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Abstract
Solanum nigrum L. is a medicinal plant of solanaceae family with distinguished therapeutic properties. Traditionally, S. nigrum. had been used as an anti-tumorgenic, antioxidant, hepatoprotective, diuretic, and antipyretic agent. The most important alkaloid member in this plant is solanine. Therefore, this study was conducted to utilize tissue culture techniques for the enhancement of solanine production in the in vitro grown cultures of this promising neglected plant. For callus growth and development experimental part, the highest callus growth parameters (callus diameter (21.4 mm) and callus fresh weight (2202.4 mg) were obtained in callus grown on Murashige & Skoog MS media supplemented with 2,4-Dichlorophenoxyacetic acid (2.0 mg·L-1) plus 1.5 mg·L-1 Thidiazuron. Similar trend was also obtained in cell suspension culture experiment, as maximum growth was recorded at similar hormone combination. Moreover, High-performance liquid chromatography analysis revealed that, solanine was affected by growth regulator type and concentration. The highest solanine levels were obtained when the explants were treated with 6-benzylaminopurine at level of 2.0 mg·L-1, as solanine content reached up to (2.61, 1.53 mg.g-1) for callus and cell suspension, respectively, while, microshoot contained the highest solanine (4.52 mg.g-1 DW) at 6-benzylaminopurine level of 1.6 mg.L-1. Additionally, carbon source had positively affected solanine level, where 0.2 M sucrose resulted in production of the highest amounts (3.13, 2.03 and 1.20 mg.g-1 DW) of solanine in microshoots, callus and cell suspension, respectively. Also, exposing microshoots and callus to light intensity of (100 µmol.m -2 s-1) yielded the highest solanine content (4.03 and 1.26 mg.g-1 DW, respectively),while the lowest solanine levels (1.50 and 0.48 mg.g-1 DW) were observed in plant material exposed to the lowest light intensity treatment (25 µmol.m -2 s-1) .
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References
- Abdelmajeed NA., Danial EN, Ayad HS (2013) The effect of environmental stress on qualitative and quantitative essential oil of aromatic and medicinal plants. Journal of Archaeological Science 66: 100-120.
- Ahmad S, Esam S, Fawaz C (2013) Qualitative and quantitative steroidal alkaloids of solanum species distributed widely in Syria by TLC and HPLC. International Journal of Pharmaceutical Sciences Review and Research 23(2): 23-27.
- Al-Hawamdeh FM, Shibli RA, Al-Qudah TS (2013) In vitro production of silymarin from Silybum marianum L. Medicinal and Aromatic Plants 1(1):2167- 0412
- Allan E (1996) Plant cell culture. In Stafford A, Warren G, ed, Plant Cell and Tissue Culture. Chichester: John Wiley and Sons, 1-23.
- Ashutosh KV, Singh RR, Seema S (2012) Improved alkaloid content in callus culture of Catharan thusroseus. Botanica Serbica 36 (2):123 -130
- Bhat MA, Mujib A, Junaid A, Mahmood U (2010) In vitro regeneration of Solanum nigrum L with enhanced solasodine production. Biological Plantarum 54: 757-760.
- Chao L, Jun C, Hung C, Sushim K, Hsiao C, Emily C, Hsin T (2011) In vitro production of benzylisoquinoline from Stephania tetrandra through callus culture under the influence of different additives. Botanical Studies 52: 285-294
- Chawla HS (2002) Introduction to plant biotechnology (2nd ed), Science publishers (INC). Enfeild (NH). USA, 57-74.
- Dewick PM (2002) Medicinal Natural Products: A Biosynthetic Approach. Second Edition. John Wiley & Sons. Ltd.
- Franca SC, Bertoni BW, Dereirea MS (1995) Antihepatotoxic agent in micropropagated plantlets of Eclipta alba. Plant Cell, Tissue and Organ Culture 40: 297-299.
- Hosakatte NM, Eun JL, Kee YP (2014) Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture 118:1–16
- Jasmin S, Daria L, Mohamed RA (2011) In vitro production of solasodine alkaloid in Solanum nigrum under salinity stress. Journal of Phytology and Phytophysiology 3:43-49
- Jayabalan S, Tamilarasan S, Muthiah J, Manikandan R (2014) Improved in vitro propagation, solasodine accumulation and assessment of clonal fidelity in regenerates of Solanum trilobatum L. by flow cytometry and SPAR methods. Plant Cell, Tissue and Organ Culture 117:125–129
- Ji YB, Gao SY, Ji CF, Zou X (2008) Induction of apoptosis in HepG2 cells by solanine and Bcl-2 protein. Journal of Ethnopharmacology 115:194-202
- Jain SM, Ochatt SJ (2010) Protocols for in vitro propagation of ornamental plants. Springer Protocols. Humana Press.
