affect the biosynthesis of secondary metabolites (Idnum and Heitman, 2005), i.e., increase contents of bioactive substances and improve nutritional and/or color quality of medicinal fungi and plants. Cordycepin (Ni et al., 2009;Cai et al., 2013;Yao et al., 2013), canotenoids (Dong et al., 2013) and polysaccharides are the main bioactive substances in C. militaris. Carotenoids also contribute to the dark golden color of its C. militaris fruiting body products. ...
... Dong et al. (2012) indicated that red light benefited mycelium growth and adenosine content, while blue light was favorable for cordycepin biosynthesis in C. militaris mycelium. Dong et al. (2012Dong et al. ( , 2013 pink light (1/3 blue + 2/3 red) increased dried matter contents and the accumulation of carotenoids and cordycepin, while red light increased adenosine content in C. militaris fruiting bodies. These results showed a potential to increase both the yield and the quality of C. militaris products through adjustment of lighting during culture. ...
... Overall, no significant differences have been identified between the LED treatments and the white fluorescent control. Dong et al. (2013) reported pink light (red:blue of 2:1) increased dried matter contents and bioefficiency of fruit bodies compared with blue light, daylight and red light. However, except for the red or far-red LED which decreased fresh and dry weights, our results demonstrated that different red and blue combinations had no ...
... The Cmwc-1, blue light receptor gene responds to light and is involved in fruiting body production (Jiao et al., 2018). The Dash gene is required for primordia during fruiting body development (Dong et al., 2013;Yang et al., 2016). The Rhf1 gene was identified in the genome sequence of C. militaris (Zheng et al., 2011a), which is involved in the hyphal branching and fruiting body production of C. militaris (Jiang and Han, 2015). ...
... polysaccharides and ergosterol) and secondary metabolite production (e.g. cordycepin) but there few genes have been characterized (Dong et al., 2013;Yin et al., 2017;Xia et al., 2017). Rhf1 is the gene that has been characterized as necessary for mycelial growth and fruiting body development (Jiang and Han, 2015). ...
... It is well known that light exposure has regulatory effects on the growth and secondary metabolism of fungi (Dong et al., 2013). Controlling darkness and daylight conditions is traditionally applied in the cultivation of cordyceps (Dong et al., 2013;Dong, Liu, Lei, Zheng & Wang, 2012;Chen, Liu & Chang, 2011). ...
... It is well known that light exposure has regulatory effects on the growth and secondary metabolism of fungi (Dong et al., 2013). Controlling darkness and daylight conditions is traditionally applied in the cultivation of cordyceps (Dong et al., 2013;Dong, Liu, Lei, Zheng & Wang, 2012;Chen, Liu & Chang, 2011). In the present study, an obvious inhibition of stromata growth in O. xuefengensis was observed under light conditions. ...
Ophiocordyceps xuefengensis is an ethnopharmacological fungus with broad pharmacological properties. Light is a critical environmental factor for the stromata formation and development of many fungi. In this study, photomorphogenesis and blue light receptor genes were studied using a strain of O. xuefengensis. Light represses vegetative growth, but conidia linked to stromata can be observed under both light and dark conditions. Light and dark conditions had little effect on the accumulation of polysaccharides and adenosine. The genes Oxwc-1 and Oxwc-2 encoding photoreceptors of O. xuefengensis were cloned and predicted to possess polypeptides of 937 and 525 amino acids, respectively. A phylogenetic analysis based on fungal WC-1/2 supported OxWC-1 and OxWC-2 were photoreceptor. The expression of both the Oxwc-1 and Oxwc-2 genes reached a maximum after receiving light stimulation for 15 min, which might relate to the inhibition of stromata growth.
... Kết quả cho thấy đèn LED hồng có hiệu suất sinh học và hàm lượng chất khô (6,77% và 40,06%) quả thể cao nhất, sau đó là ánh sáng LED xanh (4,86% và 24,44%), tiếp theo là ánh sáng huỳnh quang (4,55% và 22,69%) và cuối cùng là ánh sáng LED đỏ (4,17% và 22,06%). Đèn LED hồng được đánh giá là tối ưu nhất cho sự tích lũy hàm lượng cordycepin và carotenoid, tuy nhiên đèn LED đỏ lại phù hợp nhất cho sự lũy hàm lượng adenosine trong quả thể nấm C. militaris (Dong et al., 2013). Theo nghiên cứu của Yi và đồng tác giả (2014), trọng lượng thể quả (tươi và khô) của nấm C. militaris khi nuôi trồng ở điều kiện ánh sáng LED đỏ (630-660 nm) và LED đỏ xa (730 nm) là thấp hơn đáng kể so với nuôi trồng dưới đèn huỳnh quang, nhưng không có khác biệt nhiều giữa ánh sáng LED kết hợp (LED đỏ kết hợp với LED xanh dương theo tỷ lệ 2:1, trong đó có 10% là LED đỏ xa) so với ánh sáng huỳnh quang. ...
... Bên cạnh đó, sự kết hợp giữa ánh sáng đỏ và xanh ở tỷ lệ thích hợp là cần thiết cho sự sinh trưởng và phát triển của thực vật (Dương Tấn Nhựt, Nguyễn Bá Nam, 2014). Tương tự như ở thực vật, gần đây nhiều báo cáo cũng cho biết ánh sáng LED có bước sóng khác nhau có ảnh hưởng khác nhau đến việc tạo thành quả thể và chất lượng quả thể của nấm C. militaris (Dong et al., 2013;Yi et al., 2014;Chiang et al., 2017, Chao et al., 2019. ...
In this study, the effects of various agricultural LED lights (LED NN), including single red LED (R), single blue LED (B), and four combinations of blue, red, and warm white (W) LED (BR, BRW1, BRW2, BRW3) on the growth and development of C. militaris (Link.) Fries were evaluated in vitro. After 7 days, samples subjected to LED NN showed shorter sporocarp sprouting time and higher sprouting ratio than the control, which was subjected to T5 fluorescent light. After 2 months, LED lights with high red ratio, such as single red LED and LED BR, had suppressing effect on the growth and development of C. militaris (Link.) Fries. On the other hand, combinations of red, blue, and warm white such as LED BRW1, LED BRW2, and LED BRW3 had the positive impact on the growth and development of this fungus. Notably, samples subjected to LED BRW2 reached 5.79 cm in height, fresh biomass of 3.67 g/20 samples. Cordycepin and Adenosine levels were 64.2 and 6.37 mg/100 g fresh mass, respectively. All of studied indicators were the higher compared to those of the control and other LED lighting schemes. Therefore, it can be conlcuded that LED lighting combination with BRW2 ratio of 1:5:1 and luminous intensity of 45±2 µmol.m-2.s-1 (511,59 Lux) was suitable for the growth and development of C. militaris (Link.) Friesand a potential replacement of fluorescent light for C. militaris (Link.) Friesin vitro propagation.
... Cordyceps militaris is a well-known edible and medicinal fungus belonging to entomopathogenic fungi, which parasitizes a larvae or pupae of lepidopteran insects [1]. Since C. militaris produces a variety of bioactive compounds with functional properties, such as cordycepin (3 -deoxyadenosine), carotenoid, ergosterol and polysaccharide, it has long been used as dietary supplement and medicinal herb in East Asia, Southeast Asia and North America [2][3][4]. In addition to the commercial use of whole-cell mass or fruiting body, the bioprocess development through fermentation and downstream processes permits a prospect in the production of specific bioactive compounds as functional ingredients for diversified applications. ...
... Although the carotenoid does not play a direct physiological role in the formation of fruiting body of C. militaris, it may act as a free radical scavenger to decrease the genotoxicity of nitrogen oxide and reactive oxygen species (ROS) under light exposure [9][10][11]. In addition to the bio-pigment, the contents of bioactive compounds in C. militaris, particularly cordycepin, were altered in response to the light condition [3,12,13]. A previous study of albinism in C. militaris grown under the light condition showed that cordycepin content in the albino strain was higher than that of the wild type [14]. ...
Cordyceps militaris is currently exploited for commercial production of specialty products as its biomass constituents are enriched in bioactive compounds, such as cordycepin. The rational process development is important for economically feasible production of high quality bioproducts. Light is an abiotic factor affecting the cultivation process of this entomopathogenic fungus, particularly in its carotenoid formation. To uncover the cell response to light exposure, this study aimed to systematically investigate the metabolic responses of C. militaris strain TBRC6039 using integrative genome-wide transcriptome and genome-scale metabolic network (GSMN)-driven analysis. The genome-wide transcriptome analysis showed 8747 expressed genes in the glucose and sucrose cultures grown under light-programming and dark conditions. Of them, 689 differentially expressed genes were significant in response to the light-programming exposure. Through integration with the GSMN-driven analysis using the improved network (iRT1467), the reporter metabolites, e.g., adenosine-5′-monophosphate (AMP) and 2-oxoglutarate, were identified when cultivated under the carotenoid-producing condition controlled by light-programming exposure, linking to up-regulations of the metabolic genes involved in glyoxalase system, as well as cordycepin and carotenoid biosynthesis. These results indicated that C. militaris had a metabolic control in acclimatization to light exposure through transcriptional co-regulation, which supported the cell growth and cordycepin production in addition to the accumulation of carotenoid as a photo-protective bio-pigment. This study provides a perspective in manipulating the metabolic fluxes towards the target metabolites through either genetic or physiological approaches.
... In addition, while colour is conspicuous at short distances, it tends to blur and match the background at longer distances (Fleishman and Persons, 2001;Barnett et al., 2018). Because of UV light scattering more in the air than light of longer wavelengths (Lythgoe, 1980;Loew, 1996;Dong et al., 2013), the signal active space of these patches may be limited to shorter distances than those of chromatic signals in the human-visible spectrum, underscoring the role of these chromatic signals in closerange communication. ...
Visual acuity, the ability to discern static spatial details, is crucial for understanding how organisms perceive their environment. Lacertid lizards are diurnal, feed on small prey, and communicate using multiple visual signals, including postures, dynamic displays and conspicuous colour patches. However, their visual acuity remains unstudied, leaving a critical gap in our understanding of their visual capabilities. Visual acuity is often estimated behaviourally using an optomotor apparatus, which quantifies a reflex-orienting movement called the optomotor response (OMR), or histologically by evaluating retinal ganglion cell (RGC) densities. We combined these two techniques to estimate visual acuity in a lacertid lizard, Podarcis muralis. OMR assays estimated an acuity of 2.05 cpd, while RGC counts revealed a peak density (>7000 cells mm−2) within the area centralis, corresponding to 1.56 cpd. RGC topographic maps revealed additional areas of high cell density in the dorso-nasal and dorso-temporal regions, while the area centralis was located slightly ventrally in the centro-temporal retina. This RGC distribution suggests adaptations to resolve stimuli in the centre and lower parts of the lizard's visual field, which may enhance predator and prey detection on the ground. Using our estimated acuities, we calculated that a lizard can detect a conspecific from 10 m and a 2 mm-long object from 40 cm away. In addition, we generated AcuityView images (R package) showing how this species might discern conspecific colour patches at different distances. These images suggest that beyond 10 cm, the surroundings become increasingly blurred, suggesting a role for static visual signals specifically in close-range communication.
