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      Overexpression of the class I homeodomain transcription factor TaHDZipI‐5 increases drought and frost tolerance in transgenic wheat

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          Summary

          Characterization of the function of stress‐related genes helps to understand the mechanisms of plant responses to environmental conditions. The findings of this work defined the role of the wheat Ta HDZipI‐5 gene, encoding a stress‐responsive homeodomain–leucine zipper class I ( HD‐Zip I) transcription factor, during the development of plant tolerance to frost and drought. Strong induction of Ta HDZipI‐5 expression by low temperatures, and the elevated Ta HDZipI‐5 levels of expression in flowers and early developing grains in the absence of stress, suggests that Ta HDZipI‐5 is involved in the regulation of frost tolerance at flowering. The Ta HDZipI‐5 protein behaved as an activator in a yeast transactivation assay, and the Ta HDZipI‐5 activation domain was localized to its C‐terminus. The Ta HDZipI‐5 protein homo‐ and hetero‐dimerizes with related Ta HDZipI‐3, and differences between DNA interactions in both dimers were specified at 3D molecular levels. The constitutive overexpression of Ta HDZipI‐5 in bread wheat significantly enhanced frost and drought tolerance of transgenic wheat lines with the appearance of undesired phenotypic features, which included a reduced plant size and biomass, delayed flowering and a grain yield decrease. An attempt to improve the phenotype of transgenic wheat by the application of stress‐inducible promoters with contrasting properties did not lead to the elimination of undesired phenotype, apparently due to strict spatial requirements for Ta HDZipI‐5 overexpression.

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          Plant responses to water deficit

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            The SHINE clade of AP2 domain transcription factors activates wax biosynthesis, alters cuticle properties, and confers drought tolerance when overexpressed in Arabidopsis.

            The interface between plants and the environment plays a dual role as a protective barrier as well as a medium for the exchange of gases, water, and nutrients. The primary aerial plant surfaces are covered by a cuticle, acting as the essential permeability barrier toward the atmosphere. It is a heterogeneous layer composed mainly of lipids, namely cutin and intracuticular wax with epicuticular waxes deposited on the surface. We identified an Arabidopsis thaliana activation tag gain-of-function mutant shine (shn) that displayed a brilliant, shiny green leaf surface with increased cuticular wax compared with the leaves of wild-type plants. The gene responsible for the phenotype encodes one member of a clade of three proteins of undisclosed function, belonging to the plant-specific family of AP2/EREBP transcription factors. Overexpression of all three SHN clade genes conferred a phenotype similar to that of the original shn mutant. Biochemically, such plants were altered in wax composition (very long fatty acid derivatives). Total cuticular wax levels were increased sixfold in shn compared with the wild type, mainly because of a ninefold increase in alkanes that comprised approximately half of the total waxes in the mutant. Chlorophyll leaching assays and fresh weight loss experiments indicated that overexpression of the SHN genes increased cuticle permeability, probably because of changes in its ultrastructure. Likewise, SHN gene overexpression altered leaf and petal epidermal cell structure, trichome number, and branching as well as the stomatal index. Interestingly, SHN overexpressors displayed significant drought tolerance and recovery, probably related to the reduced stomatal density. Expression analysis using promoter-beta-glucuronidase fusions of the SHN genes provides evidence for the role of the SHN clade in plant protective layers, such as those formed during abscission, dehiscence, wounding, tissue strengthening, and the cuticle. We propose that these diverse functions are mediated by regulating metabolism of lipid and/or cell wall components.
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              The MYB96 transcription factor regulates cuticular wax biosynthesis under drought conditions in Arabidopsis.

              Drought stress activates several defense responses in plants, such as stomatal closure, maintenance of root water uptake, and synthesis of osmoprotectants. Accumulating evidence suggests that deposition of cuticular waxes is also associated with plant responses to cellular dehydration. Yet, how cuticular wax biosynthesis is regulated in response to drought is unknown. We have recently reported that an Arabidopsis thaliana abscisic acid (ABA)-responsive R2R3-type MYB transcription factor, MYB96, promotes drought resistance. Here, we show that transcriptional activation of cuticular wax biosynthesis by MYB96 contributes to drought resistance. Microarray assays showed that a group of wax biosynthetic genes is upregulated in the activation-tagged myb96-1D mutant but downregulated in the MYB96-deficient myb96-1 mutant. Cuticular wax accumulation was altered accordingly in the mutants. In addition, activation of cuticular wax biosynthesis by drought and ABA requires MYB96. By contrast, biosynthesis of cutin monomers was only marginally affected in the mutants. Notably, the MYB96 protein acts as a transcriptional activator of genes encoding very-long-chain fatty acid-condensing enzymes involved in cuticular wax biosynthesis by directly binding to conserved sequence motifs present in the gene promoters. These results demonstrate that ABA-mediated MYB96 activation of cuticular wax biosynthesis serves as a drought resistance mechanism.
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                Author and article information

                Contributors
                maria.hrmova@adelaide.edu.au
                Journal
                Plant Biotechnol J
                Plant Biotechnol. J
                10.1111/(ISSN)1467-7652
                PBI
                Plant Biotechnology Journal
                John Wiley and Sons Inc. (Hoboken )
                1467-7644
                1467-7652
                27 December 2017
                June 2018
                : 16
                : 6 ( doiID: 10.1111/pbi.2018.16.issue-6 )
                : 1227-1240
                Affiliations
                [ 1 ] School of Agriculture, Food and Wine University of Adelaide Glen Osmond SA Australia
                [ 2 ]Present address: Institute of Molecular Biosciences Mahidol University Nakhon‐Pathom Thailand
                [ 3 ]Present address: South Australian Research and Development Institute GPO Box 397 Adelaide SA 5064 Australia
                [ 4 ]Present address: Commonwealth Scientific and Industrial Research Organisation Glen Osmond SA 5064 Australia
                [ 5 ]Present address: Rothamsted Research West Common Harpenden Hertfordshire Al5 2JQ UK
                Author notes
                [*] [* ] Correspondence (Tel +61 8 8313 7160; fax +61 8 8313 7102; email: maria.hrmova@ 123456adelaide.edu.au )
                Author information
                http://orcid.org/0000-0002-3545-0605
                Article
                PBI12865
                10.1111/pbi.12865
                5978581
                29193733
                501e44d5-0a1a-4189-8a02-062ffce7c2ea
                © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 12 September 2017
                : 29 October 2017
                : 12 November 2017
                Page count
                Figures: 8, Tables: 0, Pages: 14, Words: 10367
                Funding
                Funded by: Australian Research Council Industrial Transforming Research Hub
                Award ID: IH130200027
                Funded by: Dupont‐Pioneer
                Funded by: Australian Research Council Linkage Project
                Award ID: LP120100201
                Funded by: Australian Grains Research and Development Corporation
                Funded by: Government of South Australia
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                pbi12865
                June 2018
                Converter:WILEY_ML3GV2_TO_NLMPMC version:version=5.4.0 mode:remove_FC converted:31.05.2018

                Biotechnology
                3d protein modelling,abiotic stress,activation domain,phenotypic features,protein homo‐ and hetero‐dimerization,stress‐inducible promoters

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