- Jian Z, Wei Z, Qiu H (2001) Effects of light and plant growth regulators on the biosynthesis of vindoline and other indole alkaloids in Catharan thusroseus callus cultures. Plant Growth Regulation 33: 43–49.
- Karimi E, Hawa J, Ali G, Mohd HI (2013) Light intensity effects on production and antioxidant activity of flavonoids and phenolic compounds in leaves, stems and roots of three varieties of Labisia pumila Benth. Australian Journal of Crop Science 7(7):1016-1023
- Karolak IG, Lukasz K, Halina W (2015) The effect of cytokinins on shoot proliferation, secondary metabolite production and antioxidant potential in shoot cultures of Scutellaria alpine. Plant Cell, Tissue and Organ Culture 122:699–708
- Kingston DI (1996) Natural products as pharmaceuticals and sources for lead structures. In: Meyer P. The Practice of Medicinal Chemistry. Academic Press Limited 101-16
- Lee SJ, Lim KT (2003) Antioxidative effects of glycoprotein isolated from Solanum nigrum Linne on oxygen radicals and its cytotoxic effects on the MCF-7 cell. Journal of Food Science 68:466-470
- Lenore A, Ana R, Michael E (1985) The effect of Light on the Production of Heimia Alkaloids. Phytochemistry 24(10): 2215-2218
- Mohy-Ud-Din A, Zaheer K, Mushtaq A, Muhammad A (2010) Chemotaxonomic value of alkaloids in Solanum nigrum complex. Pakestan Juornal of Botany 42(1): 653-660
- Morales C, Cusido RM, Palazon J, Bonfill M (1993) Response of Digitalis purpurea plants to temporary salinity. Journal of Plant Nutrition 16:327–335.
- Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-479.
- Moreira C, Lima S, Esquibel M, Sato A (2010) Solasodine accumulation in regenerated plants of Solanum torvum Sw. Sociedade Brasileira de Plantas Medicinais 12(1):73-79.
- Nagella P, Murthy H, Chung I (2011) In vitro production of gymnemic acid from cell suspension cultures of Gymnemasylvestre. Research Engineering and Life Science 11:537–540
- Newman D, Cragg G, Snader K (2003) Natural products as sources of new drugs over the period 1981-2002. Journal of Natural Products 66:1022-37.
- Nino J, Correa YM, Mosquera M (2009) Biological activities of steroidal alkaloids isolated from Solanum leucocarpum. Pharmaceutical Biology 47(3): 255–259
- Pronob G, Islam M (2012) Phytochemical screening of Solanum nigrum and S. myriacanthus Dunal from districts of upper Assam, India. Journal of Pharmacy 2:455-459.
- Raj D, Adam K, Andrzej D, Maria L (2015) Effect of plant growth regulators on the accumulation of indolizidine alkaloids in Securinega suffruticosa callus cultures. Plant Cell, Tissues Organ Culture 123:39–45.
- Rajani C, Ruby K, Aastha S, Jaya D (2012) Solanum nigrum with dynamic therapeutic role: review. International Journal of Pharmaceutical Sciences Review and Research 15(1): 65-71
- Raju K, Anbuganapathi G, Gokulakrishnan V (2003) Effect of dried fruits of Solanum nigrum Linn against CCl4-induced hepatic damage in rats. Biological and Pharmaceutical Bulletin 26:1618-1619.
- Raghuveer I, Anurag K, Anumalik Y, Nitika G (2015) Metabolites in Plants and its classification. World Journal of Pharmacy and Pharmaceutical Sciences 4(1): 287-305.
- Ramya J, Anjali S, Sanjay G, Indira PS, Reema G (2011) Solanum nigrum: Current Perspectives on Therapeutic Properties. Alternative Medicine Review 16:78-85
- Robbins M, Evans T, Morries P (1996) The effect of plant growth regulators on growth, morphology and condensed tannin accumulation in transformed root cultures of Lotus corniculatus. Plant Cell, Tissues and Organs Culture 44:219–227
- Sajid Z, Faheem A (2009) Effect of Thidiazuron (TDZ) on in vitro micropropagation of Solanum tuberosum L. cvs. Desiree and Cardinal. Pakistan Journal of Botany 41(4): 1811-1815
- Sekh AN, Junaid A, Rashmi K, Saeed AK (2010) Secondary metabolites production through biotechnological intervention: A Review. Emirates Journal of Food and Agriculture 22 (3): 147-161
- Sheeba E, Palanivel S (2013) Callus Induction and Antifungal Studies on Solanum surattense Burm. F. Middle-East Journal of Scientific Research 16(1):122-126.
