Disruption in the DNA Mismatch Repair Gene MSH2by CRISPR-Cas9in Indica Rice Can Create Genetic Variability

Autor: V, Karthika, Chandrashekar, Babitha K., K, Kiranmai, AG, Shankar, Makarla, Udayakumar, Ramu, Vemanna S.
Zdroj: Journal of Agricultural and Food Chemistry; April 2021, Vol. 69 Issue: 14 p4144-4152, 9p
Abstrakt: Genetic variation is crucial for crop improvement. We adopted a gene editing approach to create variations in the rice genome by targeting the mutator locus homolog 2 (MSH2), a DNA mismatch repair gene. The hypothesis is that disruption of the MSH2gene leads to a reduced DNA mismatch repair that creates INDELs, resulting in altered phenotypes. The Indica rice (IR-64) genotype was transformed with a guide RNA targeted to the MSH2gene using an Agrobacterium-mediated in plantamethod. Many plants showed integration of Cas9and gRNAconstructs in rice plants. One of the msh2mutants showed a superior phenotype due to editing and possible INDELs in the whole genome. The stable integration of the transgene and its flanking sequence analysis confirms no disruption of any gene, and the observed phenotype is due to the mutations in the MSH2gene. Few transgenic plants showed disruption of genes due to T-DNA integration that led to altered phenotypes. The plants with altered phenotypes having more tiller number, early flowering, and robust growth with a high biomass were identified. These genetically reprogrammed rice plants could be a potential resource to create more segregating population or act as donor lines to stabilize the important agronomic traits that may help in a speed breeding process.
Databáze: Supplemental Index