İn Vitro Koşullarda Domates (Lycopersicon esculentum Mill.) Bitkisinde Hipokotil ve Kotiledon Eksplantlarından Kallus ve Sürgün Oluşumu

Bu çalışmada, domates M-28 hibrit çeşidinde kallus oluşumu ve sürgün oluşumu üzerine çeşitli bitki büyüme düzenleyicilerinin ve kültür koşullarının etkisi araştırılmıştır. Tohumlar yarı kuvvette (½) Murashige-Skoog (MS) ortamında çimlendirilmiş ve 10 günlük steril fidelerin kotiledon yaprakları (5x5 mm) ve hipokotilleri (5 mm) farklı konsantrasyon ve kombinasyonlarda bitki büyüme düzenleyicileri içeren semi-solid MS ortamında kültüre alınmıştır. Kallus oluşumu ve gelişimi bakımından en iyi sonucu, her iki eksplantta (kotiledon ve hipokotil) ve her iki kültür koşulunda (16 saat aydınlık-8 saat karanlık olarak fotoperiyodik koşul ve 25 gün karanlığı takiben fotoperiyodik koşul), MS ortamına 1 mg/l BAP+1 mg/l NAA ilavesi vermiştir (%100 kallus oluşumu). Adventif sürgün oluşumu bakımından, karanlığı takiben fotoperiyodik koşulda, 2 mg/l Kin+0.2 mg/l IAA, sadece 2 mg/l BAP ve sadece 3 mg/l TDZ ilaveli ortamlarda her iki eksplantta da en iyi sonuçlar alınmıştır (sürgün veren eksplant oranı %72 ile %100'dür). Eksplant başına düşen en yüksek sürgün sayısı (17.30±5.72 adet), kotiledon eksplantlarının TDZ'li ortamda karanlığı takiben fotoperiyodik koşuldaki kültürlerinden elde edilmiştir. En yüksek adventif sürgün boyu (33.00±0.57 mm) ve sürgündeki yaprak sayısı (3.00±0.00 adet) ise hipokotil eksplantlarının 1 mg/l BAP+1 mg/l NAA ilaveli ortamda fotoperiyodik koşuldaki kültürlerinde tespit edilmiştir.

Anahtar Kelimeler:

Domates, Bitki büyüme düzenleyicileri, Fotoperiyot, Kotiledon, Hipokotil, Kallus oluşumu, Sürgün oluşumu

In Vitro Callus Formation and Shoot Regeneration from Hypocotyl and Cotyledon Explants of Tomato (Lycopersicon esculentum Mill.)

In this study, the effects of various plant growth regulators and culture conditions on the callus induction and shoot regeneration in M-28 hybrid cultivar of tomato have been evaluated. The seeds were germinated in half strength (½) Murashige-Skoog (MS) medium and the cotyledon (5x5 mm) and hypocotyl (5 mm) segments of the 10 day old seedlings were cultured on semi-solide MS medium containing different concentrations and combinations of plant growth regulators. The best result on callus formation, both explants (cotyledon and hypocotyl) and both culture conditions (photoperiodic condition at 16 hours light8 hours dark ve 25 days dark condition after photoperiodic condition), was found MS medium supplemented with 1 mg/l BAP+1 mg/l NAA (%100 callus formation). At the adventitious shoot induction, under dark condition after photoperiodic condition, the best results were obtained from MS medium supplemented with 2 mg/l Kin+0.2 mg/l IAA, only 2 mg/l BAP and only 3 mg/l TDZ and both explants (regenerated explant from %72 to %100). The highest number of shoots regenerated per explant (17.30±5.72 number), was found under dark condition after photoperiodic condition at MS medium supplemented with TDZ of cotyledon explants. The highest adventitious shoot length (33.00±0.57 mm) and the number of leaf in shoot (3.00±0.00 number) were obtained at MS supplemented with 1 mg/l BAP+1 mg/l NAA of hypocotyl explants and photoperiodic conditions

Keywords:

Tomato, Plant growth regulators, Photoperiod, Cotyledon, Hypocotyl, Callus formation, Shoot regeneration,

