Improving large-scale biomass and total alkaloid production of Dendrobium nobile Lindl. using a temporary immersion system and MeJA

doi:10.21203/rs.3.rs-538778/v1

Download PDF

Research

Improving large-scale biomass and total alkaloid production of Dendrobium nobile Lindl. using a temporary immersion system and MeJA

https://doi.org/10.21203/rs.3.rs-538778/v1

This work is licensed under a CC BY 4.0 License

You are reading this latest preprint version

Background

Dendrobium nobile Lindl. is an important pharmacopeial plant with medicinal and ornamental value. This study sought to provide technical means for the large-scale artificial production of total alkaloids in D. nobile. Seedlings were cultured in vitro in a temporary immersion bioreactor system (TIBS). The four tested immersion frequencies (min/h; 5/2, 5/4, 5/6, and 5/8) influenced the production of biomass and total alkaloid content. In addition, to compare the effects of different concentrations of phytohormone methyl jasmonate (MeJA) and treatment time on biomass and total alkaloid accumulation, MeJA was added to the TIBS medium after 50 days. Finally, the production of total alkaloids in a semi-solid system (SSS), TIBS, and TIBS combined with the MeJA system (TIBS-MeJA) were compared.

Results

The best immersion frequency was found to be 5/6 (5 min every 6 h) that ensured good levels of biomass and total alkaloid content in plantlets. 10 µM MeJA and 20 days of culture were the most favorable for alkaloid accumulation in the seedlings cultured in TIBS. The maximum content and productivity of total alkaloid on use of TIBS-MeJA was 2.32- and 4.69- fold, respectively, higher in terms of content, and 2.04- and 10.27- fold, respectively, higher in terms of productivity than those on use of TIBS and SSS.

Conclusions

Our results show that TIBS-MeJA is suitable for the large-scale production of total alkaloids in in vitro seedlings; therefore, they provide technical means for the large-scale artificial production of total alkaloids in D. nobile.

Plant Molecular Biology and Genetics

Plant Physiology and Morphology

Dendrobium nobile Lindl.

Temporary immersion bioreactor system

MeJA

Total alkaloid

Biomass

Application of a temporary immersion bioreactor system (TIBS) for the mass propagation of nobile.
Application of a TIBS-MeJA system for the artificial large-scale production of total alkaloid of nobile.
Optimization of culture conditions for better induction of total alkaloid accumulation of nobile.
Comparative analysis on functional efficiency of TIBS and semi-solid system (SSS) culture.
The TIBS culturing significantly improves accumulation of total alkaloid and biomass of nobile.
The MeJA inducing significantly improves accumulation of total alkaloid of nobile using TIBS tissue culture.

Dendrobium, which comprises approximately 1200–1500 species, is one of the largest genera in the family Orchidaceae. There are roughly 80 species within the genus Dendrobium in China[1]; of these, Dendrobium nobile Lindl is one of the most widespread species within the genus and is an important herb that has many medically important secondary metabolites, including alkaloids, flavonoids, and bioactive polysaccharides [2, 3]. D. nobile is described as “the strong body” in the ancient Chinese medical book, Compendium of Materia Medica. Over the past few decades, the species Dendrobium communis have been collected in large quantities, because of the high medicinal and ornamental value, resulting in the species becoming increasingly rare, and active preservation orders have been implemented in numerous counties, especially D. nobile. In order to solve the shortage of D. nobile, many researchers are leaning on tissue-culture instead of wild plants [4].

Alkaloids are the most common active compounds in D. nobile and are found in all parts of the plant. Modern pharmacology studies have shown that alkaloids can relieve pain and antipyretic effects, reduce the heart rate and blood pressure, slow respiration, and alleviate barbiturate poisoning [5, 6]. Furthermore, alkaloids have numerous therapeutic activities, including hypoglycemic, anti-cataract, anti-tumor, anti-cell withering, antioxidant, and anti-Alzheimer’s disease activities [7, 8]. However, D. nobile alkaloids are mainly obtained from three-year-old plants, which are not only expensive due to their long culture duration but also require a lot of space.

Plant tissue culture technology is a rapid and mass propagation method of medicinal plants, and also has the potential to increase the yield of secondary metabolites [9]. However, diverse culture systems influence the propagation and production efficiency of bioactive compounds [10–12]. At present, use of TIBS has a better performance in plant biomass and active substance accumulation, especially in officinal plants [13–16]. TIBS is a liquid culture method that allows the explants to contact the medium intermittently, thus renewing the atmosphere and supplying the nutrients to meet the growth of plants [17]. This semi-automatic micropropagation system is considered to be an effective method to reduce production costs and labor to a greater degree as compared with those of traditional culture methods, such as the semi-solid system (SSS) [18].

