CRISPR/Cas9-mediated mutagenesis of Clpsk1 in watermelon to confer resistance to Fusarium oxysporum f.sp. niveum

Plant Cell Reports
Man ZhangLina Lou

Abstract

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. ...Continue Reading

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Citations

Oct 18, 2020·Plants·Bed Prakash Bhatta, Subas Malla
Apr 13, 2021·Journal of Advanced Research·Dangquan ZhangBaohong Zhang
Apr 10, 2021·Journal of Zhejiang University. Science. B·Chao LiBaohong Zhang
May 13, 2021·Plant Cell Reports·Sukumar BiswasJianxin Shi
Jun 19, 2021·Transgenic Research·Cláudia RatoPaula Rodrigues Oblessuc
Jun 29, 2021·Frontiers in Plant Science·Young-Cheon KimSeonghoe Jang
Oct 12, 2021·Frontiers in Plant Science·Matthew Venezia, Kate M Creasey Krainer

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Methods Mentioned

BETA
gene knockout
PCR
transgenic
targeted mutations

Software Mentioned

CRISPRdirect
SAS

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