Expression of a chitinase gene from Metarhizium anisopliae in tobacco plants confers resistance against Rhizoctonia solani.
The chit1 gene from the entomopathogenic fungus Metarhizium anisopliae, encoding the endochitinase CHIT42, was placed under the control of the CaMV 35S promoter, and the resulting construct was transferred to tobacco. Seventeen kanamycin-resistant transgenic lines were recovered, and the presence of the transgene was confirmed by polymerase chain reactions and Southern blot hybridization. The number of chit1 copies was determined to be varying from one to four. Copy number had observable effects neither on plant growth nor development. Substantial heterogeneity concerning production of the recombinant chitinase, and both general and specific chitinolytic activities were detected in leaf extracts from primary transformants. The highest chitinase activities were found in plants harboring two copies of chit1 inserts at different loci. Progeny derived from self-pollination of the primary transgenics revealed a stable inheritance pattern, with transgene segregation following a mendelian dihybrid ratio. Two selected plants expressing high levels of CHIT42 were consistently resistant to the soilborne pathogen Rhizoctonia solani, suggesting a direct relationship between enzyme activity and reduction of foliar area affected by fungal lesions. To date, this is the first report of resistance to fungal attack in plants mediated by a recombinant chitinase from an entomopathogenic and acaricide fungus.
Kern
Marcelo Fernando
MF
Graduating Program in Genetics and Molecular Biology of the Biosciences Institute, Biotechnology Center, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil. mfk500@york.ac.uk
Maraschin
Simone de Faria
Sde F
Vom Endt
Débora
D
Schrank
Augusto
A
Vainstein
Marilene Henning
MH
Pasquali
Giancarlo
G
D015139:Blotting, Southern; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism*; D004789:Enzyme Activation / Q000235:genetics; D018628:Gene Dosage; D020869:Gene Expression Profiling; D052981:Metarhizium / Q000201:enzymology* / Q000235:genetics; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology / Q000382:microbiology*; D018515:Plant Leaves / Q000235:genetics / Q000276:immunology / Q000382:microbiology*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012232:Rhizoctonia / Q000276:immunology / Q000472:pathogenicity / Q000502:physiology*; D014026:Nicotiana / Q000235:genetics / Q000276:immunology / Q000382:microbiology*; D019076:Transgenes
D011994:Recombinant Proteins; D002688:Chitinases