اثر برخی تنظیم‌کننده‌های رشد گیاهی بر القای کالوس و کربوهیدارت‌ها بر ترکیبات بیوشیمیایی گیاه کاسنی (Cichorium intybus L.) در شرایط In vitro
محورهای موضوعی :
زیست شناسی سلولی تکوینی گیاهی و جانوری ، تکوین و تمایز ، زیست شناسی میکروارگانیسم
لیلا بیک مهدوی
1
,
رویا بیشه کلایی
2
,
عباسعلی دهپور جویباری
3
,
سعید سلطانی
4
1 - گروه زیست‌شناسی، واحد قایمشهر، دانشگاه آزاد اسلامی، قایمشهر، مازندران، ایران
2 - گروه زیست شناسی، دانشکده فنی و مهندسی، دانشکاه آزاد اسلامی واحد قایمشهر، قایمشهر، ایران
3 - گروه زیست شناسی دانشگاه آزاد اسلامی واحد قائمشهر
4 - فیزیولوژی گیاهی، ریست شناسی، علوم پایه، دانشگاه آزاد اسلامی قائمشهر
تاریخ دریافت : 1401/03/02
تاریخ پذیرش : 1401/10/11
تاریخ انتشار : 1402/05/01
کلید واژه:
آنتی‌اکسیدان,
آنتوسیانین,
فنل,
فلاونوئید,
چکیده مقاله :
کاسنی (Cichorium intybus L.) گیاهی دارویی است که از همه قسمت‌های این گیاه (ریشه، برگ، دانه) استفاده می‌شود. هدف این پژوهش بررسی اثر تنظیم‌کننده‌های رشد گیاهی بر القاء کالوس حاصل از برگ و دمبرگ کاسنی و بررسی اثر کربوهیدرات‌ها (ساکارز و گلوکز) بر ترکیبات بیوشیمیایی (فلاونوئید، فعالیت آنتی‌اکسیدان، فنل، قند، آنتوسیانین) کالوس‌های تولید شده می‌باشد. برای این منظور از سیتوکنین‌های BAP و KIN در 3 سطح (5/0، 1 و 5/1 میلی‌گرم در لیتر) و NAA در 3 سطح (2/0، 5/0 و 1 میلی‌گرم در لیتر) در محیط‌کشت MS برای القاء کالوس استفاده شد. اثر سطوح مختلف ساکارز و گلوکز در 5 سطح (1، 2، 3، 4، 5 درصد) با تیمار شاهد در محیط کشت ½ MS مایع مورد بررسی قرار گرفت. در حضور 1 میلی‌گرم در لیتر از BAP و NAA بیشترین درصد کال‌زایی و وزن توده کالوس مشاهده شد. نتایج نشان داد که کربوهیدرات‌های مورد مطالعه بر ترکیبات بیوشیمیایی کاسنی اثر معنی‌دار (P<0.01) دارد. بیشترین میزان فنل در تیمار 3 درصد گلوکز (28/0 میلی‌گرم گالیک‌اسید بر گرم وزن تر) و بیشترین میزان فلاونوئید در غلظت 4 درصد گلوکز (29/0 میلی‌گرم کوئرستین بر گرم وزن تر) مشاهده شد. بیشترین مقدار فعالیت‌های آنتی‌اکسیدانی و قند به ترتیب در غلظت گلوکز 1 درصد (51/89 درصد) و گلوکز 4 درصد (97/86 میکروگرم) مشاهده شد. بیشترین میزان آنتوسیانین به‌میزان 1/7 میکرومول بر گرم وزن تر در تیمار گلوکز 5 درصد مشاهده شد. با مقایسه تیمار شاهد و سطوح غلظت ساکارز و گلوکز نتیجه‌گیری شد که ترکیبات بیوشیمیایی در حضور گلوکز مقدار بیشتری را داشتند.
