Methods and systems are provided for generating and utilizing a bacterial composition that comprises at least one genetically engineered bacterial strain that fixes atmospheric nitrogen in an agricultural system that has been fertilized with more than 20 lbs of Nitrogen per acre.

The disclosure provides a synthetic biology toolkit that enables precise and effective control of gene expression in A. tumefaciens and related Rhizobia. Inducible expression systems were constructed, characterized, and optimized to obtain an expression system regulated through amplifier introduction and promoter engineering, and cognate promoters were produced and evaluated. To establish a fine-tunability, a series of spacers and a promoter library were constructed to systematically modulate both translational and transcriptional rates. The application of the tools was demonstrated by coexpressing genes with altered expression levels using a single signal. The studies carried out provide precise expression tools, facilitating rational engineering of in A. tumefaciens and related Rhizobia bacteria for advanced plant biotechnological applications.

The invention provides three novel disarmed strains of Agrobacterium tumefaciens bacteria useful for the transformation of plants. The invention provides three engineered A. tumefaciens Chry5 strains or bacterial cells thereof which comprise the Chry5 strain chromosomal background and a disarmed pTiChry5 vector, and methods of using said bacterial strains or cells for transformation of fungal or plant cells, in particular dicot or monocot plant cells, including soybean, maize, wheat, and sugarcane cells. The invention further relates to the transgenic plants created by these methods.

Transgenic host cells, vectors useful for making transgenic host cells, and kits useful for making transgenic host cells are described. Also described are transgenic plants. In some embodiments, transgenic host cells express a 4-hydroxyphenylacetaldehyde synthase (4HPAAS). In some embodiments, transgenic host cells express a tyrosol:UDP-glucose 8-O-glucosyltransferase (T8GT). The transgenic host cells are useful for biosynthesis of one or more of salidroside, icariside D2, tyrosol, and 4-hydroxypenylacetaldehyde.

The invention provides three novel disarmed strains of Agrobacteriumtumefaciens bacteria useful for the transformation of plants. The invention provides three engineered A. tumefaciens Chry5 strains or bacterial cells thereof which comprise the Chry5 strain chromosomal background and a disarmed pTiChry5 vector, and methods of using said bacterial strains or cells for transformation of fungal or plant cells, in particular dicot or monocot plant cells, including soybean, maize, wheat, and sugarcane cells. The invention further relates to the transgenic plants created by these methods.

A peptide comprised of either a binary or a tertiary peptide, the peptide contains at least 4 amino acids and up to a maximum of 16 amino acids, comprised of 2 or 3 different regions, wherein the binary peptides have 2 different regions and the tertiary peptides have 3 different regions; wherein, the peptide can be cleaved by both an animal gut protease and an insect or nematode gut protease.

Example embodiments in accordance with the present disclosure are directed to methods comprising contacting a plant part with a nucleotide sequence encoding a gene that induces plant cell matrix (PCM) formation, and culturing the plant part under growth conditions to enhance PCM formation.

Derivatives of Agrobacterium vitis strain F2/5 are disclosed. These derivatives were generated following homologous recombination with an internal fragment of targeted genes resulting in gene disruption by insertion of a copy of suicide vector pVIK165. The genes disrupted were F-avi5813 encoding a phosphopantetheinyltransferase, F-avi4329 encoding an aminotransferase and F-avi0838 (rirA) encoding an iron responsive transcriptional regulator. Such derivatives control crown gall on grapevines. In addition, these derivatives did not induce roots necrosis but enhanced root development and callus formation. On young stem explants, it was shown as well that the F2/5 derivatives are necrosis-negative.

The present disclosure provides novel compositions and methods for the production and use of Agrobacterium tumefaciens strains (for example LBA4404) that are deficient in RecA activity relative to the parent strain. Combinations with other gene-deficient-strains of Agrobacterium tumefaciens are also disclosed. Specifically, two exemplary s recA minus strains, UIA777 where chloramphenicol resistant gene disrupting the recA gene and UIA770 where kanamycin resistant gene disrupting the recA gene are provided.

The disclosure relates to a T-DNA binary vector based on bean yellow dwarf virus (BeYDV) that reduces plant cell death and increases transgene expression. In one aspect, the T-DNA region comprise a replicon cassette comprising a rep gene or a repA gene with a mutated translation initiation region. The disclosure also relates to replicating geminiviral expression system based on BeYDV comprising with an expression cassette a sequence encoding Rep and a sequence encoding the promoter of ubiquitin-3 from potato with ubiquitin fusion; an expression cassette comprising a sequence encoding RepA and a sequence encoding the promoter of ubiquitin-3 from potato with ubiquitin fusion; and an expression cassette comprising a promoter region, a 5′ UTR, a sequence encoding a recombinant protein, and a 3′ UTR. These expression cassettes are on different T-DNA cloning vectors or on one T-DNA cloning vector.