Gök Serpil TANGOLAR, Fuat ERGENOĞLU, Saadat BÜYÜKALACA

Asma (vitis spp.) anterlerinden embriyojenik kallus ve embriyo uyartımı üzerine farklı uygulamaların etkisi

Bu çalışmada, 41 B Amerikan asma anacı (V. vinifera L. cv.‘Chasselas’ x V. berlandieri) ve Yalova incisi üzüm çeşidinin (V. vinifera L.) anterlerinden embriyojenik kallus uyartımı için farklı besi ortamları ve büyümeyi düzenleyici maddeler ile kültür koşullarının etkisi araştırılmıştır. Anterler MS (Murashige ve Skoog, 1962), NN (Nitsch ve Nitsch, 1969) ve B5 (Gambourg ve ark., 1968) ortamlarında, farklı 2,4-D (2,4-Diklorofenoksiasetik asit) (0. 0.5, 1, 2 ve 4 mg/L) ve BA (Benzil Adenin) (0, 0.1, 0.2, 1 ve 2 mg/L) kombinasyonlarında, aydınlık (fotoperyod 16 saat) ve sürekli karanlık koşullarda kültüre alınmıştır. Sonuçta 2,4-D’nin tek başına kullanıldığı ortamlardan kallus elde edilememiştir. 2,4-D ve BA’nın bazı kombinasyonlarında değişik oranlarda kallus oluşumu meydana gelmiştir. 41 B anterlerinden kallus oluşumu için en uygun ortamın B5; en uygun 2,4-D + BA kombinasyonlarının 1 mg/L + 0.2 mg/L (%70) ve 2 mg/L + 1 mg/L (%73.1); en uygun kültür koşulunun ise karanlık olduğu belirlenmiştir. Yalova incisinde kallus oluşumu yalnızca 4 mg/L 2,4-D ve 2 mg/L BA içeren B5 ortamında kültüre alınan anterlerin %4.17’sinde ve 2 mg/L 2,4-D ve 1 mg/L BA içeren NN ortamındaki anterlerin %1.52’sinde meydana gelmiştir. Denemede kullanılan tüm uygulamalardan elde edilen kallusların tamamının embriyojenik olduğu belirlenmiştir. Embriyojenik kalluslar oluştukları besi ortamının hormonsuzuna yerleştirilmiş ve ayda bir olmak üzere 3 kez alt kültüre alınmıştır. Bu süre sonunda kalluslarda herhangi bir embriyo oluşumu gözlenmemiştir.

Anahtar Kelimeler:

asma, Adana, 2, 4-D, Vitis, anterler, benziladenin, kallus, kültür ortamı, embriyogenez, üzüm, uyarılma

The effects of different applications on embryogenic callus and embryo ınduction from anters of grapevine (vitis spp.)

In this study, the effects of different media, plant growth regulators and culture conditions on embryogenic callus and embryo induction from anther explants of 41 B American rootstock (V. vinifera L. cv. „Chasselas‟ x V. berlandieri) and Yalova incisi grape (V. vinifera cv.) were investigated. Anthers from two different cultivars were cultured on MS, NN and B5 medium supplemented with various combinations of 2,4-D (2,4-Dichlorophenoxyacetic acid) (0, 0.5, 1, 2 and 4 mg/L) and BA (Benzyl Adenine) (0, 0.1 0.2, 1 and 2 mg/L ) in light (photoperiod 16 hours) and dark (continuously) conditions. Callus formation was not found from media supplemented with only in combinations containing different concentrations of 2,4-D and BA was in callus induction various ratios.The most suitable medium, combination and culture condition for callus formation from anthers of 41 B were B5, 1 mg/L 2,4-D + 0.2 mg/L BA (70%), 2 mg/L 2,4-D + 1 mg/L BA (73.1%) and dark, respectively.Callus formation from anthers of Yalova incisi was only obtained from dark conditions on B5 supplemented with 4 mg/L 2,4-D + 2 mg/L BA (4.17%) and NN medium supplemented with 2 mg/L 2,4-D + 1 mg/L BA (1.52%). All of the calluses obtained were found to be embryogenic. Embryogenic calluses were sub-cultured on hormone free-medium of the same culture medium one month intervals. At the end of three months culture period, formation of the embryos on the callus was not obtained.

