scholarly journals Determination of the T-DNA Transfer and the T-DNA Integration Frequencies upon Cocultivation of Arabidopsis thaliana Root Explants

2000 ◽  
Vol 13 (6) ◽  
pp. 658-665 ◽  
Author(s):  
Sylvie De Buck ◽  
Chris De Wilde ◽  
Marc Van Montagu ◽  
Ann Depicker
Keyword(s):  
Plant Cell ◽  
Dna Transfer ◽  
Plant Genome ◽  
Root Explants ◽  
Root Cells ◽  
Dna Integration ◽  
Recombination System ◽  
Two Factors ◽  
Selection For ◽  
Transformation Competence

Using the Cre/lox recombination system, we analyzed the extent to which T-DNA transfer to the plant cell and T-DNA integration into the plant genome determine the transformation and cotransformation frequencies of Arabidopsis root cells. Without selection for transformation competence, the stable transformation frequency of shoots obtained after cocultivation and regeneration on nonselective medium is below 0.5%. T-DNA transfer and expression occur in 5% of the shoots, indicating that the T-DNA integrates in less than 10% of the transiently expressing plant cells. A limited fraction of root cells, predominantly located at the wounded sites and in the pericycle, are competent for interaction with agrobacteria and the uptake of a T-DNA, as demonstrated by histochemical GUS staining. When selection for transformation competence is applied, the picture is completely different. Then, approximately 50% of the transformants show transient expression of a second, nonselected T-DNA and almost 50% of these cotransferred T-DNAs are integrated into the plant genome. Our results indicate that both T-DNA transfer and T-DNA integration limit the transformation and cotransformation frequencies and that plant cell competence for transformation is based on these two factors.

scholarly journals Odyssey of agrobacterium T-DNA.

2001 ◽  
Vol 48 (3) ◽  
pp. 623-635 ◽  
Author(s):  
A Ziemienowicz
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Plant Cell ◽  
Plant Pathogen ◽  
Unique Feature ◽  
Dna Transfer ◽  
Soil Bacterium ◽  
Plant Genome

Agrobacterium tumefaciens, a plant pathogen, is characterized by the unique feature of interkingdom DNA transfer. This soil bacterium is able to transfer a fragment of its DNA, called T-DNA (transferred DNA), to the plant cell where T-DNA is integrated into the plant genome leading to "genetic colonization" of the host. The fate of T-DNA, its processing, transfer and integration, resembles the journey of Odysseus, although our hero returns from its long trip in a slightly modified form.

scholarly journals Bacterial Conjugation Protein MobA Mediates Integration of Complex DNA Structures into Plant Cells

1999 ◽  
Vol 181 (18) ◽  
pp. 5758-5765 ◽  
Author(s):  
Ana María Bravo-Angel ◽  
Véronique Gloeckler ◽  
Barbara Hohn ◽  
Bruno Tinland
Keyword(s):  
Plant Cell ◽  
Plant Cells ◽  
Plant Genome ◽  
Bacterial Conjugation ◽  
Dna Structures ◽  
Dna Integration ◽  
Bacterial Proteins ◽  
Plant Enzymes ◽  
Complex Integration ◽  
Cell Genome

ABSTRACT Agrobacterium tumefaciens transfers T-DNA to plant cells, where it integrates into the genome, a property that is ensured by bacterial proteins VirD2 and VirE2. Under natural conditions, the protein MobA mobilizes its encoding plasmid, RSF1010, between different bacteria. A detailed analysis of MobA-mediated DNA mobilization byAgrobacterium to plants was performed. We compared the ability of MobA to transfer DNA and integrate it into the plant genome to that of pilot protein VirD2. MobA was found to be about 100-fold less efficient than VirD2 in conducting the DNA from the pTi plasmid to the plant cell nucleus. However, interestingly, DNAs transferred by the two proteins were integrated into the plant cell genome with similar efficiencies. In contrast, most of the integrated DNA copies transferred from a MobA-containing strain were truncated at the 5′ end. Isolation and analysis of the most conserved 5′ ends revealed patterns which resulted from the illegitimate integration of one transferred DNA within another. These complex integration patterns indicate a specific deficiency in MobA. The data conform to a model according to which efficiency of T-DNA integration is determined by plant enzymes and integrity is determined by bacterial proteins.

Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells

1984 ◽  
Vol 4 (12) ◽  
pp. 2929-2931 ◽  
Author(s):  
K Blochlinger ◽  
H Diggelmann
Keyword(s):  
Selectable Marker ◽  
Dna Transfer ◽  
Direct Selection ◽  
Regulatory Sequences ◽  
Hygromycin B ◽  
Dominant Marker ◽  
Dna Coding ◽  
Sarcoma Virus ◽  
Moloney Sarcoma Virus ◽  
Selection For

The DNA coding sequence for the hygromycin B phosphotransferase gene was placed under the control of the regulatory sequences of a cloned long terminal repeat of Moloney sarcoma virus. This construction allowed direct selection for hygromycin B resistance after transfection of eucaryotic cell lines not naturally resistant to this antibiotic, thus providing another dominant marker for DNA transfer in eucaryotic cells.

scholarly journals Optimization of Agrobacterium-mediated transformation conditions of prospective genotypes of winter bread wheat by in planta method