Many studies have confirmed that the phytohormone methyl jasmonate (MeJA) can enhance the biosynthesis of secondary metabolites in officinal plants [19–21]. MeJA, as an elicitor, plays a signal role in the biosynthesis of plant alkaloids and has been reported to promote the accumulation of metabolites in Dendrobium Plants [22–25]. In the present study, TIBS and MeJA were used to promote biomass and alkaloid accumulation in D. nobile seedlings.

Effect of immersion frequency on plantlet biomass and total alkaloid during TIBS culture

The TIBS delivers an extremely aerobic environment for plant growth, as it provides forced ventilation through aeration. The immersion frequency is the most significant parameter for system productivity, which not only affects plant growth and micropropagation, but also affects bioactive compound accumulation [26]. In this study, four different immersion frequencies (5/2, 5/4, 5/6, and 5/8) were designed using TIBS culture, and a traditional semi-solid system (SSS) culture was used as a control. After 80 days of growth in the TIBS, there were significant differences in the morphology of seedlings under different immersion frequencies, especially 5/2, and the plantlets were dwarfed and crowded in the tank. The best plantlet morphology was found at an immersion frequency of 5/6 where the plants were taller and the roots showed superior morphology; the second immersion frequency was 5/8, followed by 5/4 (Fig. 1). Figure 2a shows the biomass of seedlings cultured in the reactor for 80 d under different immersion frequencies. The maximum values of fresh weight and dry weight, 349.23 g/L and 54.48 g/L, respectively, appeared in the immersion frequency of 5/6. For these parameters, the second best immersion frequency was 5/8, followed by 5/4 and 5/2. When the immersion frequency was fixed at 5/6, the fresh weight and dry weight of the seedlings in the TIBS tank increased with increasing culture time (Fig. 2b). Moreover, as can be seen in Fig. 2, the rate of increase in fresh weight increased after 40 days of culture, while that of the dry weight increased after 60 days of culture.

Different immersion frequencies and culture times directly affect the accumulation of bioactive compounds in plants, especially medicinal plants [27, 28]. Figure 3 shows the effects of immersion frequencies and culture time on the total alkaloid content and productivity of plantlets using TIBS culture. The highest total alkaloid content was found at an immersion frequency of 5/6 for all culture times. The total alkaloid productivity at the immersion frequency 5/8 was slightly higher than 5/6 after 20 days of cultivation, but the highest productivity was observed at an immersion frequency of 5/6 at other culture times. Moreover, the total alkaloid content and productivity obtained by TIBS culture were significantly higher than those obtained by the traditional semi-solid system culture at all culture times. Therefore, TIBS culture was more conducive to the accumulation of bioactive substances in D. nobile plantlets than SSS culture, which was similar to the results of Ashraf et al. (2013) [29]. As seen in Fig. 3, total alkaloid content and productivity were positively correlated with culture time, and the highest total alkaloid content (3.20 mg/g) and productivity (174.44 mg/L) appeared under the immersion frequency 5/6 after 80 days of cultivation.

Effect of MeJA concentration during TIBS culture

MeJA treatment has significant effects on biomass accumulation during plant cell, tissue, and organ cultures [30–32]. In this research, different concentrations of MeJA were added to the liquid medium after the plantlets were cultured in TIBS for 50 days. After 30 days of induction culture, different concentrations of MeJA were shown to have significant effects on the growth and proliferation of D. nobile plantlets (Fig. 4). Treatment with MeJA resulted in the necrosis of some plantlets, especially at 20 µM MeJA, where almost half of the plantlets were necrotic. Furthermore, from the perspective of individual plants, as the concentration of MeJA was increased, plantlets became shorter, with worse root growth, and a small number of plantlets became yellow. The control group without MeJA did not show necrosis and exhibited the best growth, including of the roots. Figure 5 shows the fresh and dry weights of seedlings treated with different concentrations of MeJA for 30 days (Fig. 5a) and 10 µM MeJA for different times (Fig. 5b) cultured using TIBS. With MeJA concentrations ranging from 0 to 20 µM (interval of 5 µM), the fresh weight of plantlets decreased gradually, with the maximum value obtained at 10 µM MeJA, as shown in Fig. 4. The fresh weight and dry weight of plantlets increased with the extension of culture time, but the growth rates were different in the experimental group treated with 10 µM MeJA; the rate of increase in fresh weight was first low and then high, while the dry weight showed the opposite effect, which was similar to the results of the study by Bayraktar et al. (2016) [33].

