نوع مقاله : انگلیسی
نویسندگان
1 Ph.D. Student Horticultural Sciences, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
2 Associate Professor, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
3 Professor, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
چکیده
کشت ریشههای نابجا منبعی برای تولید متابولیتهای ثانویهی با ارزش میباشد. کاسنی از جمله گیاهان دارویی از تیرهی Asteraceae بوده و شامل ترکیبات دارویی مهمی میباشد. در این تحقیق، کشت ریشههای نابجای حاصل از ریزنمونههای برگی در محیط کشت موراشیک و اسکوگ دارای غلظتهای مختلف ایندولاستیک اسید و نفتالیناستیک اسید انجام شد. بهمنظور القای ریشه، IAA (0، 2/0، 4/0 و 6/0 میلیگرم در لیتر) و NAA (0، 5/0، 1 و 5/1 میلیگرم در لیتر) استفاده شد. القای ریشه 10 روز بعد از کشت اتفاق افتاد و در تیمار شاهد القای ریشه بسیار اندک بود. پس از گذشت 4 هفته، ریشههای پررشد جدا شدند و 100 میلیگرم از ریشههای حاصل از ریزنمونههای برگی در محیط کشت مایع حاوی IAA (0، 5/0، 1 و 5/1 میلیگرم در لیتر) و NAA (0، 5/0، 1 و 5/1 میلیگرم در لیتر) کشت شدند و در شیکر با دور rpm 100 در تاریکی قرار دادهشدند. نتایج بدست آمده نشان داد بیشترین درصد تولید ریشه و میانگین تعداد ریشه در محیط کشت حاوی 5/1 میلیگرم در لیتر NAA و بیشترین وزنتر (74/0 گرم در فلاسک)، خشک (062/0 گرم در فلاسک) و فنول (1/4 میلیگرم بر گرم وزن خشک ریشه) در تیمار 5/0 میلیگرم در لیتر NAA در ترکیب با 5/0 میلیگرم در لیتر IAA بدست آمد و بیشترین میزان فلاونوئید (26/60، 88/85 و 53/98 مایکروگرم بر گرم وزن خشک ریشه) در تیمار 1 میلیگرم در لیتر NAA و 1 میلیگرم در لیتر IAA بدست آمد.
کلیدواژهها
عنوان مقاله [English]
Adventitious Root Development and Secondary Metabolites Accumulation by Auxin in Cichorium intybus L
نویسندگان [English]
- Roghayeh Fathi 1
- Mehdi Mohebodini 2
- Esmaeil Chamani 3
1 Ph.D. Student Horticultural Sciences, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
2 Associate Professor, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
3 Professor, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
چکیده [English]
ABSTRACT
Introduction:Adventitious root cultures of medicinal plants are a source of secondary metabolites of pharmaceutical importance, and are considered as an alternative method for clonal propagation and germplasm conservation in medicinal plants.Chicory (Cichorium intybus L.) is a medicinal plant from Asteraceae and isused in traditional medicine to promote appetite and digestion. This plant contains many important metabolites including chicoric asid, inulin, scoline, coumarin and flavonoids. In the current research, an efficient protocol has been developed for adventitious root culture on MS medium supplemented with different concentrations of Indole-3-acetic acid (IAA) and α-Naphthalene acetic acid (NAA).
Materials and Methods:The seeds were surface-sterilized with 50 ml l−1 sodium hypochlorite for 20 min, subsequently with 700 ml l−1 ethanol for 90 s. The surface-sterilized seeds were inoculated against the MS medium and cultures were incubated at 25 ±2 ◦C under fluorescent light for a cycle of 16 h light and 8 h dark per day. The leaves explants of 28-day-old in vitro plantlets were used as explants. For root initiation, IAA (0, 0.2, 0.4 and 0.6 mg l-1) and NAA (0, 0.5, 1 and 1.5 mg l-1) were used. After four weeks, the well-established roots were separated. To determine the best medium of composition for growth of roots, approximately 100 mg fresh weight of adventitious roots were cultured in MS liquid medium with different concentrations of IAA (0, 0.5, 1 and 1.5 mg 1-1) and NAA (0, 0.5, 1 and 1.5 mg 1-1).
Results and Discussion:According to the results, among the different concentrations of IAA, the highest root induction (72.5 percent), root number (4.75), and root branch (10.08) were exhibited by 1.5 mg/L IAA. Among different NAA levels, the highest root induction (88.88 percent), and root number (8.04) were observed in 1.5 mg/L NAA and was not significantly different from 0.5 and 1.5 mg/L NAA. This hormone at concentration of 1.5 mg/L, induced the highest root branching (18.42 per explant). The highest fresh weight (0.74 g) and dry weight (0.062 g), growth index (6.51), and phenol (4.1 mg/g DW) were obtained in MS liquid medium containing 0.5 mg 1-1 NAA in combination with 0.5 mg 1-1 IAA, and Flavonoid content in 270, 300 and 330 nm wavelengths was higher (60.26, 85.88 and 98.53 µg g-1 DW) in the roots obtained from 1 mg l−1 of NAA in combination with 1 mg l−1 of IAA. Increasing NAA concentrations induced callus mediated root formation and produced a lower number of adventitious roots. By using IAA, adventitious roots were initiated, but the frequency and average number of roots initiated were lower when compared with NAA.