Keywords:

grapevine, Adana, 2, 4-D, Vitis, anthers, benzyladenine, callus, culture media, embryogenesis, grapes, induction,

PDF

___

Bensaad, Z.M., Hennerty, M.J., Roche, T.D., 1996. Effects of Cold Pretreatment, Carbohydrate Source and Gelling Agents on Somatic Embryogenesis from Anthers of Vitis vinifera L. cvs.'Regina'and 'Reichensteiner'.Proc.Int.Sym.Plant Production in Closed Ecosystems Acta.Hort.,440:504-509.
Berres, R., Otten, L., Tinland, B., Malgarini-Clog, E., Walter, B., 1992. Transformation of Vitis Tissue by Different Strains of Agrobacterium tumefaciens Containing the T-6b Gene.Plant Cell Reports, 11: 192-195.
Cersosimo, A., Crespan, M., Paludetti, G., Altamura, M.N., 1990. Embryogenesis, Organogenesis and Plant Regeneration from Anther Culture in Vitis. 1st Internatiol Symposium ,in Vitro Culture and Horticultural Breeding. Acta Hort., 280: 307-314.
Clog, E., Bass, P., Walter, B., 1990. Plant Regeneration by Organogenesis in Vitis Rootstock Species. Plant Cell Reports, 8: 726-728.
Colby, S.M., Juncosa, A.M., Meredith, C.P., 1991. Cellular Differences in Agrobacterium Susceptibility and Regenerative Capacity Restrict the Development of Transgenic Grapevines. J. Amer. Soc. Hort. Sci., 116(2): 356-362.
Gall, O.L., Torregrosa, L., Danglot, Y., Candresse, T., Bouquet, A., 1994. Agrobacteriummediated Genetic Transformation of Grapevine Somatic Embryos and Regeneration of Transgenic Plants. Expressing the Coat Protein of Grapevine Chrome Mosaic Nepovirus (GCMV). Plant Science, 102: 161-170.
Gamborg, O.L., Miller, R.A., Ojima, K., 1968. Nutrient Requirements of Suspension Cultures of Soybean Root Cells. Exp. Cell Res. 50: 151-158.
Gray, D.J., Mortensen J.A., 1987. Initiation and Maintenance of Long Term Somatic Embryogenesis from Anthers and Ovaries of Vitis longii „Microsperma‟. Plant Cell,Tissue and Organ Culture, 9: 73-80.
Hirabayashi, T., Kozaki, I., Akihama, T., 1976. In Vitro Differentiation of Shoot from Anther Callus in Vitis. HortScience, 11(5): 511-512.
Langenbucher, M.H., Alleweldt, G., 1993. Einfluss verchiedener Vorbehandlungen auf die Induktion somatisher Embryogenese ander Antheren der Rebsorte Riesling. Vitis, 32: 1-7.
Maitz, M., Trampler, F., Gröschl, M., Camara Machada, A. Da. and Laimer Da Camara Machada, M., 2000. Use of An Ultrasound Cell Retention System for the Size Fractionation of Somatic Embryos of Woody Species. Plant Cell Reports, 19: 1057-1063.
Martinelli, L., Gribaudo, I., Bertoldi, D., Candioli, E. and Poletti, V., 2001. High Efficiency Somatic Embryogenesis and Plant Germination in Grapevine Cultivars Chardonnay and Brachetto a Grappolo Lungo. Vitis, 40(3): 111-115.
Matsuta, N., Hirabayashi, T., 1989. Embryogenic Cell Lines from Somatic Embryos of Grape (Vitis vinifera L.). Plant Cell Reports, 7: 684-687.
