Expression of a Rice Chitinase Gene Enhances Antifungal Response in Transgenic Litchi (cv. Bedana) ()
ABSTRACT
To
enhance the antifungal response of litchi, transferring rice chitinase gene
under a maize-ubiquitin promoter along with its first intron into the zygotic
embryos via Agrobacterium tumefaciens-mediated
transformation generated transgenic plants. After co-cultivation for 2 days
with recombinant Agrobacterium,
zygotic embryos were transferred onto Murashige and Skoog (MS) medium consisted
of MS salts and Gamborg (B5) vitamins with 2 mgl-1 2, 4-dichlorophenoxyacetic
acid (2, 4-D), 50 gl-1 sucrose and 8 gl-1 agar
supplemented with 25 mgl-1 hygromycin and 400 mgl-1 cefotaxime. Embryos were selected passing through a series of MS modified media
and the antibiotic resistant transgenic plantlets were analyzed. The
integration and stability of the transgene was confirmed by PCR, RT-PCR,
Southern blotting and by Western blot analyses. The transgenic plants exhibited
higher chitinase activity than the non-transformed plants. The chitinase
activity was also examined using the native polyacrylamide in-gel assay. These
analyses indicated that the foreign gene was translated into the protein of
expected molecular weight that showed chitinase activity. Following in-vitro inoculation of die-back, leaf spots and blight pathogen (Phomopsis sp.), the
transgenic plants showed delayed onset of the disease and smaller lesions. The
transgenic plants were adapted to the greenhouse and did not show any
phenotypic alterations.
Share and Cite:
Das, D. and Rahman, A. (2018) Expression of a Rice Chitinase Gene Enhances Antifungal Response in Transgenic Litchi (cv. Bedana).
American Journal of Plant Sciences,
9, 2256-2275. doi:
10.4236/ajps.2018.911163.