The present disclosure pertains to methods for delivering a compound into a cell comprising a cell wall, including passing a cell suspension through a constriction, wherein said constriction deforms the cell comprising a cell wall, thereby causing a perturbation of the cell such that the compound enters the cell, wherein said cell suspension is contacted with the compound.

Methods and compositions are provided for the creation of a Cas endonuclease-mediated double-strand break and stable integration of a heterologous polynucleotide in the genome of a somatic plant cell, for example in leaf tissue.

Nano-sized delivery vehicles utilize carbon nanotubes (CNTs) for the generic, targeted and passive transport of biomolecules into plant cells. Plant cells are transfected by adsorbing a charged biomolecular cargo on carbon nanotubes by electrostatic grafting or by dialysis-based pi-pi stacking grafting or by probe-tip sonication of complementary nucleic acid strands; and introducing into the cell the cargo-adsorbed nanotubes.

The present invention relates to methods and compositions for modifying a target site in the genome of a plant cell. Such modifications include integration of a transgene and mutations. The present invention also relates to methods and compositions for identifying and enriching for cells which comprise a modified target site.

A method for gene introduction by which higher efficiency for gene introduction than that by the conventional Agrobacterium method may be attained simply and without injuring the tissue is disclosed. According to the method of the present invention, the efficiency of gene introduction into plant cells by a bacterium belonging to genus Agrobacterium is promoted by accompanying centrifugation of the plant cells or plant tissue.

The present technology provides a nanoparticle that includes a silica network comprising crosslinked polysiloxanes, wherein the crosslinks comprise disulfide linkages, and the nanoparticle has a surface bearing charged functional groups and a surface potential of either less than −30 mV or greater than +30 mV, and wherein the nanoparticle has an average diameter of 20 nm to 60 n. The nanoparticles may be used to efficiently deliver biomolecules to plant cells, including polynucleic acids, proteins and complexes thereof (e.g., Cas9 RNP).

The invention provides methods and compositions for detecting and/or quantifying vector backbone in a nucleic acid preparation comprising a polynucleotide of interest using amplification assays that amplify a junction located between the polynucleotide of interest and the vector backbone, under conditions whereby amplification can occur, wherein the junction comprises a recognition site for a nuclease, and detecting the absence of an amplification product, whereby the absence of the amplification product indicates low or no vector backbone and/or quantifying the amount of amplification product to determine the amount of vector backbone in the nucleic acid preparation.

The invention provides novel methods and compositions for introduction, transfer or delivery of one or more biomolecules into wounded recipient plant cell(s). Methods for production of a wounded recipient cell culture and the creation of one or more mutations, edits, transgenic insertions, or other genetic changes in the recipient cell(s) are also provided. Product cells produced by such methods, and resulting cells and regenerated plants, plant parts, and progeny plants are further provided. Molecular and genetic analyses, analysis of phenotypes and traits, and use of screenable and selection markers, are also provided to confirm transfer of the biomolecule in to the recipient cell(s) and generation of the mutation, edit, transgenic insertion, or other genetic change in the recipient cell(s), and/or progeny thereof, and in plants or plant parts developed or regenerated from the foregoing.

The present invention relates to a method for modulating plant processes said method being characterized in that a plant is fed with an eco-friendly, plant- and/or algae-derived, biostimulant composition comprising exogenous small RNA molecules. In particular, the method of the invention can be used for modulating physiological or pathological plant processes, such plant growth, plant productivity, fruit quality, quality of produce, plant yield, plant response to abiotic stress and plant resistance to diseases or to infections.

The present disclosure provides, inter alia, Thraustochytrium and relevant methods and reagents, including engineered regulatory sequences from and/or operative in Thraustochytrid or Thraustochytrium, selectable markers useful for engineering microorganisms such as Thraustochytrids, means for mutagenizing microorganisms, novel strains produced by mutagenesis, and methods and compositions related to production of particular compounds in microorganisms.