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Transgene expression in the vegetative tissues of apple driven by the vascular-specific rolC and CoYMV promoters

Transgene expression in the vegetative tissues of apple driven by the vascular-specific rolC and CoYMV promoters
Transgene expression in the vegetative tissues of apple driven by the vascular-specific rolC and CoYMV promoters
The ability of the heterologous promoters, rolCP and CoYMVP, to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill.) has been studied using transgenic plants produced by Agrobacterium-mediated transformation. Replicate plants of each transgenic clone were propagated in soil to a uniform size and samples of leaf, petiole, stem, and root were taken for the measurement of β-glucuronidase (GUS) activity by fluorometric assay. The levels of expression were compared with those in tissues of a representative clone containing the CaMV 35S promoter. These quantitative GUS data were related to the copy number of transgene loci assessed by Southern blotting. The CoYMV promoter was slightly more active than the rolC promoter, although both expressed gusA at a lower level than the CaMV 35S promoter. In clones containing the rolC promoter with multiple transgene loci, expression values were generally among the highest or lowest in the range. The precise location of GUS activity in each tissue was identified by staining of whole leaves and tissue sections with a chromogenic substrate. This analysis demonstrated that with both the rolC and CoYMV promoters the reporter gene activity was primarily localised to vascular tissues, particularly the phloem. Our results indicate that both promoters would be suitable to drive the expression of transgenes to combat pests and diseases of apple that are dependent on interaction with the phloem.
apple, CoYMV, gusA expression, promoters, rolC, vascular-specific
0962-8819
391-402
Gittins, John R.
c4d269cc-aae0-4182-bc81-78dc724f7d95
Pellny, Till K.
25771847-55af-4089-8b1a-db386c74f8ea
Biricolti, Stefano
215b7d13-f709-4655-9dc4-8501a50e9776
Hiles, Elizabeth R.
99c5e1c8-334f-449a-bfdc-6e299d65a4f9
Passey, Andrew J.
04c8d08a-a304-4e33-9271-e2c4fa24da25
James, David J.
2c41d97c-8486-4379-b253-e5896572e81a
Gittins, John R.
c4d269cc-aae0-4182-bc81-78dc724f7d95
Pellny, Till K.
25771847-55af-4089-8b1a-db386c74f8ea
Biricolti, Stefano
215b7d13-f709-4655-9dc4-8501a50e9776
Hiles, Elizabeth R.
99c5e1c8-334f-449a-bfdc-6e299d65a4f9
Passey, Andrew J.
04c8d08a-a304-4e33-9271-e2c4fa24da25
James, David J.
2c41d97c-8486-4379-b253-e5896572e81a

Gittins, John R., Pellny, Till K., Biricolti, Stefano, Hiles, Elizabeth R., Passey, Andrew J. and James, David J. (2003) Transgene expression in the vegetative tissues of apple driven by the vascular-specific rolC and CoYMV promoters. Transgenic Research, 12 (4), 391-402. (doi:10.1023/A:1024286405310).

Record type: Article

Abstract

The ability of the heterologous promoters, rolCP and CoYMVP, to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill.) has been studied using transgenic plants produced by Agrobacterium-mediated transformation. Replicate plants of each transgenic clone were propagated in soil to a uniform size and samples of leaf, petiole, stem, and root were taken for the measurement of β-glucuronidase (GUS) activity by fluorometric assay. The levels of expression were compared with those in tissues of a representative clone containing the CaMV 35S promoter. These quantitative GUS data were related to the copy number of transgene loci assessed by Southern blotting. The CoYMV promoter was slightly more active than the rolC promoter, although both expressed gusA at a lower level than the CaMV 35S promoter. In clones containing the rolC promoter with multiple transgene loci, expression values were generally among the highest or lowest in the range. The precise location of GUS activity in each tissue was identified by staining of whole leaves and tissue sections with a chromogenic substrate. This analysis demonstrated that with both the rolC and CoYMV promoters the reporter gene activity was primarily localised to vascular tissues, particularly the phloem. Our results indicate that both promoters would be suitable to drive the expression of transgenes to combat pests and diseases of apple that are dependent on interaction with the phloem.

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More information

Published date: August 2003
Keywords: apple, CoYMV, gusA expression, promoters, rolC, vascular-specific
Organisations: Ocean and Earth Science

Identifiers

Local EPrints ID: 380342
URI: http://eprints.soton.ac.uk/id/eprint/380342
ISSN: 0962-8819
PURE UUID: 18addb65-94ab-46f6-abaf-0e34208f9d6d

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Date deposited: 12 Aug 2015 15:06
Last modified: 08 Jan 2022 06:20

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Contributors

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

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