- Sheeba E, Palanivel S, Parvathi S (2013) Effect of plant growth regulators on callus induction in Physalis minima Linn. International Journal of Innovative Research in Science, Engineering and Technology 2(9):4847-4851.
- Shibli RA, Ajlouni MM (2000) Somatic embryogenesis in the endemic black iris (Iris nigricans). Plant Cell, Tissue and Organ Culture 61: 15-21.
- Shibli RA, Shatnawi MA, Subaih W, Ajlouni M (2006) In vitro conservation and cryopreservation of plant genetic resources: A Review. World Journal of Agricultural Sciences 2:372-382.
- Slazak B, Elwira S, Marta S, Michał R, Justyna B, Aneta S, Ulf G, Elzbieta K (2015) Micropropagation of Viola uliginosa (Violaceae) for endangered species conservation and for somaclonal variation-enhanced cyclotide biosynthesis. Plant Cell, Tissue Organs Culture 120:179–190.
- Song GR, Ekrem GR, Zeki K (2002) Establishment of cell suspension cultures and plant regeneration in sugar beet (Beta vulgaris L.). Turkish Journal of Botany 26: 197-205
- Sundari M, Benniamin A, Manickam V (2010) Micropropagation and in vitro flowering in Solanum nigrum a medicinal plant. International Journal of Biological Technology 1(1):29-32.
- Sridhar TM, Naidu CV (2011) An efficient callus induction and plant regeneration of Solanum nigrum L. An important antiulcer medicinal plant. Journal of Phytology 3: 23-28.
- Tahtamouni RW, Shibli RA, Al- Abdallat AM., Al- Qudah TS, Younis L, Al-Baba H, Al- Ruwaiei H (2015) In vitro Conservation and Cryopreservation of Medicinal and Aromatic Plants: A Review. Jordan Journal of Agricultural Sciences 11 (1):147-167.
- Tahtamouni RW, Shibli RA, Al- Abdallat AM, Al- Qudah TS (2016) Analysis of growth, oil yield, and carvacrol in Thymbra spicata L. after slow-growth conservation. TUBİTAK 40: 213-221
- Taiz L, Zeiger E (2004) Plant Physiology, third edition, Sinauer Associates
- Verpoorte R, Contin A, Memelink J (2002) Biotechnology for the production of plant secondary metabolites. Phytochemistry Review 1:13–25.
- Wang Y, Weathers P (2007) Sugars proportionately affect artemisinin production. Plant Cell Report 26:1073–1081
- Warren MJ, Bassman J, Fellman JK, Mattinson DS, Eigenbrode S (2003) Ultraviolet-B radiation of Populustricho carpata leaves. Tree Physiology 23: 527-535
- Yogananth N, Bhakyaraj R, Chanthuru A, Parvathi S, Palanivel S (2009) Comparative analysis of solasodine from In vitro and In vivo culture of Solanum nigrum L. Kathmandu University. Journal of Science, Engineering and Technology 5: 99- 103.
References
Abdelmajeed NA., Danial EN, Ayad HS (2013) The effect of environmental stress on qualitative and quantitative essential oil of aromatic and medicinal plants. Journal of Archaeological Science 66: 100-120.
Ahmad S, Esam S, Fawaz C (2013) Qualitative and quantitative steroidal alkaloids of solanum species distributed widely in Syria by TLC and HPLC. International Journal of Pharmaceutical Sciences Review and Research 23(2): 23-27.
Al-Hawamdeh FM, Shibli RA, Al-Qudah TS (2013) In vitro production of silymarin from Silybum marianum L. Medicinal and Aromatic Plants 1(1):2167- 0412
Allan E (1996) Plant cell culture. In Stafford A, Warren G, ed, Plant Cell and Tissue Culture. Chichester: John Wiley and Sons, 1-23.
Ashutosh KV, Singh RR, Seema S (2012) Improved alkaloid content in callus culture of Catharan thusroseus. Botanica Serbica 36 (2):123 -130
Bhat MA, Mujib A, Junaid A, Mahmood U (2010) In vitro regeneration of Solanum nigrum L with enhanced solasodine production. Biological Plantarum 54: 757-760.
