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تأثیر محرک‌های مختلف زیستی و غیرزیستی بر رشد ریشه مویین در گیاه دارویی خشخاش (Papaver somniferum L.)

نوع مقاله : علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناس ارشد بیوتکنولوژی، دانشکده‌ کشاورزی، ‌دانشگاه ‌لرستان، خرم‌آباد، ایران

2 دانشیار، گروه زراعت ‌و اصلاح ‌نباتات، دانشکده‌ کشاورزی، ‌دانشگاه ‌لرستان، خرم‌آباد، ایران

3 استاد، گروه زراعت ‌و اصلاح ‌نباتات، دانشکده‌ کشاورزی، ‌دانشگاه ‌لرستان، خرم‌آباد، ایران

چکیده

گیاه خشخاش (Papaver somniferum L.)، از مهم‌ترین گیاهان در صنعت دارویی جهان و منشأ تولید آلکالوئیدها می‌باشد. یکی از تکنیک‌های تولید آلکالوئید، استفاده از کشت ریشه مویین می‌باشد؛ امّا در بیشتر مواقع تولید آلکالوئیدها در مقیاس تجاری کم است و برای افزایش آن‌ها، نیاز به تحریک تولید با روش‌های مختلفی از جمله استفاده از محرک‌های زیستی و غیرزیستی است. در این پژوهش، به‌ منظور افزایش زیست‌توده ریشه‌های مویین، اثر 4 نوع محرک (سالیسیلیک اسید، نیترات نقره، سولفات مس و عصاره مخمّر) به‌صورت آزمایش‌های جداگانه و در قالب طرح کاملاً تصادفی با سه تکرار مورد مطالعه قرار گرفت. جهت انجام این آزمایش‌ها، ریشه‌های مویین اولیه تولید شده در نهایت درون ارلن‌های حاوی 30 میلی‌لیتر محیط کشت  ½MSاضافه شدند و هر ارلن به عنوان یک تکرار در نظر گرفته شد. تأیید تراریختگی ریشه‌های مویین با تکثیر اختصاصی ژن‌های rolC، انجام شد و باندی با اندازه‌ای مطابق انتظار، روی ژل الکتروفورز مشاهده گردید. نتایج نشان داد که غلظت 5/0 میلی‌گرم در میلی‌لیتر از محرک عصاره مخمّر بیش‌ترین تأثیر را بر صفات مورفولوژیکی داشت. تجزیه و تحلیل داده‌ها نشان داد که محرک‌های زیستی، نقش مؤثرتری نسبت به تیمارهای غیرزیستی دارند و غلظت‌های بالای محرک‌ها، اعم از زیستی و غیرزیستی، بر زیست‌توده ریشه‌های مویین، تأثیر منفی دارند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Effect of Different Elicitor Treatments on Hairy Root of Medicinal Plant Poppies (Papaver somniferum L.)

نویسندگان [English]

  • Sh. Siahmansour 1
  • A. Ismaili 2
  • F. Nazarian Firouzabadi 3

1 M.Sc. Student of Biotechnology, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

2 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

3 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

چکیده [English]

Abstract
Background and Objectives
As a source of alkaloids, Poppy (Papaver somniferum L.), is one of the most important plants in the field of pharmaceutical industry. Various methods including use of elicitors, adding precursor, optimizing culture, culture of hairy roots and its metabolites engineering are used to increase the production of secondary metabolites in plant cell cultures. In recent years, cultivation of hairy roots, to the production of valuable metabolites in a number of species of medicinal plants, has taken on commercial scale; but in most cases, this technique produces low amount of alkaloid in commercial scale. Hence, it is needed to optimize the production rate using biological and non-biological elicitors.
Materials and Methods
This experiment was conducted in 2015 in biotechnology laboratory of Lorestan University, Khorramabad, Iran. Sterile explants were prepared from seedling and were then inoculated with Agrobacterium. Inoculated explants were kept in a dark condition for 2 days and then washed and transferred to glasses containing½ MS medium complemented with cefotaxime. Finally, the glasses were kept in tissue culture room until production of hairy roots. The produced hairy roots were used as explants for elicitors' experiments in flask containers. In this study, in order to increase biomass of hairy roots, 4 types of elicitor treatments (salicylic acid, silver nitrate, copper sulphate and yeast extract) were evaluated in separate experiments based on a completely randomized design with 3 replicates.
Results
Confirmation of transgenic hairy roots was tested by PCR using the rolC gene primers to ensure the removal of bacteria from the produced roots. Results of PCR products were separated by electrophoresis and expected fragments were observed in gel. The results showed that the concentration of 5.0 mg/mL of yeast extract elicitor had the most impact on morphological traits. Totally, results showed that biotic elicitors had better effects in comparison to abiotic elicitors. Also, high concentrations of elicitors in all biotic and abiotic cases had a negative impact on biomass of hairy roots.
Discussion
It was shown that biotic elicitors had better effects than abiotic elicitors. The remarkable result of this study was that the high concentration of elicitors had negative effects on biomass hairy root production in poppies plants. It seems that bio-active compounds elicitors induce the plant responses but high concentration of biotic and abiotic elicitors may cause cell death and browning of hairy root tissues.

