Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens
An efficient genetic transformation procedure has been developed via Agrobacterium tumefaciens on Hevea brasiliensis friable calli, and transgenic callus lines were established and characterised at the molecular level. Embryogenic friable calli precultured a fortnight on a CaCl2-free maintenance medium were cocultivated with agrobacteria, which were induced with acetosyringone and resuspended in CaCl2-free medium. These conditions enhanced dramatically the transient activity of the reporter gene encoding [bêta]-glucuronidase (GUS) [2]. Cocultivated friable calli were directly transferred as tissue aggregates of 1-2 mm on a maintenance medium supplemented with a bacterial static agent, the ticarcillin, in order to favour plant tissue proliferation and to inhibit the Agrobacterium overgrowth [3]. After two phases of proliferation for 3 weeks, GUS-positive cell clusters were observed, indicating that transgenic cells could proliferate. The medium was then supplemented with a selective agent, the paromomycin, at a concentration increasing at each subculture, until complete growth inhibition of non-transformed cells. DNA amplification of the transgenes was performed on the paromomycin-resistant and GUS-positive callus lines. Four transgenic callus lines have thus been established and characterised [1]. The availability of this transgenic material represents a way for clonal propagation of genetically engineered rubber trees.
Main Authors: | , , , , , , , , |
---|---|
Format: | conference_item biblioteca |
Language: | eng |
Published: |
CIRAD
|
Subjects: | F30 - Génétique et amélioration des plantes, |
Online Access: | http://agritrop.cirad.fr/513749/ http://agritrop.cirad.fr/513749/1/ID513749.pdf |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
id |
dig-cirad-fr-513749 |
---|---|
record_format |
koha |
spelling |
dig-cirad-fr-5137492023-07-19T13:27:36Z http://agritrop.cirad.fr/513749/ http://agritrop.cirad.fr/513749/ Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens. Montoro Pascal, Rattana Wiparat, Teinseree Nongluk, Tadakittisarn Sukuntaros, Pujade-Renaud Valérie, Michaux-Ferrière Nicole, Monkolsook Yupa, Kanthapura Reena, Adunsadthapong Saisunee. 2001. In : Biotechnology and rubber tree : Proceedings of IRRDB symposium, 25-28 September 2001, Montpellier, France. Sainte-Beuve Jérôme (ed.). IRRDB, CIRAD-CP-HEVEA. Montpellier : CIRAD IRRDB Symposium, Montpellier, France, 25 Septembre 2001/28 Septembre 2001. Researchers Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens Montoro, Pascal Rattana, Wiparat Teinseree, Nongluk Tadakittisarn, Sukuntaros Pujade-Renaud, Valérie Michaux-Ferrière, Nicole Monkolsook, Yupa Kanthapura, Reena Adunsadthapong, Saisunee eng 2001 CIRAD Biotechnology and rubber tree : Proceedings of IRRDB symposium, 25-28 September 2001, Montpellier, France F30 - Génétique et amélioration des plantes An efficient genetic transformation procedure has been developed via Agrobacterium tumefaciens on Hevea brasiliensis friable calli, and transgenic callus lines were established and characterised at the molecular level. Embryogenic friable calli precultured a fortnight on a CaCl2-free maintenance medium were cocultivated with agrobacteria, which were induced with acetosyringone and resuspended in CaCl2-free medium. These conditions enhanced dramatically the transient activity of the reporter gene encoding [bêta]-glucuronidase (GUS) [2]. Cocultivated friable calli were directly transferred as tissue aggregates of 1-2 mm on a maintenance medium supplemented with a bacterial static agent, the ticarcillin, in order to favour plant tissue proliferation and to inhibit the Agrobacterium overgrowth [3]. After two phases of proliferation for 3 weeks, GUS-positive cell clusters were observed, indicating that transgenic cells could proliferate. The medium was then supplemented with a selective agent, the paromomycin, at a concentration increasing at each subculture, until complete growth inhibition of non-transformed cells. DNA amplification of the transgenes was performed on the paromomycin-resistant and GUS-positive callus lines. Four transgenic callus lines have thus been established and characterised [1]. The availability of this transgenic material represents a way for clonal propagation of genetically engineered rubber trees. conference_item info:eu-repo/semantics/conferenceObject Conference info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/513749/1/ID513749.pdf text Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html http://agritrop.cirad.fr/513714/ |