MeJA treatment has remarkable effects on the accumulation of bioactive compounds in plant tissue culture [34, 35]. Bioactive compound accumulation in D. nobile plantlets cultured using TIBS was significantly affected by MeJA treatment. The total alkaloid content and productivity in the MeJA treatment groups were markedly lower than those in the control group after 10 days of TIBS culture. However, the accumulation rates of total alkaloid in plantlets were significantly higher in the treatment groups than in the control group. In particular, in the 10 µM MeJA treatment group, the content and productivity were significantly higher than other treatment groups and control group after 20 and 30 days of TIBS culture, and the maximum content (7.41 mg/g DW) and productivity (316.59 mg/L DW) were observed after 20 days of culture (Fig. 6). Thus, we can surmise that MeJA enhances the synthesis of alkaloids in in vitro propagated seedlings, as has been shown in previous studies [36, 37].

Comparison of SSS, TIBS and TIBS-MeJA

The biomass and total alkaloid content of in vitro seedlings grown under the three culture modes were compared in order to provide the most suitable scheme for the artificial production of alkaloids by D. nobile. Table 1 shows that the total alkaloid content, fresh weight, and dry weight of TIBS cultured seedlings were all significantly higher than those in the traditional semi-solid culture system. Specifically, TIBS cultured seedlings treated with MeJA (TIBS-MeJA) contained the maximum total alkaloid content, which was 2.32- and 4.69- fold higher than TIBS and SSS cultures without MeJA. However, the maximum fresh and dry weights were obtained in MeJA-free TIBS culture, because many plant tissue culture systems have diminished biomass after MeJA treatment [38]. Table 1 also shows that the dry weight of plantlets cultured using the TIBS-MeJA system was 0.88- and 2.19- fold higher than that of plants grown in TIBS and SSS, respectively; thus, the maximum total alkaloid productivity appeared at TIBS-MeJA and was 2.04- and 10.27- fold higher than that of the other two culture systems. Therefore, we propose that the combination of TIBS and MeJA is an ideal method for the artificial production of alkaloids from D. nobile plant tissue culture seedlings.

Table 1

Total alkaloid and biomass production of *D. nobile* after 80 days of culture in different systems
Culture system	Total alkaloid content (mg/g DW)	Fresh weight (g/L)	Dry weight (g/L)
SSS	1.58 ± 0.03c	185.02 ± 6.75c	21.80 ± 0.68c
TIBS	3.20 ± 0.05b	349.23 ± 11.63a	54.48 ± 3.69a
TIBS-MeJA	7.41 ± 0.17a	283.68 ± 6.76b	47.73 ± 0.85b
TIBS-MeJA means seedlings are cultured with MeJA using TIBS. Values in each column followed by different letters are significantly different at p ≤ 0.05 as of Post Hoc Multiple Comparisons Test.

TIBS is designed using liquid medium to intermittently contact with plant tissue to provide nutrition, which accords to the natural plant growth conditions. This system provides an advantageous growth environment for plantlets in liquid culture, including suitable nutrients and effective gas exchange, to ensure the healthy growth of seedlings [39, 40]. Immersion frequency is an important culture parameter of TIBS, which plays a decisive role in plant growth and accumulation of metabolic components. The results of this study show that a low immersion frequency (5/6 and 5/8) was beneficial to the accumulation of total alkaloids in plantlets, while a high immersion frequency (5/2 and 5/4) had the opposite effect (Fig. 3), which was similar to the findings of Ivanov et al. (2011) [41] and Malik et al. (2018) [42]. The reason for this may be that a low immersion frequency provided a drought environment, triggering plant stress and resulting in the accumulation of secondary metabolites.

It is known that MeJA has a high effect on plant growth and accumulation of active substances. However, the optimal concentration and treatment time of MeJA for maximum yield of active substances varied with the culture systems, e.g. 100 µM MeJA showed a maximum level of saponin content in cell suspension culture of Leucas aspera Spreng over a period of 18 days [43], treatment with 150 µM MeJA for 72 hours enhanced camptothecin production in tissue cultures of Ophiorrhiza mungos L. var. angustifolia (Thw.) Hook. f. [44], and induction of 100 µM MeJA for 7 days promoted the production of valerian acid in valerian hairy root cultures of Valeriana officinalis [45]. Therefore, it is necessary to screen MeJA concentration and treatment time in order to obtain the maximum accumulation of bioactive compounds. In this study, we added different concentrations of MeJA to the TIBS liquid medium that had been cultured for 30 d. Results showed that the maximum total alkaloid content of plantlets was found at 10 µM MeJA after culture for 20 more days.

