Differential activity of the mannopine synthase and the CaMV 35S promoters during development of transgenic rapeseed plants

Iván Stefanov, Sándor Fekete, László Bögre, János Pauk, A. Fehér, D. Dudits

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Fusions of the promoter of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) gene to the β-glucuronidase (GUS) reporter gene have been introduced into various cultivars of Brassica napus via Agrobacterium-mediated transformation. Transgenic rapeseed plants have been also regenerated from winter cultivars (Santana, Arabella) by shoot induction from kanamycin-resistant callus tissues on the medium supplemented with AgNO3. Transformation was confirmed by Southern hybridization of genomic DNA from primary transformants and PCR analysis of DNA from second generation seedlings. β-glucuronidase activity analyzed by fluorometric assay or histochemical staining indicated a differential expression pattern for the two promoters. Organogenesis from in vitro cultured callus tissues was coupled with a relative increase of CaMV 35S promoter activity and reduction of mas promoter function. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was the most active in the cotyledons. Etiolated seedlings cultured in dark, showed reduced activity of the mas promoter. At rosette stage, both promoters were more active in elder plant parts than in younger ones. The highest activity values were recorded in cotyledons. After bolting, reduced promoter function was detected in upper parts of the transformed plants. Histological staining showed that the CaMV 35S promoter was active in the cortex, the phloem and the vascular cambium, while the mas promoter directed gene expression in the phloem. In conclusion, both promoters were found to be functional in majority of the studied organs of transgenic rapeseed plants, however the promoter activity varied considerably between organs and tissues at various developmental stages.

Original languageEnglish
Pages (from-to)175-186
Number of pages12
JournalPlant Science
Volume95
Issue number2
DOIs
Publication statusPublished - 1994

Fingerprint

Caulimovirus
Brassica rapa
Cauliflower mosaic virus
Genetically Modified Plants
messenger RNA
rapeseed
Viruses
promoter regions
genetically modified organisms
RNA
Seedlings
Phloem
Cotyledon
Glucuronidase
Bony Callus
Tissue
Sambucus
Cambium
Genes
Staining and Labeling

Keywords

  • Agrobacterium transformation
  • Brassica napus
  • CaMV 35S promoter
  • Expression
  • mas promoter

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry
  • Biotechnology

Cite this

Differential activity of the mannopine synthase and the CaMV 35S promoters during development of transgenic rapeseed plants. / Stefanov, Iván; Fekete, Sándor; Bögre, László; Pauk, János; Fehér, A.; Dudits, D.

In: Plant Science, Vol. 95, No. 2, 1994, p. 175-186.

Research output: Contribution to journalArticle

Stefanov, Iván ; Fekete, Sándor ; Bögre, László ; Pauk, János ; Fehér, A. ; Dudits, D. / Differential activity of the mannopine synthase and the CaMV 35S promoters during development of transgenic rapeseed plants. In: Plant Science. 1994 ; Vol. 95, No. 2. pp. 175-186.
@article{9423033b9dd7496bbdd6eee8ca46524e,
title = "Differential activity of the mannopine synthase and the CaMV 35S promoters during development of transgenic rapeseed plants",
abstract = "Fusions of the promoter of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) gene to the β-glucuronidase (GUS) reporter gene have been introduced into various cultivars of Brassica napus via Agrobacterium-mediated transformation. Transgenic rapeseed plants have been also regenerated from winter cultivars (Santana, Arabella) by shoot induction from kanamycin-resistant callus tissues on the medium supplemented with AgNO3. Transformation was confirmed by Southern hybridization of genomic DNA from primary transformants and PCR analysis of DNA from second generation seedlings. β-glucuronidase activity analyzed by fluorometric assay or histochemical staining indicated a differential expression pattern for the two promoters. Organogenesis from in vitro cultured callus tissues was coupled with a relative increase of CaMV 35S promoter activity and reduction of mas promoter function. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was the most active in the cotyledons. Etiolated seedlings cultured in dark, showed reduced activity of the mas promoter. At rosette stage, both promoters were more active in elder plant parts than in younger ones. The highest activity values were recorded in cotyledons. After bolting, reduced promoter function was detected in upper parts of the transformed plants. Histological staining showed that the CaMV 35S promoter was active in the cortex, the phloem and the vascular cambium, while the mas promoter directed gene expression in the phloem. In conclusion, both promoters were found to be functional in majority of the studied organs of transgenic rapeseed plants, however the promoter activity varied considerably between organs and tissues at various developmental stages.",
keywords = "Agrobacterium transformation, Brassica napus, CaMV 35S promoter, Expression, mas promoter",
author = "Iv{\'a}n Stefanov and S{\'a}ndor Fekete and L{\'a}szl{\'o} B{\"o}gre and J{\'a}nos Pauk and A. Feh{\'e}r and D. Dudits",
year = "1994",
doi = "10.1016/0168-9452(94)90091-4",
language = "English",
volume = "95",
pages = "175--186",
journal = "Plant Science",
issn = "0168-9452",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

