Table 1.

Some examples of the examples of in vitro culture of somatic embryos for the production of specialized metabolites

Species	Specialized metabolite	Main use	Culture conditions			References
			Nutrient medium	Culture type	Culture conditions (Temperature, light, photoperiod)
Aralia elata	Triterpene glycosides: saponins	Cosmetic ingredients, antidiabetic and hepatitis drug	Callus induction: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo initiation and proliferation: MS medium supplemented with IAA (3.0 mg L−1) + BAP (0.5 mg L−1) + 3% sucrose	Callus cultures	25 ± 2 °C, 12 h light and dark photoperiod, fluorescent light, 27 µmol m−2 s−1	Cheng et al. (2021)
Catharanthus roseus	Alkaloid: Vinblastine	Anticancer	Callus induction: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo initiation and proliferation: MS medium supplemented with NAA (1.0 mg L−1) + BAP (0.5 mg L−1) + 3% sucrose Somatic embryo maturation and germination: MS medium GA3 (1 mg L−1) + 3% maltose	Callus cultures	25 °C day/20 °C dark, 16 h photoperiod, fluorescent light at PPFD of 100 µmol m−2 s−1	Aslam et al. (2010)
Citrus pardisi	Flavone glycosides: Naringin	Sweeteners	Induction embryogenic callus: Murashige and Tucker medium supplemented with 280 mM glycerol or 79 mM sucrose	Callus cultures	26 ± 1 °C, 16 h photoperiod, fluorescent light, 14 µmol m−2 s−1	Gavish et al. (1989)
Eleutherococcus chiisanensis	Triterpenoid glycosides: Eleutheroside A or Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures in Erlenmeyer flasks/bioreactors: Murashige and Skoog medium + 3%. Embryo germination: Murashige and Skoog medium + 3% sucrose and 20 µM GA3.	Shake flask cultures: 250 mL Erlenmeyer flasks containing 50 mL of medium were maintained on a gyratory shaker at 110 rpm. and 10 L bioreactors Balloon-type bubble bioreactors containing 5 L medium were aerated with 0.1 vvm sterile air	24 ± 1 °C, 16 h photoperiod, fluorescent light, 40 µmol m−2 s−1	Jeong et al. (2005)
Eleutherococcus koreanum	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium of 1/3, 1/2, full and double strength supplemented with 2,4-D (0, 0.5, 1 and 2 mg L−1), TDZ (0, 0.01, 0.1, 1.0 mg L−1) or without growth regulators and 30 g L−1 sucrose were tested. In another set of experiments effect of 1/2 strength Murashige and Skoog medium supplemented with 15, 30, 60, and 90 g L−1 sucrose was tested on embryo induction. Somatic embryo suspension cultures in Erlenmeyer flasks/bioreactors: 1/3 strength Murashige and Skoog medium + 6% sucrose	Shake flask culture: 300 mL Erlenmeyer flasks containing 100 mL of medium were maintained on a gyratory shaker at 110 rpm Bioreactor cultures: 20 L capacity balloon-type bubble/airlift bioreactors containing 18 L of 1/3 MS medium supplemented with 60 g L−1 sucrose were used. Bioreactor cultures were aerated at 0.1 vvm	Cultures maintained at 23 °C under continuous dark conditions	Park et al. (2005)
Eleutherococcus senticosus	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures in Erlenmeyer flasks/bioreactors: Murashige and Skoog medium + 3% sucrose	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.1 vvm sterile air	During induction of embryos: 24 ± 1 °C, 16 h photoperiod, fluorescent light, 24 µmol m−2 s−1 Bioreactor cultures: Maintained at 12, 18, 24, and 30 °C in the dark	Shohael et al. (2006b)
Eleutherococcus senticosus	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures in bioreactors: Murashige and Skoog medium + 3% sucrose	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.1 vvm sterile air	24 ± 1 °C, 16 h photoperiod, 50 µmol m−2 s−1, and radiations were tested dark (control), fluorescent, monochromatic red LED (peak emission 660 nm), monochromatic blue LED (peak emission 470 nm), blue plus far-red LED (1:1)	Shohael et al. (2006a)
Eleutherococcus senticosus	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures bioreactors: Murashige and Skoog medium + 3% sucrose	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.1 vvm sterile air	Bioreactor cultures: 25 °C 16 h photoperiod, fluorescent light, 35 µmol m−2 s−1; The cultures were elicited at 0, 50, 100, 150, 200, 300 and 400 µM methyl jasmonate	Shohael et al. (2007)
Eleutherococcus senticosus	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures in Erlenmeyer flasks/bioreactors: Murashige and Skoog medium + 3% sucrose	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.1 vvm sterile air	During induction of embryos: 24 ± 1 °C, 16 h photoperiod, fluorescent light, 24 µmol m−2 s−1 Bioreactor cultures: 24 ± 1 oC and dark incubation. Effect of inoculum density (1,3, 5, 7, 10 g cells L−1), aeration volume (0.05, 0.1, 0.2, and 0.3 vvm) Pilot scale bioreactor cultures: 500 L capacity balloon and drum type bioreactors	Shohael et al. (2014a, b)
