Patlıcanda In vitro Rejenerasyon Ortamında Farklı Orizalin ve Kolhisin Konsantrasyonlarının Tetraploid Bitki Üretimine Etkisi

Bu çalışmanın amacı, Faselis F1 ve Karnaz F1 patlıcan çeşitlerinde kolhisin ve orizalinin in vitro rejenerasyon ortamında kullanılmasıyla tetraploid bitki elde etmektir. Araştırmada, yaprak eksplantları 10 µM BA ve 1 µM IAA içeren katı MS rejenerasyon ortamında 5 ve 7 gün bekletilmiştir. Ardından bu eksplantlara kolhisinin 2,5 ve 3,75 mM konsantrasyonları 8, 16 ve 32 saat süreyle; orizalinin ise 28,8 ve 43,2 µM konsantrasyonları 12, 24 ve 36 saat süreyle uygulanmış ve tekrar kolhisinsiz ve orizalinsiz rejenerasyon ortamına aktarılmıştır. Rejenerasyon ortamında oluşan kallus, tomurcuk veya kısa sürgünler 0,5 µM BA ile desteklenmiş MS ortamına alınarak bitkilerin oluşması sağlanmış ve ploidi seviyesi flow sitometri ile belirlenmiştir. Karnaz F1 çeşidinin eksplantlarına 3,75 mM kolhisin dozunun uygulaması sonucu, 2,5 mM’a göre daha yüksek tetraploid bitki oluşumu sağlanmıştır. Bu çeşitte uygulanan orizalin denemesinde ise en yüksek tetraploid bitki oluşumu, eksplantların 7 gün rejenerasyon ortamında bekletilmesinden sonra 43,2 µM orizalin konsantrasyonunun 24 saat uygulamasından elde edilmiştir. Karnaz F1 çeşidinde kolhisin ve orizalin uygulamasından elde edilen tetraploid bitkilerde çiçek tozu canlılığı %76,99 ve %81,19, çimlenme %19,14 ve %17,98 olarak gerçekleşmiştir. Faselis F1 çeşidinin rejenerasyon ortamında 7 gün bekletilen yaprak eksplantları 2,5 mM kolhisin konsantrasyonunda 8 ve 32 saat inkübe edildiğinde, diğer uygulamalara göre daha yüksek tetraploid bitki elde edilmiştir. Aynı çeşidin orizalin denemesinde, eksplantların 5 gün rejenerasyon ortamında bekletilmesi sonucu daha yüksek tetraploid bitki üretilmiştir. Faselis F1 çeşidinde kolhisin ve orizalin uygulamaları sonucu üretilen tetraploid bitkilerde çiçek tozu canlılığı %86,41 ve %95,68, çimlenme ise %26,54 ve %28,47 olarak belirlenmiştir.

The Effect of Different Oryzalin and Colchicine Concentrations Supplemented to In vitro Regeneration Medium on Tetraploid Plant Production in Eggplant

The objective of this study is to obtain tetraploid plants in eggplant cultivars, Faselis (F1) and Karnaz (F1), by applying colchicine and oryzalin in in vitro regeneration medium (RM: MS with 10 µM BA and 1 µM IAA). In the study, leaf explants which have been incubated on the solidified RM for 5 or 7 days were cultured on the same medium with 2.5 or 3.75 mM colchicine for 8, 16 or 32 hours; or with 28.8 or 43.2 µM oryzalin for 12, 24 or 36 hours. Then the explants were transferred to RM without colchicine and oryzalin. Callus, buds or short shoots formed in the RM were transferred to MS medium with 0.5 μM BA to obtain long plants. The ploidy levels of regenerants were determined by flow cytometry. In Karnaz, higher tetraploid plant formation was achieved from 3.75 mM colchicine compared to 2.5 mM colchicine. The highest tetraploid plant rate was obtained by applying of 43.2 µM oryzalin for 24 hours to the explants after incubation on the RM for 7 days. Pollen viability of the tetraploid plants produced by application of colchicine and oryzalin were 76.99% and 81.19% and germination were 19.14% and 17.98%. In Faselis, tetraploid plants were obtained by applying 2.5 mM colchicine to the explants for 8 or 32 hours after incubation on RM for 7 days. However, in the oryzalin experiment, the higher tetraploid plants were produced when the explants were incubated for 5 days in the RM. Pollen viability of the tetraploid plants obtained from applications of colchicine and oryzalin were 86.41% and 95.68% and germination were 26.54% and 28.47%.

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Al Hakimi A, Monneveaux P, Nachit MM. 1998. Direct and indirect selection for drought tolerance in alien tetraploid wheat x durum wheat crosses. Euphytica, 100: 287-294. DOI: https://doi.org/10.1007/978-94-011-4896-2_49.
