Callus Induction and Plant Regeneration of Chrysanthemum morifolium and C. coccineum via Direct and Indirect Organogenesis and Genetic Fidelity Analysis Using IRAP, ISSR and SCoT Molecular Markers
محورهای موضوعی : مجله گیاهان زینتیFardin Nasri 1 , Hedayat Zakizadeh 2 , Yavar Vafaee 3 , Ali Akbar Mozafari 4
1 - Department of Horticultural Sciences, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran
2 - Department of Horticultural Sciences, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran
3 - Department of Horticultural Science, Agriculture Faculty, University of Kurdistan, Sanandaj, Iran
4 - Department of Horticultural Science, Agriculture Faculty, University of Kurdistan, Sanandaj, Iran
کلید واژه: Callus induction, Embryogenesis, Cultivar response, Direct regeneration, Genetic fidelity,
چکیده مقاله :
In vitro propagation of C. morifolium cv. ‘Homa’ and cv. ‘Delkash’ and wild C. coccineum via direct and indirect organogenesis and somatic embryogenesis were investigated. BAP at 0, 1, 2 and 3 mg l-1 or NAA at 0, 0.05, 0.1 and 0.2 mg l-1 concentrations were used to induce direct and indirect organogenesis of shoot tip explants. To study the callus induction and somatic embryogenesis, the young leaf explants were cultured on MS medium containing BAP (0, 1, 2 or 3 mg l-1) and 2,4-D (0, 1, 2 or 3 mg l-1). Direct shoot regeneration was achieved from shoot tip explants of ‘Homa’ and ‘Delkash’ as well as C. coccineum. The highest number of shoots through direct regeneration (13.78 and 8.89 shoots per explant for C. coccineum and C. morifoilum ‘Homa’, respectively) were observed in the treatment with 2 mg l-1 BAP and 0.05 mg l-1 NAA. In both species, the highest frequency of callus formation and embryogenesis were obtained on medium containing 2.0 mg l-1 2,4-D and 2 mg l-1 BAP. Genetic fidelity of 10 acclimatized plants derived from direct regeneration of each species was confirmed using six inter-retrotransposon amplified polymorphism (IRAP), inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) primers. A total of 56, 56 and 39 fragments were amplified for IRAP, ISSR, and SCoT, respectively. In general, our results showed that finding a better response of explants to embryogenesis or organogenesis in a specific cultivar and with special PGRs combinations and concentrations play an important role in the in vitro propagation efficiency of chrysanthemum species.
در این تحقیق، تکثیر درون شیشهای داوودی گونه C. morifolium ارقام ’هما‘ و ’دلکش‘ و گونه بومی (C. coccineum) از طریق اندام زایی مستقیم، غیر مستقیم و جنین زایی غیرجنسی گزارش شده است. BAP در غلظتهای صفر، 1، 2 و 3 میلیگرم در لیتر و NAA در غلظتهای صفر، 05/0، 1/0 و 2/0 میلیگرم در لیتر جهت القای اندامزایی مستقیم و غیر مستقیم در ریزنمونههای نوک شاخههای انتهایی استفاده شد. جهت مطالعه القای کالوس و جنین زایی غیرجنسی، ریزنمونههای جوان برگ بر روی محیط کشت MS حاوی BAP (صفر، 1، 2 و 3 میلیگرم در لیتر) و 2و4-دی (صفر، 1، 2 و 3 میلیگرم در لیتر) کشت شدند. باززایی مستقیم شاخه از ریزنمونههای نوک شاخههای انتهایی ارقام ’هما‘ و ’دلکش‘ و همچنین در گونه بومی C. coccineum مشاهده شد. بالاترین تعداد شاخهها از طریق اندامزایی مستقیم (78/13 و 89/8 شاخه در هر ریزنمونه به ترتیب برای C. coccineumو ’هما‘) با 2 میلی گرم در لیتر BAP و 05/0 میلیگرم در لیتر NAA مشاهده شد. در هر دو گونه، بالاترین فراوانی تشکیل کالوس و جنینزایی روی محیط کشت حاوی 2 میلیگرم در لیتر 2و4-دی و 2 میلیگرم در لیتر BAP بدست آمد. پایداری ژنتیکی 10 گیاه سازگار یافته حاصل از اندامزایی مستقیم از هر گونه با استفاده از6 پرایمر از نشانگرهای مولکولی (IRAP, ISSR, SCoT) به اثبات رسید. در کل 56، 56 و 39 قطعه برای IRAP، ISSR و SCoT بهترتیب تکثیر شدند. در کل، نتایج ما نشان داد که یافتن عکسالعمل بهتر ریزنمونهها به جنینزایی یا اندامزایی در یک رقم ویژه و با یک ترکیب هورمونی و غلظتهای آنها نقش مهمی را در کارآیی تکثیر درون شیشهای گونههای داوودی، بازی میکند.