Eleutherococcus sessiliflorus	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures in Erlenmeyer flasks/bioreactors: Murashige and Skoog medium + 3% sucrose + 4 mg L−1 GA3.	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.1 vvm sterile air; Temporary immersion and continuous immersion methods were tested.	During induction of embryos: 24 ± 1 °C, 16 h photoperiod, fluorescent light, 24 µmol m−2 s−1 Bioreactor cultures: 25 °C 16 h photoperiod, fluorescent light, 35 µmol m−2 s−1	Shohael et al. (2005)
Eleutherococcus sessiliflorus	Phenylpropanoid glycosides: Eleutheroside B, Eleutheroside E Coumarin glycoside: Eleutheroside E1	Immunostimulants, anti-inflammatory and antiviral	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + 3% sucrose Somatic embryo suspension cultures in bioreactors: Murashige and Skoog medium + 3% sucrose + 4 mg L−1 GA3.	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.1 vvm sterile air	Bioreactor cultures: 25 °C 16 h photoperiod, fluorescent light, 35 µmol m−2 s−1; The cultures were elicited at 0, 50, 100, 150, 200, 300 and 400 µM methyl jasmonate	Shohael et al. (2008)
Peucedanum japonicum	Phenolics: Chlorogenic acid	Antioxidant, food additive, and nutraceutical agent	Callus induction: Murashige and Skoog medium supplemented with 2,4-D (0.1-5.0 mg L−1) + 3% sucrose Somatic embryo initiation and proliferation: MS medium supplemented with ABA (1.0 mg L−1) + 3% sucrose	Callus cultures	Callus induction: 25 ± 2 °C and dark condition; Induction of somatic embryos: The effect of blue (450 nm), green (525 nm), red (660 nm), and far-red (730 nm) LED in different combinations (at 54–64 µmol m−2 s−1) have been studied.	Chen et al. (2016)
Plumbago rosea	Naphthoquinone: Plumbagin	Anticoagulant and antineoplastic agent	Somatic embryo suspension cultures in Erlenmeyer flasks: Murashige and Skoog medium + 3% sucrose + 5.06 µM IAA + 2.68 µM NAA + 1.33 µM BA. Effect of different concentrations of NH+ 4 (1–15 µM) and KNO3 (1–15 µM); acetylsalicylic acid (ASA, 0, 2.77, 5.54, 8.32, 11.09 µM) in combination with IAA (2.53, 5.06 µM) was tested.	Shake flask cultures: 250 mL Erlenmeyer flasks containing 60 mL medium on a gyratory shaker at 100 rpm.	25 ± 2 °C, 12 h light and dark photoperiod, fluorescent light, 30 µmol m−2 s−1	Komaraiah et al. (2004)
Psoralea corylifolia	Furanocoumarins: Psoralen	Anti-vitiligo and anti-psoriasis agent	Induction of somatic embryos: Murashige and Skoog’s medium supplemented with 6 µM NAA and 30 µM glutamine + 3% sucrose Somatic embryo suspension cultures in Erlenmeyer flasks: Murashige and Skoog medium + 3% sucrose + 1 µM NAA + 4 µM BA. Studied the effect of 10–50 g L−1 glucose or fructose or sucrose or maltose; or 1 µM NAA + 4 µM BA, 30 g L−1 sucrose, and 5–25 µM glutamic acid alone and 1–5 µM ABA alone or combination of glutamine plus ABA	Shake flask cultures: 250 mL Erlenmeyer flasks containing 50 mL medium on a gyratory shaker at 100–110 rpm.	25 ± 2 °C, 16 h light and dark photoperiod, fluorescent light, 12 µmol m−2 s−1	Baskaran and Jayabalan (2009)
Rosa rugosa	Phenolics and Flavonoids	Antioxidants and cosmetic ingredient	Induction of somatic embryos: Murashige and Skoog’s medium supplemented with 45.0 µM 2,4-D + 3% sucrose Somatic embryo suspension cultures in Erlenmeyer flasks: Murashige and Skoog medium + 3% sucrose.	Shake flask cultures: 250 mL Erlenmeyer flasks containing 100 mL medium on a gyratory shaker at 100 rpm.	24 ± 2 °C, continuous fluorescent light, 40 µmol m−2 s−1	Jang et al. (2016)
Santalum album	Sesquiterpenoids: α-santalol and β-santalol	Natural medicine, Perfume; Anticancer agent	Induction of somatic embryos: Woody plant medium supplemented with 4.52 µM 2,4-D + 100 mg L−1 myoinositol + 3% sucrose Somatic embryo suspension cultures in Bioreactors: WPM supplemented with 100 mg L−1 myoinositol + 0.5 100 mg L−1 IAA + 0.5 100 mg L−1 BA + 3% sucrose	Bioreactors: 2 L capacity airlift bioreactors and 10 L Nelagene culture vessel cultures were aerated with 4 L min−1 airflow	25 ± 2 °C, 16 h photoperiod, fluorescent light, 40 µmol m−2 s−1	Misra and Dey (2013)
Vitis amurensis	Stilbene: Resveratrol	Dietary supplement	Induction of somatic embryos: Murashige and Skoog medium supplemented with 2,4-D (1 mg L−1) + BA (2 mg L−1) + 3% sucrose Somatic embryo suspension cultures in bioreactors: B5 medium + 0.3 mg L−1 + 0.01 mg L−1 NAA and 3% sucrose.	Bioreactors: 3 L capacity balloon-type bubble bioreactors containing 2 L medium and cultures were aerated with 0.2 vvm sterile air	During induction of embryos: 25 °C, 16 h photoperiod, fluorescent light, 30 µmol m−2 s−1 Bioreactor cultures: 25 °C in the dark	Sun et al. (2016)

ABA Abscisic acid; 2,4-D: 2,4-dichlorophenoxyacetic acid; BA N⁶-benzyladenine; BAP N⁶-beznylaminopurine; GA3 Gibberellic acid; IAA Indole-3-acetic acid; IBA Indole-3-butyric acid; LED Light emitting diodes; MS Murashige and Skoog (1962) medium; MT Murashige and Tucker (1969) medium; NAA α-naphthalene acetic acid; PPFD Photosynthetic photon flux density; TDZ Thidiazuron; vvm air volume/medium volume/minute; WPM Woody plant medium (Lloyd and McCown 1981)