TY - JOUR

T1 - Differential activity of the mannopine synthase and the CaMV 35S promoters during development of transgenic rapeseed plants

AU - Stefanov, Iván

AU - Fekete, Sándor

AU - Bögre, László

AU - Pauk, János

AU - Fehér, A.

AU - Dudits, D.

PY - 1994

Y1 - 1994

N2 - Fusions of the promoter of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) gene to the β-glucuronidase (GUS) reporter gene have been introduced into various cultivars of Brassica napus via Agrobacterium-mediated transformation. Transgenic rapeseed plants have been also regenerated from winter cultivars (Santana, Arabella) by shoot induction from kanamycin-resistant callus tissues on the medium supplemented with AgNO3. Transformation was confirmed by Southern hybridization of genomic DNA from primary transformants and PCR analysis of DNA from second generation seedlings. β-glucuronidase activity analyzed by fluorometric assay or histochemical staining indicated a differential expression pattern for the two promoters. Organogenesis from in vitro cultured callus tissues was coupled with a relative increase of CaMV 35S promoter activity and reduction of mas promoter function. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was the most active in the cotyledons. Etiolated seedlings cultured in dark, showed reduced activity of the mas promoter. At rosette stage, both promoters were more active in elder plant parts than in younger ones. The highest activity values were recorded in cotyledons. After bolting, reduced promoter function was detected in upper parts of the transformed plants. Histological staining showed that the CaMV 35S promoter was active in the cortex, the phloem and the vascular cambium, while the mas promoter directed gene expression in the phloem. In conclusion, both promoters were found to be functional in majority of the studied organs of transgenic rapeseed plants, however the promoter activity varied considerably between organs and tissues at various developmental stages.

AB - Fusions of the promoter of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) gene to the β-glucuronidase (GUS) reporter gene have been introduced into various cultivars of Brassica napus via Agrobacterium-mediated transformation. Transgenic rapeseed plants have been also regenerated from winter cultivars (Santana, Arabella) by shoot induction from kanamycin-resistant callus tissues on the medium supplemented with AgNO3. Transformation was confirmed by Southern hybridization of genomic DNA from primary transformants and PCR analysis of DNA from second generation seedlings. β-glucuronidase activity analyzed by fluorometric assay or histochemical staining indicated a differential expression pattern for the two promoters. Organogenesis from in vitro cultured callus tissues was coupled with a relative increase of CaMV 35S promoter activity and reduction of mas promoter function. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was the most active in the cotyledons. Etiolated seedlings cultured in dark, showed reduced activity of the mas promoter. At rosette stage, both promoters were more active in elder plant parts than in younger ones. The highest activity values were recorded in cotyledons. After bolting, reduced promoter function was detected in upper parts of the transformed plants. Histological staining showed that the CaMV 35S promoter was active in the cortex, the phloem and the vascular cambium, while the mas promoter directed gene expression in the phloem. In conclusion, both promoters were found to be functional in majority of the studied organs of transgenic rapeseed plants, however the promoter activity varied considerably between organs and tissues at various developmental stages.

KW - Agrobacterium transformation

KW - Brassica napus

KW - CaMV 35S promoter

KW - Expression

KW - mas promoter

UR - http://www.scopus.com/inward/record.url?scp=0028008224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028008224&partnerID=8YFLogxK

U2 - 10.1016/0168-9452(94)90091-4

DO - 10.1016/0168-9452(94)90091-4

M3 - Article

VL - 95

SP - 175

EP - 186

JO - Plant Science

JF - Plant Science

SN - 0168-9452

IS - 2

ER -