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  • 1 Center for Genetic Engineering and Biotechnology of Sancti Spiritus, P.O. Box 83, P.C. 60200, Sancti Spiritus, Cuba
  • 2 National Institute of Agricultural Sciences, Carretera La Francia km 1 ½, Pinar del Rio, Cuba
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In this work it was developed marker-free transgenic indica rice plants (cv J-104) by biolistic co-transformation and segregation approach. We attempted to express the NmDef02 antifungal defensin. Primary transformants were regenerated from embryogenic callus on culture medium with 50 mg/L hygromycin. Screening of hpt-marker-free transgenic lines was made by PCR in T1 progeny lines, germinated on semisolid medium without hygromycin. Relative expression of NmDef02 mRNA was examined by quantitative RT-PCR in marker-free T1 plants. In vitro antifungal test was performed by disk diffusion assay against Sarocladium oryzae. PCR assay verified that 15.12% of T1 plants were marker-free (NmDef02+/hpt−). RT-PCR analysis indicated that NmDef02 gene was successfully transcribed and the transgenic lines displayed different expression levels of the NmDef02 cDNA. Protein extracts of marker-free lines with high relative expression of NmDef02 inhibited fungus mycelial growth around disks. In contrast, it was confirmed fungus proliferation on disks impregnated with protein extracts of non-transgenic plants. The results of the present work demonstrated that the expression of the NmDef02 defensin in transgenic rice plants is effective against the phytopathogenic fungus Sarocladium oryzae under in vitro conditions. Thus, NmDef02 defensin could be a useful tool for J-104 rice improvement.

  • Bradford, M. M. (1976): A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.

    • Search Google Scholar
    • Export Citation
  • Campbell, B. T., Baenziger, P. S., Mitra, A., Soto, S. and Clemente, Campbell (2000): Inheritance of multiple transgenic in wheat. Crop Science 40, 11331141.

    • Search Google Scholar
    • Export Citation
  • Chong-Pérez, B. and Angenon, G. (2013): Strategies for generating marker-free transgenic plants. In: I. Sithole-Niang (ed.): Genetic Engineering. InTech., pp. 1748.

    • Search Google Scholar
    • Export Citation
  • Chu, C. C., Wang, C. C., Sun, C. S., Chen, H., Yin, K. C. and Chu, C. Y. (1975): Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Scientia Sinica 18, 659668.

    • Search Google Scholar
    • Export Citation
  • Coca, M., Peñas, M., Gómez, J., Campo, S., Bortolotti, C., Messeguer, J. and San Segundo, B. (2006): Enhanced resistance to the rice blast fungus Magnaporthe grisea conferred by expression of a cecropin A gene in transgenic rice. Planta 223, 392406.

    • Search Google Scholar
    • Export Citation
  • De Bondt, A., Zaman, S., Broekaert, W., Cammue, B. and Keulemans, De Bondt (1999): Genetic transformation of apple (Malus pumila Mill.) for increased fungal resistance: in vitro antifungal activity in protein extracts of transgenic apple expressing Rs-AFP2 or Ace-AMP1. Acta Horticulturae 484, 565570.

    • Search Google Scholar
    • Export Citation
  • Dellaporta, S. L., Wood, J. and Hicks, J. B. (1983): A plant DNA minipreparation: Version II. Plant Molecular Biology Reporter 1, 1921.

    • Search Google Scholar
    • Export Citation
  • Devi, S., Mishra, M. K. and Elliott, M. (2012): Regeneration of transgenic rice with bacterial ipt gene driven by senescence specific (SAG12) promoter by particle bombardment. Tropical Life Sciences Research 23, 3948.

    • Search Google Scholar
    • Export Citation
  • Jackson, M. A., Anderson, D. J. and Birch, R. G. (2013): Comparison of Agrobacterium and particle bombardment using whole plasmid or minimal cassette for production of high-expressing, low-copy transgenic plants. Transgenic Research 22, 143151.

    • Search Google Scholar
    • Export Citation
  • Jha, S. and Chattoo, B. B. (2010): Expression of a plant defensin in rice confers resistance to fungal phytopathogens. Transgenic Research 19, 373384.

    • Search Google Scholar
    • Export Citation
  • Jha, S., Tank, H. G., Prasad, B. D. and Chattoo, B. B. (2009): Expression of Dm-AMP1 in rice confers resistance to Magnaporthe oryzae and Rhizoctonia solani. Transgenic Research 18, 5969.

