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Trasformazione biolistica del sorgo mediante l'impiego di un gene della chitinasi del riso [Sorghum bicolor (L.) Moench - Oryza sativa L.]  [sep1998]

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Zhu, H.
Jeoung, J.M.
Liang, G.H. (Kansas State Univ., Manhattan (USA). Dept. of Agronomy)
Muthukrishnan, S.
Krishnaveni, S. (Kansas State Univ., Manhattan (USA). Dept. of Biochemistry)
Wilde, G. (Kansas State Univ., Manhattan (USA). Dept. of Entomology)

Abstract:
A rice chitinase gene, G11, which may have a protective role against fungal pathogens, was introduced into a sorghum inbred "Tx430". Calli derived from immature zygotic embryos were bombardaded with tungsten particles coated with a plasmid DNA containing this gene and the bar gene as a selectable marker. After transformation, six fertile transgenic plants were obtained from a total of 1,100 bombarded calli. Molecular analyses by phosphinothricin acetyltransferase (PAT) acitivity assay, Southern blotting, and western blotting confirmed the presence and expression of the selectable bar gene and the rice chitinase gene in primary transgenic plants (T0). However, gene silencing occurred in few T0 plants at certain growth stages. Progeny analysis showed that the introduced chitinase gene was inherited and segregated among T1 plants (progeny of selfed T0 plants). Western blot analysis showed a 3:1 Mendelian segregation of the rice chitinase gene in T1 progeny, suggesting that the active chitinase gene(s) was inserted in a single locus in the T0 plants





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