Many medicinal plants, such as Dendrobium species, grow for a long time in nature, requiring growth periods of around three years; hence, the period of extraction of bioactive compounds from plants is very long. In order to quickly obtain medicinal plant compounds plant tissue culture technologies which can effectively shorten the growth cycle have been designed and adopted. For example, plant tissue culture for sustainable valorization of secondary metabolites of Bryophyllum sp. [46], and in vitro shoot culture of Rhododendron fortunei used for the commercial production of raw materials for extracting bioactive phytochemicals [47]. Previous studies have reported that the use of bioreactor culture systems and MeJA induction could promote the accumulation of plant bioactive compounds [36, 37, 48, 49]. In the present study, we combined the bioreactor systems with MeJA to more efficiently produce alkaloids from D. nobile tissue culture seedlings. The results show that the TIBS-MeJA system was more conducive to the synthesis of alkaloids and could effectively shorten the production cycle, thus reducing production costs.

TIBS can be applied for the large-scale biomass of D. nobile, and total alkaloid accumulation can be improved by selecting appropriate TIBS immersion frequencies. Moreover, treatment with MeJA has a high elicitation effect on bioactive compound accumulation in in vitro-cultured D. nobile seedlings, and about 10 µM MeJA benefited the production of total alkaloids. Thus, we propose that TIBS and MeJA are necessary for the large-scale production of alkaloids. In this study, to obtain the highest total alkaloid content and productivity, 10 µM MeJA was added to the TIBS liquid medium after plantlets were cultured for 50 days, and the culture was continued for 20 more days. Large-scale biomass and total alkaloid production of D. nobile were successfully accomplished using TIBS and MeJA.

Plant materials and preparation

D. nobile seeds were collected from Yunnan Province, China, and grown in a greenhouse at Nanjing Normal University. Mature capsules of D. nobile obtained by artificial pollination were surface sterilized using 75% alcohol and 10% hydrogen peroxide, then the sterile seeds were seeded on ½MS (Murashige and Skoog, 1962) medium (pH 6.0), replenished with 25 g·L^− 1 sucrose, 80 g·L^− 1 CW (coconut water), 0.5 mg·L^− 1 NAA (α-naphthaleneacetic acid), and 7.2 g·L^− 1 agar. Seeds were cultured in a photoperiod of 10 h light/14 h dark at 25 ± 1°C for 30 days after 5 days of dark culture, and plantlets (stem length: 2–3 cm) from the seeds were used as plant material for further experiments.

TIBS and SSS culture of D. nobile

The temporary immersion bioreactor system (TIBS) was provided by Biofunction Co. Ltd. (Nanjing, China) with culture volume of 6.6 L, which includes controller, culture tank, Connecting tube, air filter (0.22 µ m), etc. (Fig. 7). The liquid ½MS medium (pH 6.0, 1 L) containing 25 g·L^− 1 sucrose, 0.5 mg·L^− 1 NAA, and 80 g·L^− 1 CW, and approximately 300 D. nobile plantlets were placed in each container for all treatments. Four immersion frequencies, 5/2, 5/4, 5/6, and 5/8, were compared for biomass and alkaloid content of D. nobile seedlings in TIBS. Here, the immersion frequencies indicate the time immersed in liquid culture and the interval; “5/2” for example, indicates that the plants were immersed in liquid medium for 5 min every 2 hours.

Semi-solid system (SSS): 300 explants were grown in 20 bottles (each bottle volume 0.5 L) of semi-solid medium averagely. SSS medium was added to 7.2 g·L^− 1 agar as support. All cultures were maintained in 10 h photoperiod under cool white light (1800 lx) at 25 ± 1°C for 80 days.