... This relationship has been observed in other studies, where light was found to be an essential environmental factor for C. militaris primordial initiation and fruiting body growth [18]. Although carotenoids may not be directly involved in fruiting body formation, they could act as free radical scavengers, helping protect the fungus from oxidative stress during this crucial developmental stage [19]. C. militaris may use a light-induced stress response mechanism during fruiting body formation to optimize the production of valuable secondary metabolites, such as cordycepin. ...
Cordyceps militaris, a fungus widely used in traditional Chinese medicine and pharmacology, is recognized for its abundant bioactive compounds, including cordycepin and carotenoids. The growth, development, and metabolite production in various fungi are influenced by the complex interactions between regulatory cascades and light-signaling pathways. However, the mechanisms of gene regulation in response to light exposure in C. militaris remain largely unexplored. This study aimed to identify light-responsive genes and potential transcription factors (TFs) in C. militaris through an integrative transcriptome analysis. To achieve this, we reconstructed an expanded gene regulatory network (eGRN) comprising 507 TFs and 8662 regulated genes using both interolog-based and homolog-based methods to build the protein–protein interaction network. Aspergillus nidulans and Neurospora crassa were chosen as templates due to their relevance as fungal models and the extensive study of their light-responsive mechanisms. By utilizing the eGRN as a framework for comparing transcriptomic responses between light-exposure and dark conditions, we identified five key TFs—homeobox TF (CCM_07504), FlbC (CCM_04849), FlbB (CCM_01128), C6 zinc finger TF (CCM_05172), and mcrA (CCM_06477)—along with ten regulated genes within the light-responsive subnetwork. These TFs and regulated genes are likely crucial for the growth, development, and secondary metabolite production in C. militaris. Moreover, molecular docking analysis revealed that two novel TFs, CCM_05727 and CCM_06992, exhibit strong binding affinities and favorable docking scores with the primary light-responsive protein CmWC-1, suggesting their potential roles in light signaling pathways. This information provides an important functional interactive network for future studies on global transcriptional regulation in C. militaris and related fungi.
... Factors such as light quality and intensity can influence growth, pigmentation, conidia formation induction, and germination (Minussi et al., 1997), as well as the shape and size of the conidia in many fungal species (Mathur & Neergaard, 1973). Similar to plants, fungi exhibit a circadian rhythm, with the ability to discern day from night, due to a wide array of photoreceptors within their cells that are sensitive to a broad spectrum of electromagnetic waves, ranging from ultraviolet to infrared (Purschwitz et al., 2006;Dong et al., 2013;Ohm et al., 2013). ...
The procurement of inoculum for bioassays with target spot, a significant disease in cotton and soybeans, caused by Corynespora cassiicola can be hindered by the low production of conidia on artificial culture media. The study aimed to determine whether mechanical stress on the mycelium and exposure to ultraviolet-C (UV-C) radiation for varying durations could enhance conidial production in C. cassiicola isolates. Eight isolates from cotton and soy were used, grown in V8 juice medium. After five days of incubation in a climate-controlled chamber, each isolate either underwent mycelium scraping or remained unscraped and was subsequently exposed to UV-C radiation for either 1.0, 1.5 or 2.0 min, in comparison to a control group with zero UV exposure time. Following these procedures, conidial suspensions from each isolate were obtained and quantified (conidia per mL) using a Neubauer chamber The ISO 3S and ISO 4S isolates were found to produce more conidia than the other isolates, regardless of whether they were subjected to mycelium scraping or exposure to UV-C radiation. For most isolates, exposure to UV-C radiation for 1.0-1.5 min led to increased conidium production. Generally, it was not feasible to discern differences in conidial production with respect to the mycelium scraping process. Nevertheless, exposure to UV-C radiation for 1.0 min can be used to induce conidium production in C. cassiicola isolates.
... Compared to liquid fermentation, solid-state cultivation requires a longer duration for fruiting body formation, and high-value compounds are obtained from the fruiting bodies. Therefore, high-quality fruiting bodies are commonly acquired through solid-state cultivation with growth cycles under optimal light wavelengths and light-darkness rhythms (Dong et al., 2013a). ...
... Wu et al. (7) showed that red and yellow light promoted the mycelial growth of Pleurotus eryngii in solid culture compared with the white light. Dong et al. (8) showed that pink light increased the dry matter content of Cordyceps militaris fruiting body compared with the blue light. Yu et al. (9) investigated the effects of light quality on the growth of mycelium and fruiting bodies of Volvariella volvacea, and found that blue-green light was beneficial for the growth of mycelium, and the number and yield of fruiting bodies were also the highest under blue-green light. ...
Edible fungi has certain photo-sensitivity during the mushroom emergence stage, but there has been few relevant studies on the responses of Lyophyllum decastes to different light quality. L. decastes were planted in growth chambers with different light qualities that were, respectively, white light (CK), monochromatic red light (R), monochromatic blue light (B), mixed red and blue light (RB), and the mixture of far-red and blue light (FrB). The photo-sensitivity of L. decastes was investigated by analyzing the growth characteristics, nutritional quality, extracellular enzymes as well as the light photoreceptor genes in mushroom exposed to different light treatments. The results showed that R led to mycelium degeneration, fungal skin inactivation and failure of primordial formation in L. decastes. The stipe length, stipe diameter, pileus diameter and the weight of fruiting bodies exposed to RB significantly increased by 8.0, 28.7, 18.3, and 58.2% respectively, compared to the control (p < 0.05). B significantly decreased the stipe length and the weight of fruiting body, with a decrease of 8.5 and 20.2% respectively, compared to the control (p < 0.05). Increased color indicators and deepened simulated color were detected in L. decastes pileus treated with B and FrB in relative to the control. Meanwhile, the expression levels of blue photoreceptor genes such as WC-1, WC-2 and Cry-DASH were significantly up-regulated in mushroom exposed to B and FrB (p < 0.05). Additionally, the contents of crude protein and crude polysaccharide in pileus treated with RB were, respectively, increased by 26.5 and 9.4% compared to the control, while those in stipes increased by 5.3 and 58.8%, respectively. Meanwhile, the activities of extracellular enzyme such as cellulase, hemicellulase, laccase, manganese peroxidase, lignin peroxidase and amylase were significant up-regulated in mushroom subjected to RB (p < 0.05), which may promote the degradation of the culture materials. On the whole, the largest volume and weight as well as the highest contents of nutrients were all detected in L. decastes treated with RB. The study provided a theoretical basis for the regulation of light environment in the industrial production of high quality L. decastes.
... C. militaris contains several active components, such as cordycepin (3 ′ -deoxyadenosine), adenosine, mannitol, polysaccharide, ergosterol, and carotenoid [2,3]. These substances possess biological activities, including anti-oxidant, anti-fungal, anti-bacterial, and immunomodulatory effects and anti-inflammatory activities [4][5][6][7][8][9][10]. ...
Simple Summary
Cordyceps militaris is an entomopathogenic fungus with potential health benefits. These benefits have made this fungus marketable. Employing the transcriptome-integrated genome-scale metabolic model approach using gene inactivity moderated by metabolism and expression framework (iPS1474-tiGSMM), this work reveals metabolic fluxes in correlation with the expressed genes involved in the cordycepin and carotenoid biosynthetic pathways of C. militaris under light exposure. Additionally, an analysis of reporter metabolites emphasizes central carbon, purine, and fatty acid metabolisms, uncovering crucial processes for C. militaris adaptation to light exposure.
Abstract
The genome-scale metabolic model (GSMM) of Cordyceps militaris provides a comprehensive basis of carbon assimilation for cell growth and metabolite production. However, the model with a simple mass balance concept shows limited capability to probe the metabolic responses of C. militaris under light exposure. This study, therefore, employed the transcriptome-integrated GSMM approach to extend the investigation of C. militaris’s metabolism under light conditions. Through the gene inactivity moderated by metabolism and expression (GIMME) framework, the iPS1474-tiGSMM model was furnished with the transcriptome data, thus providing a simulation that described reasonably well the metabolic responses underlying the phenotypic observation of C. militaris under the particular light conditions. The iPS1474-tiGSMM obviously showed an improved prediction of metabolic fluxes in correlation with the expressed genes involved in the cordycepin and carotenoid biosynthetic pathways under the sucrose culturing conditions. Further analysis of reporter metabolites suggested that the central carbon, purine, and fatty acid metabolisms towards carotenoid biosynthesis were the predominant metabolic processes responsible in light conditions. This finding highlights the key responsive processes enabling the acclimatization of C. militaris metabolism in varying light conditions. This study provides a valuable perspective on manipulating metabolic genes and fluxes towards the target metabolite production of C. militaris.
... When cultivating edible mushrooms, blue light is employed to stimulate the growth of fruiting bodies. The research has been conducted to investigate the photobiological effects of LED light and the biosynthetic activity, as well as the formation of fruiting bodies of Cordyceps militaris fungi [26,27]. Arjona et al. (2009) reported that blue light induces morphogenesis of Pleurotus ostreatus [28]. ...
The findings of the study demonstrate the impact of low-intensity laser and quasi-monochromatic light on the biosynthetic
activity of the edible medicinal fungus L. edodes during submerged cultivation. An artificial lighting installation based on
matrices of light-emitting diodes (LED) emitting light at 470 nm (blue), 530 nm (green), 650 nm (red), and argon gas laser
(488 nm) was used. Irradiation with blue and red LED and laser led to a shortening of the lag phase by 2 days and an increase
in the mycelial mass. Irradiation with laser light resulted in the highest mycelial mass yield (14.1 g/L) on the 8th day of
cultivation. Irradiation in all used wavelength ranges caused an increase in the synthesis of both extracellular and intracellular
polysaccharides. Laser light at 488 nm and LED at 470 nm proved to be the most effective. Irradiation with red, green,
and blue laser light caused an increase in the total amount of fatty acids in the mycelial mass compared to the control. A
significant distinction in qualitative composition was observed: short-chain acids C6‒
C12 compounds were produced under
red light irradiation, whereas long-chain C20‒
C24 were formed under green light irradiation. The most significant changes
in the aromatic profile of the mycelial mass and culture liquid were recorded upon irradiation with green light. The content
of aromatic components increased 24.6 times in the mycelial mass and 38.5 times in the culture liquid. The results suggest
the possibility of using low-intensity quasi-monochromatic light for targeted regulation of L. edodes biosynthetic activity.
