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      Varying the expression pattern of the strigolactone receptor gene DAD2 results in phenotypes distinct from both wild type and knockout mutants

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          Abstract

          The action of the petunia strigolactone (SL) hormone receptor DAD2 is dependent not only on its interaction with the PhMAX2A and PhD53A proteins, but also on its expression patterns within the plant. Previously, in a yeast-2-hybrid system, we showed that a series of a single and double amino acid mutants of DAD2 had altered interactions with these binding partners. In this study, we tested the mutants in two plant systems, Arabidopsis and petunia. Testing in Arabidopsis was enabled by creating a CRISPR-Cas9 knockout mutant of the Arabidopsis strigolactone receptor (AtD14). We produced SL receptor activity in both systems using wild type and mutant genes; however, the mutants had functions largely indistinguishable from those of the wild type. The expression of the wild type DAD2 from the CaMV 35S promoter in dad2 petunia produced plants neither quite like the dad2 mutant nor the V26 wild type. These plants had greater height and leaf size although branch number and the plant shape remained more like those of the mutant. These traits may be valuable in the context of a restricted area growing system such as controlled environment agriculture.

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          A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures

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            Floral dip: a simplified method forAgrobacterium-mediated transformation ofArabidopsis thaliana

            The Agrobacterium vacuum infiltration method has made it possible to transform Arabidopsis thaliana without plant tissue culture or regeneration. In the present study, this method was evaluated and a substantially modified transformation method was developed. The labor-intensive vacuum infiltration process was eliminated in favor of simple dipping of developing floral tissues into a solution containing Agrobacterium tumefaciens, 5% sucrose and 500 microliters per litre of surfactant Silwet L-77. Sucrose and surfactant were critical to the success of the floral dip method. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at the highest rate. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary, and Agrobacterium could be applied to plants at a range of cell densities. Repeated application of Agrobacterium improved transformation rates and overall yield of transformants approximately twofold. Covering plants for 1 day to retain humidity after inoculation also raised transformation rates twofold. Multiple ecotypes were transformable by this method. The modified method should facilitate high-throughput transformation of Arabidopsis for efforts such as T-DNA gene tagging, positional cloning, or attempts at targeted gene replacement.
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              Strigolactone inhibition of shoot branching.

              A carotenoid-derived hormonal signal that inhibits shoot branching in plants has long escaped identification. Strigolactones are compounds thought to be derived from carotenoids and are known to trigger the germination of parasitic plant seeds and stimulate symbiotic fungi. Here we present evidence that carotenoid cleavage dioxygenase 8 shoot branching mutants of pea are strigolactone deficient and that strigolactone application restores the wild-type branching phenotype to ccd8 mutants. Moreover, we show that other branching mutants previously characterized as lacking a response to the branching inhibition signal also lack strigolactone response, and are not deficient in strigolactones. These responses are conserved in Arabidopsis. In agreement with the expected properties of the hormonal signal, exogenous strigolactone can be transported in shoots and act at low concentrations. We suggest that endogenous strigolactones or related compounds inhibit shoot branching in plants. Furthermore, ccd8 mutants demonstrate the diverse effects of strigolactones in shoot branching, mycorrhizal symbiosis and parasitic weed interaction.
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                Author and article information

                Contributors
                URI : https://loop.frontiersin.org/people/40221Role: Role: Role: Role: Role: Role: Role: Role: Role:
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                URI : https://loop.frontiersin.org/people/31865Role: Role:
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                URI : https://loop.frontiersin.org/people/26537Role: Role: Role: Role: Role: Role: Role: Role:
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                11 October 2023
                2023
                : 14
                : 1277617
                Affiliations
                [1] The New Zealand Institute for Plant and Food Research Limited , Auckland, New Zealand
                Author notes

                Edited by: Weiwei Zhang, Purdue University, United States

                Reviewed by: Francois Fabien Barbier, The University of Queensland, Australia; Mark Waters, University of Western Australia, Australia

                *Correspondence: Kimberley C. Snowden, kimberley.snowden@ 123456plantandfood.co.nz ; Revel S. M. Drummond, revel.drummond@ 123456plantandfood.co.nz
                Article
                10.3389/fpls.2023.1277617
                10600376
                37900765
                646d54db-279c-4e59-bad6-507e2c650b7a
                Copyright © 2023 Drummond, Lee, Luo, Dakin, Janssen and Snowden

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 14 August 2023
                : 22 September 2023
                Page count
                Figures: 5, Tables: 0, Equations: 0, References: 42, Pages: 11, Words: 6186
                Funding
                Funded by: Marsden Fund , doi 10.13039/501100009193;
                The authors declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the Marsden Fund from Royal Society Te Apārangi Contract PAF1301, and by the Growing Futures Fund from The New Zealand Institute for Plant and Food Research Limited.
                Categories
                Plant Science
                Original Research
                Custom metadata
                Plant Physiology

                Plant science & Botany
                strigolactone,branch,development,gene expression,mutation,receptor,plant,petunia
                Plant science & Botany
                strigolactone, branch, development, gene expression, mutation, receptor, plant, petunia

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