Mauro, M. Cl., Nef, C., Fallot, J., 1986. Stimulation of Somatic Embryogenesis and Plant Regeneration from Anther Culture of Vitis vinifera cv. Cabernet Sauvignon. Plant Cell Reports, 5: 377-380.
Mozsar, J., Viczian, O., 1996. Genotype Effect on Somatic Embryogenesis and Plant Regeneration of Vitis spp. Vitis, 35 (4): 155-157.
Mozsar, J., Süle, S., 1994. A Rapid Method for Somatic Embryogenesis and Plant Regeneration from Cultured Anthers of Vitis riparia. Vitis, 33: 245-246.
Mullins, M.G., Srinivasan, C., 1976. Somatic Embryos and Plantlets From an Ancient Clone of the Grapevine (cv. Cabernet Sauvignon) by Apomixis In Vitro. J. of Exp. Botany,27(100): 1022-1030.
Murashige, T., Skoog, F.M., 1962. A Revised Medium for Rapid Growth and Bioassays with Tabacco Tissue Cultures. Pysiol. Plant., 15: 473-497.
Nakano, M., Sakakibara T., Watanabe Y., Mn, M., 1997. Establishment of Embryogenic Cultures in Several Cultivars of Vitis vinifera and V. labruscana. Vitis, 36 (3): 141-145.
Nitsch, J., Nitsh, C., 1969. Haploid Plants From Pollen Grains. Science, 163: 85-87.
Perl, A., Gollop, R., Lipsky, A., Holland, D., Sahar, N., Or, E., Elyasi, R., 1996. Regeneration and Transformation of Grape (Vitis vinifera L.). Plant Tissue Culture and Biotechnology, 2(4): 187-193.
Perl, A., Saad, S., Sahar, N., Holland, D., 1995. Establishment of Long-Term Embryogenic Cultures of Seedless Vitis vinifera Cultivars- A Synergistic Effect of Auxins and the Role of Abscisic Acid. Plant Science 104: 193-200.
Rajasekaran, K., Mullins, M.G., 1979. Embryos and Plantlets from Cultured Anthers of Hybrid Grapevines. J. of Experimental Botany, 30(116): 399-407.
Rajasekaran, K., Mullins, M.G., 1983. Influence of Genotype and Sex-expresssion on Formation of Plantlets by Cultured Anthers of Grapevines. Agronomie, 3(3): 233-238.
Robacker, C., 1993. Somatic Embryogenesis and Plant Regeneration From Muscadine Grape Leaf Explants. HortScience, 28(1): 53-55.
Salunkhe, C.K., Rao, P.S., Mhatre. M., 1997. Induction of Somatic Embryogenesis and Plantlets in Tendrils of Vitis vinifera L. Plant Cell Reports, 17: 65-67.
Salunkhe, C.K., Rao, P.S., Mhatre. M., 1999. Plantlet Regeneration via Somatic Embryogenesis in Anter Callus of Vitis latifolia L. Plant Cell Reports, 18: 670-673.
Scorza, R., Cordts, J. M., Rammıng, D.W., Emershad, R. L., 1995. Transformation of Grape (Vitis vinifera L.) Zygotic-derived Somatic Embryos and Regeneration of Transgenic Plants. Plant Cell Reports, 14: 589-592.
Scorza, R., Cordts, J.M., Gray, D.J., Gonsalves, D., Emershad, R.L., Rammıng, D.W., 1996.Producing Transgenic “Thompson Seedless” Grape (Vitis vinifera L.) Plants. J.Amer.Soc. Hort. Sci., 121(4): 616-619.
Stamp, J.A., Meredith, C.P., 1988a. Proliferative Somatic Embryogenesis from Zygotic Embryos of Grapevine. J. Amer. Soc. Hort. Sci., 113(6): 941-945.
Stamp, J.A., Meredith, C.P., 1988b. Somatic Embryogenesis from Leaves and Anthers of Grapevine. Scientia Horticulturae, 35: 235-250.