Conclusion: Adventitious roots obtained by different concentration of auxins are a suitable tool for the production of plant secondary metabolites due to their genetic stability, and generally, show a fast growth rate. This study describes the protocol for adventitious root induction which could further be useful for the production of secondary metabolites and biomass
کلیدواژهها [English]
- Chicory
- Indole-3-aceticacid
- Naphthalene acetic acid
- Phenolic content
Impacts of hormonal elicitors and photoperiod regimes on elicitation of bioactive secondary volatiles in cell cultures of Ajugabracteosa.Journal of Photochemistry and Photobiology B: Biology, 183, 242-250. doi: 10.1016/j.jphotobiol.2018.04.044
salt strength induced changes in growth, physiology and secondary metabolite content in adventitious roots of Morindacitrifolia: the role of antioxidant enzymes and phenylalanine ammonia lyase. Plant Cell Reports, 29(7), 685-694. doi: 10.1007/s00299-010-0854-4
A. (2013). Identification of phenolic constituents in
red chicory salads (Cichoriumintybus) by high-performance liquid chromatography with diode array detection and electrospray ionisation tandem mass spectrometry. Food Chemistry, 138(2-3), 1062-1071. doi: 10.1016/j.foodchem.2012.11.060
of Horticultural Science, 49(3), 393-405. doi: 10.30479/ijgpb.2017.1491 [In Farsi with English abstract]
by Methyl jasmonate and salicylic acid. Journal
of Plant Biotechnology, 40(3), 178-183. doi: 10.5010/JPB.2013.40.3.178
compound and its protective effects against homocysteine-induced cytotoxicity and oxidative stress. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 202, 314-318. doi: 10.1016/j.saa.2018.05.058
Products, 105, 124-132. doi: 10.1016/j.indcrop. 2017.05.010
I., Choi, K. B., Morishige, T., ..., & Yamada,
Y. (2001). Metabolic engineering of plant alkaloid
biosynthesis. Proceedings of the National Academy of Sciences, 98(1), 367-372. doi: 10.1016/S1872-2075(08)60035-7
as signaling molecules. Current Opinion in Plant biology, 2(5), 410-418. doi: 10.1016/S1369-5266(99)00014-X
F. (2018). Effect of different elicitor treatments
on hairy root of medicinal plant poppies (Papaver somniferum L.). Plant Productions, 41(1), 29-42.
doi: 10.22055/ppd.2018.13548 [In Farsi with English abstract]
S., Tanaka, M., Endo, T., & Kaji, A. (1990). Lobeline production by hairy root culture of Lobelia inflata L. Plant Cell Reports, 9(6), 307-310. doi: 10.1007/BF00232857
Impacts of hormonal elicitors and photoperiod regimes on elicitation of bioactive secondary volatiles in cell cultures of Ajugabracteosa.Journal of Photochemistry and Photobiology B: Biology, 183, 242-250. doi: 10.1016/j.jphotobiol.2018.04.044
salt strength induced changes in growth, physiology and secondary metabolite content in adventitious roots of Morindacitrifolia: the role of antioxidant enzymes and phenylalanine ammonia lyase. Plant Cell Reports, 29(7), 685-694. doi: 10.1007/s00299-010-0854-4
A. (2013). Identification of phenolic constituents in
red chicory salads (Cichoriumintybus) by high-performance liquid chromatography with diode array detection and electrospray ionisation tandem mass spectrometry. Food Chemistry, 138(2-3), 1062-1071. doi: 10.1016/j.foodchem.2012.11.060
of Horticultural Science, 49(3), 393-405. doi: 10.30479/ijgpb.2017.1491 [In Farsi with English abstract]
by Methyl jasmonate and salicylic acid. Journal
of Plant Biotechnology, 40(3), 178-183. doi: 10.5010/JPB.2013.40.3.178
compound and its protective effects against homocysteine-induced cytotoxicity and oxidative stress. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 202, 314-318. doi: 10.1016/j.saa.2018.05.058
Products, 105, 124-132. doi: 10.1016/j.indcrop. 2017.05.010
I., Choi, K. B., Morishige, T., ..., & Yamada,
Y. (2001). Metabolic engineering of plant alkaloid
biosynthesis. Proceedings of the National Academy of Sciences, 98(1), 367-372. doi: 10.1016/S1872-2075(08)60035-7
as signaling molecules. Current Opinion in Plant biology, 2(5), 410-418. doi: 10.1016/S1369-5266(99)00014-X
F. (2018). Effect of different elicitor treatments
on hairy root of medicinal plant poppies (Papaver somniferum L.). Plant Productions, 41(1), 29-42.
doi: 10.22055/ppd.2018.13548 [In Farsi with English abstract]
S., Tanaka, M., Endo, T., & Kaji, A. (1990). Lobeline production by hairy root culture of Lobelia inflata L. Plant Cell Reports, 9(6), 307-310. doi: 10.1007/BF00232857