... Genomic studies have determined a model based on the genetics, physiology, and metabolism of C. militaris, where a C:N ratio of 8:1 in synthetic media optimizes the growth and production of cordycepin (metabolite indicator of the quality of Cordyceps species) . Light is another factor that influences fruiting body development: in artificial rice culture, 500-1000 lux is used (Shrestha et al. 2004a(Shrestha et al. , 2012Sung et al. 2006;Hong et al. 2010), while 20-500 lux allows the development of stromata in culture on insects (Chen and Ichida 2002) and pink light (1/3,450-460 + 2/3,620-630 nm) facilitates the highest production of dry matter, carotenes and nucleotides such as cordycepin and adenosine (Dong et al. 2013). ...
Distributed in Mexico, Cordyceps mexicana is a phylogenetic sister species of the Cordyceps militaris group. The present study evaluated biomass production of this species in in vitro cultures and fruiting bioassays to determine the capacity and time for primordia development of different strains of the species in different substrates. The mycelial growth of the strains of C. mexicana showed cottony, convex, circular, or irregular growth, with a fimbriated and lobulate-fimbriated margin and whitish and light yellow surface coloration in PDA (potato dextrose agar) medium. Biomass production in PD (Potato Dextrose) medium differed among strains (P<0.05), with CIRB-Cm_c14, CIRB-Cm_c16, CIRB-Cm_c24, and CIRB-Cm_c25 presenting the highest dry weight values (0.42, 0.48, 0.493 and 0.487 g/100 ml, respectively). In solid culture media, the highest biomass production was presented in Sabouraud dextrose agar with yeast (medium rich), intermediate production in PDA (rich medium) and malt extract agar (general medium), and lowest production in maize agar and wheat agar. In fruiting bioassays conducted at 500–1000 lux and 90% humidity on substrates of brown rice, rice with added mealworm (Tenebrio molitor; Coleoptera) larvae, and rice with added “chinicuil” (Comadia redtenbacheri: Lepidoptera) larvae, C. mexicana demonstrated the ability to completely invade the substrates; however, the time taken for completely invade the substrates and primordia development differed, being shorter in brown rice and brown rice with added “chinicuils”. This study is the first to describe mycelial growth in Cordyceps mexicana in vitro and solid cultures, as well as the production of primordia in the species.
... For most herb species, red combined with blue light significantly increased plant yield than monochromatic red or blue light. The yield of herb plants decreased as blue light proportion increased, but this trend varied among species [19] . Some studies also showed that the concentrations of essential oils and phenolic compounds in various herbs were enhanced, and the antioxidant capacities were improved by employing red, blue, and/or ultraviolet light compared with white light or sunlight [2] . ...
The objective of this study was to evaluate the growth and volatile oil concentrations of thyme (Thymus vulgaris L.) plants under different red to blue light ratios of 4:1 (R4B1), 2:1 (R2B1), 1:1 (R1B1), and 1:2 (R1B2) with light-emitting diodes (LEDs) and fluorescent light (FL) in a plant factory. Thyme plants were sampled at three intervals of 12, 24, and 36 d after treatment. The results showed that the growth and medicinal components accumulation of thyme plants were significantly affected by different light qualities. The significant higher biomass, leaf area, and volatile oil concentrations of thyme plants were obtained under treatment R4B1 compared with treatment FL, regardless of the cultivation period. When analyzing the volatile oil constituents of thyme plants, thymol, γ-terpinene, p-cymene and α-terpinene were detected as the main constituents. However, the response of these different constituents varied with different light qualities. The above results indicated that the targeted constituent concentrations could be manipulated by employing different light qualities according to various purposes. Based on the above results, R4B1 can be considered as the optimal light treatment for thyme plants growth and volatile oil production in a closed production system with LEDs.
... These optimization strategies in solid medium culture led to a 68% (1.73 g/bottle) increase in fruiting body yield and a 63% (9.17 mg/g) increase of COR yield in fruiting body. Light is an important factor in the production of fruiting bodies and bioactive compounds in C. militaris [79]. Chiang et al. studied the effects of LED wavelengths on the production of bioactive compounds in C. militaris cultivated on brown rice [80]. ...
C. militaris is an insect-born fungus that belongs to Ascomycota and Cordyceps. It has a variety of biological activities that can be applied in medicine, health-care products, cosmeceuticals and other fields. Cordycepin (COR) is one of the major bioactive components identified from C. militaris. Thus, C. militaris and COR have attracted extensive attention. In this study, chemical synthetic methods and the biosynthesis pathway of COR were reviewed. As commercially COR was mainly isolated from C. militaris fermentation, the optimizations for liquid and solid fermentation and genetic modifications of C. militaris to increase COR content were also summarized. Moreover, the research progress of genetic modifications of C. militaris and methods for separation and purification COR were introduced. Finally, the existing problems and future research direction of C. militaris were discussed. This study provides a reference for the production of COR in the future.
... According to Yousef and coauthors showed that a combination of R and B LED light with a high R portion was effective in producing vigorous grafted tomato seedlings compared to blue and red light alone 20,47,48 . Also, Dong et al. 49 , as compared to blue light alone, red light alone, and sunshine, combined red and blue light (1/3 blue light at 450-460 nm + 2/3 red light at 620-630 nm, at 400 lx and 12 h photoperiod for 60 days) improved the DW and bio efficiency of Cordyceps militaris mushroom. Plant production was considerably improved for most species when combined light wavelengths with a considerable proportion of red light supplemented by blue light were used 50 . ...
Light is one of the most important limiting factors for photosynthesis and the production of
plants, especially in the regions where natural environmental conditions do not provide sufficient sunlight, and there is a great dependence on artificial lighting to grow plants and produce food.
The influence of light intensity, quality, and photoperiod on photosynthetic pigments content and some biochemical and growth traits of cucumber seedlings grown under controlled conditions was investigated. An orthogonal design based on a combination of different light irradiances, ratio of
LEDs and photoperiods was used. Treaments consisted of three light irradiance regimes (80, 100, and 150 μmol m−2 s−1) provided by light‐emitting diodes (LEDs) of different ratios of red and blue (R:B) (30:70, 50:50, and 70:30) and three different photoperiods (10/14, 12/12, and 14/10 h). The white light was used as a control/reference. Plant height, hypocotyl length, stem diameter, leaf area, and soluble sugar content were highest when exposed to LM9 (150 μmol m−2 s−1; R70:B30; 12/12 h) light mode, while the lowest values for the above parameters were obtained under LM1 (80 μmol m−2 s−1; R30:B70; 10/14 h). Higher pigments contents (chlorophyll a, chlorophyll b, and carotenoid) were obtained when light regime LM9 (150 μmol m−2 s−1; R70:B30; 12/12 h) was applied. In general, cucumber seedlings grown under the LM9 regime showed a significant increase in growth as well as photosynthetic capacity. It seems that the content of photosynthetic pigments is the key factor responsible for
the performance of cucumber seedlings grown under different lighting modes, compared to other traits studied. We recommend monitoring the content of chlorophyll a, b, and their ratio value when studying the light requirement of cucumber plants.
... A study by Yang et al. [12] revealed that the fungi without the blue-light receptor gene (wc-1) showed a significant reduction in both cordycepin and carotenoid. Other evidence also suggested that the carotenoid content of C. militaris is enhanced by light [6,13], although light stress during the late growth stage may reduce carotenoid production. [19]. ...
Simple Summary
Cordyceps militaris is an edible fungus that has been long used in traditional medicine. A large body of research has provided evidence of the medicinal properties of C. militaris extract and its demand has been increasing over the years. This study aims to construct and understand the light-responsive gene regulatory network of the fungi by combining the transcription factor (TF)-target gene interactions with the transcriptomic analysis of C. militaris under a light-programming condition. The study identified several key TFs and their gene targets that regulate growth, development and secondary metabolite production in the fungi under specific light conditions.
Abstract
Cordyceps militaris is an edible fungus that produces many beneficial compounds, including cordycepin and carotenoid. In many fungi, growth, development and secondary metabolite production are controlled by crosstalk between light-signaling pathways and other regulatory cascades. However, little is known about the gene regulation upon light exposure in C. militaris. This study aims to construct a gene regulatory network (GRN) that responds to light in C. militaris. First, a genome-scale GRN was built based on transcription factor (TF)-target gene interactions predicted from the Regulatory Sequence Analysis Tools (RSAT). Then, a light-responsive GRN was extracted by integrating the transcriptomic data onto the genome-scale GRN. The light-responsive network contains 2689 genes and 6837 interactions. From the network, five TFs, Snf21 (CCM_04586), an AT-hook DNA-binding motif TF (CCM_08536), a homeobox TF (CCM_07504), a forkhead box protein L2 (CCM_02646) and a heat shock factor Hsf1 (CCM_05142), were identified as key regulators that co-regulate a large group of growth and developmental genes. The identified regulatory network and expression profiles from our analysis suggested how light may induce the growth and development of C. militaris into a sexual cycle. The light-mediated regulation also couples fungal development with cordycepin and carotenoid production. This study leads to an enhanced understanding of the light-responsive regulation of growth, development and secondary metabolite production in the fungi.
... This was better than Rhodotorula glutinis, a fungal elicitor, which stimulated the accumulation of carotenoids by 13.72% [37]. According to a previous study, the TCC of C. militaris fruiting bodies was significantly increased through stimulation by light [38]. On the other hand, low concentrations of fluoride having no effect [39] or positively stimulating photosynthetic pigments have also been reported [25]. ...
Cordyceps militaris possesses several compounds with medicinal properties, and is commonly used in traditional Chinese functional food and medicine for a variety of health benefits. Because of its rare occurrence in nature, the market demand for artificial C. militaris is on the rise. Furthermore, efforts to increase its bioactive ingredients have also been considered in research. In this study, we aimed to investigate the effect of fluoride on the growth and enrichment of bioactive compounds in C. militaris. A wide range of potassium fluoride concentrations (0, 0.001, 0.01, 0.1, and 1 mM) were added to the culture media as a source of fluoride during the cultivation of C. militaris fruiting bodies. The contents of fluorine and bioactive substances of the fruiting bodies in normal (NM) and fluorine-supplemented (FM) media were measured and compared. C. militaris raised in the growth medium supplemented with 0.01 mM potassium fluoride led to a 44.86% (1.55 ± 0.14 g/bottle) increase in biomass and a 23.43% (3161.38 ± 35.71 µg/g) increase in total carotenoid content in the fruiting bodies. Furthermore, a remarkable increase in superoxide dismutase-like activity (84.75 U/mg) and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (IC50 = 2.59 mg/mL) was recorded. In human cancer cell-based assays, C. militaris raised in FM caused stronger cytotoxicity, apoptosis, and cell cycle arrest in human osteosarcoma cells. These results demonstrated that a low dose of fluoride could stimulate the growth of C. militaris fruiting bodies and enhance the production of bioactive ingredients that possess useful antioxidant and anticancer activities.
... It has been reported that C. militaris pigments were carotenoids (Dong et al., 2013a;Yi et al., 2014). Therefore, all unigenes derived from RNA-seq data were searched against the carotenoid biosynthetic pathway in the KEGG database, and the results showed that only the CCM_06728 (CAO-2) and CCM_09155 (YLO-1) genes were found to be involved in the biosynthesis of carotenoids (Supplementary Figure S2). ...
