با همکاری انجمن علمی گیاهان دارویی ایران

نوع مقاله : مقاله علمی پژوهشی

نویسندگان

1 دانشجوی دکتری اصلاح نباتات گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی دانشگاه زنجان، زنجان، ایران

2 دانشیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی دانشگاه زنجان، زنجان، ایران

3 دانشیار، گروه علوم باغبانی، دانشکدگان کشاورزی و منابع طبیعی، دانشگاه تهران، تهران، ایران

4 استادیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی دانشگاه زنجان، زنجان، ایران

چکیده

زعفران زراعی Crocus sativus L. گیاهی دارویی، پرکاربرد و گران‌قیمت بوده دارای مواد موثره مهم: کروسین، پیکروکروسین و سافرانال است که ویژگی های رنگ، طعم و بو به آن می‌دهد. جستجوی منابع جدید تولید کننده آپوکارتنوئیدهای مذکور اهمیت تحقیق روی آن را روز به روز آشکارتر می‌کند. از آنجایی که کشور ایران دارای گونه های وحشی دیپلوئید با ویژگی‌هایی نزدیک به گونه زراعی تری‌پلوئید عقیم است، در تحقیق حاضر دو گونه زعفران خزر C.caspius و زیباC.Speciosus درکنار گونه زراعی، بررسی و 6 ژن فعال در مسیر تولید سه ماده موثره مذکور، CCD2,ALDH,PSY,LCY,BCH,UGT74 (و یک ژن خانه دار TUB به عنوان ژن کنترل داخلی با بیان ثابت در تمام بافت‌ها) در دو بافت گل کلاله و گلپوش (تپال) و در پنج مرحله (زمان) گلدهی شامل مرحله تشکیل غنچه (غنچه نارس)، غنچه آماده شکفتن،گل تازه باز شده، گل بالغ و گل پیر مورد تحقیق قرار گرفت. با کمک Real Time PCR نسبت بیان (Fold change) به ژن کنترل داخلی (با لگاریتم برپایه 2) در ژن‌های مختلف بررسی و الگو های بیان متفاوت و گاهاً متشابه با بیان ژن در گونه زراعی در دو بافت تپال و کلاله مشاهده شد. براساس نتایج حاصل در این مطالعه و افزیش و یا کاهش بیان برخی ژن‌های دخیل در مسیرهای تولید آپوکارتنوئیدها، همزمان با رشد مراحل از غنچه تا مرحله پیری گل، انتظار می رود از سه ماده پیش‌گفته و یا از سایر آپوکارتنوئیدها در متابولوم گل‌ها، مواردی در گونه های غیر تری‌پلوئید کشف و رهیافت‌هایی برای استفاده تجاری از گونه‌های وحشی زعفران ایرانی ایجاد و مسیر تجاری سازی آن هموارگردد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Investigating the expression of some genes related to the production of apocarotenoids in the growth stages of stigma and tepal of cultivated saffron and two wild species

نویسندگان [English]

  • Mehdi Hassanlou 1
  • Mohammad Reza Azimi Moghaddam 2
  • Seyed Alireza Salami 3
  • Ehsan Mohsenifard 4

1 Ph.D. Student of Plant Breeding, Department of Plant Production and Genetics, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.

2 Associate Professor, Department of Plant Production and Genetics, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

3 Associate Professor, Department of Plant College of Agriculture and Natural resources, University of Tehran, Karaj, Iran.

4 Assistant Professor, Department of Plant Production and Genetics, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.

چکیده [English]

Agricultural saffron Crocus sativus L. is a medicinal, widely used and expensive plant and has important effective substances: crocin, picrocrocin and safranal, which gives it the characteristics of color, taste and smell. The search for new sources producing the aforementioned apocarotenoids makes the importance of research on it more obvious day by day. Since the country of Iran has diploid wild species with characteristics close to the sterile triploid cultivated species, in the present research, two species of Caspian saffron C. caspius and Zeiba C. Speciosus, along with the cultivated species, were investigated and 6 genes active in the production of the three mentioned effective substances , CCD2, ALDH, PSY, LCY, BCH, UGT74 (and a housekeeping gene TUB as an internal control gene with constant expression in all tissues) in two flower tissues, stigma and perianth (tepal) and in five stages (time) of flowering including: The stages of bud formation (immature bud), bud ready to bloom, newly opened flower, mature flower and old flower were investigated. Using of Real Time PCR, the expression ratio (fold change) to the internal control gene (with logarithm based on 2) was investigated in different genes and different and sometimes similar expression patterns were observed with the gene expression in the cultivated species in both the stigma and stigma tissues. Based on the results obtained in this study and the increase or decrease in the expression of some genes involved in the pathways of apocarotenoids production, at the same time as the growth of the stages from the bud to the senescence stage of the flower, it is expected from the three aforementioned substances or from other apocarotenoids in the metabolom of flowers, cases have been found in the non-triploid species and create approaches for the commercial use of Iranian wild saffron species and pave the way for its commercialization.

