In vitro Selection for Resistans to Head Blight (Fusarium Spp.) in Wheat (Triticumaestivum L.) and Plant Regeneratıon

Year 2008, Volume: 21 Issue: 1, 55 - 64, 01.06.2008

Ş. E. Arıcı N. K. Koç Namık Kemal Koç

Abstract

In this study, it was aimed to get resistance wheat lines against to Fusarium Head Blight via somaclonal variation by using in vitro selection. For this purpose, MS media were prepared including lethal doses of culture filtrate of F. graminearum, F. culmorum and fusaric acid (30%, 0.3 mM). Embriogenic calli of Adana 99, Genc 99, and Seri 82 wheat cultivars were cultured on this medium. Thus 19 plants were regenerated from three wheat genotypes after the selection of embriogenic calli. DNA isolation of 19 plants were done and then RAPD-PCR analysis were applied using six various primers. As regard to amplification results by electrophoresis, bands were detected when the primer OPA-05, OPA-04, OPD-05 were used while polymorphism were not detected. To reveal genetic variation among the wheat genotypes further studies, such as various and more primers should applied.

Keywords

Wheat, RAPD-PCR, resistance, Fusarium Head Blight, in vitro selection

IN VITRO SELEKSİYON TEKNİĞİ İLE BUĞDAY (Triticum aestivum L.)’DA FUSARIUM (Fusarium spp)’A DAYANIKLI HÜCRE HATLARININ ELDE EDİLMESİ VE BİTKİ REGENERASYONU

Abstract

Çalışmada somaklonal varyasyondan yararlanarak in vitro seleksiyonla Buğday Başak Yanıklığı Hastalığına dayanıklı hatlar elde etme olanakları araştırılmıştır. Bu amaçla F.graminearum ve F. culmorum izolatlarının kültür filtratı ve Fusarik asit için belirlenen lethal dozları (%30;0.30mM) içeren MS ortamı üzerinde kültüre alınan Adana 99, Genç 99, Seri 82 buğday çeşitlerine ait embriyogenik kalluslardan seleksiyon sonucu 19 bitki regenere edilmiştir. Bu bitkilerden izole edilen DNA örnekleri 6 farklı primer ile RAPD-PCR analizine tabi tutulmuş, kullanılan OPA05, OPA-04 ve OPD-05 primerleri, amplifikasyon sonucunda oluşan PCR ürünlerinin elektroforezinde bantlar oluştururken, polimorfizimin gerçekleşmediği, diğer 3 primerin ise amplifikasyona hiç reaksiyon göstermediği belirlenmiştir. Bu bitkilerde olası genetik farklılığın ortaya çıkartılması için farklı primerlerin denenmesinin gerekliliği ortaya konulmuştur

