Hybrid genes in the analysis of transformation conditions

Negrutiu, Ioan; Shillito, Raymond D.; Potrykus, Ingo; Biasini, G.; Sala, F.

doi:10.1007/bf00015814

Cited by 484 publications

(185 citation statements)

References 23 publications

Supporting

Mentioning

182

Contrasting

Order By: Relevance

“…However, it is possible that not all the cells in the protoplast population are equally competent to transformation. In addition, N. plumbaginifolia protoplasts are 10 times less competent than N. tabacum protoplasts using either polyethylene glycol (PEG) transformation or Ds transposition from plasmid into plant genome 63 electroporation (Negrutiu et aL, 1987). Not all excision events result in integration, as shown by Dooner and Belachew (1989) at the bz locus in maize.…”

Section: Discussionmentioning

confidence: 99%

“…Viviani grown in greenhouse (one batch) or in a controlled culture room (8 h light 75 liE m -2 sec -1 at 25°C 16 h dark at 17°C and a constant 90% relative humidity, the other batch), according to a previously described procedure (Bourgin et al, 1979). Ca(NO3)2-polyethylene glycol-mediated DNA transfer was done as reported by Negrutiu et al (1987). Plasmids were isolated in E. coil strains MC1061 or HB101.…”

Section: Protoplasts Transformation and Plant Regenerationmentioning

confidence: 99%

See 1 more Smart Citation

Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast‐derived cells

1994

View full text Add to dashboard Cite

SummaryNicotiana plurnbaginifolia haploid protoplasts were co-transformed with two plasmids, one with a NPT-II/Ds element and one with a gene encoding an amino-terminal truncated Ac transposese. It is shown that Ds can efficiently transpose from extrachromosomal DNA to IV. plurnbaginifolia chromosomes when the Ac transposese gene is present in trans. Ds has been shown to have transposed into the plant genome in a limited number of copies (1.9 copies per genome), for 21/32 trensgenic lines tested. The flanking sequences present in the original plasmid are missing in these 21 plants. In only two of 21 plants was pert of the transposese construct integrated. By segregation analysis of tranegenic progeny, Ds was shown to be present in the heterozygous state in 10 lines even though haploid protoplaste had been originally transformed. This observation could indicate that integration occurred after or during DNA replication that leads to protoplast diploidization.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Protoplasts Transformation and Plant Regenerationmentioning

confidence: 99%

Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast‐derived cells

1994

View full text Add to dashboard Cite

show abstract

“…Mesophyll protoplasts were isolated from in vitro-grown tobacco plants, as in Negrutiu et al (1987). Freshly isolated protoplasts (4 · 10 4 ppt ll )1 ) were transfected with 10 lg of plasmid DNA by polyethylene glycol-mediated DNA uptake (Walden et al, 1994).…”

Section: Intracellular Protein Localizationmentioning

confidence: 99%

The Brassica juncea BjCdR15, an ortholog of Arabidopsis TGA3, is a regulator of cadmium uptake, transport and accumulation in shoots and confers cadmium tolerance in transgenic plants

et al. 2009

View full text Add to dashboard Cite

Summary• A bZIP transcription factor from Brassica juncea (BjCdR15) was isolated by the cDNA-amplified fragment length polymorphism technique after cadmium treatment. Sequence analysis indicated high similarity between BjCdR15 and Arabidopsis TGA3. In Arabidopsis, TGA3 transcription is also induced by cadmium; hence, we investigated whether BjCdR15 is involved in cadmium tolerance and whether it can functionally replace TGA3 protein in Arabidopsis tga3-2 mutant plants.• BjCdR15 expression was detected mainly in the epidermis and vascular system of cadmium-treated plants, and increased in roots and leaves after cadmium treatment. The overexpression of BjCdR15 in Arabidopsis and tobacco enhanced cadmium tolerance: overexpressing plants showed high cadmium accumulation in shoots. Conversely, Arabidopsis tga3-2 mutant plants showed high cadmium content in roots and inhibition of its transport to the shoot.• We demonstrated that BjCdR15 can functionally replace TGA3: in 35S:: BjCdR15-tga3-2 plants, the long-distance transport of cadmium from root to shoot was restored and these plants showed an increased cadmium content in shoots compared with all other assays. In addition, BjCdR15 ⁄ TGA3 regulated the synthesis of phytochelatin synthase and the expression of several metal transporters.• The results indicate that BjCdR15 ⁄ TGA3 transcription factors play a crucial role in the regulation of cadmium uptake by roots and in its long-distance root to shoot transport. BjCdR15 ⁄ TGA3 may thus be considered as useful candidates for potential biotechnological applications in the phytoextraction of cadmium from polluted soils.

