Table 1 Editing efficiency of different targeted NCR genes in nodules harvested from hairy roots mediated by A. rhizogenes transformation.

From: Targeted mutagenesis of Medicago truncatula Nodule-specific Cysteine-Rich (NCR) genes using the Agrobacterium rhizogenes-mediated CRISPR/Cas9 system

 

NCR169

NCR068

NCR089

NCR128

NCR161

Number of sequenced nodules

3 white

3 pink

15

13

10

20

Number nodules carrying max. 50% WT allele

3

2

15

13

9

19

Number of fully mutant nodules (mismatch/indel)

3

0*

14

13

9

12

Number of fully mutant nodules (only indel)

3

0*

7

13

9

12

Nodule samples carrying only indel alleles

100%

0%

47%

100%

90%

60%

  1. Randomly selected nodules were harvested 3–5 wpi with S. medicae WSM419 and total DNA was isolated from fixed nodule sections. The PCR amplified target regions were sequenced on Illumina platform and the reads were analysed by using the CRISPResso2 tool. The gene NCR169 was targeted for gene editing as a control. Three pinkish elongated nodules were selected randomly from hairy roots and also three small white nodules were picked randomly from plants which did not form any pink nodules on their roots.
  2. *One of the pinkish nodules carried mutant alleles on large proportion (94%). This mutation was a three bp deletion that resulted in an in-frame deletion of the first Glu residue of the mature peptide NCR169. The lack of this single residue probably did not abolish the function of peptide NCR169.
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