Figure 1 | Scientific Reports

Figure 1

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

Figure 1The alternative text for this image may have been generated using AI.

Plant gross and nodulation phenotype of Agrobacterium rhizogenes-mediated transgenic Medicago truncatula Jemalong 2HA hairy-roots mutagenized with CRISPR/Cas9 gene editing. No sign of nitrogen deficiency was observed on plants transformed with empty vector (a) or the construct targeting NCR068 (c) 4 weeks post inoculation with S. medicae WSM419. Transgenic roots were identified by fluorescence of DsRed protein. Elongated pink nodules, showing the characteristic features of indeterminate nodules, developed on transgenic roots. These nodules were fully colonized by rhizobia visualized by the fluorescent dye SYTO13 (d,f). (b) Targeted mutagenesis of NCR169 resulted in transgenic plants showing the symptoms of nitrogen starvation. Small, spherical and white ineffective nodules were formed on NCR169-targeted roots. (e) ncr169 mutant nodules did not contain infected cells in the region corresponding to the nitrogen fixation zone of wild-type plants. Inset images in (d–f) show the higher magnification of cells in nodule regions indicated by white rectangles. (g,h) The targeted regions were amplified and sequenced from the transgenic nodules. The sequence analysis revealed biallelic mutations causing frameshifts in edited NCR169 (g) and NCR068 (h) genes. ZII, infection zone; IZ, interzone; ZIII, nitrogen fixation zone; EV, empty vector.

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