Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions

In order to introduce currently-available genes with agronomical value into banana, two genetic transformation protocols have been optimized. Firstly, regenerable protoplasts isolated from embryogenic cell suspensions of the cultivar Bluggoe have been used for the introduction of several chimaeric u...

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Main Authors: Sagi, L., Remy, S., Verelst, B., Panis, Bartholomeus, Cammue, B.P.A., Volckaert, G., Swennen, Rony L.
Format: Journal Article
Language:Inglés
Published: 1995
Subjects:
Online Access:https://hdl.handle.net/10568/101089
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author Sagi, L.
Remy, S.
Verelst, B.
Panis, Bartholomeus
Cammue, B.P.A.
Volckaert, G.
Swennen, Rony L.
author_browse Cammue, B.P.A.
Panis, Bartholomeus
Remy, S.
Sagi, L.
Swennen, Rony L.
Verelst, B.
Volckaert, G.
author_facet Sagi, L.
Remy, S.
Verelst, B.
Panis, Bartholomeus
Cammue, B.P.A.
Volckaert, G.
Swennen, Rony L.
author_sort Sagi, L.
collection Repository of Agricultural Research Outputs (CGSpace)
description In order to introduce currently-available genes with agronomical value into banana, two genetic transformation protocols have been optimized. Firstly, regenerable protoplasts isolated from embryogenic cell suspensions of the cultivar Bluggoe have been used for the introduction of several chimaeric uidA gene constructs by electroporation. With the inclusion of polyethylene glycol and heat shock, the frequency of transiently expressing protoplasts reached 1.8% as shown by an in situ β-glucuronidase assay. A duplicated 35S promoter with an alfalfa mosaic virus leader sequence (pBI-426) induced the highest expression rate among the constructs tested. Embryogenic cell suspensions of cv. Bluggoe have also been bombarded with accelerated particles coated with a high expression uidA gene construct (pEmuGN) using a biolistic gun. After a partial optimization of the procedure, transient GUS assays reproducibly demonstrated the presence of 400 blue foci in 30 μl of settled cell volume (approximately 25 mg cells). Selection and characterization of antibiotic-resistant transformed cultures is in progress.
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spelling CGSpace1010892023-09-25T09:16:55Z Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions Sagi, L. Remy, S. Verelst, B. Panis, Bartholomeus Cammue, B.P.A. Volckaert, G. Swennen, Rony L. genetics bananas protoplasts embryos (plant) In order to introduce currently-available genes with agronomical value into banana, two genetic transformation protocols have been optimized. Firstly, regenerable protoplasts isolated from embryogenic cell suspensions of the cultivar Bluggoe have been used for the introduction of several chimaeric uidA gene constructs by electroporation. With the inclusion of polyethylene glycol and heat shock, the frequency of transiently expressing protoplasts reached 1.8% as shown by an in situ β-glucuronidase assay. A duplicated 35S promoter with an alfalfa mosaic virus leader sequence (pBI-426) induced the highest expression rate among the constructs tested. Embryogenic cell suspensions of cv. Bluggoe have also been bombarded with accelerated particles coated with a high expression uidA gene construct (pEmuGN) using a biolistic gun. After a partial optimization of the procedure, transient GUS assays reproducibly demonstrated the presence of 400 blue foci in 30 μl of settled cell volume (approximately 25 mg cells). Selection and characterization of antibiotic-resistant transformed cultures is in progress. 1995 2019-04-24T12:29:52Z 2019-04-24T12:29:52Z Journal Article https://hdl.handle.net/10568/101089 en Limited Access Sagi, L., Remy, S., Verelst, B., Panis, B., Cammue, B.P., Volckaert, G. & Swennen, R. (1995). Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions. Euphytica, 85(1-3), 89-95.
spellingShingle genetics
bananas
protoplasts
embryos (plant)
Sagi, L.
Remy, S.
Verelst, B.
Panis, Bartholomeus
Cammue, B.P.A.
Volckaert, G.
Swennen, Rony L.
Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions
title Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions
title_full Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions
title_fullStr Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions
title_full_unstemmed Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions
title_short Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions
title_sort transient gene expression in transformed banana musa cv bluggoe protoplasts and embryogenic cell suspensions
topic genetics
bananas
protoplasts
embryos (plant)
url https://hdl.handle.net/10568/101089
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