Cordyceps militaris, a valuable edible and medicinal fungus, has attracted increasing attention because of its various bioactive ingredients. However, the biosynthetic pathway of C. militaris carotenoids is still unknown due to lack of transcriptome information. To uncover genes related to the biosynthesis of C. militaris carotenoids, the transcriptomes of mycelia CM10_D cultured under dark conditions and mycelia CM10_L cultured under light exposure conditions were sequenced. Compared with mycelia CM10_D, 866 up-regulated genes and 856 down-regulated genes were found in mycelia CM10_L. Gene ontology (GO) analysis of differentially expressed genes (DEGs) indicated that DEGs were mainly classified into the “metabolic process,” “membrane,” and “catalytic activity” terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs suggested that DEGs were mainly enriched in “metabolic pathways,” “MAPK signaling pathway-yeast,” and “biosynthesis of secondary metabolites.” In addition, the carotenoid content of the Cmtns gene deletion mutant (ΔCmtns) was significantly lower than that of the wild-type C. militaris CM10, while the carotenoid content of the complementary strain (ΔCmtns-c) of the Cmtns gene was not significantly different from that of C. militaris CM10, suggesting that the Cmtns gene significantly affected the biosynthesis of carotenoids in C. militaris. These results potentially pave the way for revealing the biosynthetic pathway of carotenoids and improving carotenoids production in C. militaris.
... There are many studies on the cultivation of caterpillar fungi, in order to characterize the physiological and biochemical responses under different conditions. Effect of temperature (Hung, Keawsompong, Hanh, Sivichai, & Hywel-Jones, 2009), light (Dong et al., 2013;Yang & Dong, 2014) and nutrients on the growth of cordyceps fungi and the production of their metabolites were investigated. These medicinal fungi were also subjected to heat-shock, frozenshock, osmotic or oxidative stresses, and it seems they have strong physiological adaptations to environmental stress (Ren & Yao, 2013). ...
Ophiocordyceps xuefengensis is a kind of caterpillar fungus and used as ethnopharmacological invigorant in China. A transcriptomic approach was applied to reveal the differential gene expression of O. xuefengensis under breathable and airtight conditions, and the targeted metabolites were determined by HPLC-QTOF-MS/MS technology. Growth was inhibited during 28 d cultivation and a total of 170 differentially expressed genes were obtained under airtight condition. The differentially expressed genes were mainly significantly enriched into amino acid metabolic pathways, namely tryptophan metabolism, phenylalanine metabolism, arginine and proline metabolism. These significantly enriched metabolic pathways were all involved in the enzyme amidase. Meanwhile, the content of tryptophan, phenylalanine, arginine and proline were increased under airtight condition. Consequently, the metabolism of these amino acids was supposed to be suppressed in O. xuefengensis under airtight condition. The detailed knowledge of this transcriptional program and biochemical analysis will allow for a systematic understanding of cordyceps fungus cultivation.
... Another study found that the fewest C. militaris fruiting bodies formed under red light. 39 Synnemata can grow under blue, green, and white light, and the dry weight of samples grown under the various light wavelengths was not significantly different. Blue light seems to promote conidia production and white light, HEA production. ...
Isaria cicadae (syn. Cordyceps cicadae) is one of the most valued edible and medicinal fungi and has been used in Asia as a substitute for Ophiocordyceps sinensis. Wild I. cicadae is limited and seasonal, and its cultivation is deserved. In this investigation we studied synnema formation by and nucleoside production in cicada flower under different environmental conditions. I. cicadae produced an asexual structure and mitospores instead of meiotic ascospores; this indicates that the term “synnema” is more suitable than “fruiting body” for this species. The optimal temperature was 25°C for growth of I. cicadae mycelia on potato dextrose agar plates but was 20°C for synnema formation on wheat medium. Synnemata can grow well under blue, green, and white light, and the dry weight of samples grown under these 3 light wavelengths is not significantly different. However, neither primordia nor synnemata formed under red light. Blue light promotes conidia production and white light promotes N⁶-(2-hydroxyethyl)-adenosine (HEA) production. Weak white light at 50 and 150 lux was more suitable for synnema production than strong-intensity light at 850 lux. The growth curve showed that HEA content has the same trend as synnema production over the entire cultivation period. The optimal harvesting time for I. cicadae cultivated on wheat medium is 35 days after inoculation. HEA content in the synnemata cultivated on wheat medium under the optimal conditions was significantly higher than that of the wild species and of synnemata cultivated on pupae, suggesting that synnemata cultivated on wheat medium may have potential as a substitute for wild resources. The results presented herein provide a new strategy for producing superior-quality synnemata of I. cicadae and further elucidate the effects of environmental conditions on metabolite accumulation in fungi.
... After exposure to light, the hyphae color changes from white to yellow or orange, and then the primordia begin to develop. Light affects biomass growth (Kho et al., 2016) and metabolism, such as cordycepin and carotenoid formation (Dong et al., 2013b;Lian, Dong, Yang, & Sun, 2014). Fluorescent lamps (Wu, Liang, Tseng, & Hu, 2016) and light-emitting diode (LED) (Chiang, Liang, Wang, & Liang, 2017;Kho et al., 2016) can be used to cultivate C. militaris fruiting bodies. ...
Cordyceps militaris is widely cultivated as a functional food and traditional medicine in China and Southeast Asia. In this study, C. militaris hybrid strains were isolated from different parts of hybrid fruiting body cultivated through mating-based sexual reproduction of ascospores. We investigated the effect of strains, solid-state substrates and light condition on production of yield and contents of bioactive compounds (cordycepin and adenosine) using HPLC in fruiting bodies. The results indicated that yield and production of bioactive compounds exist distinct differences on strain separated part, solid-substrates and light conditions. The middle strain obtained the highest yield on wheat substrate. The yield was higher on wheat substrate, the concentration of cordycepin and adenosine in fruiting bodies were higher on rice substrate. The red light is beneficial to high yield, and white light is conducive to the accumulation of cordycepin in fruiting bodies, the light condition has little effect on the adenosine content.
... Our results were similar to those of the previous reports, but no orange color was observed, which could be caused by dark cultivation in this study. Light condition could influence the synthesis of pigments, resulting in different color appearance (Dong et al. 2013a). For in-stance, the content of xanthophylls, a kind of novel carotenoids, in C. militaris is significantly increased with the treatment of pink light (Dong et al. 2013b). ...
Caterpillar fungi have numerous pharmacological and therapeutic applications in traditional medicine, due to a variety of active chemical constituents, such as cordycepin and adenosine. It is imperative to discover new resource for artificial cultivation and biometabolite production since the traditional natural species are endangered. In this study, a new strain HACM 001 was isolated and identified as Ophiocordyceps xuefengensis by rDNA-ITS sequencing. This strain showed the potential of artificial infection to caterpillar larvae leading to mummification, as well as fermentation mycelia in liquid culture and cultivation stromata in solid medium. Eight nucleosides and nucleobases, especially cordycepin and adenosine, were determined and analyzed with HPLC-DAD-Q-TOF-MS/MS technology. Cordycepin was detected in all forms of present O. xuefengensis strain at different contents, among which the highest content (37.1 μg/g) appeared in the stromata cultivated on solid medium. The content of adenosine in mycelia and stromata, respectively, reached 1155 μg/g and 1470 μg/g. Therefore, O. xuefengensis might be an alternative source for obtaining artificial fungus-caterpillar-larvae complex and producing cordycepin and adenosine.
... Biologically active constituents in medicinal mushrooms have been of great interest since long. Dong et al. [41] reported that environmental conditions affected the accumulation of active components and the biological efficiency of Cordyceps militaris. Lin et al. [42] showed that the substrate not only affects the yield of C. militaris, but also the production of medicinally active components. ...
Ganoderma lucidum is a medicinal mushroom that has been widely used in East Asia for the treatment of various diseases. The pharmacological activity of this fungus is primarily attributable to the polysaccharides and triterpenoids. In this study, to obtain the fruit bodies with improved content of active constituents, we examined the effect of salicylic acid (SA) and calcium ion on the biosynthesis of polysaccharides and triterpenoids by spraying the chemicals during the fruiting. To explore the underlying mechanisms for the variation, the transcripts of related genes involved in the polysaccharide and triterpenoid biosynthesis were measured. Results showed that Ca²⁺ had no effect on production of polysaccharides and triterpenoids, whereas SA increased triterpenoid content by 23.32%, compared to the control, but it had little influence on polysaccharide production. Interestingly, the combined induction increased polysaccharide and triterpenoid content by 9.02% and 13.61%, respectively, compared to the control. Under Ca²⁺ induction, the transcript of ugp gene in the polysaccharide biosynthetic pathway up-regulated in all three stages (mycelium, primordium, and fruit body), while pgm and gls gave no response in the mycelium and primordium stages, and up-regulated in the fruit body stage. Differently, six key triterpenoid biosynthetic genes including hmgr, hmgs, mvd, fps, sqs, and ls did not respond to the induction. In the case of SA and combined induction, pgm and ugp were up-regulated in all three stages, while gls showed an increased expression in the primordium stage and no response in other stages. The six triterpenoid biosynthetic genes were up-regulated in all three stages. The present study provides a useful approach to producing G. lucidum fruit bodies with high polysaccharide and triterpenoid content. This is important to the G. lucidum industry.
... These may be extracted from fruiting bodies or mycelia and are currently studied as therapeutic agents [7-8, 10, 12-13]. Nucleosides can be differentially induced by intrinsic or environmental factors, such as the developmental stage of the fungus, light exposure and wavelength, the composition of the culture media, among other factors [7][8], [12][13][14][15]. Traditionally, the nucleosides cordycepin (3'-deoxyadenosine), adenosine, inosine, guanosine, and uridine have been used as markers to assess the quality of natural and cultured species of Cordyceps [4,[16][17]. ...
Cordyceps sensu lato is a genus of arthropod-pathogenic fungi, which have been used traditionally as medicinal in Asia. Within the genus, Ophiocordyceps sinensis is the most coveted and expensive species in China. Nevertheless, harvesting wild specimens has become a challenge given that natural populations of the fungus are decreasing and because large-scale culture of it has not yet been achieved. The worldwide demand for products derived from cultivable fungal species with medicinal properties has increased recently. In this study, we propose a new species, Cordyceps nidus, which parasitizes underground nests of trapdoor spiders. This species is phylogenetically related to Cordyceps militaris, Cordyceps pruinosa, and a sibling species of Cordyceps caloceroides. It is found in tropical rainforests from Bolivia, Brazil, Colombia and Ecuador. We also investigated the medicinal potential of this fungus based on its biochemical properties when grown on four different culture media. The metabolic profile particularly that of nucleosides, in polar and non-polar extracts was determined by UPLC, and then correlated to their antimicrobial activity and total phenolic content. The metabolome showed a high and significant dependency on the substrate used for fungal growth. The mass intensities of nucleosides and derivative compounds were higher in natural culture media in comparison to artificial culture media. Among these compounds, cordycepin was the predominant, showing the potential use of this species as an alternative to O. sinensis. Furthermore, methanol fractions showed antimicrobial activity against gram-positive bacteria, and less than 3.00 mg of gallic acid equivalents per g of dried extract were obtained when assessing its total phenolic content by modified Folin-Ciocalteu method. The presence of polyphenols opens the possibility of further exploring the antioxidant capacity and the conditions that may enhance this characteristic. The metabolic composition and biochemical activity indicate potential use of C. nidus in pharmaceutical applications.
