Through the use of electroporation and a soybean (Glycine max L.) protoplast system, we generated stably transformed cell lines expressing a number of foreign genes (neomycin phosphotransferase,β-glucuronidase, chloramphenicol acetyl transferase, and phosphinothricin acetyl transferase). Selected and unselected marker genes were cointroduced either linked on a single plasmid or as separate plasmids. Calli expressing multiple genes were recovered, and Cotransformation frequencies were established for both cases. Our results show a 50% cotransformation frequency in the case of linked genes. In situations in which two genes are introduced on independent plasmids, cotransformation frequencies are 18%-27%. Similar rates of cotransformation were observed among various marker pairs.
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Front Plant Sci
March 2024
Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
Doubled haploid (DH) technology provides an effective way to generate homozygous genetic and breeding materials over a short period of time. We produced three types of homozygous gene-edited mutants (, , and ) by CRISPR/Cas9 in durum wheat. PCR restriction enzymes and sequencing confirmed that the editing efficiency was up to 53.
View Article and Find Full Text PDFBiology (Basel)
January 2024
State Key Laboratory of Biocontrol and Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
CRISPR/Cas9 systems are commonly used for plant genome editing; however, the generation of homozygous mutant lines in remains challenging. Here, we present a CRISPR/Cas9-based protocol that allows the efficient generation of mutants. Gene editing was performed for the LysM receptor kinase gene and two major facilitator superfamily transporter genes.
View Article and Find Full Text PDFAppl Microbiol Biotechnol
January 2024
College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
Monascus pilosus has been used to produce lipid-lowering drugs rich in monacolin K (MK) for a long period. Genome mining reveals there are still many potential genes worth to be explored in this fungus. Thereby, efficient genetic manipulation tools will greatly accelerate this progress.
View Article and Find Full Text PDFInt J Biol Macromol
January 2024
Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, China; Ministry of Agriculture and Rural Affairs-Key Laboratory for Crop Pest Monitoring and Green Control, College of Plant Protection, China Agricultural University, Beijing 100193, China.
Colletotrichum higginsianum causes anthracnose disease in brassicas. The availability of the C. higginsianum genome has paved the way for the genome-wide exploration of genes associated with virulence/pathogenicity.
View Article and Find Full Text PDFPlant Cell Rep
March 2023
The Genome Center, University of California, Davis, Davis, CA, USA.
GRF-GIF chimeric proteins from multiple source species enhance in vitro regeneration in both wild and cultivated lettuce. In addition, they enhance regeneration in multiple types of lettuce including butterheads, romaines, and crispheads. The ability of plants to regenerate in vitro has been exploited for use in tissue culture systems for plant propagation, plant transformation, and genome editing.
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