کلیدواژه‌ها [English]

  • Abiotic
  • Agrobacterium rhizogenesis
  • Biomass
  • Biotic
  • Transgenic
مراجع
Ahmadian Chashmi, N., Sharifi, M., Karimi, F. and Rahnama, H. (2010). Comparative study of tropane alkaloids production in hairy roots and plantlet cultures of (Atropa belladonna L.) by salicylic acid treatments. Iranian Journal of Plant Biology, 2(3), 63-76. [In Farsi]
Akbari, Z., Qaderi, A., Kalate-Jari, S., Mehrafarin, A. and Naghdi Badi, H. A. (2012). Changes of Trigonelline Biosynthesis under Nitrogenous Compounds in Hairy-root Culture of Iranian Fenugreek (Trigonella foenum-graecum L.). Journal of Medicinal Plants, 11(2), 128-135. [In Farsi].
Alderete, L. G. S., Talano, M. A., Ibanez, S. G., Purro, S., Agostini, E., Milrad, S. R. and Medina, M. I. (2009). Establishment of transgenic tobacco hairy roots expressing basic peroxidases and its application for phenol removal. Journal of Biotechnology, 139(4), 273-279.
Bustin, S. and Dorudi, S. (1998). Molecular assessment of tumour stage and disease recurrence using PCR-based assays. Molecular Medicine Today, 4(9), 389-396.
Echevarria-Machado, I., Escobedo-G, M. R. M. and Larque-Saavedra, A. (2007). Responses of transformed (Catharanthus roseus) roots to femtomolar concentrations of salicylic acid. Plant Physiology and Biochemistry, 45(6-7), 501-507.
Facchini, P. J. (2001). Alkaloid biosynthesis in plants: Biochemistry, cell biology, molecular regulation, and metabolic engineering applications. Annual Review of Plant Biology, 52, 29-66.
Furner, I. J. Huffman, G. A. Amasino, R. M. Garfinkel, D. J. Gordon, M. P. and Nester, E. W. (1986). An Agrobacterium transformation in the evolution of the genus Nicotiana. Nature, 319, 422-427.
Kang, S. M., Jung, H. Y., Kang, Y. M., Yun, D. J., Bahk, J. D., Yang, J. K. and Choi, M. S. (2004). Effects of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of (Scopolia parviflora). Plant Science, 166(3), 745-751.
Kim, H. J., Chen, F., Wang, X. and Rajapakse, N. C. (2006). Effect of methyl jasmonate on secondary metabolites of sweet basil (Ocimum basilicum L.). Journal of Agricultural and Food Chemistry, 54(6), 2327-2332.
Lee, E., Park, S. and Paek, K. (2015). Enhancement strategies of bioactive compound production in adventitious root cultures of Nakai subjected to methyl jasmonate and salicylic acid elicitation through airlift bioreactors. Plant Cell, Tissue and Organ Culture, 120(1), 1-10.
Maksymiec, W. and Krupa, Z. (2006). The effects of short-term exposition to Cd, excess Cu ions and jasmonate on oxidative stress appearing in (Arabidopsis thaliana). Environmental and Experimental Botany, 57(1-2), 187-194.
Maksymiec, W., Wianowska, D., Dawidowicz, A. L., Radkiewicz, S., Mardarowicz, M. and Krupa, Z. (2005). The level of jasmonic acid in (Arabidopsis thaliana) and (Phaseolus coccineus) plants under heavy metal stress. Journal of Plant Physiology, 162(12), 1338-1346.
Memelink, J., Verpoorte, R. and Kijne, J. W. (2001). ORC Anization of jasmonate-responsive gene expression in alkaloid metabolism. Trends in Plant Science, 6(5), 212-219.