Ali, N., Kafi, M., Mirmasoumi, M. and Babalar, M. 2005. Micropropagation of damask rose (Rosa damascene Mill) cvs. ‘Azaran’ and ‘Ghamsar’.International Journal of Agricultural and Biology,7: 535-538.
Atul Babu, G.A., Vinoth, A. and Ravindhran, R. 2018. Direct shoot regeneration and genetic fidelity analysis in finger millet using ISSR markers. Plant Cell, Tissue and Organ Culture, 132 (1): 157-164.
Benson, E.E. 2000. In vitro plant recalcitrance: An introduction. In vitro Cellular and Develomental Biology Plant, 36: 141-148.
Bhatia, R., Singh, K.P., Sharma, T.R. and Jhang, T. 2011. Evaluation of the genetic fidelity of in vitro-propagated gerbera (Gerbera jamesonii Bolus) using DNA-based markers. Plant Cell, Tissue and Organ Culture, 104 (1): 131-135.
Bhattacharyya, P., Kumaria, S., Diengdoh R. and Tandon, P. 2014. Genetic stability and phytochemical analysis of the in vitro regenerated plants of Dendrobium nobile Lindl, an endangered medicinal orchid. Meta gene, 2: 489-504.
Chaudhury, A. and Rongda, Q.U. 2000. Somatic embryogenesis and plant regeneration of turf type Bermuda grass: Effect of 6-benzyadenine in callus induction medium. Plant Cell, Tissue and Organ Culture, 60 (2): 113-120.
Collard, B.C.Y. and Mackill, D.J. 2009. Start codon targeted (SCoT) polymorphism: A simple, novel DNA marker technique for generating genetargeted markers in plants. Plant Molecular and Biology Reporter, 27: 86-93.
Deyi, Z., You-yin, Z., Qian, L., Ti, Z. and De-gang, Z. 2011. Production of embryogenic callus and plant regeneration from elite guizhou waxy maize inbred lines. Agricultural Sciences in China, 10 (4): 490-498.
Doyle, J.J. and Doyle, J.L. 1990. Isolation of plant DNA from fresh tissue. Focus,12: 13-15.
Emami, A., Shabanian, N., Rahmani, M.S., Khadivi, A. and Mohammad-Panah, N. 2018. Genetic characterization of the Crataegus genus: Implications for in situ conservation. Scientia Horticulturae,231: 56-65.
Feher, A., Pasternack, T.P. and Dudits, D. 2003. Transition of somatic plant cells to an embryogenic state. Plant Cell, Tissue and Organ Culture, 74: 201-228.
Florence, M.W. and Rangan, T.S. 1981. In vitro clonal multiplication of pyrethrum (Chrysanthemum cinerariaefolium Vis.) by micropropagation. Plant Science Letters, 22: 219-226.
Francis, D. and Sorrell, D.A. 2001. The interface between the cell cycle and plant growth regulators: A mini review. Plant Growth Regulation, 33: 1–12.
Gamborg, O.L., Murashige, T.A. and Vasil, L.K. 1976. Plant tissue culture media. In Vitro, 12: 473-81.
Gao, Y., Bo, Z., Guoxun, D. and Qixiang, Z. 2001. Shoot regeneration from stem and leaf explants of Dendrathema grandiflorum. Journal of Beijing Forestry University, 23 (1): 32-33.
Gaspar, T., Kevers, C., Faivre-Rampant, O., Crèvecoeur, M., Penel, C., Greppin, H. and Dommes, J. 2003. Changing concepts in plant hormone action. In vitro Cellular and Develomental Biology Plant, 39: 85-105.
Himstedt, J.P., Jacobsen, H.J. and Fisher-Kluver, G. 2001. Shoot regeneration from stem and leaf explants of chrysanthemum (Dendranthema x grandiflorum). Acta Horticulturae, 560: 421-424.
Hitmi, A., Barthomeuf, C. and Sallanon, H. 1998. Rapid mass propagation of Chrysanthemum cinerariaefolium Vis. by callus culture and ability to synthesise pyrethrins. Plant Cell Reporters, 19: 156-160.