چکیده انگلیسی:
Cichorium intybus L. is a biennial plant that all parts of this plant (roots, leaves, seeds) are used medicinally. The aim of this study was to investigate the effect of growth regulators on the induction of calluses from chicory leaves and petioles and to investigate the effect of carbohydrates (sucrose and glucose) on the biochemical composition (flavonoid, Antioxidant activity, phenol, sugar, anthocyanin) of the produced Calli. For this purpose, BAP and KIN cytokinins at three levels (0.5, 1 and 1.5 mg/l) and NAA auxin at three levels (0.2, 0.5 and 1 mg/l) in MS medium were used to induction callus. Also, the effect of different levels of sucrose and glucose was investigated on five levels (1, 2, 3, 4, 5%) with control treatment ½ MS medium. The highest percentage of callus formation and callus weight was observed in culture medium containing 1 mg / l of BAP and NAA. The results showed that the studied carbohydrates had a significant effect on the biochemical composition of chicory (P<0.01). The highest amount of phenol was observed in the treatment of 3% glucose (0.28 mg gallic acid / g fresh weight) and the highest amount of flavonoids was observed in the concentration of 4% glucose (0.29 mg quercetin / g fresh weight). Also, the highest amount of antioxidant and sugar activities were observed in glucose concentration of 1% (89.51%) and glucose 4% (86.97 µg), respectively. The highest amount of anthocyanin (7.1 µM per gram of fresh weight) was observed in 5% glucose treatment.
منابع و مأخذ:
Norbak, R., Nielsen, K. and Kondo, T., 2002. Anthocyanins from flowers of Cichorium intybus. Phytochemistry, 60(4), pp.357-359.
Mozaffarian V. Recognition of medicinal and aromatic plants of Iran. Tehran: Farhang Mo'aser. 2012.
Al-Snafi AE. Medical importance of Cichorium intybus–A review. IOSR Journal of Pharmacy. 2016;6(3):41-56.
Sigaroudi, F, Jaroudi, S, Taqizadeh, M. Therapeutic applications of medicinal plants. Arjmand Press. 2017.
Mulabagal V, Tsay HS. Plant cell cultures-an alternative and efficient source for the production of biologically important secondary metabolites. International journal of applied science and engineering. 2004 Mar;2(1):29-48.
Ebrahimi MA, Payan A. Induction of callus and somatic embryogenesis from cotyledon explants of Fagonia indica Burm. Journal of Medicinal Plants and By-Products. 2013 Jan 1;2(2):209-14.
Khan T, Abbasi BH, Zeb A, Ali GS. Carbohydrate-induced biomass accumulation and elicitation of secondary metabolites in callus cultures of Fagonia indica. Industrial Crops and Products. 2018 Dec 15;126:168-76.
Ghasemzadeh A, Ghasemzadeh N. Flavonoids and phenolic acids: Role and biochemical activity in plants and human. Journal of medicinal plants research. 2011 Dec 16;5(31):6697-703.
Morkunas I, Ratajczak L. The role of sugar signaling in plant defense responses against fungal pathogens. Acta Physiologiae Plantarum. 2014 Jul;36(7):1607-19.
Giri A, Narasu ML. Transgenic hairy roots: recent trends and applications. Biotechnology advances. 2000 Mar 1;18(1):1-22.
Bellettre A, Couillerot JP, Vasseur J. Effects of glycerol on somatic embryogenesis in Cichorium leaves. Plant cell reports. 1999 Nov;19(1):26-31.
Bolouri‐Moghaddam MR, Le Roy K, Xiang L, Rolland F, Van den Ende W. Sugar signalling and antioxidant network connections in plant cells. The FEBS journal. 2010 May;277(9):2022-37.
Amir M, Aqil M, Ismail MV, Akhtar M, Khan AH, Mujeeb M. Effect of carbon source and inubation temperature on total content of secondary metabolites of callus culture of Solanum nigrum. World Journal of Pharmaceutical Research. 2017 May 25;6(8):905-22.
Dantas LA, Faria PS, Dário BM, Arantes AL, Silva FG, Avila RG, Pereira PS, Neto AR. The impact of carbon source on cell growth and the production of bioactive compounds in cell suspensions of Hancornia speciosa Gomes. Scientific Reports. 2021 Dec 21;11(1):1-4.
