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CRISPR/Cas9-mediated mutagenesis of Clpsk1 in watermelon to confer resistance to Fusarium oxysporum f.sp. niveum
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CRISPR/Cas9-mediated mutagenesis of Clpsk1 in watermelon to confer resistance to Fusarium oxysporum f.sp. niveum
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CRISPR/Cas9-mediated mutagenesis of Clpsk1 in watermelon to confer resistance to Fusarium oxysporum f.sp. niveum
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Journal Article
CRISPR/Cas9-mediated mutagenesis of Clpsk1 in watermelon to confer resistance to Fusarium oxysporum f.sp. niveum
2020
Overview
Key message
CRISPR/Cas9-mediated editing of
Clpsk1
enhanced watermelon resistance to
Fusarium oxysporum
.
The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has proven to be an effective genome-editing tool for crop improvement. Previous studies described that Phytosulfokine (PSK) signalling attenuates plant immune response. In this work, we employed the CRISPR/Cas9 system to knockout
Clpsk1
gene, encoding the PSK precursor, to confer enhanced watermelon resistance to
Fusarium oxysporum
f.sp.
niveum
(
FON
). Interactions between PSK and
FON
were analysed and it was found that transcript of
Clpsk1
was significantly induced upon
FON
infection. Meanwhile, application of exogenous PSK increased the pathogen growth. Then, one sgRNA, which targeted the first exon of
Clpsk1
, was selected for construction of pRGEB32-CAS9-gRNA-Clpsk1 expression cassette. The construct was then transformed to watermelon through
Agrobacterium tumefaciens
-mediated transformation method. Six mutant plants were obtained and three types of mutations at the expected position were identified based on Sanger sequencing. Resistance evaluation indicated that
Clpsk1
loss-of-function rendered watermelon seedlings more resistant to infection by
FON
. These results indicate that CRISPR/Cas9-mediated gene modification is an effective approach for watermelon improvement.
Publisher
Springer Berlin Heidelberg,Springer Nature B.V
