A to D, Root growth of E. prostrata (A), A. retroflexus (B), S. viridis (C), and E. indica (D) in the absence (left) or presence of 1 µm (middle) and 5 µm C17 (right).
4), indicating that C17 administration increased the sensitivity to indaziflam and isoxaben. Having established that base editing was occurring in Arabidopsis T1 plants transformed with one of our base-editing vectors, additional T1 seeds were directly sown on medium supplemented with 1 μm C17. At low concentrations of C17 (50 nm), isoxaben (1 nm), or indaziflam (100 pm), no growth inhibition occurred (Fig. Moreover, the ability to mimic C17 resistance through clustered regularly interspaced short palindromic repeats (CRISPR)-mediated base editing suggests the possibility to transfer CBI multiresistance to any crop of interest.
C17 structure-function analysis. Isoxaben-resistant mutants are sensitive to C17. Statistically significant differences compared with plants grown in the absence of C17 are indicated; *P < 0.01 (two-sided Student's t test).
We accept the following cards through Worldpay. To circumvent this restriction, we have successfully applied the CRISPR base editor system BE3 (Komor et al., 2016). Brassica oleracea (collaboration with Harwood lab at John Innes Centre; Lawrenson et al, 2015). What does international collaboration mean for you?
Work is ongoing in the Ajioka Lab to construct yeast-based systems for genome scale DNA assembly, editing and shuttling to plant systems.
Patron, 2016; Raitskin and Patron, 2016), Chloroplasts by BASF on Flickr, licensed under CC-BY-NC-ND 2.0. These data demonstrate that introducing a CESA mutant allele can bring C17 tolerance to plants. Editing DNA can lead to changes in physical traits, like eye color, and disease risk.
Marchantia polymorpha (Haseloff Lab and Schornack Lab). Because only three C17-resistant T1 plants were initially identified, additional lines were selected on kanamycin. Genome Editing-Based Engineering of CESA3 Dual Cellulose-Inhibitor-Resistant Plants, Real-time imaging of cellulose reorientation during cell wall expansion in, Chemical genetics to examine cellulose biosynthesis, Indaziflam herbicidal action: A potent cellulose biosynthesis inhibitor, Reduced cellulose synthesis invokes lignification and defense responses in, The AID/APOBEC family of nucleic acid mutators, Effects of isoxaben on sensitive and tolerant plant cell cultures: I. Metabolic fate of isoxaben, Nonmotile cellulose synthase subunits repeatedly accumulate within localized regions at the plasma membrane in, Deciphering the evolution of herbicide resistance in weeds, Resistance against herbicide isoxaben and cellulose deficiency caused by distinct mutations in same cellulose synthase isoform CESA6, Organization of cellulose synthase complexes involved in primary cell wall synthesis in, A simple and rapid method for the preparation of plant genomic DNA for PCR analysis, Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in, Efficient genome editing in plants using a CRISPR/Cas system, Identification of cell-wall stress as a hexose-dependent and osmosensitive regulator of plant responses, Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1, Global perspective of herbicide-resistant weeds, Mitochondrial defects confer tolerance against cellulose deficiency, Immunogold labeling of rosette terminal cellulose-synthesizing complexes in the vascular plant, Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage, GreenGate—a novel, versatile, and efficient cloning system for plant transgenesis, Cellulose synthase complexes: Composition and regulation, Efficacy of preemergence herbicides for controlling a dinitroaniline-resistant goosegrass (, A revised medium for rapid growth and bio assays with tobacco tissue cultures, Visualization of cellulose synthase demonstrates functional association with microtubules, A cheminformatics review of auxins as herbicides, Structural and immunocytochemical characterization of the walls of dichlobenil‐habituated BY‐2 tobacco cells, Preemergence and postemergence control of artilleryweed (, Modifications of cellulose synthase confer