Asensio IC, Prohens J, Gisbert C. 2014. Vigor for in vitro culture traits in S. melongena x S. aethiopicum hybrids with potential as rootstocks for eggplant. Sci. World J., ID 702071:1-8. DOI: http://dx.doi.org/10.1155/2014/702071.
Bletsos F, Thanassoulopoulos C, Roupakias D. 2003. Effect of grafting on growth, yield and Verticillium wilt of eggplant. HortScience, 38: 183-186.
Chavez R, Brown CR, Iwanaga M. 1988. Application of interspecific sesquiploidy to introgression of PLRV resistance from non-tuber-bearing Solanum etuberosum to cultivated potato germplasm. Theor. Appl. Genet., 76: 497- 500. DOI: 10.1007/BF00260898.
Collonnier C, Fock I, Kashyap V, Rotino GL, Daunay MC, Lian Y, Mariska IK, Rajam MV, Servaes A, Ducreux G, Sihachakr D. 2001. Applications of biotechnology in eggplant. Plant Cell, Tissue and Organ Culture, 65: 91-107. DOI: https://doi.org/10.1023/A:1010674425536.
Collonnier C, Fock I, Mariska I, Servaes A, Vedel F, SiljakYakovlev S, Souvannavong V, Sihachakr D. 2003. GISH confirmation of somatic hybrids between Solanum melongena and S. torvum: assessment of resistance to both fungal and bacterial wilts. Plant Physiol. Biochem., 41: 459- 470. DOI: https://doi.org/10.1016/S0981-9428(03)00054-8.
Çağlar G, Bağcı S. 2004. Bazi Cucumis türleri arasindaki melezlemelerde embriyo kurtarma yoluyla in vitro hibrit bitki regenerasyonu. Akdeniz Üniversitesi Ziraat Fakültesi Dergisi, 17(2): 175-182.
Çürük S, Dayan A, Tekeli FÖ. 2014a. İnterspesifik melez patlıcan bitkilerinde in vitro rejenerasyon. 10. Sebze Tarım Sempozyomu. Tekirdağ, 2-4/09/2014. ss: 302-309.
Çürük S, Doksöz S, Külahlıoğlu İ. 2014b. Diploid ve tetraploid interspesifik hibrit patlıcan (Solanum melongena x Solanum torvum) genotiplerinde aşının çiçek tozu verimliliği ve bitki morfolojisi üzerine etkisi. Kesin sonuç raporu. Proje No:112O751 (TÜBITAK-TOVAG).
Daunay MC. 2008. Eggplant. In: Prohens J, Nuez F (Eds). Handbook of crop breeding vegetables II: Fabaceae, Liliaceae, Umbelliferae and Solanacaea. New York, USA: Springer. pp:163-220. eBook ISBN: 978-0-387-74110-9.
Daunay MC, Hazra P. 2012. Eggplant. In: Peter KV, Hazra P (Eds.). Handbook of Vegetables. Houston, TX, USA: Studium Press. Pp: 257-322.
Dhooghe E, Van Laere K, Eeckhaut T, Leus L, Van Huylenbroeck J. 2011. Mitotic chromosome doubling of plant tissues in vitro. Plant Cell, Tissue and Organ Culture, 104: 359–373. DOI: https://doi.org/10.1007/s11240-010-9786-5.
FAO 2018. Food and Agriculture Organization of the United Nations. Erişim Adresi: http://faostat3.fao.org/faostatgateway/go/to/download/Q/QC/E. Erişim: [5/4/2018].
Gousset C, Collonnier C, Mulya K, Mariska I, Rotino GL, Besse P, Servaes A, Sihachakr D. 2005. Solanum torvum, as a useful source of resistance against bacterial and fungal diseases for improvement of eggplant (S. melongena L.). Plant Sci., 168: 319-327. DOI: https://doi.org/10.1016/j.plantsci. 2004.07.034.
Hanudin H, Hanafiah Goas MA. 1992. Screening of eggplant accessions for resistance to bacterial wilt. In: Hartman GL, Hayward AC (Eds.). Bacterial Wilt. Taiwan: ACIAR Proceedings, 45: 191-192. ISBN:1-86320-087-8.
Jansky S. 2006. Overcoming hybridization barriers in potato. Plant Breeding, 125: 1-12. DOI: 10.1111/j.1439- 0523.2006.01178.x.
Kabaş A, Arı E, Zengin S, İlbi H, Aysan Y, Oğuz A. 2016. Development of resistant tomato population with bacterial canker resistance genes from interspecific hybrids by the support of embryo rescue. Derim, 33(1): 27-46. DOI:10.16882/derim.2016.07605.
