scholarly journals 635 Stability of Herbicide Resistance and GUS Expression in Transgenic Hybrid Poplars during Several Years of Field Trials and Vegetative Propagation

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 557B-557
Author(s):  
Richard Meilan ◽  
Caiping Ma ◽  
Steven H. Strauss
Keyword(s):  
Herbicide Resistance ◽  
Transgene Expression ◽  
Marker Gene ◽  
Field Trials ◽  
Gus Expression ◽  
Gene Encoding ◽  
Hybrid Poplars ◽  
Transgenic Lines ◽  
Single Field ◽  
The Stability

We assessed the stability of transgene expression in 79 transgenic lines (i.e., transformation events) of hybrid poplars during several years of field trials. The transgenic lines were comprised of 40 lines of hybrid cottonwoods (P. trichocarpa × P. deltoides) that were grown at three field sites, and 39 lines of hybrid aspens (section Leuce, P. alba × P. tremula) that were grown at a single field site. All the lines were transformed with a binary construct that included two genes that confer tolerance to glyphosate (GOX and CP4), a gene encoding resistance to the antibiotic kanamycin (nptII), and a visible marker gene (GUS). Agrobacterium tumefaciens was used for transformation; callogenesis and organogenesis occurred under kanamycin selection. In addition to repeated applications of herbicide to test stability of transgene expression, for the first time, we challenged ramets of 40 lines that had not previously been tested for herbicide resistance in their fourth season of vegetative growth. We report on the stability of herbicide resistance and GUS expression and evidence for somaclonal variation in growth and leaf morphology.

A novel T-DNA vector design for selection of transgenic lines with simple transgene integration and stable transgene expression

2005 ◽  
Vol 32 (8) ◽  
pp. 671 ◽  
Author(s):  
Song Chen ◽  
Christopher A. Helliwell ◽  
Li-Min Wu ◽  
Elizabeth S. Dennis ◽  
Narayana M. Upadhyaya ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Marker Gene ◽  
Direct Repeat ◽  
Transgene Silencing ◽  
Selective Marker ◽  
Hairpin Rna ◽  
Transgenic Lines ◽  
Flanking Sequences ◽  
Dna Vector ◽  
Selection Of

Plants transformed with Agrobacterium frequently contain T-DNA concatamers with direct-repeat (d / r) or inverted-repeat (i / r) transgene integrations, and these repetitive T-DNA insertions are often associated with transgene silencing. To facilitate the selection of transgenic lines with simple T-DNA insertions, we constructed a binary vector (pSIV) based on the principle of hairpin RNA (hpRNA)-induced gene silencing. The vector is designed so that any transformed cells that contain more than one insertion per locus should generate hpRNA against the selective marker gene, leading to its silencing. These cells should, therefore, be sensitive to the selective agent and less likely to regenerate. Results from Arabidopsis and tobacco transformation showed that pSIV gave considerably fewer transgenic lines with repetitive insertions than did a conventional T-DNA vector (pCON). Furthermore, the transgene was more stably expressed in the pSIV plants than in the pCON plants. Rescue of plant DNA flanking sequences from pSIV plants was significantly more frequent than from pCON plants, suggesting that pSIV is potentially useful for T-DNA tagging. Our results revealed a perfect correlation between the presence of tail-to-tail inverted repeats and transgene silencing, supporting the view that read-through hpRNA transcript derived from i / r T-DNA insertions is a primary inducer of transgene silencing in plants.

scholarly journals The Maternal Chromosome Set Is the Target of the T-DNA in the in Planta Transformation of Arabidopsis thaliana

Genetics ◽  
2000 ◽  
Vol 155 (4) ◽  
pp. 1875-1887
Author(s):  
Nicole Bechtold ◽  
Bénédicte Jaudeau ◽  
Sylvie Jolivet ◽  
Bruno Maba ◽  
Daniel Vezon ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Marker Gene ◽  
Gus Expression ◽  
Insertion Mutant ◽  
In Planta Transformation ◽  
In Planta ◽  
Gene Encoding ◽  
Agrobacterium Strain ◽  
Resistance Markers ◽  
Transformation Methods