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Aazami, M.A., Torabi, M., Shekari, F., 2010. Response of Some Tomato Cultivars to Sodium Chloride Stress Under In Vitro Culture Condition. African Journal of Biotechnology, 5(18), 2589-2592.
Ali, A.A., Yossef, T.R., El-Banna, A., 2012. Cytokinin- Cytokinin Interaction Ameliorates the Callus Induction and Plant Regeneration of Tomato (Solanum lycopersicum Mill.). Acta Agronomica Hungarica, 60(1), 47-55.
Babaoğlu, M., Yorgancılar, M., Akbudak, M.A., 2001. Doku Kültürü: Temel Laboratuvar Teknikleri, Bölüm 1, s.1-35. Bitki Biyoteknolojisi, Doku Kültürü ve Uygulamaları, Eds. Babaoğlu, M., Gürel, E., Özcan, S., Selçuk Üniversitesi Vakfı Yayınları, Konya, 374s.
Bhatia, P., Ashwath, N., 2005. Effect of Medium pH on Shoot Regeneration from the Cotyledonary Explants of Tomato. Biotechnology, 4(1), 7-10.
Bhatia, P., Ashwath, N., Midmore, D.J., 2005. Effects of Genotype, Explant Orientation, and Wounding on Shoot Regeneration in Tomato. In Vitro Cell Development in Biology, 41, 457-464.
Busse, J.S., Figueroa-Cabanas, M., Stimart, D.P., 2005. Developmental Anatomy of Adventitious Shoot Formation on Snapdragon (Antirrhinum majus L.) Hypocotyls In Vitro. J. Amer. Soc. Hort. Sci, 130(2), 147-151.
Chaudhry, Z., Abbas, S., Yasmin, A., Rashid, H., Ahmed, H., Anjum, M.A., 2010. Tissue Culture Studies in Tomato (Lycopersicon esculentum) var. Moneymaker. Pakistan Journal of Botany, 42(1), 155-163.
El-Bakry, A., 2002. Effect Genotype, Growth Regulators, Carbon Source and pH on Shoot Induction and Plant Regeneration in Tomato. In Vitro Cell Development in Biology, 38, 501-507.
El-Meleigy, E.-S.A., Gabr, M.F., Mohamed, F.H. ve Ismail, M.A., 2004. Responses to NaCl Salinity of Tomato Cultivated and Breeding Lines Differing in Salt Tolerance in Callus Cultures. Int J Agri Biol, 6 (1), 19-26.
Gubis, J., Lajchova, Z., Farago, J., Jurekova, Z., 2004. Effect of Growth Regulators on Shoot Induction and Plant Regeneration in Tomato (Lycopersicon esculentum Mill.). Biologia, 59/3, 405-408.
Harish, M.C., Rajeevkumar, S., Sathishkumar, R., 2010. Efficient In Vitro Callus Induction and Regeneration of Different Tomato Cultivars of India. Asian Journal of Biotechnology, 2(3), 178-184.
Ishag, S., Osman, M.G., Khalafalla, M.M., 2009. Effects of Growth Regulators, Explant and Genotype on Shoot Regeneration in Tomato (Lycopersicon esculentum cv. Omdurman). International Journal of Sustainable Crop Production, 4(6), 7-13.
Moghaieb, R.E.A., Saneoka, H., Fujita, K., 1999. Plant Regeneration from Hypocotyl and Cotyledon Explant of Tomato (Lycopersicon esculentum Mill.). Soil Science and Plant Nutrition, 45(3), 639-646.
Mohamed, A.N., Ismail, M.R., Rahman, M.H., 2010. In Vitro Response from Cotyledon and Hypocotyls Explants
Benzylaminopurine. African Journal of Biotechnology, 9(30), 4802-4807. by Inducing
6- Murashige, T., Skoog, F., 1962. A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Physiology Plantarum, 15, 473-497.
Namitha, K.K., Negi, P.S., 2013. Morhogenetic Potential of Tomato (Lycopersicon esculentum) cv. 'Arka Ahuti' to Plant Growth Regulators. Noyulae Scientia Biologicae, 5(2), 220-225.
Osman, M.G., Elhadi, E.A., Khalafalia, M.M., 2010. Callus Formation and Organogenesis of Tomato (Lycopersicon esculentum Mill, c.v. Omdurman) Induced by Thidiazuron. African Journal of Biotechnology, 9(28), 4407-441.
Plana, D., Alvarez, M., Lara, R.M., Florido, M., Alvarez, F., Moya, C., 2005. A New In Vitro Regeneration Protocol in Tomato (Lycopersicon esculentum Mill.). Cultivos Tropicales, 26(2), 17-20.
Rashid, R., Bal, S.S., 2010. Effect of Hormones on Direct Shoot Regeneration in Hypocotyl Explants of Tomato. Notulac Scientia Biologicae, 2(1), 70-73.
Sheeja, T.E., Mondal, A.B., Rathore, R.K.S., 2004. Efficient Plantlet Regeneration in Tomato (Lycopersicon esculentum Mill.). Plant Tissue Culture, 14(1), 45-53.
Vikram, G., Madhusudhan, K., Srikanth, K., Laxminarasu, M, Swamy, N.R., 2011. Effect of Plant Growth Regulators on In Vitro Organogenesis in Cultivated Tomato (Solanum lycopersicum L.). Journal of Research in Biology, 4, 263-268.
Yokaş, İ., Tuna, A.L., Bürün, B., Altunlu, H., Altan, F., Kaya, C., 2008. Responses of the Tomato Different Salt Forms and Rates. Turkish Journal of Agriculture and Forestry, 32, 319-329. Semboller % ºC l cm mm µm g mg atm atmosfer