... The combination of red and blue light provided better excitation of photoreceptors including phytochromes, cryptochromes, and phototropins, and resulted in higher photosynthetic activity than plants under either monochromatic red or blue light [36]. Dong et al. [37] reported that combined red and blue light (1/3 blue light at 450−460 nm + 2/3 red light at 620−630 nm, at 400 lx and 12-h photoperiod for 60 days) increased the DW and bioefficiency of Cordyceps militaris mushroom compared with blue light alone, red light alone, and sunlight. For most species, combined light wavelengths with a large proportion of red light supplemented with blue light significantly increased plant yield [16]. ...
In recent years, consumption of herb products has increased in daily diets, contributing to the prevention of cardiovascular diseases, chronic diseases, and certain types of cancer owing to high concentrations of phytonutrients such as essential oils and phenolic compounds. To meet the increasing demand for high quality herbs, controlled environment agriculture is an alternative and a supplement to field production. Light is one of the most important environmental factors influencing herb quality including phytonutrient content, in addition to effects on growth and development. The recent development and adoption of light-emitting diodes provides opportunities for targeted regulation of growth and phytonutrient accumulation by herbs to optimize productivity and quality under controlled environments. For most herb species, red light supplemented with blue light significantly increased plant yield. However, plant yield decreased when the blue light proportion (BP) reached a threshold, which varied among species. Research has also shown that red, blue, and ultraviolet (UV) light enhanced the concentration of essential oils and phenolic compounds in various herbs and improved antioxidant capacities of herbs compared with white light or sunlight, yet these improvement effects varied among species, compounds, and light treatments. In addition to red and blue light, other light spectra within the photosynthetically active region—such as cyan, green, yellow, orange, and far-red light—are absorbed by photosynthetic pigments and utilized in leaves. However, only a few selected ranges of light spectra have been investigated, and the effects of light quality (spectrum distribution of light sources) on herb production are not fully understood. This paper reviews how light quality affected the growth and phytonutrient accumulation of both culinary and medicinal herbs under controlled environments, and discusses future research opportunities to produce high quantity and quality herbs.
... 12 Pink light proved to be optimal for cordycepin accumulation. 18 There are many reports on improving cordycepin production by various factors. Ferrous sulfate addition could improve cordycepin production. ...
Background:
In commercial production of Cordyceps militaris (a famous Chinese medicine), usually cereal grains were utilized as cultivation substrates. This study is aimed to evaluate the efficiency of agricultural wastes as substitute materials in the low cost production of C. militaris. Cottonseed shells (CS), corn cob particles (CCP), Italian poplar sawdusts (IPS) and substrates spent by Flammulina velutipes (SS) were employed to cultivate C. militaris, using rice medium as control.
Results:
CS and CCP were suitable for the fruit body formation of C. militaris, with yields of 22 g bottle(-1) and 20 g bottle(-1) respectively. The fruit bodies grown on CCP showed the highest levels of cordycepin and adenosine up to 9.45 mg g(-1) and 5.86 mg g(-1) , respectively. The content of D-mannitol in fruit body obtained on CS was 120 mg g(-1) (80% of the control group), followed by that on CCP, 100 mg g(-1) . The fruit bodies cultivated on CCP displayed a high crude polysaccharide level of 26.9 mg g(-1) , which was the mostly close to the control group (34.5 mg g(-1) ).
Conclusion:
CS and CCP are effective substrates for the production of fruit body and bioactive compounds by C. militaris. This study provides a new approach of decreasing the cost of C. militaris cultivation and dealing with these agricultural wastes.
Macrofungi have been considered for centuries as healthy foods. In addition, they are also widely used as the main component and traditional medicines in different cultures. Of these mushrooms, strains belonging to Cordyceps sp. are considered very precious candidates based on the wide range of their bioactive compounds, which are used in the treatment of different diseases. Based on the increased market demand, many attempts are now being made to cultivate this type of mushroom on an industrial scale using both solid and liquid cultivation systems. These in vitro cultivation strategies are now undergoing different optimization strategies to maximize the desired product quality and yield, shorten the cultivation time, and reduce the production cost. This chapter will cover the recent development of cultivation strategies applied for Cordyceps sp. cultivation in general, with more focus on liquid culture cultivation systems for both surface and submerged cultures for the production of the key important metabolites: cordycepin, adenosine, and polysaccharides.
Edible fungi have certain photo-sensitivity during the mushroom emergence stage, but there have been few relevant studies on the responses of Ganoderma lucidum to different light irradiation conditions. Ganoderma lucidum were planted in an environmentally controllable mushroom room with different light supply modes that were, respectively, continuous white light (CK), red light (R), green light (G), blue light (B), and intermittent red light (R-), green light (G-), and blue light (B-), with a total light intensity of 15 μmol·m⁻²·s⁻¹ and a light/dark (L/D) period of 12 h/12 h for each treatment. The interval in intermittent light treatments was 30 min. The optimal light supply mode suitable for the growth of Ganoderma lucidum was explored by analyzing the characteristics, nutritional quality, and extracellular enzyme activity in mushrooms exposed to different light treatments. The results showed that red light (whether in continuous or intermittent supply modes) inhibited the fruiting body differentiation of Ganoderma lucidum, showing delayed differentiation or complete undifferentiation. The highest stipe length and pileus diameter of fruiting bodies were detected under G- treatment, which were, respectively, increased by 71.3% and 3.2% relative to the control. The highest weight of fruiting bodies was detected under G treatment, which was significantly increased by 21.4% compared to the control (p < 0.05). Intermittent light mode seemed to be more conducive to the size development of the fruiting body, while continuous light mode was beneficial for increasing the weight. The highest contents of crude protein and total triterpenes in pileus were detected under G treatment (significantly 14.9% and 28.1% higher than the control, respectively), while that of the crude polysaccharide was detected under G- treatment (significantly 35.7% higher than the control) (p < 0.05). The highest activities of extracellular enzymes such as cellulase, hemicellulase, laccase, lignin peroxidase, and amylase were detected in fruiting bodies subjected to G treatment, which were significantly increased by 11.9%~30.7% in the pileus and 9.5%~44.5% in the stipe. Green light might increase the weight and nutrient accumulation in the pileus of Ganoderma lucidum via up-regulating the extracellular enzyme activities. This study provides an effective light supply strategy for regulating the light environment in the industrial production of Ganoderma lucidum.
Abstract The findings of the study demonstrate the impact of low-intensity laser and quasi-monochromatic light on the biosynthetic activity of the edible medicinal fungus L. edodes during submerged cultivation. An artificial lighting installation based on matrices of light-emitting diodes (LED) emitting light at 470nm (blue), 530nm (green), 650nm (red), and argon gas laser (488nm) was used. Irradiation with blue and red LED and laser led to a shortening of the lag phase by 2days and an increase in the mycelial mass. Irradiation with laser light resulted in the highest mycelial mass yield (14.1g/L) on the 8th day of cultivation. Irradiation in all used wavelength ranges caused an increase in the synthesis of both extracellular and intracel-lular polysaccharides. Laser light at 488nm and LED at 470nm proved to be the most effective. Irradiation with red, green, and blue laser light caused an increase in the total amount of fatty acids in the mycelial mass compared to the control. A significant distinction in qualitative composition was observed: short-chain acids C6‒C12 compounds were produced under red light irradiation, whereas long-chain C20‒C24 were formed under green light irradiation. The most significant changes in the aromatic profile of the mycelial mass and culture liquid were recorded upon irradiation with green light. The content of aromatic components increased 24.6 times in the mycelial mass and 38.5 times in the culture liquid. The results suggest the possibility of using low-intensity quasi-monochromatic light for targeted regulation of L. edodes biosynthetic activity.
This manuscript deals with cordycepin, an interesting secondary compound produced from entomopathogenic fungus, Cordyceps. It has attracted commercial interest due to its immense pharmacological importance beneficial to human health. In this study, the contents of cordycepin and its derivatives, like adenine and adenosine, were evaluated through solid-state fermentation using combinations of various grains as substrate. Treatment with grain combination numbers 2, 7, 8, and 9 exhibited higher cordycepin content (1.621, 1.929, 1.895, and 1.996 mg/g cordycepin, respectively) than control (rice). The grain combination number 7 exhibited significantly higher adenine content (700 mg/g) than the control and all other combinations. Treatments with grain combination numbers 2, 5, and 7 exhibited higher adenosine content (2.719, 2.938, and 3.392 mg/g, respectively); however, no significant increase in adenosine content was noted in any treatments. The biomass including fresh mycelium and fruit body was found higher in grain combination numbers 7 and 9, leading to enhanced cordycepin content. Overall, the increase in the fresh biomass significantly enhanced cordycepin accumulation. The level of cordycepin was recorded as higher than that of its derivatives, adenosine and adenine. The grain combination of rice, wheat, jowar, bajra, and sugarcane bagasse added to basal medium exhibited the highest cordycepin content and was found suitable for solid-state fermentation of Cordyceps militaris. To our understanding, the present study is the first to use combinations of cereals for the production of cordycepin from C. militaris.
Cordyceps militaris is an entomopathogenic ascomycete with similar pharmacological importance to that of the wild caterpillar fungus Ophiocordyceps sinensis. C. militaris has attracted significant research and commercial interest due to its content in bioactive compounds beneficial to human health and the relative ease of cultivation under laboratory conditions. However, room for improvement exists in the commercial-scale cultivation of C. militaris and concerns issues principally related to appropriate strain selection, genetic degeneration of cultures, and substrate optimization. In particular, culture degeneration—usually expressed by abnormal fruit body formation and reduced sporulation—results in important economic losses and is holding back investors and potential growers (mainly in Western countries) from further developing this highly promising sector. In the present review, the main factors that influence the generation of biomass and metabolites (with emphasis on cordycepin biosynthesis) by C. militaris are presented and evaluated in conjunction with the use of a wide range of supplements or additives towards the enhancement of fungal productivity in large-scale cultivation processes. Moreover, physiological and genetic factors that increase or reduce the manifestation of strain degeneration in C. militaris are outlined. Finally, methodologies for developing protocols to be used in C. militaris functional biology studies are discussed.
