Removing the mustard oil bomb from seeds: transgenic ablation of myrosin cells in oilseed rape (Brassica napus) produces MINELESS seeds
Identifieur interne : 000D67 ( Main/Exploration ); précédent : 000D66; suivant : 000D68Removing the mustard oil bomb from seeds: transgenic ablation of myrosin cells in oilseed rape (Brassica napus) produces MINELESS seeds
Auteurs : Birgit Hafeld Borgen [Norvège] ; Ole Petter Thangstad [Norvège] ; Ishita Ahuja [Norvège] ; John Trevor Rossiter [Royaume-Uni] ; Atle Magnar Bones [Norvège]Source :
- Journal of Experimental Botany [ 0022-0957 ] ; 2010-03-10.
Abstract
Many plant phytochemicals constitute binary enzymeglucoside systems and function in plant defence. In brassicas, the enzyme myrosinase is confined to specific myrosin cells that separate the enzyme from its substrate; the glucosinolates. The myrosinase-catalysed release of toxic and bioactive compounds such as isothiocyanates, upon activation or tissue damage, has been termed the mustard oil bomb and characterized as a toxic mine in plant defence. The removal of myrosin cells and the enzyme that triggers the release of phytochemicals have been investigated by genetically modifying Brassica napus plants to remove myrosinase-storing idioblasts. A construct with the seed myrosin cell-specific Myr1.Bn1 promoter was used to express a ribonuclease, barnase. Transgenic plants ectopically expressing barnase were embryo lethal. Co-expressing barnase under the control of the Myr1.Bn1 promoter with the barnase inhibitor, barstar, under the control of the cauliflower mosaic virus 35S promoter enabled a selective and controlled death of myrosin cells without affecting plant viability. Ablation of myrosin cells was confirmed with light and electron microscopy, with immunohistological analysis and immunogold-electron microscopy analysis showing empty holes where myrosin cells normally are localized. Further evidence for a successful myrosin cell ablation comes from immunoblots showing absence of myrosinase and negligible myrosinase activity, and autolysis experiments showing negligible production of glucosinolate hydrolysis products. The plants where the myrosin defence cells have been ablated and named MINELESS plants. The epithiospecifier protein profile and glucosinolate levels were changed in MINELESS plants, pointing to localization of myrosinases and a 35kDa epithiospecifier protein in myrosin cells and a reduced turnover of glucosinolates in MINELESS plants.
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DOI: 10.1093/jxb/erq039
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ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract">Many plant phytochemicals constitute binary enzymeglucoside systems and function in plant defence. In brassicas, the enzyme myrosinase is confined to specific myrosin cells that separate the enzyme from its substrate; the glucosinolates. The myrosinase-catalysed release of toxic and bioactive compounds such as isothiocyanates, upon activation or tissue damage, has been termed the mustard oil bomb and characterized as a toxic mine in plant defence. The removal of myrosin cells and the enzyme that triggers the release of phytochemicals have been investigated by genetically modifying Brassica napus plants to remove myrosinase-storing idioblasts. A construct with the seed myrosin cell-specific Myr1.Bn1 promoter was used to express a ribonuclease, barnase. Transgenic plants ectopically expressing barnase were embryo lethal. Co-expressing barnase under the control of the Myr1.Bn1 promoter with the barnase inhibitor, barstar, under the control of the cauliflower mosaic virus 35S promoter enabled a selective and controlled death of myrosin cells without affecting plant viability. Ablation of myrosin cells was confirmed with light and electron microscopy, with immunohistological analysis and immunogold-electron microscopy analysis showing empty holes where myrosin cells normally are localized. Further evidence for a successful myrosin cell ablation comes from immunoblots showing absence of myrosinase and negligible myrosinase activity, and autolysis experiments showing negligible production of glucosinolate hydrolysis products. The plants where the myrosin defence cells have been ablated and named MINELESS plants. The epithiospecifier protein profile and glucosinolate levels were changed in MINELESS plants, pointing to localization of myrosinases and a 35kDa epithiospecifier protein in myrosin cells and a reduced turnover of glucosinolates in MINELESS plants.</div></front></TEI><affiliations><list><country><li>Norvège</li><li>Royaume-Uni</li></country></list><tree><country name="Norvège"><noRegion><name sortKey="Borgen, Birgit Hafeld" sort="Borgen, Birgit Hafeld" uniqKey="Borgen B" first="Birgit Hafeld" last="Borgen">Birgit Hafeld Borgen</name></noRegion><name sortKey="Ahuja, Ishita" sort="Ahuja, Ishita" uniqKey="Ahuja I" first="Ishita" last="Ahuja">Ishita Ahuja</name><name sortKey="Bones, Atle Magnar" sort="Bones, Atle Magnar" uniqKey="Bones A" first="Atle Magnar" last="Bones">Atle Magnar Bones</name><name sortKey="Bones, Atle Magnar" sort="Bones, Atle Magnar" uniqKey="Bones A" first="Atle Magnar" last="Bones">Atle Magnar Bones</name><name sortKey="Thangstad, Ole Petter" sort="Thangstad, Ole Petter" uniqKey="Thangstad O" first="Ole Petter" last="Thangstad">Ole Petter Thangstad</name></country><country name="Royaume-Uni"><noRegion><name sortKey="Rossiter, John Trevor" sort="Rossiter, John Trevor" uniqKey="Rossiter J" first="John Trevor" last="Rossiter">John Trevor Rossiter</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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