Plant Cell Reports

, Volume 36, Issue 3, pp 399–406

Efficient CRISPR/Cas9-based gene knockout in watermelon

  • Shouwei Tian
  • Linjian Jiang
  • Qiang Gao
  • Jie Zhang
  • Mei Zong
  • Haiying Zhang
  • Yi Ren
  • Shaogui Guo
  • Guoyi Gong
  • Fan Liu
  • Yong Xu
Original Article

Abstract

Key message

CRISPR/Cas9 system can precisely edit genomic sequence and effectively create knockout mutations in T0 generation watermelon plants.

Abstract

Genome editing offers great advantage to reveal gene function and generate agronomically important mutations to crops. Recently, RNA-guided genome editing system using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) has been applied to several plant species, achieving successful targeted mutagenesis. Here, we report the genome of watermelon, an important fruit crop, can also be precisely edited by CRISPR/Cas9 system. ClPDS, phytoene desaturase in watermelon, was selected as the target gene because its mutant bears evident albino phenotype. CRISPR/Cas9 system performed genome editing, such as insertions or deletions at the expected position, in transfected watermelon protoplast cells. More importantly, all transgenic watermelon plants harbored ClPDS mutations and showed clear or mosaic albino phenotype, indicating that CRISPR/Cas9 system has technically 100% of genome editing efficiency in transgenic watermelon lines. Furthermore, there were very likely no off-target mutations, indicated by examining regions that were highly homologous to sgRNA sequences. Our results show that CRISPR/Cas9 system is a powerful tool to effectively create knockout mutations in watermelon.

Keywords

CRISPR/Cas9 Genome editing PDS Watermelon 

Supplementary material

299_2016_2089_MOESM1_ESM.pdf (11 kb)
Supplementary material 1 (PDF 11 kb)
299_2016_2089_MOESM2_ESM.pdf (5.9 mb)
Supplementary material 2 (PDF 6031 kb)
299_2016_2089_MOESM3_ESM.pdf (10.8 mb)
Supplementary material 3 (PDF 11016 kb)
299_2016_2089_MOESM4_ESM.pdf (102 kb)
Supplementary material 4 (PDF 102 kb)
299_2016_2089_MOESM5_ESM.pdf (114 kb)
Supplementary material 5 (PDF 114 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Shouwei Tian
    • 1
  • Linjian Jiang
    • 2
  • Qiang Gao
    • 3
  • Jie Zhang
    • 1
  • Mei Zong
    • 1
  • Haiying Zhang
    • 1
  • Yi Ren
    • 1
  • Shaogui Guo
    • 1
  • Guoyi Gong
    • 1
  • Fan Liu
    • 1
  • Yong Xu
    • 1
  1. 1.National Engineering Research Center for VegetablesBeijing Academy of Agriculture and Forestry Sciences, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China), Beijing Key Laboratory of Vegetable Germplasm ImprovementBeijingChina
  2. 2.Department of Plant PathologyChina Agricultural UniversityBeijingChina
  3. 3.Beijing University of AgricultureBeijingChina

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