MeJA elicitation experiments

MeJA (Sigma-Aldrich, 392707) was dissolved in ethanol to prepare a stock solution and filter-sterilized through a 0.22 µm nylon filter. When the plantlets were cultured in TIBS for 50 days, MeJA was added through aseptic replacement of all liquid media, and the immersion frequency was 5/6. MeJA was used as the elicitor at final concentrations of 5, 10, 15 and 20 µΜ (µmol·L^− 1), and liquid medium without MeJA was used as control group. All TIBS cultures were maintained under cool white light (1800 lx) at 25 ± 1°C for a 10 hr photoperiod with, and plantlets were harvested after 10, 20, and 30 days to determine fresh weight, dry weight, and alkaloid accumulation.

Determination of biomass and total alkaloid content

The tissue culture seedlings of D. nobile were taken out from the culture containers and washed with tap water. The fresh weight (FW, g/L) was weighed after the water on the plant surface was absorbed with absorbent paper using analytical balance. The dry weight (DW, g/L) was weighed after plant dried in the oven at 60°C for 36–48 hr to absolute dryness. 0.5 g dried plant was grinded into powder using mortar, then added ammonia solution and let stand for 0.5 hr. Following this, the mixed liquids were poured into 50 mL flask and then added 25 mL chloroform for extraction. The chloroform in the flask was dried with a rotary evaporator after maintained in a water bath at 70℃ for 2.5 hr, after which 5 mL of chloroform was poured into the flask to dissolve the dry residue and then 2 mL of chloroform extract was aspirated and added chloroform to 10 mL. Then, 5 mL of pH 4.5 potassium hydrogen phthalate buffer and 2 mL of 0.04% (w/v) bromocresol green solution were mixed and poured into the chloroform extract. Afterwards, shook the mixture violently for 3 min, then let it stand for 30 min, and then added 1 ml alkaline alcohol (0.01 mol·L^− 1 NaOH ) to 5 ml lower fractions for analysis. The absorbance value was determined by spectrophotometer at 620nm, and then the total alkaloid content was calculated by a standard curve equation: y = 0.063x + 0.027, (R² = 0.996) (Y and X are the absorbance and content of dendrobine) which was obtained with dendrobine as the reference standard. The content and productivity of total alkaloid were calculated by the following formula: content (mg·g^− 1 DW) = dendrobine (mg) × 5/0.5 g and productivity (mg·L-1) = DW (g·L^− 1) × content (mg·g^− 1 DW) [23, 38].

Acknowledgments

The authors thank to Ou Jiangtao and Hu Yanhua for their excellent technical help.

Authors’ contributions

BHZ and XYD conceived and designed research. BHZ, ZTN and CL conducted experiments. QYX and WL contributed analytical tools. XJL and XYD analyzed data and wrote the manuscript. All authors read and approved the final manuscript.

Funding

This work was supported by the financial support of the Jiangsu Agriculture Science and Technology Innovation Fund (No. CX(18)3063).

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding authors on reasonable request.

