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Biblioteca (s) : |
INIA Treinta y Tres. |
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Fecha : |
22/12/2020 |
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Actualizado : |
11/03/2021 |
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Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
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Autor : |
CASTILLO, A.; LOPEZ, V.; TAVARES, E.; SANTIÑAQUE, F.; DALLA RIZZA, M. |
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Afiliación : |
ALICIA MARIA CASTILLO SALLE, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; VICTOR JULIAN LOPEZ DEL PUERTO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; E. TAVARES, Forestal Oriental, Paysandú Uruguay.; F. SANTIÑAQUE, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.; MARCO DALLA RIZZA VILARO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay. |
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Título : |
Polyploid induction of Eucalyptus dunnii Maiden to generate waviability in breeding programs. [Polyploid induction of Eucalyptus dunnii Maiden to generate variability in breeding programs]. |
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Complemento del título : |
Special Issue X Encuentro Latinoamericano y del Caribe de Biotecnología Agropecuaria; XII Simposio REDBIO Argentina. Roca, William, Ed. 12 al 15 de noviembre de 2019, Montevideo, Uruguay. |
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Fecha de publicación : |
2020 |
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Fuente / Imprenta : |
Agrociencia Uruguay 2020, v. 24, no. NE2, Article 381. DOI: 10.31285/AGRO.24.413 |
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Páginas : |
9 p. |
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ISSN : |
2301-1548 |
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DOI : |
10.31285/AGRO.24.381 |
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Idioma : |
Inglés |
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Notas : |
Article history: Received 29 Jun. 2020 // Accepted 28 Sep. 2020 // Published 17 Dec 2020. Comité científico editor: Dra. Marisa López-Bilbao (INTA, Hurlingham, Provincia de Buenos Aires, Argentina); Dra. Sandra Sharry (Universidad Nacional de la Plata, La Plata, Buenos Aires, Argentina); Dra. Alicia Castillo (Investigadora Principal, Unidad de Biotecnología de INIA, Uruguay) ; Dr. Juan Izquierdo (Profesor Libre Facultad de Agronomía, Udelar, Presidente Academia Chilena Ciencias Agronómicas) ; Dr. Gerardo Gallego (Coordinador del Laboratorio de Biotecnología Vegetal del Centro Internacional de Agricultura Tropical-CIAT Cali, Colombia) ; Dra. Elizabeth Hodson (Profesora emérita de la Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia); Dr. Paul Chavarriaga (líder de la plataforma de transformación genética y edición de genomas del Centro Internacional de Agricultura Tropical-CIAT Cali, Colombia) ; Dr. Marco Dalla-Rizza (Coordinador de la Unidad de Biotecnología, Investigador principal referente, presidente REDBIO 2016-2020). |
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Contenido : |
Eucalyptus dunnii Maiden produces good quality cellulose pulp, showing good frost tolerance. However, in Uru-guay, it needs more than six years to reach the reproductive stage. Genome duplication was proposed as a strategy to obtain useful variability. The application of mitosis inhibitors for chromosomal duplication, in explants growing in vitro, was evaluated. Two antimitotic agents were used: colchicine and oryzalin in different concen-trations and exposure times, in two types of explants: explants growing in vitro and pre-germinated seeds. The number of chloroplasts was used as a rapid ploidy estimator and confirmed by flow cytometry. For chloroplast count, fluorescein diacetate (FDA) applied to in vitro leaves was used for staining. Oryzalin was effective for inducing plant duplication in E. dunnii from in vitro explants. In pre-germinated seeds, both antimitotic agents induced polyploids. The average number of chloroplasts was 5.5 in diploid control plants and more than 7 in tetraploids. Obtained plantlets were successfully cloned in the greenhouse. This is the first report on artificial polyploidy obtained in E. dunnii. |
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Palabras claves : |
COLCHICINA; COLCHICINE; CULTIVO IN VITRO; EUCALIPTO; IN VITRO CULTURE; ORYZALIN; ORYZALINE; POLIPLOIDIA; POLYPLOIDY. |
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Asunto categoría : |
F30 Genética vegetal y fitomejoramiento |
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URL : |
http://www.ainfo.inia.uy/digital/bitstream/item/14914/1/Castillo-Agrociencia-2020.pdf
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Marc : |
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008 2020 bl uuuu u00u1 u #d
022 $a2301-1548
024 7 $a10.31285/AGRO.24.381$2DOI
100 1 $aCASTILLO, A.
245 $aPolyploid induction of Eucalyptus dunnii Maiden to generate waviability in breeding programs. [Polyploid induction of Eucalyptus dunnii Maiden to generate variability in breeding programs].$h[electronic resource]
260 $c2020
300 $a9 p.