Abstract In planta transformation methods are now commonly used to transform Arabidopsis thaliana by Agrobacterium tumefaciens. The origin of transformants obtained by these methods has been studied by inoculating different floral stages and examining gametophytic expression of an introduced β-glucuronidase marker gene encoding GUS. We observed that transformation can still occur after treating flowers where embryo sacs have reached the stage of the third division. No GUS expression was observed in embryo sacs or pollen of plants infiltrated with an Agrobacterium strain bearing a GUS gene under the control of a gametophyte-specific promoter. To identify the genetic target we used an insertion mutant in which a gene essential for male gametophytic development has been disrupted by a T-DNA bearing a Basta resistance gene (BR). In this mutant the BR marker is transferred to the progeny only by the female gametes. This mutant was retransformed with a hygromycin resistance marker and doubly resistant plants were selected. The study of 193 progeny of these transformants revealed 25 plants in which the two resistance markers were linked in coupling and only one plant where they were linked in repulsion. These results point to the chromosome set of the female gametophyte as the main target for the T-DNA.

scholarly journals Morpho-Physiological Testing of NaCl Sensitivity of Tobacco Plants Overexpressing Choline Oxidase Gene

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1102
Author(s):  
Galina N. Raldugina ◽  
Sergey V. Evsukov ◽  
Liliya R. Bogoutdinova ◽  
Alexander A. Gulevich ◽  
Ekaterina N. Baranova
Keyword(s):  
Choline Oxidase ◽  
Culture Methods ◽  
Gene Encoding ◽  
Oxidase Gene ◽  
Bacterial Enzyme ◽  
Transgenic Lines ◽  
High Regeneration ◽  
Whole Plants ◽  
Implicit Response

In this study the transgenic lines (TLs) of tobacco (Nicotianatabacum L.), which overexpress the heterologous gene encoding the bacterial enzyme choline oxidase were evaluated. The goal of our work is to study the effect of choline oxidase gene expression on the sensitivity of plant tissues to the action of NaCl. The regenerative capacity, rhizogenesis, the amount of photosynthetic pigments and osmotically active compounds (proline and glycine betaine) were assessed by in vitro cell culture methods using biochemical and morphological parameters. Transgenic lines with confirmed expression were characterized by high regeneration capacity from callus in the presence of 200 mmol NaCl, partial retention of viability at 400 mmol NaCl. These data correlated with the implicit response of regenerants and whole plants to the harmful effects of salinity. They turned out to be less sensitive to the presence of 200 mmol NaCl in the cultivation medium, in contrast to the WT plants.

Yeast ribosomal protein L1 is required for the stability of newly synthesized 5S rRNA and the assembly of 60S ribosomal subunits

1993 ◽  
Vol 13 (5) ◽  
pp. 2835-2845
Author(s):  
M Deshmukh ◽  
Y F Tsay ◽  
A G Paulovich ◽  
J L Woolford
Keyword(s):  
Ribosomal Protein ◽  
Ribosomal Subunit ◽  
Single Copy ◽  
5S Rrna ◽  
Gene Encoding ◽  
Ribosomal Subunits ◽  
Ribosomal Protein L1 ◽  
The Stability ◽  
And Function

Ribosomal protein L1 from Saccharomyces cerevisiae binds 5S rRNA and can be released from intact 60S ribosomal subunits as an L1-5S ribonucleoprotein (RNP) particle. To understand the nature of the interaction between L1 and 5S rRNA and to assess the role of L1 in ribosome assembly and function, we cloned the RPL1 gene encoding L1. We have shown that RPL1 is an essential single-copy gene. A conditional null mutant in which the only copy of RPL1 is under control of the repressible GAL1 promoter was constructed. Depletion of L1 causes instability of newly synthesized 5S rRNA in vivo. Cells depleted of L1 no longer assemble 60S ribosomal subunits, indicating that L1 is required for assembly of stable 60S ribosomal subunits but not 40S ribosomal subunits. An L1-5S RNP particle not associated with ribosomal particles was detected by coimmunoprecipitation of L1 and 5S rRNA. This pool of L1-5S RNP remained stable even upon cessation of 60S ribosomal subunit assembly by depletion of another ribosomal protein, L16. Preliminary results suggest that transcription of RPL1 is not autogenously regulated by L1.

scholarly journals Robust, But Transient Expression of Adeno-Associated Virus-Transduced Genes During Human T Lymphopoiesis

Blood ◽  
1997 ◽  
Vol 90 (12) ◽  
pp. 4854-4864 ◽  
Author(s):  
Jason P. Gardner ◽  
Haihong Zhu ◽  
Peter C. Colosi ◽  
Gary J. Kurtzman ◽  
David T. Scadden
Keyword(s):  
Cell Differentiation ◽  
T Cell ◽  
Gene Transfer ◽  
Transgene Expression ◽  
Marker Gene ◽  
T Cell Differentiation ◽  
Target Cells ◽  
Hematopoietic Stem ◽  
The Impact ◽  
T Lymphopoiesis