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Zhu SY, Niu ZT, Xue QY, Wang H, Xie XZ, Ding XY. Accurate authentication of Dendrobium officinale and its closely related species by comparative analysis of complete plastomes. Acta Pharm. Sin. B. 2018; 8(6):969-980.
Cheng XF, Chen W, Zhou ZH, Liu JJ, Wang HZ. Functional characterization of a novel tropinone reductase-like gene in Dendrobium nobile Lindl. J. Plant Physiol. 2013; 170(10):958-964.
Wen ZZ, Lin Y, Liu YQ, Wang M, Wang YQ, Liu W. Effects of paclobutrazol in vitro on transplanting efficiency and root tip development of Dendrobium nobile. Biol. Plant. 2013; 57(3):576-580.
Bhattacharyya P, Kumaria S, Tandon P. High frequency regeneration protocol for Dendrobium nobile: A model tissue culture approach for propagation of medicinally important orchid species. S. Afr. J. Bot. 2016; 104:232-243.
Nie J, Jiang LS, Zhang Y, Tian Y, Li LS, Lu YL, Yang WJ, Shi JS. Dendrobium nobile Lindl. Alkaloids Decreases the Level of Intracellular beta-Amyloid by Improving Impaired Autolysosomal Proteolysis in APP/PS1 Mice. Front. Pharmacol. 2018; 9:01479.
Huang S, Wu Q, Liu H, Ling H, He YQ, Wang CH, Wang ZT, Lu YL, Lu YF. Alkaloids of dendrobium nobile lindl. Altered hepatic lipid homeostasis regulation of bile acids. J. Ethnopharmacol. 2019; 241:111976.
Liu B, Huang B, Liu J, Shi JS. Dendrobium nobile Lindl alkaloid and metformin ameliorate cognitive dysfunction in senescence-accelerated mice via suppression of endoplasmic reticulum stress. Brain Res. 2020; 1741:146871.
Lv LL, Liu J, Li LS, Jin F, Xu YY, Wu Q, Liu J, Shi JS. Dendrobium nobile Lindl. Alkaloids Ameliorate Cognitive Dysfunction in Senescence Accelerated SAMP8 Mice by Decreasing Amyloid-beta Aggregation and Enhancing Autophagy Activity. J. Alzheimer's Dis. 2020; 76(2):657-669.
Kunakhonnuruk B, Kongbangkerd A, Inthima P. Improving large-scale biomass and plumbagin production of Drosera communis A.St.-Hil. by temporary immersion system. Ind. Crop. Prod. 2019; 137:197-202.
Kuo CL, Agrawal DC, Chang HC, Chiu YT, Huang CP, Chen YL, Huang SH, Tsay HS. In vitro culture and production of syringin and rutin in Saussurea involucrata (Kar. et Kir.) - an endangered medicinal plant. Bot. Stud. 2015; 56:8.
Malik S, Mirjalili MH, Fett-Neto AG, Mazzafera P, Bonfill M. Living between two worlds: two-phase culture systems for producing plant secondary metabolites. Crit. Rev. Biotechnol. 2013; 33(1):1-22.
Swaraz AM, Sumi SK, Sultana F, Hasan M, Islam MM, Bari MW, Islam MA, Satter MA, Ahmed KS, Hossain MH. Bioactive compound and bioactivity fidelitous micropropagation method of Blumea lacera (Burm. f.) DC.: A large scale production potential. Ind. Crop. Prod. 2020; 151:112370.
Jang HR, Lee HJ, Shohael AM, Park BJ, Paek KY, Park SY. Production of biomass and bioactive compounds from shoot cultures of Rosa rugosa using a bioreactor culture system. Hortic., Environ. Biotechnol. 2016; 57(1):79-87.
Kunakhonnuruk B, Inthima P, Kongbangkerd A. In Vitro Propagation of Rheophytic Orchid, Epipactis flava Seidenf.-A Comparison of Semi-Solid, Continuous Immersion and Temporary Immersion Systems. Biology-Basel. 2019; 8(4):8.
Ptak A, Moranska E, Skrzypek E, Warchol M, Spina R, Laurain-Mattar D, Simlat M. Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA (R) bioreactor. PeerJ. 2020; 8:e8688.
Szopa A, Kokotkiewicz A, Bednarz M, Jafernik K, Luczkiewicz M, Ekiert H. Bioreactor type affects the accumulation of phenolic acids and flavonoids in microshoot cultures of Schisandra chinensis (Turcz.) Baill. Plant Cell, Tissue Organ Cult. 2019; 139(1):199-206.
Ramírez-Mosqueda MA, Iglesias-Andreu LG, Ramírez-Madero G, Hernández-Rincón EU. Micropropagation of Stevia rebaudiana Bert. in temporary immersion systems and evaluation of genetic fidelity. S. Afr. J. Bot. 2016; 106:238-243.
Baque MA, Moh SH, Lee EJ, Zhong JJ, Paek KY. Production of biomass and useful compounds from adventitious roots of high-value added medicinal plants using bioreactor. Biotechnol. Adv. 2012; 30(6):1255-1267.
