Heat‐shock tagging: a simple method for expression and isolation of plant genome DNA flanked by T‐DNA insertions

Matsuhara, Shio; Jingu, Fumito; Takahashi, Taku; Komeda, Yoshibumi

doi:10.1046/j.1365-313x.2000.00716.x

Cited by 57 publications

(49 citation statements)

References 26 publications

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“…However, we observed that heat-shock induction was more efficient when applied on in vitro cultured young plantlets, than on mature plants. As previously reported by Matsuhara et al (2000), this suggests that HSP18.2 promoter induction requires a high humidity environment as it is found for in vitro cultures. As pointed out in earlier reviews (Gatz and Lenk, 1998;Padidam, 2003), a good inducible system should have a null or low basal expression.…”

Section: Discussionsupporting

confidence: 85%

“…The HSP18.2 promoter presents a weak basal expression at a growth temperature of 23°C, except in vascular bundles and in siliques where a higher expression can be detected. These expression levels are strongly increased by incubation at a temperature of about 37°C (Takahashi et al, 1992;Matsuhara et 18S al, 2000). Despite the basal expression, the heat-inducible gene silencing should be suitable in numerous and various purposes.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies on the promoter of the HSP18.2 gene from Arabidopsis, encoding a heat shock protein, indicated that it functions as a strong inducible system in plants (Takahashi and Komeda, 1989;Takahashi et al, 1992, Matsuhara et al, 2000. Heat-shock inducible promoters have been used with success in Caenorhabditis elegans to control RNAi constructs showing that it was possible to inactivate specific genes for the duration of the expression period (Tavernarakis et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

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Gene silencing using a heat-inducible RNAi system in Arabidopsis

Masclaux

Charpenteau

Takahashi

et al. 2004

Biochemical and Biophysical Research Communications

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Controlling gene expression during plant development is an efficient tool to explore gene function. In this paper, we describe a gene expression system driven by a heath shock gene promoter (HSP18.2), to trigger the expression of an intron-containing inverted repeat. RNA interference became a powerful way for gene functional analysis by reverse genetic approaches. However, constitutive gene silencing cannot be used with genes involved in fundamental processes such as embryo viability. Inducible promoters provide an alternative approach for temporal and spatial gene expression control. And we described here a new, system, complementary to those using chemical gene inducers. To evaluate the efficiency of this system, RNA corresponding to the phytoene desaturase gene of Arabidopsis thaliana was used as a reporter gene in transgenic plants and a comparative study was performed using either the CaMV35S constitutive promoter or the HSP18.2 inducible promoter.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Gene silencing using a heat-inducible RNAi system in Arabidopsis

Masclaux

Charpenteau

Takahashi

et al. 2004

Biochemical and Biophysical Research Communications

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“…The Alc gene expression system, which is based on a regulon from Aspergillus nidulans (Felenbok et al 1988;Pateman et al 1983;Sealy-Lewis and Lockington 1984), and the heat-shock inducible promoter (Matsuhara et al 2000;Takahashi et al 1992) were used as inducible promoters. The alcR transcriptional regulator is expressed with the CaMV35S promoter such that, in the presence of ethanol, ALCR induces expression of any gene fused to a modified alcR promoter (Caddick et al 1998).…”

Section: Induction Of Double Flowers In Pharbitis Nil Using a Class-cmentioning

confidence: 99%

“…The alcR transcriptional regulator is expressed with the CaMV35S promoter such that, in the presence of ethanol, ALCR induces expression of any gene fused to a modified alcR promoter (Caddick et al 1998). On the other hand, the heat-shock inducible promoter HSP18.2 from Arabidopsis, encoding a heatshock protein, is indicated to function as a strong inducible system in plants (Takahashi et al 1992;Matsuhara et al 2000). These inducible promoters have been used to construct controllable gene expression systems in plant cells.…”

Section: Induction Of Double Flowers In Pharbitis Nil Using a Class-cmentioning

confidence: 99%

Induction of double flowers in Pharbitis nil using a class-C MADS-box transcription factor with Chimeric REpressor gene-Silencing Technology

