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      Drought Resistance by Engineering Plant Tissue-Specific Responses

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          Abstract

          Drought is the primary cause of agricultural loss globally, and represents a major threat to food security. Currently, plant biotechnology stands as one of the most promising fields when it comes to developing crops that are able to produce high yields in water-limited conditions. From studies of Arabidopsis thaliana whole plants, the main response mechanisms to drought stress have been uncovered, and multiple drought resistance genes have already been engineered into crops. So far, most plants with enhanced drought resistance have displayed reduced crop yield, meaning that there is still a need to search for novel approaches that can uncouple drought resistance from plant growth. Our laboratory has recently shown that the receptors of brassinosteroid (BR) hormones use tissue-specific pathways to mediate different developmental responses during root growth. In Arabidopsis, we found that increasing BR receptors in the vascular plant tissues confers resistance to drought without penalizing growth, opening up an exceptional opportunity to investigate the mechanisms that confer drought resistance with cellular specificity in plants. In this review, we provide an overview of the most promising phenotypical drought traits that could be improved biotechnologically to obtain drought-tolerant cereals. In addition, we discuss how current genome editing technologies could help to identify and manipulate novel genes that might grant resistance to drought stress. In the upcoming years, we expect that sustainable solutions for enhancing crop production in water-limited environments will be identified through joint efforts.

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          Most cited references158

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          Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting

          TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease, TALENs bind and cleave DNA in pairs. Binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. We present a method and reagents for efficiently assembling TALEN constructs with custom repeat arrays. We also describe design guidelines based on naturally occurring TAL effectors and their binding sites. Using software that applies these guidelines, in nine genes from plants, animals and protists, we found candidate cleavage sites on average every 35 bp. Each of 15 sites selected from this set was cleaved in a yeast-based assay with TALEN pairs constructed with our reagents. We used two of the TALEN pairs to mutate HPRT1 in human cells and ADH1 in Arabidopsis thaliana protoplasts. Our reagents include a plasmid construct for making custom TAL effectors and one for TAL effector fusions to additional proteins of interest. Using the former, we constructed de novo a functional analog of AvrHah1 of Xanthomonas gardneri. The complete plasmid set is available through the non-profit repository AddGene and a web-based version of our software is freely accessible online.
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            Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements.

            Prokaryotes contain short DN repeats known as CRISPR, recognizable by the regular spacing existing between the recurring units. They represent the most widely distributed family of repeats among prokaryotic genomes suggesting a biological function. The origin of the intervening sequences, at present unknown, could provide clues about their biological activities. Here we show that CRISPR spacers derive from preexisting sequences, either chromosomal or within transmissible genetic elements such as bacteriophages and conjugative plasmids. Remarkably, these extrachromosomal elements fail to infect the specific spacer-carrier strain, implying a relationship between CRISPR and immunity against targeted DNA. Bacteriophages and conjugative plasmids are involved in prokaryotic population control, evolution, and pathogenicity. All these biological traits could be influenced by the presence of specific spacers. CRISPR loci can be visualized as mosaics of a repeated unit, separated by sequences at some time present elsewhere in the cell.
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              Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin.

              Numerous prokaryote genomes contain structures known as clustered regularly interspaced short palindromic repeats (CRISPRs), composed of 25-50 bp repeats separated by unique sequence spacers of similar length. CRISPR structures are found in the vicinity of four genes named cas1 to cas4. In silico analysis revealed another cluster of three genes associated with CRISPR structures in many bacterial species, named here as cas1B, cas5 and cas6, and also revealed a certain number of spacers that have homology with extant genes, most frequently derived from phages, but also derived from other extrachromosomal elements. Sequence analysis of CRISPR structures from 24 strains of Streptococcus thermophilus and Streptococcus vestibularis confirmed the homology of spacers with extrachromosomal elements. Phage sensitivity of S. thermophilus strains appears to be correlated with the number of spacers in the CRISPR locus the strain carries. The authors suggest that the spacer elements are the traces of past invasions by extrachromosomal elements, and hypothesize that they provide the cell immunity against phage infection, and more generally foreign DNA expression, by coding an anti-sense RNA. The presence of gene fragments in CRISPR structures and the nuclease motifs in cas genes of both cluster types suggests that CRISPR formation involves a DNA degradation step.
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                Author and article information

                Contributors
                URI : https://loop.frontiersin.org/people/667209
                URI : https://loop.frontiersin.org/people/817323
                URI : https://loop.frontiersin.org/people/26426
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                22 January 2020
                2019
                : 10
                : 1676
                Affiliations
                [1] Department of Molecular Genetics, Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB , Barcelona, Spain
                Author notes

                Edited by: Oscar Lorenzo, University of Salamanca, Spain

                Reviewed by: Parvathi Madathil Sreekumar, Kerala Agricultural University, India; Pablo Albertos, Technical University of Munich, Germany

                *Correspondence: Ana I. Caño-Delgado, ana.cano@ 123456cragenomica.es

                This article was submitted to Plant Physiology, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2019.01676
                6987726
                32038670
                bb7f74da-0141-46e3-a0b8-c59144029025
                Copyright © 2020 Martignago, Rico-Medina, Blasco-Escámez, Fontanet-Manzaneque and Caño-Delgado

                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
                : 08 July 2019
                : 28 November 2019
                Page count
                Figures: 3, Tables: 0, Equations: 0, References: 189, Pages: 19, Words: 11597
                Funding
                Funded by: European Research Council 10.13039/501100000781
                Award ID: ERC-2015-CoG – 683163
                Funded by: Agencia Estatal de Investigación 10.13039/501100011033
                Award ID: BIO2016-78150-P
                Funded by: European Regional Development Fund 10.13039/501100008530
                Award ID: BIO2016-78150-P
                Funded by: Generalitat de Catalunya 10.13039/501100002809
                Award ID: 2017SGR718
                Funded by: Ministerio de Economía y Competitividad 10.13039/501100003329
                Award ID: BIO2016-78150-P
                Funded by: Fundación Tatiana Pérez de Guzmán el Bueno 10.13039/501100010805
                Categories
                Plant Science
                Review

                Plant science & Botany
                drought,arabidopsis,cereals,genome editing,cell-specific regulation
                Plant science & Botany
                drought, arabidopsis, cereals, genome editing, cell-specific regulation

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