Chao L, Jun C, Hung C, Sushim K, Hsiao C, Emily C, Hsin T (2011) In vitro production of benzylisoquinoline from Stephania tetrandra through callus culture under the influence of different additives. Botanical Studies 52: 285-294
Chawla HS (2002) Introduction to plant biotechnology (2nd ed), Science publishers (INC). Enfeild (NH). USA, 57-74.
Dewick PM (2002) Medicinal Natural Products: A Biosynthetic Approach. Second Edition. John Wiley & Sons. Ltd.
Franca SC, Bertoni BW, Dereirea MS (1995) Antihepatotoxic agent in micropropagated plantlets of Eclipta alba. Plant Cell, Tissue and Organ Culture 40: 297-299.
Hosakatte NM, Eun JL, Kee YP (2014) Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture 118:1–16
Jasmin S, Daria L, Mohamed RA (2011) In vitro production of solasodine alkaloid in Solanum nigrum under salinity stress. Journal of Phytology and Phytophysiology 3:43-49
Jayabalan S, Tamilarasan S, Muthiah J, Manikandan R (2014) Improved in vitro propagation, solasodine accumulation and assessment of clonal fidelity in regenerates of Solanum trilobatum L. by flow cytometry and SPAR methods. Plant Cell, Tissue and Organ Culture 117:125–129
Ji YB, Gao SY, Ji CF, Zou X (2008) Induction of apoptosis in HepG2 cells by solanine and Bcl-2 protein. Journal of Ethnopharmacology 115:194-202
Jain SM, Ochatt SJ (2010) Protocols for in vitro propagation of ornamental plants. Springer Protocols. Humana Press.
Jian Z, Wei Z, Qiu H (2001) Effects of light and plant growth regulators on the biosynthesis of vindoline and other indole alkaloids in Catharan thusroseus callus cultures. Plant Growth Regulation 33: 43–49.
Karimi E, Hawa J, Ali G, Mohd HI (2013) Light intensity effects on production and antioxidant activity of flavonoids and phenolic compounds in leaves, stems and roots of three varieties of Labisia pumila Benth. Australian Journal of Crop Science 7(7):1016-1023
Karolak IG, Lukasz K, Halina W (2015) The effect of cytokinins on shoot proliferation, secondary metabolite production and antioxidant potential in shoot cultures of Scutellaria alpine. Plant Cell, Tissue and Organ Culture 122:699–708
Kingston DI (1996) Natural products as pharmaceuticals and sources for lead structures. In: Meyer P. The Practice of Medicinal Chemistry. Academic Press Limited 101-16
Lee SJ, Lim KT (2003) Antioxidative effects of glycoprotein isolated from Solanum nigrum Linne on oxygen radicals and its cytotoxic effects on the MCF-7 cell. Journal of Food Science 68:466-470
Lenore A, Ana R, Michael E (1985) The effect of Light on the Production of Heimia Alkaloids. Phytochemistry 24(10): 2215-2218
Mohy-Ud-Din A, Zaheer K, Mushtaq A, Muhammad A (2010) Chemotaxonomic value of alkaloids in Solanum nigrum complex. Pakestan Juornal of Botany 42(1): 653-660
Morales C, Cusido RM, Palazon J, Bonfill M (1993) Response of Digitalis purpurea plants to temporary salinity. Journal of Plant Nutrition 16:327–335.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-479.
Moreira C, Lima S, Esquibel M, Sato A (2010) Solasodine accumulation in regenerated plants of Solanum torvum Sw. Sociedade Brasileira de Plantas Medicinais 12(1):73-79.
Nagella P, Murthy H, Chung I (2011) In vitro production of gymnemic acid from cell suspension cultures of Gymnemasylvestre. Research Engineering and Life Science 11:537–540
Newman D, Cragg G, Snader K (2003) Natural products as sources of new drugs over the period 1981-2002. Journal of Natural Products 66:1022-37.
Nino J, Correa YM, Mosquera M (2009) Biological activities of steroidal alkaloids isolated from Solanum leucocarpum. Pharmaceutical Biology 47(3): 255–259
Pronob G, Islam M (2012) Phytochemical screening of Solanum nigrum and S. myriacanthus Dunal from districts of upper Assam, India. Journal of Pharmacy 2:455-459.
Raj D, Adam K, Andrzej D, Maria L (2015) Effect of plant growth regulators on the accumulation of indolizidine alkaloids in Securinega suffruticosa callus cultures. Plant Cell, Tissues Organ Culture 123:39–45.
