Transgene expression driven by heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill.) |
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Authors: | John R Gittins Till K Pellny Elizabeth R Hiles Cristina Rosa Stefano Biricolti David J James |
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Institution: | (1) Plant Breeding and Biotechnology, Horticulture Research International, East Malling, West Malling, Kent, ME19 6BJ, UK,;(2) Dipartimento di Ortoflorofrutticoltura, Università degli Studi di Firenze, 50144 Firenze, Italy, IT |
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Abstract: | It is desirable that the expression of transgenes in genetically modified 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 β-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 influence 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.
Received: 15 June 1999 / Accepted: 12 August 1999 |
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Keywords: | : Gene promoter – β -Glucuronidase – Malus (transgenic) – Ribulose-1 5-bisphosphate carboxylase/oxygenase(SSU) – Transgene expression (gusA) – Transgenic apple |
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