500 $aArticle history: Received 29 Jun. 2020 // Accepted 28 Sep. 2020 // Published 17 Dec 2020. Comité científico editor: Dra. Marisa López-Bilbao (INTA, Hurlingham, Provincia de Buenos Aires, Argentina); Dra. Sandra Sharry (Universidad Nacional de la Plata, La Plata, Buenos Aires, Argentina); Dra. Alicia Castillo (Investigadora Principal, Unidad de Biotecnología de INIA, Uruguay) ; Dr. Juan Izquierdo (Profesor Libre Facultad de Agronomía, Udelar, Presidente Academia Chilena Ciencias Agronómicas) ; Dr. Gerardo Gallego (Coordinador del Laboratorio de Biotecnología Vegetal del Centro Internacional de Agricultura Tropical-CIAT Cali, Colombia) ; Dra. Elizabeth Hodson (Profesora emérita de la Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia); Dr. Paul Chavarriaga (líder de la plataforma de transformación genética y edición de genomas del Centro Internacional de Agricultura Tropical-CIAT Cali, Colombia) ; Dr. Marco Dalla-Rizza (Coordinador de la Unidad de Biotecnología, Investigador principal referente, presidente REDBIO 2016-2020).
520 $aEucalyptus dunnii Maiden produces good quality cellulose pulp, showing good frost tolerance. However, in Uru-guay, it needs more than six years to reach the reproductive stage. Genome duplication was proposed as a strategy to obtain useful variability. The application of mitosis inhibitors for chromosomal duplication, in explants growing in vitro, was evaluated. Two antimitotic agents were used: colchicine and oryzalin in different concen-trations and exposure times, in two types of explants: explants growing in vitro and pre-germinated seeds. The number of chloroplasts was used as a rapid ploidy estimator and confirmed by flow cytometry. For chloroplast count, fluorescein diacetate (FDA) applied to in vitro leaves was used for staining. Oryzalin was effective for inducing plant duplication in E. dunnii from in vitro explants. In pre-germinated seeds, both antimitotic agents induced polyploids. The average number of chloroplasts was 5.5 in diploid control plants and more than 7 in tetraploids. Obtained plantlets were successfully cloned in the greenhouse. This is the first report on artificial polyploidy obtained in E. dunnii.
653 $aCOLCHICINA
653 $aCOLCHICINE
653 $aCULTIVO IN VITRO
653 $aEUCALIPTO
653 $aIN VITRO CULTURE
653 $aORYZALIN
653 $aORYZALINE
653 $aPOLIPLOIDIA
653 $aPOLYPLOIDY
700 1 $aLOPEZ, V.
700 1 $aTAVARES, E.
700 1 $aSANTIÑAQUE, F.
700 1 $aDALLA RIZZA, M.
773 $tAgrociencia Uruguay 2020$gv. 24, no. NE2, Article 381. DOI: 10.31285/AGRO.24.413
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Registro original : |
INIA Treinta y Tres (TT) |
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Registro completo
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Biblioteca (s) : |
INIA Tacuarembó. |
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Fecha actual : |
13/03/2018 |
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Actualizado : |
13/03/2018 |
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Tipo de producción científica : |
Abstracts/Resúmenes |
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Autor : |
NIKICHUK, N.; TORRES, D. |
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Afiliación : |
NATALIA ISABEL NIKICHUK BELL, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; DIEGO GABRIEL TORRES DINI, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay. |
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Título : |
Protocol: fast and high-performance Eucalyptus DNA extraction in 96-well plate method. |
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Fecha de publicación : |
2014 |
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Fuente / Imprenta : |
In: IUFRO Forest Tree Breeding Conference, August 25-29, Prague, Czech Republic, 2014. Book of Abstracts. |
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Páginas : |
p. 54 |
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Idioma : |
Inglés |
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Contenido : |
Forestry genetics and genomics require a great number of samples to produce accurate results. This is a simple method with high-throughput DNA extraction that combined CTAB protocol with fast rupture tissue and centrifuge steps in plates to obtain good yield, quality and low price. Approximately 576 samples can be processed for day. The obtained DNA is suitable for use in PCR-based study like Microsatellites. This method employs 96 Racked Collection Microtubes (Qiagen). Eucalyptus tissue 15mg was placed in each well together with a tungsten bead and 420ul of CTAB2X buffer distributed with a multichannel pipette, the samples were disrupted to full power in a TissueLyser (Qiagen) for 3min. Then we added 360ul of CIA24:1 to each tube