کلیدواژه‌ها [English]

  • Crocin
  • Khazar
  • Picrocrocin
  • Safranal
  • Ziba
Ahrazem, O., Argandona, J., Fiore, A., Aguado, C., Lujan, R., Rubio-Moraga, A., Marro, M., Araujo-Andrade, C., Loza-Alvarez, P., Diretto, G., and Gomez-Gomez L. 2018. Transcriptome analysis in tissue sectors with contrasting crocins accumulation provides novel insights into apocarotenoid biosynthesis and regulation during chromoplast biogenesis. Scientific Reports 8 (1): 1-17. https://doi.org/10.1038/s41598-018-21225-z.
Ahrazem, O., Argandona, J., Fiore, A., Rujas, A., Rubio-Moraga, A., Castillo, R., and Gomez-Gomez L. 2019. Multi-species transcriptome analyses for the regulation of crocins biosynthesis in Crocus 20 (1): 1-15. https://doi.org/10.1186/s12864-019-5666-5.
Ahrazem, O., Lopez, A.J., Argandona, J., Castillo, R., Rubio-Moraga, A., and Gomez-Gomez, L. 2020. Differential interaction of proteins with the PSY enzymes in saffron. Scientific Reports 10 (1): 1-11. https://doi.org/10.1038/s41598-020-57480-2.
Alsayied, N.F., Fernandez, J.A., Schwarzacher, T., and Heslop-Harrison, J.S. 2015. Diversity and relationships of Crocus sativus and its relatives analysed by inter-retroelement amplified polymorphism (IRAP). Annals of Botany 116: 359-368. https://doi.org/10.1093/aob/mcv 103.
AlaviSiney, S.M. 2015. Evaluation of saffron (Crocus sativus) ecotypes diversity in Zanjan conditions. PhD Dissertation, Faculty of Agriculture, University of Zanjan, Iran. (In Persian with English Summary).
Baba, S.A., Mohiuddin, T., Basu, S., Swarnkar, M.K., Malik, A.H., Wani, Z.A., Abbas, N., Singh, A.K., and Ashraf, N. 2015. Comprehensive transcriptome analysis of Crocus sativus for discovery and expression of genes involved in apocarotenoid biosynthesis. BMC Genomics 16: 698-712. https://doi.org/10.1186/s12864-015-1894-5.
Bustin, S.A., Benes, V., Garson, J.A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pifaffl, M.W., Shipley, G.L., Vandesompele, J., and Wittwer, C.T. 2009. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55 (4): 611-622. https://doi.org/10.1373/clinchem.2008 .112797.
Castillo, R., Fernandez, J.A., and Gomez-Gomez L. 2005. Implications of carotenoid biosynthetic genes in apocarotenoid formation during the stigma development of Crocus sativus and its closer relatives. Plant Physiology 139: 674-689. Www. Plantphysiol.org/cgi/doi/10. 1104/ pp.105.067827
Ebrahimzadeh, H., Saboora, A., Noori-Dalooii, M.R., and Ghaffari, S.M. 1998. Chromosomal studies on four Iranian crocus species (Iridaceae). Iran Journal Botany 7 (2): 179-192.
Fernandez, J.A. 2004. Biology, biotechnology, and biomedicine of saffron. Recent Research Development Plant Science 127-159.
Ghaffari, S.M., and Djavadi, S.B. 2007. Chromosome study on Crocus cancellatus subsp. Damascenus from Iran. Iranian Journal Botany 13 (1): 1-3.
Golshani, F., Fakheri B.A., Solouki, M., Mahdinezhad, N., and Kiani Feriz, M.R. 2019. Study of phylogenetic relationships of some wild and crop species of Iranian Crocus by ITS nuclear loci. BIOCELL 43 (3): 225-232.