Hispidin is a secondary metabolite with pharmacological properties that is produced by the medicinal mushroom Phellinus linteus. In this study, we evaluated the effect of metal ion addition to grain media while determining the optimal illumination and temperature conditions for hispidin production in P. linteus 03 using solid-state fermentation. Six metal ions were added to these media and P. linteus 03 grown in these media were for hispidin production under optimal incubation temperature and illumination conditions. The highest yield and specific productivity of hispidin, and total hispidin content were achieved at 25 °C in a dark environment which was maintained throughout the entire fermentation period for brown rice and pearl barley media. Addition of Zn²⁺ ions to brown rice medium resulted in the highest yield and specific productivity of hispidin and total hispidin content at 6 weeks of fermentation; these were 2.53-, 2.62-, and 2.43-fold, respectively higher than those observed in the control. This study shows that hispidin production by P. linteus 03 can be enhanced by the addition of Zn²⁺ ions to the grain media.
The effects of light factors affecting growth and quality of vegetables and fruits at mature stages have been often reported while for that of microgreens is still limited. This study was to determine the optimum light spectrum, light intensity and photoperiod that enhanced growth and nutritional values of amaranth microgreens by using light-emitting diodes (LEDs). Two amaranth cultivars, red amaranth and leafy vegetable amaranth, were cultured for 8 days under different light spectra, light intensities and photoperiods provided by LEDs. Three consecutive experiments in the split-plot design were conducted by which, for each experiment, light treatment was a main-plot factor and amaranth cultivar was a sub-plot factor. The first experiment, amaranth microgreens were irradiated by different light spectra: red, blue, red plus blue (70R:30B) and white with photosynthetic photon flux density (PPFD) of 130 μmol m⁻² s⁻¹. The second experiment, the plants were illuminated under various light intensities: 130, 180, 230, 280 μmol m⁻² s⁻¹ PPFD with red plus blue (70R:30B) spectrum. The third experiment, the microgreens were treated with different photoperiods: 8, 12, 16, 20 h (h) under red plus blue (70R:30B) spectrum and 280 μmol m⁻² s⁻¹ PPFD. The results indicated that red plus blue (70R:30B) spectrum, 280 μmol m⁻² s⁻¹ PPFD and 16 h photoperiod enhanced the fresh yield, chlorophyll a, b, total carotenoids, anthocyanin, and vitamin C contents and total antioxidant capacity for both red amaranth and leafy vegetable amaranth microgreens. The cultivar differences in the responses to light spectra and photoperiods were found.
Cordyceps is a traditional Chinese medicinal herb well-known in China, Korea and Japan since B.C. 2,000. The original entomopathogenic fungus, Cordyceps sinensis belonging to the genus Cordyceps could not be found inside Korean peninsula due to the absence of the host insect for the corresponding entomogenous fungus. The development of artificial production methods of Korean type Cordyceps using the silkworm Bombyx mori as in vivo culture medium for the the entomopathogenic fungus Paecilomyces tenuipes is the first, and wonderful occasion in the research history of insect industry of this global world. The aim of this article is to review the historical research background, mass-production methods, and pharmacological effects of the silkworm-dongchunghacho (Paecilomyces tenuipes) which is a newly developed Korean medicinal insect-borne mushroom, and another non-insect-borne medicinal mushroom (Cordyceps militaris and Cordyceps pruinosa). Their biological actions include anti-tumor, immunostimulating, anti-fatigue, anti-stress, anti-oxidant, anti-aging, anti-diabetic, anti-inflammatory, anti-thrombosis, hypolipidaemic and insecticidal effects. The bioactive principles are protein-bound polysaccharides (hexose, hexosamin), cordycepin, D-manitol, acidic polysaccharide etc. Protein-bound polysaccharides and n-butanol fractions were demonstrated to show a significant anti-tumor activities but did not show a cytotoxicities. D-mannitol exhibited a significant prolongation of the life span in tumor bearing mice. Ergosterol did not show an efficient anti-tumor activity, but showed a significant phagocytosis enhancing activity. Anti-tumor activity of silkworm-dongchunghacho might be attributed to immuno-stimulating activities rather than cytotoxic effects [164]. Also this review comprises the breeding of Dongchunghacho varieties, optimization of culture conditions, improvement of learning and memory by Dongchunghacho, application of them as foods and chemical constituents.
Response surface methodology (RSM) with a Box–Behnken design (BBD) was used to optimize the extraction conditions of carotenoid antioxidants from Cordyceps militaris. An enzyme-assisted extraction (EAE) method was developed and optimized to enhance 2,2-diphenyl-1-picrylhydrazil (DPPH) radical-scavenging activity of carotenoid antioxidants from C. militaris. The optimum conditions were as follows: a pH of 4.1, an extraction time of 40.4 min, an extraction temperature of 48.7 °C and 0.43 % enzyme (cellulose: pectinase, 1:1, w/w) concentration. Under these conditions, the DDPH radical-scavenging activity of C. militaris extracts was 81.62±2.26 %, indicating that the BBD is an efficient approach to develop mathematical models for predictingcarotenoids antioxidant extraction from C. militaris. The successful extraction of carotenoid antioxidants from C. militaris provides a basis for the development and utilization of C. militaris resources.
We investigated the effects of light intensity in the 3 cultivation stages separately—the mycelium colonization stage, the primordial initiation stage, and the fruiting stage (in order)—on fruiting body and bioactive compound production by Cordyceps militaris. In the mycelium colonization stage, rice substrates were incubated in a spawn running room at 23°C. During the primordial initiation stage, C. militaris was grown at 18°C and illuminated 12 hours/day. In the fruiting stage the temperature was 23°C, with illumination provided 12 hours/day. The highest fruiting body yield and biological efficiency were 4.06 g dry weight/bottle and 86.83%, respectively, under 1750 ± 250 lux during the second and third stages. The cordycepin content was highest during the second and third stages under 1250 ± 250 lux. The mannitol and polysaccharide contents were highest under 1250 ± 250 and 1750 ± 250 lux during the primordial initiation stage and the fruiting stage, respectively. Thus, with controlled lighting, C. militaris can be cultivated in rice-water medium to increase fruiting body yield and bioactive compound production.
Light-emitting diodes (LEDs) with varying light wavelengths (blue, green, yellow, red and white) were used to study the effects of light source on the biomass and exopolysaccharide (EPS) production of Cordyceps militaris. In a solid-state culture, green and red light sources gave the best hyphal growth. Logistic rate equations were used to calculate the kinetic behavior of the biomass growth and EPS production of a submerged culture under various light sources. According to the results, a maximal biomass concentration of 17.06 g/L was obtained using red light, with a specific growth rate of 1.47 day-1. In contrast, the highest EPS production of 2404.2 mg/L was obtained under blue light, giving a growth- and non-growth-associated product formation coefficient of 17.32 mg/g and 10.83 mg/g/day, respectively. According to the model fitting, a mixed-growth-associated pattern emerged when the culture was illuminated with LEDs, which is quite different from the non-growth-associated mode under dark conditions. The extraordinarily high EPS production of C. militaris under these conditions offers a good source of polysaccharides for further academic and medical applications.
The content of carotenoids and retinoids was compared in the eyes of two Finnish populations of the opossum shrimp, Mysis relicta, which have been reproductively isolated for at least 9000 years: one from the deep, dark, Lake Pääjärvi, the other from the Baltic Sea (Pojoviken Bay). The eyes of the lake population (LP) are highly susceptible to light damage, while those of the sea population (SP) are more resistant. Carotenoids are known to act as antioxidants protecting cells against free radicals and reactive oxygen species; retinoids, on the contrary, may be phototoxic in certain conditions. Analyzed by spectrophotometry and HPLC, the carotenoid content was broadly similar in the eyes of the two populations as regards both total amount and relative proportions of more than 20 components. Noteworthy differences were found in only three of the major components, among these astaxanthin, which was two times higher in SP compared with LP. The most interesting finding was the 1.6-fold higher content of retinoids in LP compared with SP, with retinol as the dominant component (40% of total) in both populatiosns. Retinol is a precursor of the visual-pigment chromophore retinal. The result is consistent with the idea that animals inhabiting extremely dim light environments, where very little photoregeneration of metarhodopsin to rhodopsin can occur, need a large store of chromophore (or precursors) for effective "dark" regeneration of visual pigment. We suggest that almost all the rhodopsin is then in the native state and massive pigment activation following exposure to stronger light may trigger photoreceptor damage. If such animals are handled without due light protection, e.g., when transferred to a new habitat or collected for biological experiments, their vision will be severely impaired.
Characteristic growth patterns of Cordyceps militaris isolates on various media, under varying light conditions and at varying incubation periods were examined. Light was found to be the most critical single factor in determining the density, texture, and pigmentation of the mycelial culture of the fungus. However, under the light condition, the degree of pigmentation and mycelial density were found to be affected by the incubation period and type of medium. Irrespective of the variations in medium type or incubation period, there was no pigmentation of the mycelium under dark condition. Radial growth of the mycelium was faster under dark incubation rather than under light incubation. Abundant mycelial density and darkest pigmentation of C. militaris isolates were produced in nutritionally rich media like SDAY, SMAY and CZYA, suggesting that these media may fulfill all the requirements for vegetative growth of the fungus. Growth characteristics of C. militaris isolates could be easily observed by the simple agar culture method, which would be useful to characterize the phenotypic characteristics of large number of pure cultures of the fungus under given conditions of growth factors such as medium, light and temperature.
Mycelial sensitivity of higher Basidiomycetes medicinal Ling Zhi or Reishi mushroom, Ganoderma lucidum, to the light of various spectral compositions, beam geometry, and polarization has been studied. G. lucidum cultures showed the greatest sensitivity to blue and red light. Light processing of the inoculum has lowered the quantity needed for inoculation and reduced the duration of the fermentation process. The influence of irradiation on polysaccharide synthesis, carbohydrate composition of exopolisaccharides, and mycelial microstructures of G. lucidum has been investigated. The more pronounced stimulating effect of coherent and pulsed light in comparison with noncoherent and continuous light has been observed.
The Cordyceps mushrooms have a long history as medicinal fungi. In Traditional Chinese Medicine, Cordyceps have been used to treat several conditions including cancers for thousand of years. Extracts from both mycelium and fruiting bodies of C. sinensis, C. militaris and other Cordyceps species showed significant anticancer activities by various mechanisms such as, modulating immune system and inducing cell apoptosis. Some polysaccharide components and cordycepin (3'-deoxyadenosine) have been isolated from C. sinensis and C. militaris, which acted as potent anticancer components. This review article aims to further elucidate the importance of Cordyceps mushrooms by summarizing the findings of some of the important research works concerning possible mechanism of anticancer activity of this mushroom.