Sage-Ono

Ozeki

Hiyama

et al. 2011

Plant Biotechnology

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The Chimeric REpressor gene-Silencing Technology (CRES-T) system is a novel reverse genetic method using a chimeric transcriptional repressor fusing an EAR transcriptional repression domain called SRDX. We sought to change the flower shape of Pharbitis nil, a model ornamental flower, using an Arabidopsis transcription factor fused with SRDX. For the first trial modulating flower shape we transformed with the class-C MADS-box transcription factor AGAMOUS (AG) fused with SRDX (AGSRDX). Defects in class-C genes cause double flowers in Arabidopsis and Pharbitis. However, when AGSRDX was expressed under the CaMV 35S promoter (p35S), the transgenic Pharbitis bore a malformed flower with a protruding pistil. We then used DUPLICATED (DP), one of the class-C genes in Pharbitis. The p35S :: DPSRDXintroduced callus were difficult to regenerate during transgenic steps, but occasionally made a perfect double flower bud showing severe growth defects. The flower buds never developed to flower opening stage. These results indicate that CRES-T is functional in Pharbitis but even using a conserved transcription factor, some species-specific variation might exist. To avoid these unwanted effects, we recruited inducible promoters to control expression of the chimeric transcription repressors in combination with the DNA-binding domain of GAL4 in yeast fused with SRDX and the GAL4 upstream activator sequence (UAS). Normal regeneration was observed by inducible repression of DPSRDX during in vitro redifferentiation, and the double-flowered Pharbitis was generated. We successfully induced a non-transformant (NT)-like flower in DPSRDX-expressing double-flowered transformants. Our approach will enable us to breed transgenic horticultural plants with inducible fertility.

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Plant Transformation Technology: Agrobacterium ‐Mediated Transformation

Komari

Ishida

Hiei

2004

Handbook of Plant Biotechnology

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The soil phytopathogen Agrobacterium tumefaciens induces tumours, known as crown galls, mainly on dicotyledonous plants. Such tumours are generated by a complex, multi‐step transformation process. Another species, A. rhizogenes , causes hairy root disease in higher plants via an identical process. Agrobacterium has been utilised for the transfer of genes to dicotyledonous plants. Now, monocotyledonous plants are routinely transformed by Agrobacterium despite the fact that these plants, including important cereals, were thought until recently to be outside the range of this technology. Most of the common transformation methods heavily depend on tissue culture technology and we refer to such methods as in vitro transformation. The only case of a routine tissue culture free method is the ‘ in planta ’ transformation of Arabidopsis thaliana . Numerous critical factors are involved in both approaches. In transformation in vitro , key factors include choice of vectors and bacterial strains, types of plant tissues to be infected, procedures of preparation the tissues, protocols of infection and co‐cultivation, methods for subsequent culture and selection of transformed cells, antibiotics to remove infecting bacteria, techniques for regeneration of transgenic plants and genotypes of plants. It is our opinion that the type and quality of the starting material is the most important one among them. The capacity to serve as a host plant for crown gall disease is no longer a prerequisite for a host of vector systems based on Agrobacterium . The real prerequisite for in vitro transformation is the availability of technology for dedifferentiation of tissues and regeneration of plants in a given species. On the other hand, the biological processes involved in in planta transformation are yet to be elucidated. Recent evidence suggested that ovules are the primary target, and further basic understanding is likely to help extend the number of species transformed by the method. The plant species that are routinely transformed by Agrobacterium are expanding rapidly. Some gymnosperms, several forest trees and fruit trees, various legumes, and cereal and non‐cereal monocotyledons, which were once considered very recalcitrant, are now in a long list of transformable plants. Arabidopsis , tobacco and rice are the top three species that were transformed during the last two years. Although it is only half a dozen years since the current procedure of rice transformation mediated by Agrobacterium was published, the economic importance and the accumulation of genomic information has made rice the species of focus in plant biotechnology. The advantages of Agrobacterium ‐mediated transformation include the transfer of pieces of DNA with defined ends and minimal rearrangement, the transfer of relatively large segments of DNA, the integration of small numbers of copies of genes into plant chromosomes and the high quality and fertility of transgenic plants. However, transformation does not always produce such ‘clean’ events. Formation of repeats of transgenes, certain rearrangements, integration of non‐target DNA segments and unstable expression of transgenes are among the complications. Although the majority of transgenic plants usually appear ‘good’ in a particular test, accumulation of ‘dropped’ plants is significant after multiple rounds of characterisation and screening. Therefore, further improvement in each of steps is highly desired. Application of the gene transfer mediated by Agrobacterium is further expanding. Transient expression of genes delivered by Agrobacterium is now a useful tool in the study of promoters and gene function. Vectors specifically designed to carry very large segments of DNA have been developed and extensively tested. Insertional mutagenesis by DNA transferred by Agrobacterium is a routine technique in genomics study in Arabidopsis and rice. Methods for targeted integration of transgenes to genomes of higher plants have been drawing considerable attention. Various technologies for the production of ‘selection marker free’ transgenic plants are now in place for better public acceptance of biotechnology products.

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Heat‐shock tagging: a simple method for expression and isolation of plant genome DNA flanked by T‐DNA insertions

Cited by 57 publications

References 26 publications

Gene silencing using a heat-inducible RNAi system in Arabidopsis

Gene silencing using a heat-inducible RNAi system in Arabidopsis

Induction of double flowers in Pharbitis nil using a class-C MADS-box transcription factor with Chimeric REpressor gene-Silencing Technology

Plant Transformation Technology: Agrobacterium ‐Mediated Transformation

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