Kashyap V, Kumar SV, Collonier C, Fusari F, Haciour R, Rotino GL, Sihachakr D, Rajam MV. 2003. Biotecnology of eggplant. Sci. Hort., 97: 1-25. https://doi.org/10.1016/S0304- 4238(02)00140-1.
Khan MMR, Isshiki S. 2008. Development of a male sterile eggplant by utilizing the cytoplasm of Solanum virginianum and a biparental transmission of chloroplast DNA in backcrossing. Scientia Horticulturae, 117: 316-320. https://doi.org/10.1016/j.scienta.2008.05.006.
Kulkarni M, Borse T. 2010. Induced polyploidy with gigas expression for root traits in Capsicum annuum (L.). Plant Breeding, 129(4): 461-464. DOI: 10.1111/j.1439- 0523.2009.01696.x.
Kumchai J, Wei YC, Lee CY, Chen FC, Chin SW. 2013. Production of interspecific hybrids between commercial cultivars of the eggplant (Solanum melongena L.) and its wild relative S. torvum. Genet. Mol. Res., 12 (1): 755-764. DOI: 10.4238/2013.
Leon JR, Reyes-Valdes MH, Mendoza-Rodriguez DV, RamirezGodina F, Robledo-Torres V, Gomez-Martinez M, Hernandez-Guzman G. 2015. Meiotic analysis of four crosspollinated generations in a synthetic autotetraploid population of husk tomato (Physalis ixocarpa). Journal of Experimental Botany, 84(1): 101-106.
Mears JA. 1980. Chemistry of polyploids: a summary with comments on Parthenium (Asteraceae-Ambrosiinae). In: Lewis WH (ed.) Polyploidy: biological relevance. New York: Plenum Press, 13: 77-102. DOI: https://doi.org/10.1007/978- 1-4613-3069-1_5.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum, 15: 473-497. https://doi.org/10.1111/j.1399- 3054.1962.tb08052.x.
Osborn TC, Pires, JC, Birchler JA, Auger DL, Chen ZJ, Lee HS, Comai L, Madlung A, Doerge RW, Colot V, Martienssen RA. 2003. Understanding mechanisms of novel gene expression in polyploids. Trends Genet., 19: 141–147. DOI: 10.1016/S0168-9525(03)00015-5.
Plazas M, Vilanova S, Gramazio P, Rodriguez-Burruezo A, Fita A, Herraiz FJ, Ranil R, Fonseka R, Niran L, Fonseka H. 2016. Interspecific hybridization between eggplant and wild relatives from different genepools. J. Am. Soc. Hortic. Sci., 141: 34-41.
Ramanna MS, Hermsen JGTH. 1981. Structural hybridity in the series Etuberosa of the genus Solanum and its bearing on crossability. Euphytica, 30: 15-31. DOI: 10.1007/BF00033655.
Riddle NC, Kato A, Birchler JA. 2006. Genetic variation for the response to ploidy change in Zea mays L. Theor. Appl. Genet., 114:101-111. DOI: 10.1007/s00122-006-0414-z.
Sakhanokho HF, Islam-Faridi MN. 2014. Spontaneous autotetraploidy and its impact on morphological traits and pollen viability in Solanum aethiopicum. HortScience, 49(8): 997-1002.
Singh AK, Singh M, Singh AK, Singh R, Kumar S, Kallo G. 2006. Genetic diversity within the genus Solanum (Solanaceae) as revealed by RAPD markers. Curr. Sci., 90 (5): 711-716.
Song P, Kang W, Ellen BP. 1997. Chromosome doubling of Allium fistulosum x A. cepa interspesific F1 hybrids through colchicine treatment of regenerating callus. Euphytica, 93: 257–262.
Stanys V, Weckman A, Staniene G, Duchovskis P. 2006. In vitro induction of polyploidy in japanese quince (Chaenomeles japonica). Plant Cell, Tissue and Organ Culture, 84: 263-268. DOI: 10.1007/s11240-005-9029-3.
Tal M. 1980. Physiology of polyploids. In: Lewis WH (ed.). Polyploidy: biological relevance. New York: Plenum Press. 13: 61-76. PMID: 550845.
Tuna M. 2014. Flow sitometri ve tarımsal araştırmalarda kullanımı. II. Flow Sitometri ve Tarımsal Araştırmalarda Kullanımı Eğitim Programı Notları, Tekirdağ, 16-17 Ocak 2014.
Xiong YC, Li FM, Zhang T. 2006. Performance of wheat crops with different chromosome ploidy: root-sourced signals, drought tolerance, and yield performance. Planta, 224: 710- 718. DOI: 10.1007/s00425-006-0252-x.
Xu C, Huang Z, Liao T, Li Y, Kang X. 2016. In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus. Plant Cell Tissue and Organ Culture, 125: 1-9. DOI: https://doi.org/10.1007/s11240-015-0922-0.