3'-Deoxyadenosine, also known as cordycepin, is a known polyadenylation inhibitor with a large spectrum of biological activities, including anti-proliferative, pro-apoptotic and anti-inflammatory effects. In this study we confirm that cordycepin reduces the length of poly(A) tails, with some mRNAs being much more sensitive than others. The low doses of cordycepin that cause poly(A) changes also reduce the proliferation of NIH3T3 fibroblasts. At higher doses of the drug we observed inhibition of cell attachment and a reduction of focal adhesions. Furthermore, we observed a strong inhibition of total protein synthesis that correlates with an inhibition of mammalian target of rapamycin (mTOR) signaling, as observed by reductions in Akt kinase and 4E-binding protein (4EBP) phosphorylation. In 4EBP knock-out cells, the effect of cordycepin on translation is strongly reduced, confirming the role of this modification. In addition, the AMP-activated kinase (AMPK) was shown to be activated. Inhibition of AMPK prevented translation repression by cordycepin and abolished 4EBP1 dephosphorylation, indicating that the effect of cordycepin on mTOR signaling and protein synthesis is mediated by AMPK activation. We conclude that many of the reported biological effects of cordycepin are likely to be due to its effects on mTOR and AMPK signaling.
WHITE COLLAR-1 (WC-1) and WC-2, the two PAS domain-containing transcription factors, are the positive elements of the circadian
feedback loops in Neurospora. In addition, both proteins are essential components for the light input of various blue light responses, including the light
entrainment of the circadian clock. Recently, we identified WC-1 as the blue light photoreceptor responsible for these light
responses. In this study, we show that the formation of the FRQ-WC complex in vivo, a step critical in closing the circadian negative feedback loop, requires WC-1. In addition, we show that WC-1 negatively
regulates the expression ofwc-2 at the level of the transcription, forming another interacting loop. In a wc-1 mutant, we demonstrate that there is alternative protein initiation of WC-1, and the requirements of WC-1 for the light induction
of frq and other genes differ significantly, suggesting the existence of different WC complexes in the cell. Consistent with this
interpretation, our results show that there are at least two different types of WC-1/WC-2 complexes in vivo, and that the larger WC-1/WC-2 complex contains more than one WC-1 molecule. Using a series ofwc-1 mutants, we show that the WC-1 PASC domain and its C-terminal region are essential for the formation of the WC-1/WC-2 complex.
Functional analyses reveal that the DNA-binding domain of WC-1 is required only for the activation of frq in the dark and not for the light function of the protein, confirming that WC-1 is a multifunctional protein with separable
protein domains.
Phycomyces blakesleeanus is a filamentous zygomycete fungus that produces striking elongated single cells that extend up to 10 cm into the air, with each such sporangiophore supporting a sphere containing the spores for dispersal. This organism has served as a model for the detection of environmental signals as diverse as light, chemicals, touch, wind, gravity, and adjacent objects. In particular, sporangiophore growth is regulated by light, and it exhibits phototropism by bending toward near-UV and blue wavelengths and away from far-UV wavelengths in a manner that is physiologically similar to plant phototropic responses. The Phycomyces madA mutants were first isolated more than 40 years ago, and they exhibit reduced sensitivity to light. Here, we identify two (duplicated) homologs in the White Collar 1 family of blue-light photoreceptors in Phycomyces. We describe that the madA mutant strains contain point mutations in one of these genes and that these mutations cosegregate with a defect in phototropism after genetic crosses. Thus, the phototropic responses of fungi through madA and plants through phototropin rely on diverse proteins; however, these proteins share a conserved flavin-binding domain for photon detection.
• light, oxygen, or voltage domain
• Max Delbrück
• photosensor
• White Collar 1
• evolution
The growth responses of nine human intestinal bacteria to liquid culture of Cordyceps militaris Link. Pt. (Ascomycotina: Clavicipitaceae) collected from a pupa of Bombyx mori L. (Lepidoptera: Bombycidae) were examined using spectrophotometric and impregnated paper disk methods and compared to those of tetracycline and chloramphenicol, as well as those of Coptis japonica root-derived berberine chloride. The biologically active constituent of the cultures was characterized as cordycepin (3'-deoxyadenosine) by spectroscopic analysis. This compound revealed potent growth-inhibiting activity toward Clostridium paraputrificum and Clostridium perfringens at 10 mu g/disk without adverse effects on the growth of Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Lactobacillus casei, whereas tetracycline and chloramphenicol inhibited the growth of these lactic acid-producing bacteria, clostridia and Escherichia coli. However, C. militaris-derived materials revealed no growth stimulation on the bifidobacteria and lactobacilli. These results may be an indication of at least one of the pharmacological actions of C. militaris. As a naturally occurring antibacterial agent, cordycepin could be useful as a new preventive agent against various diseases caused by clostridia.
A water-soluble polysaccharide(named MCMP) was isolated from the mycelium with high yield mutation Cordyceps militaris by hot-water extraction, deproteinization by sevage, alcohol precipitation, anion-exchange and gel filtration chromatography CL-6B. The polysaccharide contained mannose, rhamnose, galactose and glucose in a molar ratio of 59.36:1:8.31:39.50, of which the average molecular weight is 8100. In our research, Hep-G2 cells, Hela cells and mesangial cells were chosen to determine the anti-tumor activity of the polysaccharide. The results of MTT assay show that polysaccharides of the mutant strain presented inhibitory activity on the cells proliferation after 48 h incubation.
The influence of initial pH value, various nitrogen sources, plant oils, and modes of propagation (shake-flask and static culture) on the production of biomass, exopolysaccharide (EPS), adenosine and, in particular, cordycepin, by Cordyceps militaris CCRC 32219 were investigated. Optimal conditions for mycelial growth, EPS and cordycepin production were observed at relatively low pH. Amongst organic sources, yeast extract (YE) was favorable for EPS and cordycepin production, while corn steep powder (CSP) was favorable for adenosine production. A lower C/N ratio was favorable for adenosine and cordycepin production; however, too low a C/N ratio led to diminished production. All plant oils tested stimulate mycelial growth and EPS production of C. militaris, but they did not show much effect on the adenosine and cordycepin production. A two-stage fermentation process by combining shake-flask fermentation with static culture significantly enhanced cordycepin production. A Box–Behnken experimental design was employed to optimize the production of cordycepin, which showed that the optimum conditions to produce cordycepin by C. militaris CCRC 32219 were at pH 6, YE concentration of 45 g/l and 8.0 day of the shake culture followed by 16 days of the static culture. Under the optimized conditions, the maximum production (2214.5 mg/l) of cordycepin was obtained, which is much higher than those reported up to date.
To investigate the effects of selenium and light wavelengths on the growth of liquid-cultured Cordyceps militaris and the main active components’ accumulation, culture conditions as selenium selenite concentrations and light of different wavelengths were studied. The results are: adenosine accumulation proved to be significantly selenium dependent (R
2 = 0.9403) and cordycepin contents were determined to be not significantly selenium dependent (R
2 = 0.3845) but significantly enhanced by selenium except for 20 ppm; there were significant differences in cordycepin contents, adenosine contents, and mycelium growth caused by light wavelengths: cordycepin, blue light > pink light > daylight, darkness, red light; adenosine, red light > pink light, darkness, daylight, blue light; and mycelium growth, red light > pink light, darkness, daylight > blue light. In conclusion, light wavelength had a significant influence on production of mycelia, adenosine, and cordycepin, so lightening wavelength should be changed according to target products in the liquid culture of C. militaris.
To investigate the effects of selenium on the main active components of Cordyceps militaris fruit bodies, selenium-enriched cultivation of C. militaris and the main active components of the fruit bodies were studied. Superoxide dismutase (SOD) activity and contents of cordycepin, cordycepic acid, and organic selenium of fruit bodies were sodium selenite concentration dependent; contents of adenosine and cordycep polysaccharides were significantly enhanced by adding sodium selenite in the substrates, but not proportional to sodium selenite concentrations. In the cultivation of wheat substrate added with 18.0 ppm sodium selenite, SOD activity and contents of cordycepin, cordycepic acid, adenosine, cordycep polysaccharides, and total amino acids were enhanced by 121/145%, 124/74%, 325/520%, 130/284%, 121/145%, and 157/554%, respectively, compared to NS (non-selenium-cultivated) fruit bodies and wild Cordyceps sinensis; organic selenium contents of fruit bodies reached 6.49 mg/100 g. So selenium-enriched cultivation may be a potential way to produce more valuable medicinal food as a substitute for wild C. sinensis.
The basis for the vivid color of carotenoids and their antioxidant activity is the multiple conjugated double bonds, which are characteristic for these phytonutrients. Moreover, the cleavage of these oxidation-prone double bonds leads to the formation of apocarotenoids. A large number of carbonyl-containing oxidation products are expected to be produced as a result of carotenoid oxidation and these can be further metabolized into the corresponding acids and alcohols. As discussed in this review, many, but not all, of these potential products have been detected and identified in plants as well as in human and animal plasma and tissues. Some of these compounds were found to be biologically active as anticancer agents. In addition to the inhibition of cancer cell proliferation, several carotenoid metabolites were shown to modulate the activity of various transcription systems. These include ligand-activated nuclear receptors, such as the retinoic acid receptor, retinoid X receptor, peroxisome proliferator-activated receptor and estrogen receptor, as well as other transcription systems that have an important role in cancer, such as the electrophile/antioxidant response element pathway and nuclear factor-κB. Therefore, apocarotenoids can be considered as natural compounds with multifunctional, rather than monofunctional, activity and, thus, can be useful in the prevention of cancer and other degenerative diseases.
The nucleoside analogue cordycepin (3'-deoxyadenosine, 3'-dA) is substantially more cytotoxic to terminal deoxynucleotidyl transferase positive (TdT+) leukemic cells than to TdT leukemic cells in vitro in the presence of an adenosine deaminase inhibitor, deoxycoformycin (dCF), and has been considered as a therapeutic agent for TdT+ leukemia. The intracellular metabolism of 3'-dA was examined with HPLC, and the mechanism of its anti-TdT+ leukemic activity was analyzed. In the presence of dCF (2.5 microM), TdT+ leukemic cells (N = 5) were sensitive to the cytotoxic effect of 3'-dA, whereas TdT (N = 6) cells were not. A high level of 3'-dA-5'-triphosphate (3'-dATP) formation was detected in TdT+ NALM-6 cells (67 pmol/10(6) cells) and TdT- K562 cells (49 pmol/10(6) cells) when cultured with 1 microM [3'-3H]-labeled 3'-dA. A substantial level of 3'-dATP was detected in TdT HUT-102 cells (27 pmol/10(6) cells), whereas the level of 3'-dATP in TdT+ MOLT-4 cells was low (0.3 pmol/10(6) cells). The mean IC50 values of 3'-dA against phytohemagglutinin (PHA)-activated and resting peripheral blood mononuclear cells (PBM) (N = 5) were 8 and 32 microM, respectively. There was a modest level of 3'-dATP (7 pmol/10(6) cells) in PHA-PBM, whereas a lower level of 3'-dATP was detected in resting PBM (2.5 pmol/10(6) cells). These data suggest that the presence of 3'-dATP is not sufficient for the antileukemic effect of 3'-dA, but that TdT positivity is essential, and that PBM are significantly less sensitive to the cytotoxicity of 3'-dA in vitro. Further development of 3'-dA as a potential antileukemic agent to treat patients with TdT+ leukemia is warranted.