Vermeulen A, Desprez B, Lenoir P, Chupeau MC, Chupeau Y. Isolation and culture of mesophyll protoplasts from two cytoplasmic albino mutants of Cichorium intybus L. Journal of plant physiology. 1993 Sep 1;142(3):377-80.
Eskandari, H. Optimization of regeneration of chicory plant (Cichorium intybus L.) under the influence of different hormonal timers in MS and B5 cultures. Master Thesis, Faculty of Basic Sciences, Shahed University. 2010.
Velayutham P, Ranjithakumari BD, Baskaran P. An efficient in vitro plant regeneration system for Cichorium intybus L.–an important medicinal plant. Organogenesis. 2003;2000(2001).
Sagharyan M, Ganjeali A, Cheniany M, Kouhi SM. Optimization of callus induction with enhancing production of phenolic compounds production and antioxidants activity in callus cultures of Nepeta binaloudensis Jamzad (Lamiaceae). Iranian Journal of Biotechnology. 2020 Oct;18(4):e2621.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia plantarum. 1962 Jul;15(3):473-97.
McCready RM, Guggolz J, Silviera V, Owens HS. Determination of starch and amylose in vegetables. Analytical chemistry. 1950 Sep 1;22(9):1156-8.
Wagner GJ. Content and vacuole/extravacuole distribution of neutral sugars, free amino acids, and anthocyanin in protoplasts. Plant physiology. 1979 Jul;64(1):88-93.
Slinkard K, Singleton VL. Total phenol analysis: automation and comparison with manual methods. American journal of enology and viticulture. 1977 Jan 1;28(1):49-55.
Chang CC, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of food and drug analysis. 2002 Jul 1;10(3).
Ebrahimzadeh MA, Pourmorad F, Hafezi S. Antioxidant activities of Iranian corn silk. Turkish Journal of biology. 2008 Feb 19;32(1):43-9.
Mix-Wagner G, Gailung O. The influence of different carbon sources on callus formation and shoot proliferation of chicory (Cichorium intybus L. var. sativum). Landbauforschung Volkenrode. 1996 Jan 1;46(1):1-4.
Rehman RU, Israr M, Srivastava PS, Bansal KC, Abdin MZ. In vitro regeneration of witloof chicory (Cichorium intybus L.) from leaf explants and accumulation of esculin. In Vitro Cellular & Developmental Biology-Plant. 2003 Mar;39(2):142-6.
Nikolaeva TN, Zagoskina NV, Zaprometov MN. Production of phenolic compounds in callus cultures of tea plant under the effect of 2, 4-D and NAA. Russian Journal of Plant Physiology. 2009 Jan;56(1):45-9.
Amir M, Aqil M, Ismail MV, Akhtar M, Khan AH, Mujeeb M. Effect of carbon source and inubation temperature on total content of secondary metabolites of callus culture of Solanum nigrum. World Journal of Pharmaceutical Research. 2017 May 25;6(8):905-22.
Baque MD, Elgirban A, Lee EJ, Paek KY. Sucrose regulated enhanced induction of anthraquinone, phenolics, flavonoids biosynthesis and activities of antioxidant enzymes in adventitious root suspension cultures of Morinda citrifolia (L.). Acta Physiologiae Plantarum. 2012 Mar;34(2):405-15.
Amorim HV, Dougall DK, Sharp WR. The effect of carbohydrate and nitrogen concentration on phenol synthesis in Paul's Scarlet Rose cells grown in tissue culture. Physiologia Plantarum. 1977 Jan;39(1):91-5.
Khan T, Abbasi BH, Khan MA, Shinwari ZK. Differential effects of thidiazuron on production of anticancer phenolic compounds in callus cultures of Fagonia indica. Applied biochemistry and biotechnology. 2016 Apr;179(1):46-58.
