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    • Development of transgenic finger millet (Eleusine coracana (L.) Gaertn.) resistant to leaf blast disease

      S Ignacimuthu S Antony Ceasar

      Articles Volume 37 Issue 1 March 2012 pp 135-147

    • Fulltext

       

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      Permanent link:
      https://www.ias.ac.in/article/fulltext/jbsc/037/01/0135-0147

    • Keywords

       

      Agrobacterium-mediated transformation; fungal resistance; leaf blast; millet; rice chitinase

    • Abstract

       

      Finger millet plants conferring resistance to leaf blast disease have been developed by inserting a rice chitinase (chi11) gene through Agrobacterium-mediated transformation. Plasmid pHyg-Chi.11 harbouring the rice chitinase gene under the control of maize ubiquitin promoter was introduced into finger millet using Agrobacterium strain LBA4404 (pSB1). Transformed plants were selected and regenerated on hygromycin-supplemented medium. Transient expression of transgene was confirmed by GUS histochemical staining. The incorporation of rice chitinase gene in R0 and R1 progenies was confirmed by PCR and Southern blot analyses. Expression of chitinase gene in finger millet was confirmed by Western blot analysis with a barley chitinase antibody. A leaf blast assay was also performed by challenging the transgenic plants with spores of Pyricularia grisea. The frequency of transient expression was 16.3% to 19.3%. Stable frequency was 3.5% to 3.9%. Southern blot analysis confirmed the integration of 3.1 kb chitinase gene. Western blot analysis detected the presence of 35 kDa chitinase enzyme. Chitinase activity ranged from 19.4 to 24.8. In segregation analysis, the transgenic R1 lines produced three resistant and one sensitive for hygromycin, confirming the normal Mendelian pattern of transgene segregation. Transgenic plants showed high level of resistance to leaf blast disease compared to control plants. This is the first study reporting the introduction of rice chitinase gene into finger millet for leaf blast resistance.

    • Author Affiliations

       

      S Ignacimuthu1 S Antony Ceasar1

      1. Division of Plant Biotechnology, Entomology Research Institute, Loyola College, Chennai 600 034

    • Dates

       
      Published
      March 2012
      Manuscript received
      16 September 2011
      Accepted
      7 December 2011

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