Akhgari A, Laakso I, Maaheimo H, Choi YH, Seppanen-Laakso T, Oksman-Caldentey KM, Rischer H. Methyljasmonate Elicitation Increases Terpenoid Indole Alkaloid Accumulation in Rhazya stricta Hairy Root Cultures. Plants-Basel. 2019; 8(12): 534.
Gao WJ, Meng QR, Luo H, Chen F, Zhou YW, He M. Transcriptional responses for biosynthesis of flavor volatiles in methyl jasmonate-treated Chrysanthemum indicum var. aromaticum leaves. Ind. Crop. Prod. 2020; 147:112254.
Luo WY, Yang F, Piao XC, Jin MY, Tian W, Gao Y, Lian ML. Promising strategy to efficiently improve the kinsenoside and polysaccharide production of rhizome cultures of Anoectochilus roxburghii (Wall.) Lindl. Ind. Crop. Prod. 2018; 125:269-275.
Commault AS, Fabris M, Kuzhiumparambil U, Adriaans J, Pernice M, Ralph PJ. Methyl jasmonate treatment affects the regulation of the 2-C-methyl-D-erythritol 4-phosphate pathway and early steps of the triterpenoid biosynthesis in Chlamydomonas reinhardtii. Algal Res. 2019; 39: 101462.
Jiao CY, Song C, Zheng SY, Zhu YP, Jin Q, Cai YP, Lin Y. Metabolic Profiling of Dendrobium officinale in Response to Precursors and Methyl Jasmonate. Int. J. Mol. Sci. 2018; 19(3):728.
Pei TL, Ma PD, Ding K, Liu SJ, Jia YY, Ru M, Dong JE, Liang ZS. SmJAZ8 acts as a core repressor regulating JA-induced biosynthesis of salvianolic acids and tanshinones in Salvia miltiorrhiza hairy roots. J. Exp. Bot. 2018; 69(7):1663-1678.
Yi SY, Kuang TD, Miao YY, Xu Y, Wang Z, Dong LB, Tan NH. Discovery and characterization of four glycosyltransferases involved in anthraquinone glycoside biosynthesis in Rubia yunnanensis. Org. Chem. Front. 2020; 7(17):2442-2448.
McAlister B, Finnie J, Watt MP, Blakeway F. Use of the temporary immersion bioreactor system (RITA^®) for production of commercial Eucalyptus clones in Mondi Forests (SA). Plant Cell, Tissue Organ Cult. 2005; 81(3):347-358.
Jesionek A, Kokotkiewicz A, Wlodarska P, Zabiegala B, Bucinski A, Luczkiewicz M. Bioreactor shoot cultures of Rhododendron tomentosum (Ledum palustre) for a large-scale production of bioactive volatile compounds. Plant Cell, Tissue Organ Cult. 2017; 131(1):51-64.
Ruta C, De Mastro G, Ancona S, Tagarelli A, De Cillis F, Benelli C, Lambardi M. Large-Scale Plant Production of Lycium barbarum L. by Liquid Culture in Temporary Immersion System and Possible Application to the Synthesis of Bioactive Substance. Plants-Basel. 2020; 9(7):844.
Ashraf MF, Abd Aziz M, Stanslas J, Kadir MA. Optimization of immersion frequency and medium substitution on microtuberization of Chlorophytum borivilianum in RITA system on production of saponins. Process Biochem. 2013; 48(1):73-77.
Li C, Wang P, Menzies NW, Lombi E, Kopittke PM. Effects of methyl jasmonate on plant growth and leaf properties. J. Plant Nutr. Soil Sci. 2018; 181(3):409-418.
See KS, Bhatt A, Keng CL. Effect of sucrose and methyl jasmonate on biomass and anthocyanin production in cell suspension culture of Melastoma malabathricum (Melastomaceae). Rev. Biol. Trop. 2011; 59(2):597-606.
Zhao YX, Chen YC, Gao M, Yin HF, Wu LW, Wang YD. Overexpression of geranyl diphosphate synthase small subunit 1 (LcGPPS.SSU1) enhances the monoterpene content and biomass. Ind. Crop. Prod. 2020; 143:111269.
Bayraktar M, Naziri E, Akgun IH, Karabey F, Ilhan E, Akyol B, Bedir E, Gurel A. Elicitor induced stevioside production, in vitro shoot growth, and biomass accumulation in micropropagated Stevia rebaudiana. Plant Cell, Tissue Organ Cult. 2016; 127(2):289-300.
Baenas N, Garcia-Viguera C, Moreno DA. Elicitation: A Tool for Enriching the Bioactive Composition of Foods. Molecules. 2014; 19(9):13541-13563.
Hidalgo D, Sanchez R, Lalaleo L, Bonfill M, Corchete P, Palazon J. Biotechnological Production of Pharmaceuticals and Biopharmaceuticals in Plant Cell and Organ Cultures. Curr. Med. Chem. 2018; 25(30):3577-3596.
Paeizi M, Karimi F, Razavi K. Changes in medicinal alkaloids production and expression of related regulatory and biosynthetic genes in response to silver nitrate combined with methyl jasmonate in Catharanthus roseus in vitro propagated shoots. Plant Physiol. Biochem. 2018; 132:623-632.