Zahedzadeh F, Kakavand F, Mahna N. Effects of carbohydrate, light, nitrogen and magnesium on in vitro production of anthocyanin in apple. Int J Biosci. 2015;6(5):250-60.
Rajendran L, Ravishankar GA, Venkataraman LV, Prathiba KR. Anthocyanin production in callus cultures ofDaucus carota as influenced by nutrient stress and osmoticum. Biotechnology letters. 1992 Aug;14(8):707-12.
Mizukami H, Tomita K, Ohashi H. Anthocyanin accumulation and changes in activities of phenylalanine ammonia-lyase and chalcone synthase in roselle (Hibiscus sabdariffa L.) callus cultures. Plant cell reports. 1989 Dec;8(8):467-70.
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Norbak, R., Nielsen, K. and Kondo, T., 2002. Anthocyanins from flowers of Cichorium intybus. Phytochemistry, 60(4), pp.357-359.
Mozaffarian V. Recognition of medicinal and aromatic plants of Iran. Tehran: Farhang Mo'aser. 2012.
Al-Snafi AE. Medical importance of Cichorium intybus–A review. IOSR Journal of Pharmacy. 2016;6(3):41-56.
Sigaroudi, F, Jaroudi, S, Taqizadeh, M. Therapeutic applications of medicinal plants. Arjmand Press. 2017.
Mulabagal V, Tsay HS. Plant cell cultures-an alternative and efficient source for the production of biologically important secondary metabolites. International journal of applied science and engineering. 2004 Mar;2(1):29-48.
Ebrahimi MA, Payan A. Induction of callus and somatic embryogenesis from cotyledon explants of Fagonia indica Burm. Journal of Medicinal Plants and By-Products. 2013 Jan 1;2(2):209-14.
Khan T, Abbasi BH, Zeb A, Ali GS. Carbohydrate-induced biomass accumulation and elicitation of secondary metabolites in callus cultures of Fagonia indica. Industrial Crops and Products. 2018 Dec 15;126:168-76.
Ghasemzadeh A, Ghasemzadeh N. Flavonoids and phenolic acids: Role and biochemical activity in plants and human. Journal of medicinal plants research. 2011 Dec 16;5(31):6697-703.
Morkunas I, Ratajczak L. The role of sugar signaling in plant defense responses against fungal pathogens. Acta Physiologiae Plantarum. 2014 Jul;36(7):1607-19.
Giri A, Narasu ML. Transgenic hairy roots: recent trends and applications. Biotechnology advances. 2000 Mar 1;18(1):1-22.
Bellettre A, Couillerot JP, Vasseur J. Effects of glycerol on somatic embryogenesis in Cichorium leaves. Plant cell reports. 1999 Nov;19(1):26-31.
Bolouri‐Moghaddam MR, Le Roy K, Xiang L, Rolland F, Van den Ende W. Sugar signalling and antioxidant network connections in plant cells. The FEBS journal. 2010 May;277(9):2022-37.
Amir M, Aqil M, Ismail MV, Akhtar M, Khan AH, Mujeeb M. Effect of carbon source and inubation temperature on total content of secondary metabolites of callus culture of Solanum nigrum. World Journal of Pharmaceutical Research. 2017 May 25;6(8):905-22.
Dantas LA, Faria PS, Dário BM, Arantes AL, Silva FG, Avila RG, Pereira PS, Neto AR. The impact of carbon source on cell growth and the production of bioactive compounds in cell suspensions of Hancornia speciosa Gomes. Scientific Reports. 2021 Dec 21;11(1):1-4.
Vermeulen A, Desprez B, Lenoir P, Chupeau MC, Chupeau Y. Isolation and culture of mesophyll protoplasts from two cytoplasmic albino mutants of Cichorium intybus L. Journal of plant physiology. 1993 Sep 1;142(3):377-80.
Eskandari, H. Optimization of regeneration of chicory plant (Cichorium intybus L.) under the influence of different hormonal timers in MS and B5 cultures. Master Thesis, Faculty of Basic Sciences, Shahed University. 2010.
