An Active<i>Ac/Ds</i>Transposon System for Activation Tagging in Tomato Cultivar M82 Using Clonal Propagation

Carter, Jared D.; Pereira, Andy; Dickerman, Allan W.; Veilleux, Richard E.

doi:10.1104/pp.113.213876

Cited by 21 publications

(13 citation statements)

References 55 publications

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“…Thus, numerous T-DNA mutant collections have been developed in Arabidopsis (Alonso et al, 2003;Campisi et al, 1999;Feldmann, 1991;Krysan et al, 1999;Qin et al, 2003;Sessions et al, 2002) and other crops like rice (Hsing et al, 2007;Jeon et al, 2000;Jeong et al, 2002;Wan et al, 2009;Wu et al, 2003). In tomato, two activation tagging collections have been generated in the cultivars Micro-Tom (Mathews et al, 2003), a dwarf genotype bearing several mutations affecting plant development (Carvalho et al, 2011;Mart ı et al, 2006), and M82 (Carter et al, 2013), a processing tomato variety with determinate growth habit.…”

Section: Introductionmentioning

confidence: 99%

A collection of enhancer trap insertional mutants for functional genomics in tomato

Pérez-Martín

Yuste‐Lisbona

Pineda

et al. 2017

Plant Biotechnology Journal

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SummaryWith the completion of genome sequencing projects, the next challenge is to close the gap between gene annotation and gene functional assignment. Genomic tools to identify gene functions are based on the analysis of phenotypic variations between a wild type and its mutant; hence, mutant collections are a valuable resource. In this sense, T‐DNA collections allow for an easy and straightforward identification of the tagged gene, serving as the basis of both forward and reverse genetic strategies. This study reports on the phenotypic and molecular characterization of an enhancer trap T‐DNA collection in tomato (Solanum lycopersicum L.), which has been produced by Agrobacterium‐mediated transformation using a binary vector bearing a minimal promoter fused to the uidA reporter gene. Two genes have been isolated from different T‐DNA mutants, one of these genes codes for a UTP‐glucose‐1‐phosphate uridylyltransferase involved in programmed cell death and leaf development, which means a novel gene function reported in tomato. Together, our results support that enhancer trapping is a powerful tool to identify novel genes and regulatory elements in tomato and that this T‐DNA mutant collection represents a highly valuable resource for functional analyses in this fleshy‐fruited model species.

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Section: Introductionmentioning

confidence: 99%

A collection of enhancer trap insertional mutants for functional genomics in tomato

Pérez-Martín

Yuste‐Lisbona

Pineda

et al. 2017

Plant Biotechnology Journal

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“…This system uses tetramer copies of cauliflower mosaic virus (CaMV) 35 S enhancers 15 and the enhancers upon integration in the recipient plant genome can function in either orientation at the place of insertion, thereby causing transcriptional activation of nearby genes resulting in dominant gain-of-function mutations 16 . There are several reports citing this technique with the development of AT lines in Arabidopsis 17 , 18 , japonica rice 16 , 19 , indica rice 20 , tomato 21 , 22 , poplar 23 , strawberry, potato 24 , barley 25 and sorghum 26 .…”

Section: Introductionmentioning

confidence: 99%

Activation-tagging in indica rice identifies a novel transcription factor subunit, NF-YC13 associated with salt tolerance

Manimaran

Reddy

Moin

et al. 2017

Sci Rep

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Nuclear factor Y (NF-Y) is a heterotrimeric transcription factor with three distinct NF-YA, NF-YB and NF-YC subunits. It plays important roles in plant growth, development and stress responses. We have reported earlier on development of gain-of-function mutants in an indica rice cultivar, BPT-5204. Now, we screened 927 seeds from 70 Ac/Ds plants for salinity tolerance and identified one activation-tagged salt tolerant DS plant (DS-16, T3 generation) that showed enhanced expression of a novel ‘histone-like transcription factor’ belonging to rice NF-Y subfamily C and was named as OsNF-YC13. Localization studies using GFP-fusion showed that the protein is localized to nucleus and cytoplasm. Real time expression analysis confirmed upregulation of transcript levels of OsNF-YC13 during salt treatment in a tissue specific manner. Biochemical and physiological characterization of the DS-16 revealed enhanced K+/Na+ ratio, proline content, chlorophyll content, enzymes with antioxidant activity etc. DS-16 also showed transcriptional up-regulation of genes that are involved in salinity tolerance. In-silico analysis of OsNF-YC13 promoter region evidenced the presence of various key stress-responsive cis-regulatory elements. OsNF-YC13 subunit alone does not appear to have the capacity for direct transcription activation, but appears to interact with the B- subunits in the process of transactivation.

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“…In plants, the Arabidopsis (Arabidopsis thaliana) T-DNA insertion lines (Berardini et al, 2015), as well as transposon collections in maize (Zea mays;McCarty et al, 2005) and Medicago truncatula (Tadege et al, 2008) and TILLING populations in wheat (Triticum aestivum; http://www.wheat-tilling.com), are powerful resources, provided an insertion or point mutation has been identified in the gene(s) of interest. In tomato (Solanum lycopersicum), several mutagenized populations are available derived from fast-neutron, ethyl methanesulfonate (EMS), TILLING, and transposontagging approaches (Menda et al, 2004;Minoia et al, 2010;Saito et al, 2011;Carter et al, 2013). In cases where mutants are not readily available, researchers may rely on transitive or stable gene-silencing approaches to study genes of interest (Smith et al, 2000;Burch-Smith et al, 2004).…”

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confidence: 99%

Generation of a Collection of Mutant Tomato Lines Using Pooled CRISPR Libraries

et al. 2017

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The high efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)-mediated mutagenesis in plants enables the development of high-throughput mutagenesis strategies. By transforming pooled CRISPR libraries into tomato (), collections of mutant lines were generated with minimal transformation attempts and in a relatively short period of time. Identification of the targeted gene(s) was easily determined by sequencing the incorporated guide RNA(s) in the primary transgenic events. From a single transformation with a CRISPR library targeting the immunity-associated leucine-rich repeat subfamily XII genes, heritable mutations were recovered in 15 of the 54 genes targeted. To increase throughput, a second CRISPR library was made containing three guide RNAs per construct to target 18 putative transporter genes. This resulted in stable mutations in 15 of the 18 targeted genes, with some primary transgenic plants having as many as five mutated genes. Furthermore, the redundancy in this collection of plants allowed for the association of aberrant T0 phenotypes with the underlying targeted genes. Plants with mutations in a homolog of an Arabidopsis () boron efflux transporter displayed boron deficiency phenotypes. The strategy described here provides a technically simple yet high-throughput approach for generating a collection of lines with targeted mutations and should be applicable to any plant transformation system.

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An ActiveAc/DsTransposon System for Activation Tagging in Tomato Cultivar M82 Using Clonal Propagation

Cited by 21 publications

References 55 publications

A collection of enhancer trap insertional mutants for functional genomics in tomato

A collection of enhancer trap insertional mutants for functional genomics in tomato

Activation-tagging in indica rice identifies a novel transcription factor subunit, NF-YC13 associated with salt tolerance

Generation of a Collection of Mutant Tomato Lines Using Pooled CRISPR Libraries

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