Jevremovic´, S. and Radojevic, L.J. 2004. Mass production of different chrysanthemum (Chrysanthemum morifolium) cultivars by culture in vitro. Journal of Science and Agricultural Research, 65: 47-54.
Joshi, P. and Dhawan, V. 2007. Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biologia Plantarum, 51 (1): 22-26.
Kalendar, R., Grob, T., Regina, M., Suoniemi, A. and Schulman, A. 1999. IRAP and REMAP: Two new retrotransposon-based DNA fingerprinting techniques. Theoretical Applied Genetics, 98: 704-711.
Kengkarj, P., Smitamana, P. and Fujime, Y. 2008. Assessment of somaclonal variation in chrysanthemum (Dendranthema grandiflora Kitam.) using RAPD and morphological analysis. Plant Tissue Culture and Biotechnology, 18(2): 139-149.
Kumar, S., Prasad, K.V. and Choudhary, M.L. 2006. Detection of genetic variability among chrysanthemum radio-mutants using RAPD markers. Current Science, 90: 1108-1113.
Kumar, A., Singh, S.P. and Bhakuni, R.S. 2005. Secondary metabolites of chrysanthemum genus and their biological activities. Current Science, 89: 1489-1501.
Lim, K.B., Kwon, S.J., Lee, S.I. and Hwang, Y.J. 2012. Influence of genotype, explant source, and gelling agent on in vitro shoot regeneration of chrysanthemum.
Horticulture, Environment, and Biotechnology, 53: 329-335.
Mandal, A.K.A. and Datta, S.K. 2005. Direct somatic embryogenesis and plant regeneration from ray florets of chrysanthemum. Biologia Plantarium, 49 (1): 29-33.
Meyer, W., Mitchell, T.G., Freedman, E.Z. and Vilgays, R. 1993. Hybridization probes for conventional DNA fingerprinting used as single primers in the polymerase chain reaction to distinguish strains of Cryptococcus neoformans. Journal of Clinical Microbiology, 31: 2274-2280.
Miler, N. and Zalewska, M. 2014. Somaclonal variation of chrysanthemum propagated in vitro from different explants types. Acta Scientiarum Polonorum Horticulture, 13 (2): 69-82.
Mozafari, A.A., Vafaee, Y. and Karami, E. 2015. In vitro propagation and conservation of Satureja avromanica Maroofi—an indigenous threatened medicinal plant of Iran. Physiology and Molecular Biology of Plants, 21 (3): 433-439.
Nahid, J.S., Shyamali, S. and Kazumi, H. 2007. High frequency shoot regeneration from petal explants of Chrysanthemum morifolium Ramat. in vitro. Pakistan Journal of Biological Sciences, 10: 3356-3361.
Naing, A.H., Kim, C.K., Yun, B.J., Jin, J.Y. and Lim, K.B. 2013. Primary and secondary somatic embryogenesis in chrysanthemum cv. Euro. Plant Cell, Tissue and Organ Culture, 112: 361-368.
Obukosia, S. D., Kimani, E., Wathaika, K., Mutitu, E. and Kimani, P.M. 2004. Effect of growth regulators and genotypes on pyrethrum in vitro. In vitro Cellular and Develomental Biology-Plant. Pl. 11: 162-166.
Petersen, G. and Seberg, O. 1998. Molecular characterization and sequence polymorphism of the alcohol dehydrogenase 1 gene in Hordeum vulgare L. Euphytica, 102: 57-63.
Petty, L.M., Harberd, N.P., Carre´, I.A., Thomas, B. and Jackson, S.D. 2003. Expression of the Arabidopsis Gai gene under its own promoter causes a reduction in plant height in chrysanthemum by attenuation of the gibberellin response. Plant Sciences, 164: 175-182.
Rahmani, M.S., Pijut, P.M. and Shabanian, N. 2016. Protoplast isolation and genetically true-to-type plant regeneration from leaf-and callus-derived protoplasts of Albizia julibrissin. Plant Cell, Tissue and Organ Culture, 127 (2): 475-488.
Rahmani, M.S., Pijut, P.M., Shabanian, N. and Nasri, M. 2015. Genetic fidelity assessment of in vitro-regenerated plants of Albizia julibrissin using SCoT and IRAP fingerprinting. In vitro Cellular and Develomental Biology-Plant, 51(4): 407-419.
Rashmi, R. and Trivedi, M.P. 2014. Effect of various growth hormone concentration and combination on callus induction, nature of callus and callogenic response of Nerium odorum. Biotechnology and Applied Biochemistry, 172 (5): 2562-2570.