Ji, A., Jia, J., Xu, Z., Li, Y., Bi, W., Ren, F., He C., Liu, J., Hu, K., and Song, J. 2017. Transcriptome-Guided Mining of Genes Involved in Crocin Biosynthesis. Frontiers in Plant Science 8 (518): 1-12. https://doi.org/10.3389/fpls.2017.00518.
Jain, M., Srivastava, P.L., Verma, M., Ghangal, R., and Garg, R. 2016. De novo transcriptome assembly and comprehensive expression profiling in Crocus sativus to gain insights into apocarotenoid biosynthesis. Scientific Reports 6: 22456-22468. https://doi.org/ 10.1038/ srep22456.
Khansarinejhad, B., Hasandokht, M.R., and Nazeri, V. 2014. Genetic study of two wild species of saffron using morphological traits and RAPD molecular markers. Breeding of Agronomic and Horticultural Crop 2 (1): 105-118.
Mathew, B., and Brighton, C.A. 1977. Four central Asian Crocus species (Liliaceae). Iran Journal Botany 1 (2): 123-135.
Mir, J.I., Ahmed, N., Khan, M.H., and Mokhdomi, T.A. 2015. Apocarotenoid gene expression in saffron (Crocus sativus L.). Scientific Research and Essays 10 (15): 482-488. https://doi.org/ 10.5897/SRE2015.6277.
Mir, J.I., Ahmed, N., Wafai, A.H., and Qadri, R.A. 2012. Relative expression of CsZCD gene and apocarotenoid biosynthesis during stigma development in Crocus sativus L. Physiology and Molecular Biology of Plants 18 (4): 371-375. https://doi.org/10.1007/s12298-012-0131-9.
Namayandeh, A., Nemati, Z., Kamelmanesh, M.M., Mokhtari, M., and Mardi. 2012. Genetic relationships among species of Iranian crocus (Crocus spp.). Crop Breeding Journal 3 (1): 61-67.
Rubio-Moraga, A., Rambla, JL., Ahrazem, O., Granell, A., and Gomez-Gomez L. 2009. Metabolite and target transcript analyses during Crocus sativus stigma development. Phytochemistry 70: 1009-1016.
Shu, H.M., GUO, S.Q., and NI, W.C. 2020. Comprehensive transcriptome analysis of different Crocus flower tissues uncovers genes involved in crocin biosynthesis. Biologica Plantarum 64: 504-511. https://doi.org/10.32615/bp.2020.068.
Taheri-Dehkordi, A., Naderi, R., Martinelli, F., and Salami, S.A. 2020. A robust workflow for indirect somatic embryogenesis and cormlet production in saffron (Crocus sativus L.) and its wild allies; C. caspius and C.speciosus. Heliyon 6: e05841. https://doi.org/ 10.1016/j.heliyon.
Tofighi, Z., Mohamadi, H., Shokrzadeh, M., Ghahremani, M.H., Noori, S., and Habibi E. 2017. Cytotoxic effect of hydro-alcoholic extract of Crocus caspius on breast cancer cell lines 27 (151): 32-40.
Vahedi, M., Salami, S.A., Shokrpour, M., and Rezadoost, H. 2019. Comparative performance of transcriptome assembly programs for saffron (Crocus sativus L.). Saffron Agronomy and Technology 7 (1): 69-80.  https://doi.org/10.220480/jsat.2017.87859.1235. (In Persian with English Summary).
Yue, J., Wang, R., Ma, X., Liu, J.,Lu, X., Balaso, Thakar, S.B., An, N., Liu, J., Xia, E., and Liu, Y. 2020. Full-length transcriptome sequencing provides insights into the evolution of apocarotenoid biosynthesis in Crocus sativus. Elsevier 18: 774-783. https://doi.org/ 10. 1016/j.csbj.2020.03.022.
Zinati, Z., Shamloo-Dashtpagerdi, R., and Behpouri A. 2016. In silico identification of miRNAs and their target genes and analysis of gene co-expression network in saffron (Crocus sativus L.) stigma. Molecular Biology Research Communications 5 (4): 233-246.