Of the total carotenoids in respective algal samples, beta-carotene in Spirulina platensis was 69.5%, astaxanthin and its esters in Haematococcus pluvialis was 81.38%, and lutein in Botryococcus braunii was 74.6%. The carotenoids were characterized by mass spectrometry. A time-course study of carotenoids in rats after administration of microalgal biomass showed peak levels in plasma, liver, and eyes at 2, 4, and 6 h, respectively. Beta-carotene accumulation in Spirulina-fed rats was maximum in eye tissues at 6 h. Similarly, levels of astaxanthin and lutein in Haematococcus- and Botryococcus-fed rats were also maximal in eye tissues. Astaxanthin from H. pluvialis showed better bioavailability than beta-carotene and lutein. The antioxidant enzymes, catalase, superoxide dismutase, peroxidase, and TBARS were significantly high in plasma at 2 h and in liver at 4 h, evidently offering protection from free radicals. This study implies that microalgae can be a good source of carotenoids of high bioavailability and nutraceutical value.
Cordyceps militaris is a potential harbour of bio-metabolites for herbal drugs and evidences are available about its applications for revitalization of various systems of the body from ancient times. Amongst all the species, C. militaris is considered as the oldest source of some useful chemical constituents. Besides their popular applications for tonic medicine by the all stairs of the community, the constituents of C. militaris are now used extensively in modern systems of medicine. The current survey records the mysterious potentials of C. militaris are boosting up the present herbal treatments, as well as gearing up the green pharmacy revolution, in order to create a friendly environment with reasonable safety. Evidence showed that the active principles of C. militaris are beneficial to act as pro-sexual, anti-inflammatory, anti-oxidant/anti-aging, anti-tumour/anti-cancer/anti-leukemic, anti-proliferative, anti-metastatic, immunomodulatory, anti-microbial, anti-bacterial, anti-viral, anti-fungal, anti-protozoal, insecticidal, larvicidal, anti-fibrotic, steroidogenic, hypoglacaemic, hypolipidaemic, anti-angiogenetic, anti-diabetic, anti-HIV, anti-malarial, anti-fatigue, neuroprotective, liver-protective, reno-protective as well as pneumo-protective, let alone their other synergistic activities, which let it be marketable in the western countries as over-the-counter medicine. A number of culture techniques for this mushroom have been noticed, for example, storage/stock culture, pre-culture, popular/indigenous culture (spawn culture, husked rice culture and saw dust culture) and, special/laboratory culture (shaking culture, submerged culture, surface liquid culture and continuous/repeated batch culture). The prospects for herbal biotechnology regarding drug discovery using C. militaris delivering what it has promised are high, as the technology is now extremely more powerful than before. This study chiefly highlights the medicinal uses of the mushroom C. militaris including its culture techniques, also aiming to draw sufficient attention of the researchers to the frontier research needs in this context.
Lutein and zeaxanthin are members of the oxygenated carotenoids found particularly in egg yolks and dark-green leafy vegetables. A great deal of research has focused on their beneficial roles in eye health. The present article summarises the current literature related to the bioactivity of these carotenoids, emphasising their effects and possible mechanisms of action in relation to human eye health. Available evidence demonstrates that lutein and zeaxanthin are widely distributed in a number of body tissues and are uniquely concentrated in the retina and lens, indicating that each has a possible specific function in these two vital ocular tissues. Most of epidemiological studies and clinical trials support the notion that lutein and zeaxanthin have a potential role in the prevention and treatment of certain eye diseases such as age-related macular degeneration, cataract and retinitis pigmentosa. The biological mechanisms for the protective effects of these carotenoids may include powerful blue-light filtering activities and antioxidant properties. Although most studies point towards significant health benefits from lutein and zeaxanthin, further large-scale randomised supplementation trials are needed to define their effects on ocular function in health and disease.
The carB gene, encoding the phytoene dehydrogenase of Mucor circinelloides, was isolated by heterologous hybridisation with a probe derived from the corresponding gene of Phycomyces blakesleeanus. The cDNA and genomic copies complemented phytoene dehydrogenase defects in Escherichia coli and in carB mutants of M. circinelloides, respectively. Fluence-response curves for transcript accumulation were constructed after different blue-light pulses. The level of carB mRNA accumulation reached values up to 150-fold higher than basal levels in darkness. Several elements in the promoter of this gene resemble a consensus sequence identified in Neurospora crassa (APE) which is essential for blue-light regulation. Comparison of the available phytoene dehydrogenase sequences from plants, fungi, algae and bacteria suggests that the two known types of phytoene dehydrogenase are more closely related to each other than previously thought.
The selective extraction of capsaicinoids and carotenoids from chili guajillo "puya" flour was studied. When ethanol was used as solvent, 80% of capsaicinoids and 73% of carotenoids were extracted, representing an interesting alternative for the substitution of hexane in industrial processes. Additionally, when the flour was pretreated with enzymes that break the cell wall and then dried, extraction in ethanol increased to 11 and 7% for carotenoid and capsaicinoid, respectively. A selective two-stage extraction process after the treatment with enzymes is proposed. The first step uses 30% (v/v) ethanol and releases up to 60% of the initial capsaicinoids, and the second extraction step with industrial ethanol permits the recovery of 83% of carotenoids present in the flour.
Commercial production of carotenoids from microorganisms competes mainly with synthetic manufacture by chemical procedures. Efficient stimulation of carotenoid biosynthesis is expected to promote accumulation of carotenoid by microbes. This review describes the variety of environmental and cultural stimulants studied during the last few decades which enhance volumetric production and cellular accumulation of commercially important carotenoids from microalgae, fungi and bacteria. Stimulation of carotenoid production by white-light illumination and temperature fluctuation is discussed along with supplementation of metal ions, salts, organic solvents, preformed precursors and several other chemicals in the culture broth. Reports on the improvements in yield are reviewed and assessed from a biotechnology point of view.
The tonic effect of Cordyceps militaris (CM), Paecilomyces japonia (PJ), Phellinus linteus (PL), Ganoderma lucidum (GL), Grifola frondosa (GF), and Panax ginseng (PG) was examined based on the forced swimming capacity and the change of biochemical parameters in ICR mice. The treatment groups were orally administered medicinal plant extracts (500 mg/kg per day), while the control group received distilled water for 4 weeks. The swimming times to exhaustion were longer in the CM, PJ, and GF groups than in the control group (P < 0.05). Plasma TG levels were lower in the treatment groups than in the control group. Plasma glucose levels were not significantly different between the control group and each treatment group except the PG group. Plasma lactate and ammonia levels of the PJ and GF groups were lower than those of the control group (P < 0.05). There were no significant differences in the content of liver and gastrocnemius muscle glycogen between the control group and each treatment group. In conclusion, PJ and GF extracts enhanced the forced swimming capacity of mice by increasing fat utilization and by delaying the accumulation of plasma lactate and ammonia.
The effect of supplementation with Phellinus linteus (P. linteus), Paecilomyces tenuipes (P. tenuipes), and Cordyceps militaris (C. militaris) mushroom water extracts on the insulin secretion and insulin resistance of 90% pancreatectomized (Px) male Sprague Dawley rats was investigated. Px rats were daily administered 0.5 g of P. linteus, P. tenuipes, and C. militaris aqueous extracts or a placebo per 1 kg body weight with a 40% fat diet for 8 weeks. Fasting serum glucose levels were lower in rats receiving C. militaris than in the control group. Insulin secretion at the elevated serum glucose levels was lowest in rats that consumed P. tenuipes in hyperglycemic clamp. Whole body glucose disposal rates increased in C. militaris but decreased in P. tenuipes compared to those in the control group in euglycemic hyperinsulinemic clamp. The GLUT4 content and fraction velocity of glycogen synthase in the soleus and quadriceps muscles increased in the rats treated with C. militaris, but P. tenuipes decreased both. In sum, a water extract of C. militaris ameliorates insulin resistance by enhancing glucose utilization in skeletal muscles.
This study aimed to elucidate pharmacological activities of Cordyceps militaris. The 70% ethanolic extracts of cultured mycelia (CME) and fruiting bodies (FBE) of Cordyceps militaris were prepared. CME was able to directly scavenge the stable free radical diphenyl-2-picrylhydrazyl (DPPH), indicating its antioxidant activity. Both CME and FBE showed topical anti-inflammatory activity in the croton oil-induced ear edema in mice. CME was found to contain acute anti-inflammatory activity, which was evaluated using the carrageenin-induced edema, and also strong antinociceptive activity in writhing test. CME and FBE contain potent inhibitory activity on the chick embryo chorioallantoic membrane (CAM) angiogenesis in a dose-dependent manner. Cordycepin, a metabolite of Cordyceps militaris, appeared to be at least partly responsible for its anti-inflammatory and anti-angiogenic activities. CME concentration-dependently inhibited the NO production and iNOS expression upon stimulation by lipoposaccharide in RAW 264.7, a murine macrophage cell line. In brief, we demontrate that Cordyceps militaris possesses anti-inflammatory and antinociceptive activites, and related antioxidant, anti-angiogenic, and NO production-inhibitory activities.
Cyanovirin-N (CV-N) is a prokaryotic protein under development as a topical anti-HIV microbicide, an urgent and necessary approach to prevent HIV transmission in at-risk populations worldwide. We have expressed recombinant CV-N as inclusion bodies in the cytoplasm of Escherichia coli. A purification scheme has been developed that exploits the physicochemical properties of this protein, in particular its stability in a harsh inclusion body purification scheme. Under the conditions developed, this system yields 140 mg of highly purified CV-N per liter of high-density cell culture, which represents a 14-fold increase over the best recombinant CV-N yield reported to date. This purification scheme results in monomeric CV-N as analyzed by SDS-PAGE, isoelectric focusing, and reverse phase- and size exclusion-HPLC. This recombinantly expressed and refolded CV-N binds to gp120 with nanomolar affinity and retains its potent anti-HIV activities in cell-based assays. The expression and purification system described herein provides a better means for the mass production of CV-N for further microbicide development.
Selective enzyme-mediated extraction of LIGHT WAVELENGTHS FOR MICROORGANISM CULTIVATION capsaicinoids and carotenoids from chili guajillo puya (Capsicum annuum L.) using ethanol as solvent
- R I Santamaria
- M D Ba 'rzana
- E J Z Dong
- Al
SANTAMARIA, R.I., REYES-DUARTE, M.D. and BA'RZANA, E. 2000. Selective enzyme-mediated extraction of J.Z. DONG ET AL. LIGHT WAVELENGTHS FOR MICROORGANISM CULTIVATION capsaicinoids and carotenoids from chili guajillo puya (Capsicum annuum L.) using ethanol as solvent. J. Agric. Food Chem. 48, 3063–3067.
Cordyceps Mushroom: A potent anticancer nutraceutical
- Khan
Pharmacopoeia of China
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THE STATE PHARMACOPOEIA COMMISSION OF THE
CHINA. 2005. Pharmacopoeia of China, p. 75, Vol. I, Chemical
Industry Press, Beijing, China.