Methods for producing in a plant a complex transgenic trait locus comprising at least two altered target sequences in a genomic region of interest are disclosed. The methods involve the use of two or more double-strand-break-inducing agents, each of which can cause a double-strand break in a target sequence in the genomic region of interest which results in an alteration in the target sequence. Also disclosed are complex transgenic trait loci in plants. A complex transgenic trait locus comprises at least two altered target sequences that are genetically linked to a polynucleotide of interest. Plants, plant cells, plant parts, and seeds comprising one or more complex transgenic trait loci are also disclosed.

This disclosure provides compositions that include a polynucleotide including or encoding a Cas12a tracrRNA, and methods for their use in sequence-specific genome editing, especially of eukaryotic genomic sequences. In particular, this disclosure provides Cas12a tracrRNA-containing compositions and methods for their use in Cas12a-mediated editing of a target sequence, wherein the editing efficiency is increased in comparison to controls lacking the Cas12a tracrRNA.

A plant cell co-expressing at least one casein protein and at least one kinase. The at least one casein protein is phosphorylated by the at least one kinase in vivo. Casein micelles comprising phosphorylated κ-casein and at least one of αS1-casein, αS2-casein, and β-casein can be made in vivo and/or in vitro. The casein micelles can be used to make food products including milk and cheese.

The present invention relates to plants having the activity of a haploid inducer comprising mutations in certain regions of the KINETOCHORE NULL2 (KNL2) protein in a plant, in particular within the SANTA domain. Provided are methods of generating haploid cells and doubled haploid cells as well as corresponding plants and plant parts. The present invention also relates to a method for identification of a plant in a plant population, wherein the plant has at least one mutation in the KNL2 protein, in particular within the SANTA domain, which confers activity of a haploid inducer. Further provided is a set of oligonucleotides for the identification of a plant having activity of a haploid inducer.

It is an object of the present invention to provide a method for introducing a protein into a plant, which is simple and extensively applicable to various types of plant cells and proteins. The above object is achieved by the present invention to provide a complex comprising a protein of interest to be introduced into a target plant cell and a carrier peptide, a method for introducing a protein of interest into a target plant cell using the complex, and a kit comprising a protein of interest to be introduced into a target plant cell and a carrier peptide.

Provided herein are methods for increasing plant cell transformation efficiency. These methods include exposing the plant cells to a liquid medium containing a surfactant. Following exposure to the surfactant-containing medium, the cells can become more amenable to transformation and may be genetically transformed using methods known in the art. Exposure of the cells to the surfactant-containing medium prior to transformation can increase plant transformation efficiency when compared to transformation efficiency of cells not exposed to the surfactant-containing medium.

The present inventors found that the efficiency of introducing a protein into a plant dramatically increases as compared with conventional methods for cultivating a bacteria-infected plant as it is when the protein of interest is expressed with bacteria having the type III secretion system, the bacteria are brought into contact with the plant, and then the infected tissues are cultured under bacteriostatic conditions for a certain period of time.

The present invention relates to an herbicide tolerance protein, an encoding gene thereof and use thereof, the herbicide tolerance protein comprising: a protein (a) having an amino acid sequence as shown in SEQ ID NO: 1, and having an alanine substitution at least at position 176 and/or having a valine substitution at position 178 of SEQ ID NO: 1; or (b) having an amino acid sequence as shown in SEQ ID NO: 3; or (c) having an amino acid sequence as shown in SEQ ID NO: 5; or (d) having an amino acid sequence as shown in SEQ ID NO: 7; or (e) being derived from (a) by means of the amino acid sequence of (a) undergoing substitution and/or deletion and/or by added one or several amino acids, and having the activity of thifensulfuron hydrolase. The herbicide tolerance protein of the present invention has a broad application prospects in plants.

The present invention relates to methods and compositions for transforming soybean, corn, cotton, or canola explants using spectinomycin as a selective agent for transformation of the explants. The method may further comprise treatment of the explants with cytokinin during the transformation and regeneration process.

The present invention relates to excision of explant material comprising meristematic tissue from seeds, and storage of such material prior to subsequent use in plant tissue culture and genetic transformation. Methods for tissue preparation, storage, and transformation are disclosed, as is transformable meristem tissue produced by such methods, and apparati for tissue preparation.