Leaf Disk Transformation Using Agrobacterium tumefaciens-Expression of Heterologous Genes in Tobacco

Gallois, Patrick; Marinho, Paulo

doi:10.1385/0-89603-321-x:39

Cited by 129 publications

(110 citation statements)

References 16 publications

Supporting

Mentioning

109

Contrasting

Unclassified

Order By: Relevance

“…Leaf disks were incubated with Agrobacterium for 30 min, washed, and placed for cocultivation in MS medium containing 2 mg/mL of kinetin and 0.8 mg/mL of indoleacetic acid. The rooting medium, contrary to Gallois and Marinho (1995), did not contain hormones. Analysis was performed on leaves of T0 plants.…”

Section: Stable Transformations Of Tobaccomentioning

confidence: 86%

“…To create AtRGP2:GFP or GONST1:YFP expressing transgenic plants, wild-type N. tabacum cv Samson were transformed by Agrobacterium strain GV3101 harboring pBinAtRGP2:GFP or pBinGONST1:YFP plasmids accordingly, using a modified leaf disk method (Gallois and Marinho, 1995). Leaf disks were incubated with Agrobacterium for 30 min, washed, and placed for cocultivation in MS medium containing 2 mg/mL of kinetin and 0.8 mg/mL of indoleacetic acid.…”

Section: Stable Transformations Of Tobaccomentioning

confidence: 99%

See 1 more Smart Citation

Class 1 Reversibly Glycosylated Polypeptides Are Plasmodesmal-Associated Proteins Delivered to Plasmodesmata via the Golgi Apparatus

et al. 2005

View full text Add to dashboard Cite

SE-WAP41, a salt-extractable 41-kD wall-associated protein that is associated with walls of etiolated maize (Zea mays) seedlings and is recognized by an antiserum previously reported to label plasmodesmata and the Golgi, was cloned, sequenced, and found to be a class 1 reversibly glycosylated polypeptide (C1RGP). Protein gel blot analysis of cell fractions with an antiserum against recombinant SE-WAP41 showed it to be enriched in the wall fraction. RNA gel blot analysis along the mesocotyl developmental axis and during deetiolation demonstrates that high SE-WAP41 transcript levels correlate spatially and temporally with primary and secondary plasmodesmata (Pd) formation. All four of the Arabidopsis thaliana C1RGP proteins, when fused to green fluorescent protein (GFP) and transiently expressed in tobacco (Nicotiana tabacum) epidermal cells, display fluorescence patterns indicating they are Golgi- and plasmodesmal-associated proteins. Localization to the Golgi apparatus was verified by colocalization of transiently expressed AtRGP2 fused to cyan fluorescence protein together with a known Golgi marker, Golgi Nucleotide Sugar Transporter 1 fused to yellow fluorescent protein (GONST1:YFP). In transgenic tobacco, AtRGP2:GFP fluorescence is punctate, is present only in contact walls between cells, and colocalizes with aniline blue–stained callose present around Pd. In plasmolyzed cells, AtRGP2:GFP remains wall embedded, whereas GONST1:YFP cannot be found embedded in cell walls. This result implies that the targeting to Pd is not due to a default pathway for Golgi-localized fusion proteins but is specific to C1RGPs. Treatment with the Golgi disrupting drug Brefeldin A inhibits Pd labeling by AtRGP2:GFP. Integrating these data, we conclude that C1RGPs are plasmodesmal-associated proteins delivered to plasmodesmata via the Golgi apparatus.

show abstract

Section: Stable Transformations Of Tobaccomentioning

confidence: 86%

Section: Stable Transformations Of Tobaccomentioning

confidence: 99%

Class 1 Reversibly Glycosylated Polypeptides Are Plasmodesmal-Associated Proteins Delivered to Plasmodesmata via the Golgi Apparatus

et al. 2005

View full text Add to dashboard Cite

show abstract

“…Both constructs (pfld and cfld, directing Fld expression to plastids and the cytosol, respectively) were excised from pDH51 plasmids with EcoRI and inserted into compatible sites of the binary vector pCAMBIA 2200 (Hajdukiewicz et al, 1994). Recombinant plasmids were introduced into the genome of tobacco (Nicotiana tabacum cv Petit Havana) through Agrobacterium tumefaciens-mediated leaf disc transformation (Gallois and Marinho, 1995).…”