2021 ◽  
Vol 28 ◽  
pp. 66-71
Author(s):  
O. V. Dubrovna ◽  
L. V. Slivka
Keyword(s):  
Transgenic Plants ◽  
Bread Wheat ◽  
Optical Density ◽  
Dna Transfer ◽  
Plant Genome ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
The Third ◽  
Winter Bread Wheat ◽  
Inoculation Medium

Aim. Optimization of conditions for genetic transformation of new promising genotypes of winter bread wheat (T. aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by in planta method using the strain AGL0 and vector construct pBi2E. Results. The influence of air temperature, optical density of cells of agrobacterial suspension, inoculation day and composition of inoculation medium on the frequency of obtaining transgenic plants of new winter wheat genotypes was studied. The dependence of the frequency obtaining of transgenic plants from environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22°C provided the largest number (4.8%) of wheat transformants, and when the temperature is reduced to 16-18°C there is a decrease in the efficiency of T-DNA transfer into the plant genome and the lowest frequency of transformation (0.7%). Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.6 op.od. and inoculation on the third day after castration of ears. Keywords: T. aestivum, Agrobacterium-mediated transformation in planta, optimization of conditions.

scholarly journals Can crop phenology and plant height be channelized to improvise wheat productivity in diverse production environments?

2020 ◽  
Author(s):  
D Mohan ◽  
H M Mamrutha ◽  
Rinki Khobra ◽  
Gyanendra Singh ◽  
GP Singh
Keyword(s):  
Grain Weight ◽  
Selection Strategy ◽  
Production Environment ◽  
Crop Phenology ◽  
Wheat Growth ◽  
Production Environments ◽  
Two Factors ◽  
Selection For ◽  
Field Indicators ◽  
Break Yield

AbstractNon-grain parameters like height, flowering and maturity should also be tried to break yield plateau in wheat. This study explores such possibilities by analysing performance of released and pre-released varieties evaluated in ten diverse production environments of India during the period 2000-2020. Regression analysis supports relevance of such non-grain determinants in grain yield under every environment but magnitude of impact can vary. Collective contribution of non-grain parameters can be high in a production environment where growth condition is most favourable for wheat growth and every factor is important in such situations. They contribute less in the environments engrossed with abiotic stress and merely one or two factors can be earmarked for selection. Besides yield, this selection strategy can also enhance grain weight in certain environments. At a time when selection for grain attributes is not providing further push; it would be worth trying to explore these non-grain field indicators as selection strategy for further advancement in productivity and grain weight of bread wheat.

Homology Recognition During T-DNA Integration into the Plant Genome

1994 ◽  
pp. 167-189 ◽  
Author(s):  
Csaba Koncz ◽  
Kinga Németh ◽  
George P. Rédei ◽  
Jeff Schell
Keyword(s):  
Plant Genome ◽  
Dna Integration ◽  
Homology Recognition

T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation

Planta ◽  
2013 ◽  
Vol 238 (6) ◽  
pp. 1025-1037 ◽  
Author(s):  
Rim Ghedira ◽  
Sylvie De Buck ◽  
Frédéric Van Ex ◽  
Geert Angenon ◽  
Ann Depicker
Keyword(s):  
Arabidopsis Thaliana ◽  
Dna Transfer ◽  
Dna Integration ◽  
Floral Dip ◽  
Root Explant

Routine Formation and Flexibility in Social and Non-Social Behaviour of Aggressive and Non-Aggressive Male Mice

Behaviour ◽  
1990 ◽  
Vol 112 (3-4) ◽  
pp. 176-193 ◽  
Author(s):  
G.A. VAN OORTMERSSEN ◽  
S. DEN DAAS ◽  
J.M. Koolhaas ◽  
R.F. Benus
Keyword(s):  
Social Situation ◽  
Phenotypic Selection ◽  
Male Mice ◽  
Y Maze ◽  
The Social ◽  
Two Factors ◽  
Aggressive Male ◽  
Selection For ◽  
Attack Latency ◽  
The Relationship

AbstractTo investigate the relationship between aggression and routine-like behaviour the response of male mice of bidirectionally selected lines for attack latency to a change in the social and non-social environment has been measured. In a non-social situation the extent of routine-like behaviour was measured in a Y-maze in which only one of the two arms gave access to the food compartments. The number of errors made in response to reversal of the arm that was blocked was taken as indicator for the degree of routine formation. Males of the short attack latency (SAL) line made significantly more errors, and hence were more routine-like in their performance, than mice of the long attack latency (LAL) line. Males of the LAL line that nevertheless had short attack latencies (i. e. aggressive LAL mice) turned out to be flexible in their behaviour; their response was similar to that of the non-aggressive LAL males. In a social situation SAL and aggressive LAL mice were used to investigate routine formation in attacking behaviour. The males were given different amounts of experience with male opponents after which their own female was introduced as opponent. The more extended the experience with male intruders was, the more SAL males subsequently attacked their female. In contrast, LAL mice appropriately changed their behaviour towards the female opponent. Thus, the attacking behaviour of SAL mice gets routine-like, whereas that of LAL males remains flexible. It is concluded that selection for attack latency generally coincides with selection for routine-like behaviour, suggesting that these two factors are influenced by many of the same genes. Regarding the fact that aggressive males of the LAL line show flexible behaviour, it may be proposed that with the phenotypic selection for attack latency there has in fact been selected for a mechanism that determines the organization (routine-like vs flexible) of behaviour.

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