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RNAi‐induced silencing of gene expression in strawberry fruit ( Fragaria × ananassa ) by agroinfiltration: a rapid assay for gene function analysis
Author(s) -
Hoffmann Thomas,
Kalinowski Gregor,
Schwab Wilfried
Publication year - 2006
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2006.02913.x
Subject(s) - chalcone synthase , fragaria , biology , agroinfiltration , gene , ripening , rna interference , gene silencing , gene expression , phenylpropanoid , chalcone isomerase , intron , rna , botany , biochemistry , biosynthesis
Summary Intron‐containing constructs encoding self‐complementary ‘hairpin’ RNA (ihpRNA) have the potential to efficiently silence genes in a range of plant species. In this study we demonstrate the silencing of a ripening‐related chalcone synthase ( CHS ) gene in strawberry fruits ( Fragaria × ananassa cv. Elsanta) by a construct (ihpRNA) containing the partial sense and corresponding antisense sequences of CHS separated by an intron obtained from a F. × ananassa quinone oxidoreductase gene. An Agrobacterium strain carrying a T‐DNA expressing the ihpRNA transgene was injected with a syringe into the receptacles of growing fruits still attached to the plant about 14 days after pollination. As a consequence of the reduced levels of CHS mRNA and enzymatic CHS activity, the levels of anthocyanins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to a large increases in levels of (hydroxy) cinnamoyl glucose esters. We anticipate that this technique in combination with metabolite profiling analysis will be useful for studying the function of unknown genes during the development and ripening of strawberry fruit.