Keywords

Buğday, RAPD-PCR, Dayanıklılık, Fusarium Başak Yanıklığı, İn Vitro Seleksiyon

References

• Aktaş, H.E., Bostancıoğlu, H., Tunalı, B., and Bayram, E., 1996. Determination of the Root Rots Disease Agents, Their Interference with Cultural Practices and Evaluation of Varieties and Lines Against to Important Ones in Sakarya Region. Plant Protect. Bull., 36: 151-167.
• Anderson, J.A., R.W. Stack, S. Liu, B.L. Waldron, A.D. Fjeld, C. Coyne, B. Moreno-Sevilla, J. Mitchell Fetch, Q.J. Song, P.B. Cregan, and R.C. Froberg. 2001. DNA markers for Fusarium head blight resistance QTLs in two wheat populations. Theor. Appl. Genet. 102:1164-1168.
• Anonim, 2001. Tarım Bakanlığı, Tarımsal Araştırmalar Genel Müdürlüğü.
• Anonim, 2002. Adana Ziraii Mücadele Araştırma Enstitüsü.
• Bai, G.H., Plattner, R., Desjardıns, A. and Kolp, F., 2001. Resistance To Fusarium Head Blight And Deoxynivalenol Accumulation in Wheat. Plant Breeding, 120: 1-6.
• Badea, M.E., Sandulescu, D., Gozıa, O., Radutoıu, S.E., Raıcu, P., and Gorenflot, R., 1997. Effets Du Filtrat De Culture De Fusarium graminearum Et De La Température Sur I’embryogenése Somatique Chez Triticum aestivum L. Revue De Cytologie Et Biologie Végétales. Le Botaniste, 20 (2): 3-10.
• Bai, G.H., and Shaner, G., 1994. Scab of Wheat: Prospect for Control. Plant Dis., 78: 760-766.
• Bruins, M.B.M., Karsai, I., Schepers, J., and Snıjders, C. H. A., 1993. Phytotoxicity Of Deoxynivalenol To Wheat Tissue With Regard To In Vitro Selection for Fusarium Head Blight Resistance. Plant Sci., 94: 195-206.Buerstmayr, H., Lemmens, M., Hart, L., Doldi, P., Steiner, B., Stier-Steiner, M., and Ruckenbauer, P., 2002. Molecular Mapping of Qtls for Fusarium Head Blight Resistance in Spring Wheat I: Resistance To Fungal Spread (Type Iı Resistance).Theor. Appl. Genet., 104: 84-91.
• Cao, W., Scoles, G.J., Hucl, P., and Chibbar, R.N., 2000. Phylogenetic Relationships Of Five Morphological Groups of Hexaploid Wheat (Triticum Aestivum L. Em Thell) Based on Rapd Analysis. Genome, 43: 724-727.
• Chawla, H.S., and Wenzel, G., 1987. In Vitro Selection for Fusaric Acid Resistance Barley Plants. Plant Breeding, 99: 159-163.
• Curir, P., Guglieri, L., Dolci, M., Capponi, A., and Aurino, G., 2000. Fusaric Acid Production by Fusarium oxysporum f.sp.Lilii and Its Role in the Lily Basal Rot Disease. European Journal Of Plant Pathology, 106: 849-856.
• Dhar A.K., Pokras, M.H., Garcia, D.K., Evers, D.C., and Gordon, Z.J., 1997. Analysis of Genetic Diversity in Common Loon Gavia İmmer Using Rapd And Mitochondrial Rflp Techniques. Molecular Ecology, 6: 581-586.
• Gargouria, S., Bernier, L., Hajlaoui, M.R., and Marrackchi, M., 2003. Genetic Variability and Population Structure of The Wheat Foot Rot Fungus, F. culmorum, In Tunisia. European J. Plant Pathology, 109: 807-815.
• Gervais, L., Dedryver, F., Morlais, J.Y., Bodusseau, V., Negre, S., Bilous, M., Groos, C. and Trottet, M., 2003. Mapping of quantitative trait loci for field resistance to Fusarium head blight in an European winter wheat. Theoret. Appl. Genet. 106, pp. 961–970. View Record in Scopus Cited by in Scopus.
• Goryunova, S. V., Kochıeva, E. Z., Chıkıda N. N., And Pukhalskyı, V. A., 2004. Hylogenetic Relationships And Intraspecific Variationof D- Genome Aegilops L. As Revealed By Rapd Analysis. Russian Journal Of Genetics, 40(5): 515–523.
• Jin, H., Hartman, G. L., Nıckell, D., and Wıdholm, J. M., 1996. Phytotoxicity of Culture Filtrate from Fusarium solani, The Causal Agent Of Sudden Death Syndrome Of Soybean. Plant Disease, 80 (8), 922-927.
• Harris, L.J., 1999. Possible Role of Trichothecene Mycotoxins in Virulence of Fusarium graminearum On Maize. Plant Disease, 83(10): 954-960.
• Hekimhan, H., Bagcı, A., Nicol, J., Arısoy, T., and Sahın, S., 2004. Dryland Root Rot: A Major Threat To Winter Cereal Production Under Sub- Optimal Growing Conditions. 4th Intl. Crop. Sci. Congress, 26-1 Oct. Brisbane, Australia. Www.Regional.Org.Au/Au.Cs
• Kasem Z. A., Mesterhazy, A., and Sagı, F., 1991. In Vitro Techniques for Selecting Wheat (Triticum aestivum L.) For Fusarium-Resistance. I. Double-Layer Culture Technique. Euphytica, 57: 251-257.