Rahimi, S. Devi, B. Khorolragchaa, A. Kim, Y. Kim, J. Jung, S. and Yang, D. (2014). Effect of salicylic acid and yeast extract on the accumulation of jasmonic acid and sesquiterpenoids in Panax. Plant Physiology, 61(6), 811-817.
Rao, S. R. and Ravishankar, G. (2002). Plant cell cultures: chemical factories of secondary metabolites. Biotechnology Advances, 20(2), 101-153.
Richards, E., Reichardt, M. and Rogers, S. (1997). Preparation of plant DNA using CTAB. Short Protocols in Molecular Biology, 3(2), 101-111.
Savitha, B. C., Thimmaraju, R., Bhagyalakshmi, N. and Ravishankar, G. (2006). Different biotic and abiotic elicitors influence betalain production in hairy root cultures of Beta vulgaris in shake-flask and bioreactor. Process Biochemistry, 41(1), 50-60.
Shahbazi, E. and Riahi Madvar, A. (2014). Forskolin production and gene expression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase in treated (Coleus forskohlii) Plant with Cu. Plant Production Technology, 6(2), 45-56. [In Farsi].
Sharp, J. M. and Doran, P. M. (2001). Strategies for enhancing monoclonal antibody accumulation in plant cell and organ cultures. Biotechnology Progress, 17(6), 979-992.
Shi, M., Kwok, K. and Wu, J. Y. (2007). Enhancement of tanshinone production in (Salvia miltiorrhiza) Bunge (red or Chinese sage) hairy‐root culture by hyperosmotic stress and yeast elicitor. Biotechnology and Applied Biochemistry, 46(4), 191-196.
Tepfer, D. (1984). Transformation of several species of higher plants by Agrobacterium rhizogenes: Sexual transmission of the transformed genotype and phenotype. Cell, 37(3), 959-967.
Tzfira, T. and Citovsky, V. (2006). Agrobacterium-mediated genetic transformation of plants: biology and biotechnology. Current opinion in Biotechnology, 17(2), 147-154.
Van Wyk, B. E. and Wink, M. (2004). Medicinal plants of the world: an illustrated scientific guide to important medicinal plants and their uses. Portland (Oregon): Timber Press.
Weid, M., Ziegler, J. and Kutchan, T. M. (2004). The roles of latex and the vascular bundle in morphine biosynthesis in the opium poppy, (Papaver somniferum). Proceedings of the National Academy of Sciences of the United States of America, 101(38), 13957-13962.
Xing, B., Yang, D., Guo, W., Liang, Z., Yan, X., Zhu, Y. and Liu, Y. (2014). Ag+ as a more effective elicitor for production of tanshinones than phenolic acids in Salvia miltiorrhiza hairy roots. Molecules, 20(1), 309-324.
Yang, C., Chen, M., Zeng, L., Zhang, L., Liu, X., Lan, X., Tang, K. and Liao, Z. (2011). Improvement of tropane alkaloids production in hairy root cultures of Atropa belladonna by overexpressing pmt and h6h genes. Plant Omics Journal, 4(1), 29-33.
Zhao, D., Fu, C., Chen, Y. and Ma, F. (2004). Transformation of (Saussurea medusa) for hairy roots and jaceosidin production. Plant Cell Reports, 23(7), 468-474.
Zhu, W. and Stefano, G. (2004). Reticuline exposure to invertebrate ganglia increases endogenous morphine levels. Neuroendocrinology Letter, 25(5), 323-330.
تولیدات گیاهی
دوره 41، شماره 1
خرداد 1397
صفحه 29-42
فایل ها
هم رسانی
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