    • Search Google Scholar
    • Export Citation
  • Jiang, Y., Sun, L., Jiang, M., Li, K., Song, Y. and Zhu, C. (2013): Production of marker-free and RSV-resistant transgenic rie using a twin T-DNA system and RNAi. J. Biosciences 38, 573581.

    • Search Google Scholar
    • Export Citation
  • Kumar, S., Arul, L. and Talwar, D. (2010): Generation of marker-free Bt transgenic indica rice and evaluation of its yellow stem borer resistance. J. Appl. Genetics 51, 243257.

    • Search Google Scholar
    • Export Citation
  • Murashige, T. and Skoog, F. (1962): A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15, 473497.

    • Search Google Scholar
    • Export Citation
  • Parashina, E. V., Serdobinski, L., Kalle, E., Lavrova, N., Avetisov, V. and Lunin, V. (2000): Genetic engineering of oilseed rape and tomato plants expressing a radish defensin gene. Russian J. Plant Physiol. 47, 417423.

    • Search Google Scholar
    • Export Citation
  • Portieles, R., Ayra, C., Gonzalez, E., Gallo, A., Rodriguez, R., Chacon, O., Lopez, Y., Rodriguez, M., Castillo, J., Pujol, M., Enriquez, G., Borroto, C., Trujillo, L., Thomma, B. and Borras, O. (2010): NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions. Plant Biotechnology 8, 678690.

    • Search Google Scholar
    • Export Citation
  • Taylor, M. G. and Vasil, I. K. (1991): Histology of and physical factors affecting transient GUS expression in pearl millet (Pennisetum glaucum L.) embryos following microprojectile bombardment. Plant Cell Report 10, 120125.

    • Search Google Scholar
    • Export Citation
  • Terras, F. R. G., Eggermont, K., Kovaleva, V., Raikel, N., Osborn, R. and Kester, A. (1995): Small cysteine-rich antifungal proteins from radish: their role in host defense. Plant Cell. 7, 573588.

    • Search Google Scholar
    • Export Citation
  • Thomma, B., Cammue, B. and Thevissen, K. (2002): Plant defensins. Planta 216, 193202.

  • Van der Vyver, C., Conradie, T., Kossmann, J. and Lloyd, J. (2013): In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase. In Vitro Cellular and Developmental Biology 49, 198206.

    • Search Google Scholar
    • Export Citation
  • Wang, D., Zhao, Q., Zhu, D., Ao, G. and Yu, Wang (2006): Particle-bombardment-mediated co-transformation of maize with a lysine rich protein gene (sb401) from potato. Euphytica 150, 7585.

    • Search Google Scholar
    • Export Citation
  • Yao, Q., Cong, l., Chang, J. L., Li, K. X., Yang, G. X. and He, G. Y. (2006): Low copy number gene transfer and stable expression in a commercial wheat cultivar via particle bombardment. J. Experimental Botany 57, 37373746.

    • Search Google Scholar
    • Export Citation
  • Yau, Y. Y. and Stewart, C. N. (2013): Less is more: strategies to remove marker genes from transgenic plants. BMC Biotechnology 13, 3659.

    • Search Google Scholar
    • Export Citation

 

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2019  
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0,177
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Insect Science Q4
Plant Science Q4
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66/103=0,6
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Insect Science 125/142 (Q4)
Plant Science 344/431 (Q4)
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Language: English

Founded in 1966
Publication: One volume of two issues annually
Publication Programme: 2020. Vol. 55.

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Editor(s)-in-Chief: Barna, Balázs

Editorial Board

      Benedek, P. (Mosonmagyaróvár)
      Hernández J.A. (Murcia, Spain)
      Érsek, T. (Mosonmagyaróvár)
      Francke, W. (Hamburg)
      Hornok, L. (Gödöllő)
      Horváth, J. (Keszthely)
      Király, Z. (Budapest)
      Kiss, L. (Budapest)
      Kogel, K.-H. (Giessen)
      Kőmíves, T. (Budapest)
      Palkovics, L. (Budapest)
      Schoelz, J. E. (Columbia, Missouri)
      Schulz, S. (Braunschweig)
      Seybold, S. (Davis)
      Skoczowski A. (Cracow)
      Szőcs, G. (Budapest)
      Tóth, M. (Budapest)
      Virányi, F. (Gödöllő)
      Diaz-Vivancos P. (Murcia, Spain)

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