Rajani C, Ruby K, Aastha S, Jaya D (2012) Solanum nigrum with dynamic therapeutic role: review. International Journal of Pharmaceutical Sciences Review and Research 15(1): 65-71
Raju K, Anbuganapathi G, Gokulakrishnan V (2003) Effect of dried fruits of Solanum nigrum Linn against CCl4-induced hepatic damage in rats. Biological and Pharmaceutical Bulletin 26:1618-1619.
Raghuveer I, Anurag K, Anumalik Y, Nitika G (2015) Metabolites in Plants and its classification. World Journal of Pharmacy and Pharmaceutical Sciences 4(1): 287-305.
Ramya J, Anjali S, Sanjay G, Indira PS, Reema G (2011) Solanum nigrum: Current Perspectives on Therapeutic Properties. Alternative Medicine Review 16:78-85
Robbins M, Evans T, Morries P (1996) The effect of plant growth regulators on growth, morphology and condensed tannin accumulation in transformed root cultures of Lotus corniculatus. Plant Cell, Tissues and Organs Culture 44:219–227
Sajid Z, Faheem A (2009) Effect of Thidiazuron (TDZ) on in vitro micropropagation of Solanum tuberosum L. cvs. Desiree and Cardinal. Pakistan Journal of Botany 41(4): 1811-1815
Sekh AN, Junaid A, Rashmi K, Saeed AK (2010) Secondary metabolites production through biotechnological intervention: A Review. Emirates Journal of Food and Agriculture 22 (3): 147-161
Sheeba E, Palanivel S (2013) Callus Induction and Antifungal Studies on Solanum surattense Burm. F. Middle-East Journal of Scientific Research 16(1):122-126.
Sheeba E, Palanivel S, Parvathi S (2013) Effect of plant growth regulators on callus induction in Physalis minima Linn. International Journal of Innovative Research in Science, Engineering and Technology 2(9):4847-4851.
Shibli RA, Ajlouni MM (2000) Somatic embryogenesis in the endemic black iris (Iris nigricans). Plant Cell, Tissue and Organ Culture 61: 15-21.
Shibli RA, Shatnawi MA, Subaih W, Ajlouni M (2006) In vitro conservation and cryopreservation of plant genetic resources: A Review. World Journal of Agricultural Sciences 2:372-382.
Slazak B, Elwira S, Marta S, Michał R, Justyna B, Aneta S, Ulf G, Elzbieta K (2015) Micropropagation of Viola uliginosa (Violaceae) for endangered species conservation and for somaclonal variation-enhanced cyclotide biosynthesis. Plant Cell, Tissue Organs Culture 120:179–190.
Song GR, Ekrem GR, Zeki K (2002) Establishment of cell suspension cultures and plant regeneration in sugar beet (Beta vulgaris L.). Turkish Journal of Botany 26: 197-205
Sundari M, Benniamin A, Manickam V (2010) Micropropagation and in vitro flowering in Solanum nigrum a medicinal plant. International Journal of Biological Technology 1(1):29-32.
Sridhar TM, Naidu CV (2011) An efficient callus induction and plant regeneration of Solanum nigrum L. An important antiulcer medicinal plant. Journal of Phytology 3: 23-28.
Tahtamouni RW, Shibli RA, Al- Abdallat AM., Al- Qudah TS, Younis L, Al-Baba H, Al- Ruwaiei H (2015) In vitro Conservation and Cryopreservation of Medicinal and Aromatic Plants: A Review. Jordan Journal of Agricultural Sciences 11 (1):147-167.
Tahtamouni RW, Shibli RA, Al- Abdallat AM, Al- Qudah TS (2016) Analysis of growth, oil yield, and carvacrol in Thymbra spicata L. after slow-growth conservation. TUBİTAK 40: 213-221
Taiz L, Zeiger E (2004) Plant Physiology, third edition, Sinauer Associates
Verpoorte R, Contin A, Memelink J (2002) Biotechnology for the production of plant secondary metabolites. Phytochemistry Review 1:13–25.
Wang Y, Weathers P (2007) Sugars proportionately affect artemisinin production. Plant Cell Report 26:1073–1081
Warren MJ, Bassman J, Fellman JK, Mattinson DS, Eigenbrode S (2003) Ultraviolet-B radiation of Populustricho carpata leaves. Tree Physiology 23: 527-535
Yogananth N, Bhakyaraj R, Chanthuru A, Parvathi S, Palanivel S (2009) Comparative analysis of solasodine from In vitro and In vivo culture of Solanum nigrum L. Kathmandu University. Journal of Science, Engineering and Technology 5: 99- 103.