TY - JOUR

T1 - CRISPR-Based Editing of the Medicago truncatula LEC1 Gene

AU - Potsenkovskaia, Elina A.

AU - Tvorogova, Varvara E.

AU - Simonova, Veronika Y.

AU - Konstantinov, Zakhar S.

AU - Kiseleva, Anna S.

AU - Matveenko, Andrew G.

AU - Brynchikova, Anna V.

AU - Lutova, Ludmila A.

PY - 2024/11/16

Y1 - 2024/11/16

N2 - Arabidopsis thaliana LEAFY COTYLEDON1 (LEC1) gene is shown to have numerous diverse functions in plant development, including the regulation of embryo morphogenesis and maturation, hypocotyl elongation, flowering transition, etc. However, the functions of LEC1 orthologs in different plant species have not been extensively studied. In this study, we obtained a line of Medicago truncatula, a model leguminous plant, carrying the loss-of-function mutation in the MtLEC1 (MtNF-YB10) gene, orthologous to LEC1, using the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated proteins (CRISPR/Cas9) genome editing system. Edited plants with loss of MtNF-YB10 function did not demonstrate any severe abnormalities during their normal growth and gave viable seeds, but their capability for somatic embryogenesis in vitro was dramatically reduced. The T1 progeny of unedited plants with a Cas9-gRNA cassette insertion was also analyzed based on the suggestion that editing could occur during seed formation. However, no edited plants were found in the T1 generation. These results suggest divergent functions of LEC1 orthologs and make it possible to investigate potential specific MtNF-YB10 functions.

AB - Arabidopsis thaliana LEAFY COTYLEDON1 (LEC1) gene is shown to have numerous diverse functions in plant development, including the regulation of embryo morphogenesis and maturation, hypocotyl elongation, flowering transition, etc. However, the functions of LEC1 orthologs in different plant species have not been extensively studied. In this study, we obtained a line of Medicago truncatula, a model leguminous plant, carrying the loss-of-function mutation in the MtLEC1 (MtNF-YB10) gene, orthologous to LEC1, using the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated proteins (CRISPR/Cas9) genome editing system. Edited plants with loss of MtNF-YB10 function did not demonstrate any severe abnormalities during their normal growth and gave viable seeds, but their capability for somatic embryogenesis in vitro was dramatically reduced. The T1 progeny of unedited plants with a Cas9-gRNA cassette insertion was also analyzed based on the suggestion that editing could occur during seed formation. However, no edited plants were found in the T1 generation. These results suggest divergent functions of LEC1 orthologs and make it possible to investigate potential specific MtNF-YB10 functions.

KW - CRISPR

KW - LEC1

KW - Medicago truncatula

KW - MtNF-YB10

KW - agrobacterial transformation

KW - regeneration

KW - somatic embryogenesis

UR - https://www.mendeley.com/catalogue/af8ebd38-3c80-323f-91bb-9ed0abcd1bba/

U2 - 10.3390/plants13223226

DO - 10.3390/plants13223226

M3 - Article

C2 - 39599434

VL - 13

JO - Plants

JF - Plants

SN - 2223-7747

IS - 22

M1 - 3226

ER -