Abstract Recombinant adeno-associated viruses (rAAV) have been proposed to be gene transfer vehicles for hematopoietic stem cells with advantages over other virus-based systems due to their high titers and relative lack of dependence on cell cycle for target cell integration. We evaluated rAAV vector containing a LacZ reporter gene under the control of a cytomegalovirus (CMV) promoter in the context of primary human CD34+CD2− progenitor cells induced to undergo T-cell differentiation using an in vitro T-lymphopoiesis system. Target cells from either adult bone marrow or umbilical cord blood were efficiently transduced, and 71% to 79% CD2+ cells expressed a LacZ marker gene mRNA and produced LacZ-encoded protein after exposure to rAAV-CMV-LacZ. The impact of transgene expression on the differentiation of T cells was assessed by sequential quantitation of immunophenotypic subsets of virus-exposed cells and no alteration was noted compared with control. The durability of transgene expression was assessed and found to decay by day 35 with kinetics dependent on the multiplicity of infection. In addition, vector DNA was absent from CD4 or CD8 subselected CD3+ cells by DNA-polymerase chain reaction. These data suggest that rAAV vectors may result in robust transgene expression in primitive cells undergoing T-cell lineage commitment without toxicity or alteration in the pattern of T-cell differentiation. However, expression is transient and integration of the transgene unlikely. Recombinant AAV vectors are potentially valuable gene transfer tools for the genetic manipulation of events during T-cell ontogony but their potential in gene therapy strategies for diseases such as acquired immunodeficiency syndrome is limited.

scholarly journals Synthetic glycolate metabolism pathways stimulate crop growth and productivity in the field

Science ◽  
2019 ◽  
Vol 363 (6422) ◽  
pp. eaat9077 ◽  
Author(s):  
Paul F. South ◽  
Amanda P. Cavanagh ◽  
Helen W. Liu ◽  
Donald R. Ort
Keyword(s):  
Quantum Yield ◽  
Crop Yield ◽  
Metabolic Pathways ◽  
Biomass Productivity ◽  
Field Trials ◽  
Agricultural Field ◽  
Bisphosphate Carboxylase ◽  
Transgenic Lines ◽  
Glycolate Metabolism ◽  
Pathway Flux

Photorespiration is required in C3 plants to metabolize toxic glycolate formed when ribulose-1,5-bisphosphate carboxylase-oxygenase oxygenates rather than carboxylates ribulose-1,5-bisphosphate. Depending on growing temperatures, photorespiration can reduce yields by 20 to 50% in C3 crops. Inspired by earlier work, we installed into tobacco chloroplasts synthetic glycolate metabolic pathways that are thought to be more efficient than the native pathway. Flux through the synthetic pathways was maximized by inhibiting glycolate export from the chloroplast. The synthetic pathways tested improved photosynthetic quantum yield by 20%. Numerous homozygous transgenic lines increased biomass productivity between 19 and 37% in replicated field trials. These results show that engineering alternative glycolate metabolic pathways into crop chloroplasts while inhibiting glycolate export into the native pathway can drive increases in C3 crop yield under agricultural field conditions.

Stability of cabergoline in fox baits in laboratory and field conditions

2007 ◽  
Vol 34 (3) ◽  
pp. 239 ◽  
Author(s):  
Stuart McLean ◽  
Susan Brandon ◽  
Roger Kirkwood
Keyword(s):  
High Performance ◽  
Acetic Acid Solution ◽  
Hydrolysis Product ◽  
Fertility Control ◽  
Field Trials ◽  
Control Agent ◽  
Field Conditions ◽  
Dry Conditions ◽  
The Stability ◽  
Fox Control

Cabergoline is a potent inhibitor of prolactin release and a potential fertility control agent for foxes. To understand how cabergoline could behave in baits deployed for fox control, we conducted laboratory and field trials to investigate the stability of cabergoline when (1) in solution, (2) injected into a bait (deep-fried liver and Foxoff®) and (3) exposed to a range of environmental conditions, including burial. Cabergoline, dissolved in a 1% acetic acid solution, and its carboxylic acid hydrolysis product can be assayed using high-performance liquid chromatography. When stored at 4°C and at room temperature, cabergoline in solution was stable for up to 36 days. When stored under cool (≤15°C), dry conditions, cabergoline (800 µg) in commercial Foxoff® and deep-fried ox-liver baits was stable for 28 and 7 days, respectively; stability was reduced by increases in temperature (tested up to 40°C) and humidity. Recovery of cabergoline from buried baits exposed to a range of field conditions decreased rapidly in the first week, but after 56 days remained detectable at levels of 6–22% of the injected amounts. This study has important implications for baiting campaigns that use cabergoline for fox control.

