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Transgene expression driven by heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill.)

Transgene expression driven by heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill.)
Transgene expression driven by heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill.)
It is desirable that the expression of transgenes in genetically modiÆed crops is restricted to the tissues requiring the encoded activity. To this end, we have studied the ability of the heterologous ribulose-1,5- bisphosphate carboxylase/oxygenase (Rubisco) small-subunit (SSU) gene promoters, RBCS3CP (0.8 kbp) from tomato (hycopersion esculentum Mill.) and SRS1P (1.5 kbp) from soybean (Glycine max [h.] Mers.), to drive expression of the b-glucuronidase (gusA) marker gene in apple (Malus pumila Mill.). Transgenic lines of cultivar Greensleeves were produced by Agrobacterium-mediated transformation and the level of gusA expression in the vegetative tissues of young plants was compared with that produced using the cauliflower mosaic virus (CaMV) 35S promoter. These quantitative GUS data were assessed for their relationship to the copy number of transgene loci. The precise location of GUS activity in leaves was identified histochemically. The heterologous SSU promoters were active primarily in the green vegetative tissues of apple, although activity in the roots was noticeably higher with the RBCS3C promoter than with the SRS1 promoter. The mean GUS activity in leaf tissue of the SSU promoter transgenics was approximately half that of plants containing the CaMV 35S promoter. Histochemical analysis demonstrated that GUS activity was localised to the mesophyll and palisade cells of the leaf. The inØuence of light on expression was also determined. The activity of the SRS1 promoter was strictly dependent on light, whereas that of the RBCS3C promoter appeared not to be. Both SSU promoters would be suitable for the expression of transgenes in green photosynthetic tissues of apple.
Gene promoter, b-Glucuronidase, Malus(transgenic), Ribulose-1, 5-bisphosphate carboxylase/ oxygenase(SSU), Transgene expression (gusA), Transgenic apple
0032-0935
232-240
Gittins, John R.
c4d269cc-aae0-4182-bc81-78dc724f7d95
Pellny, Till K.
25771847-55af-4089-8b1a-db386c74f8ea
Hiles, Elizabeth R.
99c5e1c8-334f-449a-bfdc-6e299d65a4f9
Rosa, Christina
44b5996b-0bdf-4e01-9212-b849626f9baa
Biricolti, Stefano
215b7d13-f709-4655-9dc4-8501a50e9776
James, David J.
2c41d97c-8486-4379-b253-e5896572e81a
Gittins, John R.
c4d269cc-aae0-4182-bc81-78dc724f7d95
Pellny, Till K.
25771847-55af-4089-8b1a-db386c74f8ea
Hiles, Elizabeth R.
99c5e1c8-334f-449a-bfdc-6e299d65a4f9
Rosa, Christina
44b5996b-0bdf-4e01-9212-b849626f9baa
Biricolti, Stefano
215b7d13-f709-4655-9dc4-8501a50e9776
James, David J.
2c41d97c-8486-4379-b253-e5896572e81a

Gittins, John R., Pellny, Till K., Hiles, Elizabeth R., Rosa, Christina, Biricolti, Stefano and James, David J. (2000) Transgene expression driven by heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill.). Planta, 310, 232-240.

Record type: Article

Abstract

It is desirable that the expression of transgenes in genetically modiÆed crops is restricted to the tissues requiring the encoded activity. To this end, we have studied the ability of the heterologous ribulose-1,5- bisphosphate carboxylase/oxygenase (Rubisco) small-subunit (SSU) gene promoters, RBCS3CP (0.8 kbp) from tomato (hycopersion esculentum Mill.) and SRS1P (1.5 kbp) from soybean (Glycine max [h.] Mers.), to drive expression of the b-glucuronidase (gusA) marker gene in apple (Malus pumila Mill.). Transgenic lines of cultivar Greensleeves were produced by Agrobacterium-mediated transformation and the level of gusA expression in the vegetative tissues of young plants was compared with that produced using the cauliflower mosaic virus (CaMV) 35S promoter. These quantitative GUS data were assessed for their relationship to the copy number of transgene loci. The precise location of GUS activity in leaves was identified histochemically. The heterologous SSU promoters were active primarily in the green vegetative tissues of apple, although activity in the roots was noticeably higher with the RBCS3C promoter than with the SRS1 promoter. The mean GUS activity in leaf tissue of the SSU promoter transgenics was approximately half that of plants containing the CaMV 35S promoter. Histochemical analysis demonstrated that GUS activity was localised to the mesophyll and palisade cells of the leaf. The inØuence of light on expression was also determined. The activity of the SRS1 promoter was strictly dependent on light, whereas that of the RBCS3C promoter appeared not to be. Both SSU promoters would be suitable for the expression of transgenes in green photosynthetic tissues of apple.

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More information

Published date: 2000
Keywords: Gene promoter, b-Glucuronidase, Malus(transgenic), Ribulose-1, 5-bisphosphate carboxylase/ oxygenase(SSU), Transgene expression (gusA), Transgenic apple
Organisations: Ocean and Earth Science

Identifiers

Local EPrints ID: 380351
URI: http://eprints.soton.ac.uk/id/eprint/380351
ISSN: 0032-0935
PURE UUID: c56cdcff-5fbf-43c8-aed3-4c28888b659d

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Date deposited: 12 Aug 2015 16:08
Last modified: 15 Jul 2019 21:09

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Contributors

Author: John R. Gittins
Author: Till K. Pellny
Author: Elizabeth R. Hiles
Author: Christina Rosa
Author: Stefano Biricolti
Author: David J. James

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