show abstract

“…Tobacco (Nicotiana tabacum) protoplasts were isolated from leaves of cv SR1 (Nagy and Maliga, 1976). A polyethylene glycolbased transformation protocol then was used (Negrutiu et al, 1987;Freydl et al, 1995;Di Sansebastiano et al, 1998), and the transformed protoplasts were visualized 24 hr after transformation. …”

mentioning

confidence: 99%

Demonstration in Yeast of the Function of BP-80, a Putative Plant Vacuolar Sorting Receptor

et al. 2001

View full text Add to dashboard Cite

BP-80, later renamed VSR PS-1 , is a putative receptor involved in sorting proteins such as proaleurain to the lytic vacuole, with its N-terminal domain recognizing the vacuolar sorting determinant. Although all VSR PS-1 characteristics and in vitro binding properties described so far favored its receptor function, this function remained to be demonstrated. Here, we used green fluorescent protein (GFP) as a reporter in a yeast mutant strain defective for its own vacuolar receptor, Vps10p. By expressing VSR PS-1 together with GFP fused to the vacuolar sorting determinant of petunia proaleurain, we were able to efficiently redirect the reporter to the yeast vacuole. VSR PS-1 is ineffective on GFP either alone or when fused with another type of plant vacuolar sorting determinant from a chitinase. The plant VSR PS-1 therefore interacts specifically with the proaleurain vacuolar sorting determinant in vivo, and this interaction leads to the transport of the reporter protein through the yeast secretory pathway to the vacuole. This finding demonstrates VSR PS-1 receptor function but also emphasizes the differences in the spectrum of ligands between Vps10p and its plant equivalent. INTRODUCTIONThe plant secretory system is far less easy to study than its yeast counterpart, mainly due to the lack of an equivalent mutant collection. Plant and yeast cells, in opposition to mammalian cells, do not use the mannose 6-phosphatebased lysosomal sorting tag (Kornfeld, 1992); instead, the sorting information is carried by a peptidic sequence. In yeast, the two vacuolar proteins carboxypeptidase Y (CPY; Johnson et al., 1987) and proteinase A (Klionsky and Emr, 1989) both are produced with an N-terminal propeptide responsible for the vacuolar location of the mature protein. Almost 50 mutants defective for vacuolar protein sorting ( vps ) have allowed the identification of proteins involved in the yeast vacuolar sorting pathway. One of these proteins, Vps10p, plays a central role because it is the vacuolar receptor for CPY (Marcusson et al., 1994). Vps10p also is able to send foreign or malfolded proteins to the vacuole for degradation in a process that is believed to be independent of any sorting signal (Hong et al., 1996).In plants, two main types of vacuolar sorting determinants (VSDs) have been well studied and are believed to correspond to two separate vacuolar sorting pathways. The first type could be defined as a sequence-specific VSD (ssVSD) and has been well studied for barley aleurain (Holwerda et al., 1992) and sweet potato sporamin A (Matsuoka and Nakamura, 1991). For these two examples, the VSD is located within an N-terminal propeptide and contains a conserved tetrapeptide NPIR, the Ile being essential (Matsuoka and Nakamura, 1999). The position of this category of VSD seems to be less important than its sequence specificity, as shown with sporamin A (Koide et al., 1997). The second type of VSD is found in C-terminal propeptides (Ct-VSD); it is rather short with no obvious sequence conservation but needs to be accessibl...

show abstract

Hybrid genes in the analysis of transformation conditions

Cited by 484 publications

References 23 publications

Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast‐derived cells

Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast‐derived cells

The Brassica juncea BjCdR15, an ortholog of Arabidopsis TGA3, is a regulator of cadmium uptake, transport and accumulation in shoots and confers cadmium tolerance in transgenic plants

Demonstration in Yeast of the Function of BP-80, a Putative Plant Vacuolar Sorting Receptor

Contact Info

Product

Resources

About