Satish, L., Ceasar, S.A., Shilpha, J., Rency, A.S., Rathinapriya, P. and Ramesh, M. 2015. Direct plant regeneration from in vitro derived shoot apical meristems of finger millet (Eleusine coracana L. Gaertn). In vitro Cellular and Develomental Biology-Plant, 51: 192-200.
Sauvadet, M.A., Brochard, P. and Gibod, J.B. 1990. A protoplast to plant system in chrysanthemum: Differential responses among several commercial clones. Plant Cell Reporters, 8: 692-695.
Shahzad, A., Parveen, S., Sharma, S., Shaheen, A., Saeed, T., Yadav, V., Akhtar, R., Ahmad, Z. and Upadhyay, A. 2017. Plant tissue culture: Applications in plant improvement and conservation. In: Plant Biotechnology: Principles and Applications (pp. 37-72). Springer Singapore.
Shinoyama, H., Nomura, Y., Tsuchiya, T. and Kazuma, T. 2004. A simple and efficient method for somatic embryogenesis and plant regeneration from leaves of chrysanthemum (Dendranthema grandiflora (Ramat.) Kitamura). Plant Biotechnology, 21: 25-30.
Song, J.Y., Mattson, N.S. and Jeong, B.R. 2011. Efficiency of shoot regeneration from leaf, stem, petiole, and petal explants of six cultivars of Chrysanthemum morifolium. Plant Cell, Tissue and Organ Culture, 107: 295-304.
Staden, J. and Crounch, N. 1996. Benzyladenine and derivatestheir significance and interconversion in plants. Plant Growth Regulation, 19: 153-175.
Tanaka, K., Kanno, Y., Kudo, S. and Suzuki, M. 2000. Somatic embryogenesis and plant regeneration in chrysanthemum (Dendranthema grandiflora (Ramat.) Kitamura). Plant Cell Reports, 19: 946-953.
Teixeira da Silva, J.A. 2004. Ornamental chrysanthemums: Improvement by biotechnology. Plant Cell, Tissue and Organ Culture, 79: 1-18.
Teixeira da Silva, J.A. 2014. Organogenesis from chrysanthemum (Dendranthema x grandiflora (Ramat.)) ‘Kitamura’ petals (disc and ray florets) induced by plant growth regulators. Asia-Pacific Journal of Molecular Biology and Biotechnology, 22 (1): 145-151.
Teixeira da Silva, J.A., Shinoyama, H., Aida, R., Matsushita, Y., Raj, S.K. and Chen, F. 2013. Chrysanthemum biotechnology: Quo vadis? Critical Reviews in Plant Sciences, 32 (1): 21-52.
Thakur, J., Dwivedi, M.D., Sourabh, P., Uniyal, P.L. and Pandey, A.K. 2016. Genetic homogeneity revealed using SCoT, ISSR and RAPD markers in micropropagated Pittosporum eriocarpum Royle, an endemic and endangered medicinal plant. PloS one, 11 (7): e0159050.
Thomas, T.D. and Maseena, E.A. 2006. Callus induction and plant regeneration in Cardiospermum halicacabum (L.), an important medicinal plant. Scientia Horticulturae, 108: 332-336.
Turrea, K.C. 1989. Tissue culture media, composition and preparation in tissue culture techniques for horticulture crops. van Nostard, Remhold, New York pp. 26-51.
Wandahwa, F., van Ranst, E. and van Damme, I. 1996. Pyrethrum (Chrysanthemum cinerariaefolium Vis.) cultivation in West Kenya: Origin, ecological conditions and management. Industrial Crops and Products, 5: 307-322.
Waseem, K., Jilani, M.S. and Khan, M.S. 2009. Rapid plant regeneration of chrysanthemum (Chrysanthemum morifolium L.) through shoot tip culture. African Journal of Biotechnology, 8: 1871-1877.
Waseem, K., Khan, M.Q., Jaskani, J. and Khan, M.S. 2008. Impact of different auxins on the regeneration of chrysanthemum (Dendranthema morifolium) through in vitro shoot tip culture. Pakistan Journal of Agricultural Research, 20 (1-2): 51-57.
Zia, M., Rizvi, Z.F., Rehman, U.R. and Chaudhary, M.F. 2010. Micropropagation of two Pakistani soybean (Glycine max L.) cultivars from mature seeds cotyledon nodes. Spanish Journal of Agricultural Research, 8: 448-453.