scholarly journals Stability of Slow-Rusting Resistance to Puccinia asparagi and Managing Rust in Asparagus

Plant Disease ◽  
2012 ◽  
Vol 96 (7) ◽  
pp. 997-1000 ◽  
Author(s):  
Dennis A. Johnson
Keyword(s):  
Irrigation Management ◽  
Field Trials ◽  
Disease Progress Curve ◽  
Resistant Cultivars ◽  
Disease Progress ◽  
Slow Rusting ◽  
South Central ◽  
Progress Curve ◽  
The Stability

The stability of slow-rusting resistance to Puccinia asparagi in several asparagus cultivars was evaluated in two replicated field trials. Rust epidemics were monitored in each trial for 8 years spanning a period of 13 years (1983–1990 and 1987–1995). Inoculum of P. asparagi, an autoecious macrocyclic rust, originated each year as teliospores. In the first trial, the cultivars Jersey Titan, Jersey Centennial, Jersey Giant, Delmonte-361, and UC-157 had consistently lower area under the disease progress curve (AUDPC) values than Wash T2 and WSU-1. Cultivar Mary Washington was intermediate between the two groups of resistant and susceptible cultivars in 6 of 8 years. Jersey Titan consistently ranked number 1 for resistance with the lowest AUDPC values all 8 years. In the second trial, Jersey Giant, Delmonte-361, and UC-157 had consistently lower AUDPC values than Larac, Gynlim, Cito, Largo 17-3, and Franklim in each of 8 years. Jersey Giant, Delmonte-361, and UC-157 always ranked low (1, 2, or 3) for AUDPC. A shift from rust-susceptible to rust-resistant asparagus cultivars began in central Washington around 1996. In 2011, resistant cultivars made up nearly 96% of the asparagus plantings. From 1996 to 2011, rust was not considered a problem in commercial fields with slow-rusting resistant cultivars. Use of durable, slow-rusting cultivars, along with sanitation practices that reduced levels of aecia in nonharvested nurseries and on volunteer asparagus plants and judicious irrigation management, has effectively managed asparagus rust in commercial fields for at least 29 years in south-central Washington.

scholarly journals The Rooting of Stem Cuttings and the Stability of uidA Gene Expression in Generative and Vegetative Progeny of Transgenic Pear Rootstock in the Field

Plants ◽  
2019 ◽  
Vol 8 (8) ◽  
pp. 291 ◽  
Author(s):  
Vadim Lebedev
Keyword(s):  
Adventitious Root ◽  
Transgene Expression ◽  
Root Formation ◽  
Adventitious Root Formation ◽  
Model Systems ◽  
Unintended Effects ◽  
Uida Gene ◽  
Yearly Variation ◽  
Complex Biological Process ◽  
The Stability

Adventitious rooting plays an important role in the commercial vegetative propagation of trees. Adventitious root formation is a complex biological process, but knowledge of the possible unintended effects induced by both the integration/expression of transgenes and in vitro conditions on the rooting is limited. The long-term stability of transgene expression is important both for original transformants of woody plants and its progeny. In this study, we used field-grown pear rootstock GP217 trees transformed with the reporter ß-glucuronidase (uidA) genes with and without intron and re-transformed with the herbicide resistance bar gene as model systems. We assessed the unintended effects on rooting of pear semi-hardwood cuttings and evaluated the stability of transgene expression in progeny produced by generative (seedlings) and vegetative (grafting, cutting) means up to four years. Our investigation revealed that: (1) The single and repeated transformations of clonal pear rootstocks did not result in unintended effects on adventitious root formation in cuttings; (2) stability of the transgene expression was confirmed on both generative and vegetative progeny, and no silenced transgenic plants were detected; (3) yearly variation in the gene expressions was observed and expression levels were decreased in extremely hot and dry summer; (4) the intron enhanced the expression of uidA gene in pear plants approximately two-fold compared to gene without intron. The current study provides useful information on transgene expression in progeny of fruit trees under natural environmental conditions.

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