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The Brassica napus extA promoter: a novel alternative promoter to CaMV 35S for directing transgene expression to young stem tissues and load bearing regions of transgenic apple trees (Malus pumila Mill.)

The Brassica napus extA promoter: a novel alternative promoter to CaMV 35S for directing transgene expression to young stem tissues and load bearing regions of transgenic apple trees (Malus pumila Mill.)
The Brassica napus extA promoter: a novel alternative promoter to CaMV 35S for directing transgene expression to young stem tissues and load bearing regions of transgenic apple trees (Malus pumila Mill.)


The Brassica napus extensin A gene is highly expressed in root tissue of oilseed rape. In an attempt to identify an effective root-specific promoter for biotechnological applications, we have examined the ability of the ?940 extA promoter to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill cv. Greensleeves). Transgenic apple lines were produced by Agrobacterium tumefaciens-mediated transformation and GUS activity was analysed both quantitatively and qualitatively. The extA promoter was active in all tissues of young plants in all 15 clones examined. However Southern blot data suggested that only a proportion of the population contained the entire promoter and that others had suffered deletions of unknown length. This may have contributed to the variation seen in the quantitative and qualitative expression of GUS. Specific GUS activity was highest in the stems where it approached, and in some clones, exceeded that using the constitutive CaMV 35S promoter. Histochemical analysis confirmed that GUS was localised to tissues involved in structural support of the stem. Staining was particularly intense at nodal junctions where high tensile stress is exerted on the tissues. Maturing phloem tissues showed localisation of expression to the phloem parenchyma cells and phloem fibres. Transverse sections of the root revealed staining of primary procambial tissues including the young endodermis but no staining was seen in the cortex. Although the ?940 extA promoter is clearly not root-specific in apple, it is likely to have useful biotechnological applications in tree species.
Apple, extA promoter, gusA expression, Transgenic trees, Vegetative tissues
1380-3743
51-62
Gittins, John R.
c4d269cc-aae0-4182-bc81-78dc724f7d95
Hiles, Elizabeth R.
99c5e1c8-334f-449a-bfdc-6e299d65a4f9
Pellny, Till K.
25771847-55af-4089-8b1a-db386c74f8ea
Biricolti, Stefano
215b7d13-f709-4655-9dc4-8501a50e9776
James, David J.
2c41d97c-8486-4379-b253-e5896572e81a
Gittins, John R.
c4d269cc-aae0-4182-bc81-78dc724f7d95
Hiles, Elizabeth R.
99c5e1c8-334f-449a-bfdc-6e299d65a4f9
Pellny, Till K.
25771847-55af-4089-8b1a-db386c74f8ea
Biricolti, Stefano
215b7d13-f709-4655-9dc4-8501a50e9776
James, David J.
2c41d97c-8486-4379-b253-e5896572e81a

Gittins, John R., Hiles, Elizabeth R., Pellny, Till K., Biricolti, Stefano and James, David J. (2001) The Brassica napus extA promoter: a novel alternative promoter to CaMV 35S for directing transgene expression to young stem tissues and load bearing regions of transgenic apple trees (Malus pumila Mill.). Molecular Breeding, 7 (1), 51-62. (doi:10.1023/A:1009637706570).

Record type: Article

Abstract



The Brassica napus extensin A gene is highly expressed in root tissue of oilseed rape. In an attempt to identify an effective root-specific promoter for biotechnological applications, we have examined the ability of the ?940 extA promoter to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill cv. Greensleeves). Transgenic apple lines were produced by Agrobacterium tumefaciens-mediated transformation and GUS activity was analysed both quantitatively and qualitatively. The extA promoter was active in all tissues of young plants in all 15 clones examined. However Southern blot data suggested that only a proportion of the population contained the entire promoter and that others had suffered deletions of unknown length. This may have contributed to the variation seen in the quantitative and qualitative expression of GUS. Specific GUS activity was highest in the stems where it approached, and in some clones, exceeded that using the constitutive CaMV 35S promoter. Histochemical analysis confirmed that GUS was localised to tissues involved in structural support of the stem. Staining was particularly intense at nodal junctions where high tensile stress is exerted on the tissues. Maturing phloem tissues showed localisation of expression to the phloem parenchyma cells and phloem fibres. Transverse sections of the root revealed staining of primary procambial tissues including the young endodermis but no staining was seen in the cortex. Although the ?940 extA promoter is clearly not root-specific in apple, it is likely to have useful biotechnological applications in tree species.

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More information

Published date: January 2001
Keywords: Apple, extA promoter, gusA expression, Transgenic trees, Vegetative tissues
Organisations: Ocean and Earth Science

Identifiers

Local EPrints ID: 380349
URI: https://eprints.soton.ac.uk/id/eprint/380349
ISSN: 1380-3743
PURE UUID: 84c34c70-8e20-4ebc-9e03-d9df14ae9013

Catalogue record

Date deposited: 12 Aug 2015 15:40
Last modified: 15 Jul 2019 21:08

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