Section: Construction Of Fld Expression Vectors and Tobacco Transformmentioning

confidence: 99%

Functional Replacement of Ferredoxin by a Cyanobacterial Flavodoxin in Tobacco Confers Broad-Range Stress Tolerance

et al. 2006

View full text Add to dashboard Cite

Chloroplast ferredoxin (Fd) plays a pivotal role in plant cell metabolism by delivering reducing equivalents to various essential oxidoreductive pathways. Fd levels decrease under adverse environmental conditions in many microorganisms, including cyanobacteria, which share a common ancestor with chloroplasts. Conversely, stress situations induce the synthesis of flavodoxin (Fld), an electron carrier flavoprotein not found in plants, which can efficiently replace Fd in most electron transfer processes. We report here that chloroplast Fd also declined in plants exposed to oxidants or stress conditions. A purified cyanobacterial Fld was able to mediate plant Fd-dependent reactions in vitro, including NADP 1 and thioredoxin reduction. Tobacco (Nicotiana tabacum) plants expressing Fld in chloroplasts displayed increased tolerance to multiple sources of stress, including redox-cycling herbicides, extreme temperatures, high irradiation, water deficit, and UV radiation. Oxidant buildup and oxidative inactivation of thioredoxin-dependent plastidic enzymes were decreased in stressed plants expressing plastid-targeted Fld, suggesting that development of the tolerant phenotype relied on productive interaction of this flavoprotein with Fd-dependent oxidoreductive pathways of the host, most remarkably, thioredoxin reduction. The use of Fld provides new tools to investigate the requirements of photosynthesis in planta and to increase plant stress tolerance based on the introduction of a cyanobacterial product that is free from endogenous regulation in higher plants.

show abstract

“…Agrobacterium tumefaciens strain LBA 4404 was transformed with pNtAQP1AS by triparental mating. Transformation and regeneration of transgenic tobacco (Nicotiana tabacum) was performed according to the protocol provided by Gallois and Marinho (1995) with the exception that 500 mg/L carbenicillin was substituted for cefotaxime.…”

Section: Antisense Construct and Transformationmentioning

confidence: 99%

PIP1 Plasma Membrane Aquaporins in Tobacco

Tyree²,

et al. 2002

View full text Add to dashboard Cite

The molecular functions of several aquaporins are well characterized (e.g., by analysis of aquaporin-expressing Xenopus oocytes). However, their significance in the physiology of water transport in multicellular organisms remains uncertain. The tobacco plasma membrane aquaporin NtAQP1 was used to elucidate this issue. By comparing antisense plants that were inhibited in NtAQP1 expression with control plants, we found evidence for NtAQP1 function in cellular and whole-plant water relations. The consequences of a decrease in cellular water permeability were determined by measurement of transpiration rate and stem and leaf water potential as well as growth experiments under extreme soil water depletion. Plants impaired in NtAQP1 expression showed reduced root hydraulic conductivity and lower water stress resistance. In conclusion, our results emphasize the importance of symplastic aquaporin-mediated water transport in whole-plant water relations.

show abstract

Leaf Disk Transformation Using Agrobacterium tumefaciens-Expression of Heterologous Genes in Tobacco

Cited by 129 publications

References 16 publications

Class 1 Reversibly Glycosylated Polypeptides Are Plasmodesmal-Associated Proteins Delivered to Plasmodesmata via the Golgi Apparatus

Class 1 Reversibly Glycosylated Polypeptides Are Plasmodesmal-Associated Proteins Delivered to Plasmodesmata via the Golgi Apparatus

Functional Replacement of Ferredoxin by a Cyanobacterial Flavodoxin in Tobacco Confers Broad-Range Stress Tolerance

PIP1 Plasma Membrane Aquaporins in Tobacco

Contact Info

Product

Resources

About