• Kasem, A.Z. and Sagı, F., 1993. Culture of and Fertile Plant Regeneration from Regenerable Embryogenic Suspension Cell-Derived Protoplast of Wheat (Triticum aestivum L.). Plant Cell Report, 12: 175-179.
• Kasem Z. A., Mesterhazy, A., Bartok, T., and Sagı, F., 1996. In Vitro Techniques For Selecting Wheat (Triticum Aestivum L.) For Fusarium- Resistance. Iı. Culture Filtrate Technique and Inheritance Of Fusarium-Resistance in The Somaclones. Euphytica, 91: 341-349.
• Khan, I.A., Awan, F.S, Ahmad, A., Fu, Y.B., and Iqbal, A., 2005. Genetic Diversity of Pakistan Wheat Germplasm As Revealed By Rapd Markers. Genetic Resources and Crop Evolution, 52: 239-244
• Kudryavtsev A.M., Martynov, S.P., Broggıo, M., And Pukhalskıy, V. A., 2003. Relevance Of Rapd Analysis For Revealing Phylogenetic Relationships Between Cultivars Of Durum Wheat (Triticum Durum Desf.). Russian Journal Of Genetics, 39(9): 1043–1051.
• Latunde-Dada, A.O., and Lucas, J.A., 1988. Somaclonal Variation and Resistance To Verticillium Wilt In Lucerne Medicago sativa L., Plants Regenerated From Callus. Plant Science, 58: 111-119.
• Lin F, Xue SL, Zhang ZZ, Zhang CQ, Kong ZX, Yao GQ, Tian DG, Zhu HL, Li CJ, Cao Y, Wei JB, Luo QY, Ma ZQ, 2006. Mapping QTL associated with resistance to fusarium head blight in the Nanda2419 × Wangshuibai population. II. Type I resistance. Theoretical and Applied Genetics 112, 528–35.
• Maier, F.J., and Oettler, G., 1992. Selection for the Fusarium Toxin Deoxynivalenol İn Callus Cultures Of Triticale. Ihar Radzikow, 37 (1-4): 43-49.
• Maier, F.J and Oetller, G., 1996. Genetic Variation for Head Blight Resistance in Triticale Caused by Fusarium graminearum Isolates of Different Deoxynivalenol Production. Euphytica, 89: 387- 394.
• Mcmullen, M.P., Jones, R., and Gallenberg, D. 1997. Scab of wheat and barley: a re-emerging disease of devastating impact. Plant Dis. 81: 1340-1348.
• Mesterhazy, A., Bartok, T., and Lamper, C., 2005. Influence of Wheat Cultivar, Species of Fusarium and Isolate Aggressiveness on The Efficacy Of Fungicides for Control Of Fusarium Head Blight. Plant Disease, 87(9):1107-1115.
• Miedaner, T., Reinbrecht, C., Lauber, U., Schollenberger, M., and Geiger, H.H., 2001. Effects of Genotype Enviroment Interaction On Deoxynivalenol Accumulation and Resistance to Fusarium Head Blight in Rye, Triticale, and Wheat. Plant Breeding, 120: 97-105.
• Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473-479.
• Parry, D.W., and Nıcholson, P., 1996. Development of a PCR Assay to Detect Fusarium poae in Wheat. Plant Pathology, 45: 383-391.
• Remotti, P.C., Löffler H.J.M., and Vloten-Dotıng L.V, 1997. Selection of Cell-Lines and Regeneration of Plants Resistant To Fusaric Acid From Gladiolus X Grandiflorus Cv. ‘Peter Pears’. Euphytica, 96: 237-245.
• Sharp, P.J.,Kreis, M., Shewry, P.R., and Gale, M.D., 1988. Location Of B-Amylase Sequences in Wheat and its Relatives. Theo.Apll.Genet., 75: 286-290.
• Sun, G., Bond, M., Nass, H., Martin, R., and Dong, Z., 2003. Rapd Polymorphisms in Spring Wheat Cultivars and Lines With Different Level of Fusarium Resistance. Theor. App.Gen. 106: 1059-1067.
• Turner, A.S., O’hara, R.B., Rezenoor, H.N., Nuttall, M., Sımıth, J.N., and Nicholson, P., 1999. Visial Disease And Pcr Assessment Of Stem Base Diseases in Winter Wheat. Plant Patholgy, 48: 742-748.
• Walker ,S.L., 2001. Variation Among Isolates of Fusarium graminearum Associated With Fusarium Head Blight in Nort Carolina. Plant Disease 85 (4): 404-410.
• Wilcoxson, R.D., Busch, R.H., and Ozmon, E.A., 1992. Fusarium Head Blight Resistance in Spring Wheat Cultivars. Plant Dis., 76: 658-661.
• Wolf S.J, and Earle E.D., 1990. Inhibition of Corn Callus Growth By Helminthosporium Carbonum Race 1 Toxin. Crop Sci., 30: 728–734.
• Yang, Z., Yang, X., and Huang, D., 1998. Studies on Somaclonal Variants For Resistance To Scab In Bread Wheat (Triticum aestivum L.) Through In Vitro Selection For Tolerance To Deoxynivalenol. Euphytica, 101(2): 213-219.
• Zhou WC, Kolb FLYuJ, Bai G, Larry L, Domier LL, 2004. Molecular characterization of fusarium head blight resistance in Wangshuibai with simple sequence repeat and amplified fragment length polymorphism markers. Genome 47, 1137–43.