Velayutham P, Ranjithakumari BD, Baskaran P. An efficient in vitro plant regeneration system for Cichorium intybus L.–an important medicinal plant. Organogenesis. 2003;2000(2001).
Sagharyan M, Ganjeali A, Cheniany M, Kouhi SM. Optimization of callus induction with enhancing production of phenolic compounds production and antioxidants activity in callus cultures of Nepeta binaloudensis Jamzad (Lamiaceae). Iranian Journal of Biotechnology. 2020 Oct;18(4):e2621.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia plantarum. 1962 Jul;15(3):473-97.
McCready RM, Guggolz J, Silviera V, Owens HS. Determination of starch and amylose in vegetables. Analytical chemistry. 1950 Sep 1;22(9):1156-8.
Wagner GJ. Content and vacuole/extravacuole distribution of neutral sugars, free amino acids, and anthocyanin in protoplasts. Plant physiology. 1979 Jul;64(1):88-93.
Slinkard K, Singleton VL. Total phenol analysis: automation and comparison with manual methods. American journal of enology and viticulture. 1977 Jan 1;28(1):49-55.
Chang CC, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of food and drug analysis. 2002 Jul 1;10(3).
Ebrahimzadeh MA, Pourmorad F, Hafezi S. Antioxidant activities of Iranian corn silk. Turkish Journal of biology. 2008 Feb 19;32(1):43-9.
Mix-Wagner G, Gailung O. The influence of different carbon sources on callus formation and shoot proliferation of chicory (Cichorium intybus L. var. sativum). Landbauforschung Volkenrode. 1996 Jan 1;46(1):1-4.
Rehman RU, Israr M, Srivastava PS, Bansal KC, Abdin MZ. In vitro regeneration of witloof chicory (Cichorium intybus L.) from leaf explants and accumulation of esculin. In Vitro Cellular & Developmental Biology-Plant. 2003 Mar;39(2):142-6.
Nikolaeva TN, Zagoskina NV, Zaprometov MN. Production of phenolic compounds in callus cultures of tea plant under the effect of 2, 4-D and NAA. Russian Journal of Plant Physiology. 2009 Jan;56(1):45-9.
Amir M, Aqil M, Ismail MV, Akhtar M, Khan AH, Mujeeb M. Effect of carbon source and inubation temperature on total content of secondary metabolites of callus culture of Solanum nigrum. World Journal of Pharmaceutical Research. 2017 May 25;6(8):905-22.
Baque MD, Elgirban A, Lee EJ, Paek KY. Sucrose regulated enhanced induction of anthraquinone, phenolics, flavonoids biosynthesis and activities of antioxidant enzymes in adventitious root suspension cultures of Morinda citrifolia (L.). Acta Physiologiae Plantarum. 2012 Mar;34(2):405-15.
Amorim HV, Dougall DK, Sharp WR. The effect of carbohydrate and nitrogen concentration on phenol synthesis in Paul's Scarlet Rose cells grown in tissue culture. Physiologia Plantarum. 1977 Jan;39(1):91-5.
Khan T, Abbasi BH, Khan MA, Shinwari ZK. Differential effects of thidiazuron on production of anticancer phenolic compounds in callus cultures of Fagonia indica. Applied biochemistry and biotechnology. 2016 Apr;179(1):46-58.
Zahedzadeh F, Kakavand F, Mahna N. Effects of carbohydrate, light, nitrogen and magnesium on in vitro production of anthocyanin in apple. Int J Biosci. 2015;6(5):250-60.
Rajendran L, Ravishankar GA, Venkataraman LV, Prathiba KR. Anthocyanin production in callus cultures ofDaucus carota as influenced by nutrient stress and osmoticum. Biotechnology letters. 1992 Aug;14(8):707-12.
Mizukami H, Tomita K, Ohashi H. Anthocyanin accumulation and changes in activities of phenylalanine ammonia-lyase and chalcone synthase in roselle (Hibiscus sabdariffa L.) callus cultures. Plant cell reports. 1989 Dec;8(8):467-70.