resistance to isoxaben and thiazolidinone herbicides in, Tertiary model of a plant cellulose synthase, Cellulose biosynthesis inhibitors—a multifunctional toolbox, A tandem affinity purification-based technology platform to study the cell cycle interactome in, CESA TRAFFICKING INHIBITOR inhibits cellulose deposition and interferes with the trafficking of cellulose synthase complexes and their associated proteins KORRIGAN1 and POM2/CELLULOSE SYNTHASE INTERACTIVE PROTEIN1, Acetobixan, an inhibitor of cellulose synthesis identified by microbial bioprospecting, Developmental Programming of Thermonastic Leaf Movement, BRASSINOSTEROID-SIGNALING KINASE5 Associates with Immune Receptors and Is Required for Immune Responses, Deetiolation Enhances Phototropism by Modulating NON-PHOTOTROPIC HYPOCOTYL3 Phosphorylation Status, Ureide Permease 5 (AtUPS5) Connects Cell Compartments Involved in Ureide Metabolism, The Golgi Localization of GnTI Requires a Polar Amino Acid Residue within Its Transmembrane Domain, Unusual Roles of Secretory SNARE SYP132 in Plasma Membrane H, by The American Society of Plant Biologists. Several CBIs, such as isoxaben, flupoxam, dichlobenil, and indaziflam have been developed as herbicides. In contrast, a control construct (35S-GFP) yielded 38 transformants from 20,000 T1 seeds.
As the positive control, plants were treated with isoxaben. C17 shows an additive growth inhibition with isoxaben/indaziflam. Cloning primers are listed in Supplemental Table S1.
Large-scale assemblies require specialized cloning vehicles like artificial chromosomes for construction and transfer of genetic circuits to target organisms. C17 shows an additive growth inhibition with isoxaben/indaziflam. Lawrenson T, Shorinola O, Stacey N, Liu C, Østergaard L, Patron NJ, Uauy C, Harwood, W (2015). The annealed oligo pairs 49/51 and 50/52 were ligated into a BbsI-digested pEN-Chimera vector (a gift from Holger Puchta; Fauser et al., 2014). To achieve this purpose, the cesa3 mutant allele 2C (CESA3S983F) driven by the constitutive Cauliflower mosaic virus 35S promoter (35S-2C) was transformed into Col-0 wild-type Arabidopsis plants. As a new member of the CBI family, C17 has the potential to be developed as a new herbicide as well. Hierarchical DNA assembly methods are a necessary part of genome construction and modification. As shown in Figure 7, A–D, C17 was able to effectively inhibit root elongation of dicotyledonous crops. 6).
As mentioned above, C17 may be applicable as a component of an herbicide mixture with other CBIs to slow down the incidence of herbicide-resistant weeds.
I also think that the DNA background and shapes are needed more.
B, Root length quantification of plants in A.
A to E, Root growth of 5-d-old wild type (Col-0), a C17-tolerant mutant (8P), and three isoxaben-tolerant mutants (ixr1-1, ixr1-2, and ixr2-1) grown in the absence (A) or presence of 4 nm isoxaben (B), 200 nm C17 (C), 500 nm C17 (D), and 1 µm C17 (E). The observed inability to introduce an overexpression construct of mutated CESA3 into the genome would limit the transformation with C17-resistant alleles to those plant species for which a CESA knockout or knockdown line is available. Recently, genome editing technologies have shown great potential in plants. These Cas variants have been compared for specificity and efficiency using next-generation sequencing technologies and digital droplet based PCR assays . H, Root length quantification of plants in E, F, and G. Data represent mean ± sd (n > 4 plants). Supplemental Figure S3. We made some of their presentation slides available for you to download.
Scale bar, 5 mm. ↵1 This work was supported by a grant of the Research Foundation Flanders (G.023616N) and National Key R&D Program of China (2018YFD1000706). 8B). Get free reports and resources from our world class speakers. Synthesized cellulose can crystallize into cellulose microfibrils through inter- and intramolecular hydrogen bonds and Van der Waals forces and serve as the structural reinforcement of the cell wall (McFarlane et al., 2014).
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