Zhou W, Shi M, Deng C, Lu S, Huang F, Wang Y, Kai G. The methyl jasmonate-responsive transcription factor SmMYB1 promotes phenolic acid biosynthesis in Salvia miltiorrhiza. Hortic. Res. 2021; 8(1):10.
Wang HQ, Jin MY, Paek KY, Piao XC, Lian ML. An efficient strategy for enhancement of bioactive compounds by protocorm-like body culture of Dendrobium candidum. Ind. Crop. Prod. 2016; 84:121-130.
Etienne H and Berthouly M. Temporary immersion systems in plant micropropagation. Plant Cell, Tissue Organ Cult. 2002; 69(3):215-231.
Isah T, Umar S, Mujib A, Sharma MP, Rajasekharan PE, Zafar N, Frukh A. Secondary metabolism of pharmaceuticals in the plant in vitro cultures: strategies, approaches, and limitations to achieving higher yield. Plant Cell, Tissue Organ Cult. 2018; 132(2):239-265.
Ivanov I, Georgiev V, Georgiev M, Ilieva M, Pavlov A. Galanthamine and Related Alkaloids Production by Leucojum aestivum L. Shoot Culture using a Temporary Immersion Technology. Biochem. Biotechnol. 2011; 163(2):268-277.
Malik M, Warchol M, Pawlowska B. Liquid Culture Systems Affect Morphological and Biochemical Parameters during Rosa canina Plantlets In Vitro Production. Not. Bot. Horti Agrobot. Cluj-Napoca. 2018; 46(1):58-64.
Vijendra PD, Jayanna SG, Kumar V, Sannabommaji T, Rajashekar J, Gajula H. Product enhancement of triterpenoid saponins in cell suspension cultures of Leucas aspera Spreng. Ind. Crop. Prod. 2020; 156:112857.
Krishnan JJ, Gangaprasad A, Satheeshkumar K. Exogenous methyl jasmonate acts as a signal transducer in the enhancement of camptothecin (CPT) production from in vitro cultures of Ophiorrhiza mungos L. var. angustifolia (Thw.) Hook. f. Ind. Crop. Prod. 2018; 119:93-101.
Torkamani MRD, Jafari M, Abbaspour N, Heidary R, Safaie N. Enhanced production of valerenic acid in hairy root culture of Valeriana officinalis by elicitation. Cent. Eur. J. Biol. 2014; 9(9):853-863.
Garcia-Perez P, Lozano-Milo E, Landin M, Gallego PP. From Ethnomedicine to Plant Biotechnology and Machine Learning: The Valorization of the Medicinal Plant Bryophyllum sp. Pharmaceuticals. 2020; 13(12):444.
Wei XY, Chen JJ, Zhang CY, Wang ZH. In vitro shoot culture of Rhododendron fortunei: An important plant for bioactive phytochemicals. Ind. Crop. Prod. 2018; 126:459-465.
Perez-Hernandez J, Nicasio-Torres MD, Sarmiento-Lopez LG, Rodriguez-Monroy M. Production of anti-inflammatory compounds in Sphaeralcea angustifolia cell suspension cultivated in stirred tank bioreactor. Eng. Life Sci. 2019; 19(3):196-205.
Sitarek P, Kowalczyk T, Picot L, Michalska-Hejduk D, Bijak M, Bialas AJ, Wielanek M, Sliwinski T, Skala E. Growth of Leonurus sibiricus L. roots with over-expression of AtPAP1 transcriptional factor in closed bioreactor, production of bioactive phenolic compounds and evaluation of their biological activity. Ind. Crop. Prod. 2018; 122:732-739.

Download PDF

Editorial decision: Major revision
15 Jul, 2021
Review #2 received at journal
09 Jul, 2021
Reviewer #2 agreed at journal
10 Jun, 2021
Review #1 received at journal
09 Jun, 2021
Reviews received at journal
21 May, 2021
Reviewers invited by journal
20 May, 2021
Reviewer #1 agreed at journal
20 May, 2021
Editor assigned by journal
19 May, 2021
Submission checks completed at journal
19 May, 2021
Editor invited by journal
19 May, 2021
First submitted to journal
17 May, 2021

You are reading this latest preprint version

Improving large-scale biomass and total alkaloid production of Dendrobium nobile Lindl. using a temporary immersion system and MeJA

Status:

Version 1

Abstract

Background

Results

Conclusions

Figures

Highlights

Introduction

Results

Effect of immersion frequency on plantlet biomass and total alkaloid during TIBS culture

Effect of MeJA concentration during TIBS culture

Comparison of SSS, TIBS and TIBS-MeJA

Discussion

Conclusions

Materials And Methods

Plant materials and preparation

MeJA elicitation experiments

Determination of biomass and total alkaloid content

Declarations

References

Supplementary Files

Status:

Version 1