institution |
CIRAD FR |
collection |
DSpace |
country |
Francia |
countrycode |
FR |
component |
Bibliográfico |
access |
En linea |
databasecode |
dig-cirad-fr |
tag |
biblioteca |
region |
Europa del Oeste |
libraryname |
Biblioteca del CIRAD Francia |
language |
eng |
topic |
F30 - Génétique et amélioration des plantes F30 - Génétique et amélioration des plantes |
spellingShingle |
F30 - Génétique et amélioration des plantes F30 - Génétique et amélioration des plantes Montoro, Pascal Rattana, Wiparat Teinseree, Nongluk Tadakittisarn, Sukuntaros Pujade-Renaud, Valérie Michaux-Ferrière, Nicole Monkolsook, Yupa Kanthapura, Reena Adunsadthapong, Saisunee Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens |
description |
An efficient genetic transformation procedure has been developed via Agrobacterium tumefaciens on Hevea brasiliensis friable calli, and transgenic callus lines were established and characterised at the molecular level. Embryogenic friable calli precultured a fortnight on a CaCl2-free maintenance medium were cocultivated with agrobacteria, which were induced with acetosyringone and resuspended in CaCl2-free medium. These conditions enhanced dramatically the transient activity of the reporter gene encoding [bêta]-glucuronidase (GUS) [2]. Cocultivated friable calli were directly transferred as tissue aggregates of 1-2 mm on a maintenance medium supplemented with a bacterial static agent, the ticarcillin, in order to favour plant tissue proliferation and to inhibit the Agrobacterium overgrowth [3]. After two phases of proliferation for 3 weeks, GUS-positive cell clusters were observed, indicating that transgenic cells could proliferate. The medium was then supplemented with a selective agent, the paromomycin, at a concentration increasing at each subculture, until complete growth inhibition of non-transformed cells. DNA amplification of the transgenes was performed on the paromomycin-resistant and GUS-positive callus lines. Four transgenic callus lines have thus been established and characterised [1]. The availability of this transgenic material represents a way for clonal propagation of genetically engineered rubber trees. |
format |
conference_item |
topic_facet |
F30 - Génétique et amélioration des plantes |
author |
Montoro, Pascal Rattana, Wiparat Teinseree, Nongluk Tadakittisarn, Sukuntaros Pujade-Renaud, Valérie Michaux-Ferrière, Nicole Monkolsook, Yupa Kanthapura, Reena Adunsadthapong, Saisunee |
author_facet |
Montoro, Pascal Rattana, Wiparat Teinseree, Nongluk Tadakittisarn, Sukuntaros Pujade-Renaud, Valérie Michaux-Ferrière, Nicole Monkolsook, Yupa Kanthapura, Reena Adunsadthapong, Saisunee |
author_sort |
Montoro, Pascal |
title |
Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens |
title_short |
Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens |
title_full |
Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens |
title_fullStr |
Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens |
title_full_unstemmed |
Production of transgenic callus lines in Hevea brasiliensis via Agrobacterium tumefaciens |
title_sort |
production of transgenic callus lines in hevea brasiliensis via agrobacterium tumefaciens |
publisher |
CIRAD |
url |
http://agritrop.cirad.fr/513749/ http://agritrop.cirad.fr/513749/1/ID513749.pdf |
work_keys_str_mv |
AT montoropascal productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT rattanawiparat productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT teinsereenongluk productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT tadakittisarnsukuntaros productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT pujaderenaudvalerie productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT michauxferrierenicole productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT monkolsookyupa productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT kanthapurareena productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens AT adunsadthapongsaisunee productionoftransgeniccalluslinesinheveabrasiliensisviaagrobacteriumtumefaciens |
_version_ |
1772404004509188096 |