Efficient multi-allelic genome editing via CRISPR-Cas9 ribonucleoprotein-based delivery to mesophyll protoplasts.

Canola ( L.) is a valuable oilseed crop worldwide. However, trait improvement by breeding has been limited by its low genetic diversity and polyploid genetics.

An Efficient Fluorescence-Activated Protoplast Sorting (FAPS) and Regeneration Protocol for Canola (Brassica napus).

Protoplast sorting and purification methods are powerful tools enabling the enrichment of cellular subpopulations for basic and applied studies in plant sciences. Fluorescence-activated protoplast sorting (FAPS) is an efficient method to isolate specific protoplast populations based on innate features (size and autofluorescence) or expression of fluorescent proteins. FAPS-based methods have recently been deployed in single-cell purification for single-cell RNA sequencing-based transcriptional profiling studies.

Overcoming roadblocks for nurseries in plants: induction of meiosis.

Efforts to increase genetic gains in breeding programs of flowering plants depend on making genetic crosses. Time to flowering, which can take months to decades depending on the species, can be a limiting factor in such breeding programs. It has been proposed that the rate of genetic gain can be increased by reducing the time between generations by circumventing flowering through the induction of meiosis.

Studies on Pure Mlb (Multiple Left Border) Technology and Its Impact on Vector Backbone Integration in Transgenic Cassava.

Imperfect T-DNA processing is common during -mediated transformation, which integrates vector backbone sequences into the plant genome. However, regulatory restrictions prevent such transgenic plants from being developed for commercial deployment. The binary vector pCAMBIA2300 was modified by incorporating multiple left border (Mlb) repeats and was tested in BY2 cells, tobacco, and cassava plants to address this issue.

Generation of Strains Expressing Fluorescent Proteins Suitable for Studying Host-Endophyte Interactions and Characterisation of a T-DNA Integration Event.

Methods for the identification and localisation of endophytic fungi are required to study the establishment, development, and progression of host-symbiont interactions, as visible reactions or disease symptoms are generally absent from host plants. Fluorescent proteins have proved valuable as reporter gene products, allowing non-invasive detection in living cells. This study reports the introduction of genes for two fluorescent proteins, green fluorescent protein (GFP) and red fluorescent protein, DsRed, into the genomes of two distinct perennial ryegrass ( L.

Mesophyll Protoplasts and PEG-Mediated Transfections: Transient Assays and Generation of Stable Transgenic Canola Plants.

Plant protoplasts are derived by controlled enzymatic digestion that removes the plant cell wall without damaging the cell membrane. Protoplasts represent a true single-cell system and are useful for various biochemical and physiological studies. Protoplasts from several agriculturally important crop species can be regenerated into a fertile whole plant, extending the utility of protoplasts from transient expression assays to the generation of stable transformation events.

Clovers (Trifolium spp.).

Clovers (Trifolium spp.) constitute one of the major forage legumes widely grown for its rich protein content and its major role in maintaining environmental sustainability by improving the soil fertility. Gene technology can assist plant improvement efforts in clovers (Trifolium spp.