and both racks were centrifuged at 4000 RPM for 20min (ThermoScientific SorvallST16). Afterwards the upper phases were transferred carefully to clean rack-wells and 150ul of isopropanol were added to each well and centrifuged at 4000RPM for 20min. The liquid phases were removed and the pellets were washed twice with ethanol 70% and centrifuged at 4000RPM for 5min. Finally the pellets were resuspended in 100ul of RNAse and incubated at 37ºC for 60min. To evaluate the yield and purity, this prococol was compared against results obtained with four commercial extraction kits: DNeasy Plant Handbook® (Qiagen), Power Plant DNA® Isolation Kit (MOBIO), Wizard® Genomic DNA Purification Kit (Promega), ZR Plant / Seed DNA MiniPrep® (Zymo). The
procedure of rupture was the same for all protocols and in all cases racks of 96 wells were used. The data of absorption spectrum indicate an average concentration of 400ng/ul for CTAB protocol, Qiagen 150ng/ul, Wizard 165ng/ul MOBIO 85ng/ul for Zymo. Comparing this method to the commercial is demostrated that the final concentration superior, the time os processing is lower, also it is considerable cheaper. MenosForestry genetics and genomics require a great number of samples to produce accurate results. This is a simple method with high-throughput DNA extraction that combined CTAB protocol with fast rupture tissue and centrifuge steps in plates to obtain good yield, quality and low price. Approximately 576 samples can be processed for day. The obtained DNA is suitable for use in PCR-based study like Microsatellites. This method employs 96 Racked Collection Microtubes (Qiagen). Eucalyptus tissue 15mg was placed in each well together with a tungsten bead and 420ul of CTAB2X buffer distributed with a multichannel pipette, the samples were disrupted to full power in a TissueLyser (Qiagen) for 3min. Then we added 360ul of CIA24:1 to each tube
and both racks were centrifuged at 4000 RPM for 20min (ThermoScientific SorvallST16). Afterwards the upper phases were transferred carefully to clean rack-wells and 150ul of isopropanol were added to each well and centrifuged at 4000RPM for 20min. The liquid phases were removed and the pellets were washed twice with ethanol 70% and centrifuged at 4000RPM for 5min. Finally the pellets were resuspended in 100ul of RNAse and incubated at 37ºC for 60min. To evaluate the yield and purity, this prococol was compared against results obtained with four commercial extraction kits: DNeasy Plant Han... Presentar Todo |
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Palabras claves : |
GENÉTICA FORESTAL. |
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Thesagro : |
FORESTACIÓN. |
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Asunto categoría : |
K10 Producción forestal |
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URL : |
http://www.ainfo.inia.uy/digital/bitstream/item/8912/1/Protocol-fast-and-high-performance-Eucalyptus-DNA.pdf
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Marc : |
LEADER 02654nam a2200157 a 4500
001 1058242
005 2018-03-13
008 2014 bl uuuu u01u1 u #d
100 1 $aNIKICHUK, N.
245 $aProtocol$bfast and high-performance Eucalyptus DNA extraction in 96-well plate method.$h[electronic resource]
260 $aIn: IUFRO Forest Tree Breeding Conference, August 25-29, Prague, Czech Republic, 2014. Book of Abstracts.$c2014
300 $ap. 54
520 $aForestry genetics and genomics require a great number of samples to produce accurate results. This is a simple method with high-throughput DNA extraction that combined CTAB protocol with fast rupture tissue and centrifuge steps in plates to obtain good yield, quality and low price. Approximately 576 samples can be processed for day. The obtained DNA is suitable for use in PCR-based study like Microsatellites. This method employs 96 Racked Collection Microtubes (Qiagen). Eucalyptus tissue 15mg was placed in each well together with a tungsten bead and 420ul of CTAB2X buffer distributed with a multichannel pipette, the samples were disrupted to full power in a TissueLyser (Qiagen) for 3min. Then we added 360ul of CIA24:1 to each tube and both racks were centrifuged at 4000 RPM for 20min (ThermoScientific SorvallST16). Afterwards the upper phases were transferred carefully to clean rack-wells and 150ul of isopropanol were added to each well and centrifuged at 4000RPM for 20min. The liquid phases were removed and the pellets were washed twice with ethanol 70% and centrifuged at 4000RPM for 5min. Finally the pellets were resuspended in 100ul of RNAse and incubated at 37ºC for 60min. To evaluate the yield and purity, this prococol was compared against results obtained with four commercial extraction kits: DNeasy Plant Handbook® (Qiagen), Power Plant DNA® Isolation Kit (MOBIO), Wizard® Genomic DNA Purification Kit (Promega), ZR Plant / Seed DNA MiniPrep® (Zymo). The procedure of rupture was the same for all protocols and in all cases racks of 96 wells were used. The data of absorption spectrum indicate an average concentration of 400ng/ul for CTAB protocol, Qiagen 150ng/ul, Wizard 165ng/ul MOBIO 85ng/ul for Zymo. Comparing this method to the commercial is demostrated that the final concentration superior, the time os processing is lower, also it is considerable cheaper.
650 $aFORESTACIÓN
653 $aGENÉTICA FORESTAL
700 1 $aTORRES, D.
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INIA Tacuarembó (TBO) |
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