Portal de Periódicos da CAPES

RNA silencing as a tool for exploring gene function in ascomycete fungi

2005; Academic Press; Volume: 42; Issue: 4 Linguagem: Inglês

10.1016/j.fgb.2005.01.002

1096-0937

Hitoshi Nakayashiki, Shugo Hanada, Nguyễn Bảo Quốc, Naoki Kadotani, Yukio Tosa, S. Mayama,

Fungal and yeast genetics research

We have developed a pHANNIBAL-like silencing vector, pSilent-1, for ascomycete fungi, which carries a hygromycin resistance cassette and a transcriptional unit for hairpin RNA expression with a spacer of a cutinase gene intron from the rice blast fungus Magnaporthe oryzae. In M. oryzae, a silencing vector with the cutinase intron spacer (147 bp) showed a higher efficiency in silencing of the eGFP gene than did those with a spacer of a GUS gene fragment or a longer intron (850 bp) of a chitin binding protein gene. Application of pSilent-1 to two M. oryzae endogenous genes, MPG1 and polyketide synthase-like gene, resulted in various degrees of silencing of the genes in 70–90% of the resulting transformants. RNA silencing was also induced by a pSilent-1-based vector in Colletotrichum lagenarium at a slightly lower efficiency than in M. oryzae, indicating that this silencing vector should provide a useful reverse genetic tool in ascomycete fungi.