Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tips
Identifieur interne : 002172 ( Main/Exploration ); précédent : 002171; suivant : 002173Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tips
Auteurs : Qiaochun Wang [Finlande, République populaire de Chine] ; Wilmer J. Cuellar [Finlande] ; Minna-Liisa Rajam Ki [Finlande] ; Yukimasa Hirata [Japon] ; Jari P. T. Valkonen [Finlande]Source :
- Molecular Plant Pathology [ 1464-6722 ] ; 2008-03.
Abstract
Accumulation of viruses in vegetatively propagated plants causes heavy yield losses. Therefore, supply of virus‐free planting materials is pivotal to sustainable crop production. In previous studies, Raspberry bushy dwarf virus (RBDV) was difficult to eradicate from raspberry (Rubus idaeus) using the conventional means of meristem tip culture. As shown in the present study, it was probably because this pollen‐transmitted virus efficiently invades leaf primordia and all meristematic tissues except the least differentiated cells of the apical dome. Subjecting plants to thermotherapy prior to meristem tip culture heavily reduced viral RNA2, RNA3 and the coat protein in the shoot tips, but no virus‐free plants were obtained. Therefore, a novel method including thermotherapy followed by cryotherapy was developed for efficient virus eradication. Heat treatment caused subcellular alterations such as enlargement of vacuoles in the more developed, virus‐infected cells, which were largely eliminated following subsequent cryotherapy. Using this protocol, 20–36% of the treated shoot tips survived, 30–40% regenerated and up to 35% of the regenerated plants were virus‐free, as tested by ELISA and reverse transcription loop‐mediated isothermal amplification. Novel cellular and molecular insights into RBDV–host interactions and the factors influencing virus eradication were obtained, including invasion of shoot tips and meristematic tissues by RBDV, enhanced viral RNA degradation and increased sensitivity to freezing caused by thermotherapy, and subcellular changes and subsequent death of cells caused by cryotherapy. This novel procedure should be helpful with many virus–host combinations in which virus eradication by conventional means has proven difficult.
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DOI: 10.1111/j.1364-3703.2007.00456.x
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Therefore, supply of virus‐free planting materials is pivotal to sustainable crop production. In previous studies, Raspberry bushy dwarf virus (RBDV) was difficult to eradicate from raspberry (Rubus idaeus) using the conventional means of meristem tip culture. As shown in the present study, it was probably because this pollen‐transmitted virus efficiently invades leaf primordia and all meristematic tissues except the least differentiated cells of the apical dome. Subjecting plants to thermotherapy prior to meristem tip culture heavily reduced viral RNA2, RNA3 and the coat protein in the shoot tips, but no virus‐free plants were obtained. Therefore, a novel method including thermotherapy followed by cryotherapy was developed for efficient virus eradication. Heat treatment caused subcellular alterations such as enlargement of vacuoles in the more developed, virus‐infected cells, which were largely eliminated following subsequent cryotherapy. Using this protocol, 20–36% of the treated shoot tips survived, 30–40% regenerated and up to 35% of the regenerated plants were virus‐free, as tested by ELISA and reverse transcription loop‐mediated isothermal amplification. Novel cellular and molecular insights into RBDV–host interactions and the factors influencing virus eradication were obtained, including invasion of shoot tips and meristematic tissues by RBDV, enhanced viral RNA degradation and increased sensitivity to freezing caused by thermotherapy, and subcellular changes and subsequent death of cells caused by cryotherapy. This novel procedure should be helpful with many virus–host combinations in which virus eradication by conventional means has proven difficult.</div></front></TEI><affiliations><list><country><li>Finlande</li><li>Japon</li><li>République populaire de Chine</li></country><region><li>Uusimaa</li></region><settlement><li>Helsinki</li></settlement><orgName><li>Université d'Helsinki</li></orgName></list><tree><country name="Finlande"><region name="Uusimaa"><name sortKey="Wang, Qiaochun" sort="Wang, Qiaochun" uniqKey="Wang Q" first="Qiaochun" last="Wang">Qiaochun Wang</name></region><name sortKey="Cuellar, Wilmer J" sort="Cuellar, Wilmer J" uniqKey="Cuellar W" first="Wilmer J." last="Cuellar">Wilmer J. Cuellar</name><name sortKey="Rajam Ki, Minna Iisa" sort="Rajam Ki, Minna Iisa" uniqKey="Rajam Ki M" first="Minna-Liisa" last="Rajam Ki">Minna-Liisa Rajam Ki</name><name sortKey="Valkonen, Jari P T" sort="Valkonen, Jari P T" uniqKey="Valkonen J" first="Jari P. T." last="Valkonen">Jari P. T. Valkonen</name></country><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Qiaochun" sort="Wang, Qiaochun" uniqKey="Wang Q" first="Qiaochun" last="Wang">Qiaochun Wang</name></noRegion></country><country name="Japon"><noRegion><name sortKey="Hirata, Yukimasa" sort="Hirata, Yukimasa" uniqKey="Hirata Y" first="Yukimasa" last="Hirata">Yukimasa Hirata</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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