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1

In vitro culture and biotechnology ? fulfilled expectations?

100%
Malepszy S., Burza W.
Biotechnologia
|
2010
|
issue 2
10-17
EN
The methods for in vitro culture of plant cells, tissues and organs had focused much attention in the beginning of the last century, which resulted in setting up the first commercial laboratories over 60 years ago. These laboratories concentrated their activity on clonal propagation and their economical importance has been permanently growing. However, some of the applications of in vitro methods are still not realistic, whereas introduction of other is not satisfactory. Plant breeding is an example where application of tissue culture techniques is below expectations. There are several reasons for such situation. Two of them are of biological nature (genotypic effect, somaclonal variation), and the third reason results from the development of other molecular methods providing alternative solutions. We suggest that the main limitations in more effective usage of in vitro methods should be minimized by the development of efficient plant stem cells' culture procedures.
2

Transient expression assay for the optimization of direct gene transfer into cucumber meristem protoplasts by electroporation

81%
Burza W., Wochniak P., Wroblewski T., Malepszy S.
Journal of Applied Genetics
|
1995
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vol. 36
|
issue 1
1-10
EN
The paper obtained a new way of viable and very homogenous cucumber protoplasts.Protoplasts from cells formed in the shoot tip meristem culture were isolated from suspension.Plasmid pBI121 was introduced using imulse electric field.Effectiveness of transformation process was determinated by transient expression of ?-glucoronidase (GUS) gene, controlled by promotor 35S.The ativity of ?-glucoronidase enzyme as a product of GUS reporter gene was estimated by fluorometric method (Jefferson 1987).Parameters of electroporatian process were optimized.The transient expression of GUS gene was measured 24 h after electroporation.The highest effectiveness of transformation process was achieved using three electric impulses at the initial voltage of 250-350V at 10-sec.intervals as a results of discharging a 140 ?F capacitor and 50-70 ?g x cm/1000 plasmid DNA in the presence of 50 ?g x cm/1000 carrier DNA.The system presented is an effective method of exogenic DNA transfer, which is indicated by a high transient expression of the reporter gene.In comparison to Agrobacterium tumefaciens and A.rhizogenes, this alternative method of gene transfer can be used for obtaining transgenic cucumber plants.
3

Characterization of a cucumber (Cucumis sativus L.) somaclonal variant with paternal inheritance

81%
Malepszy S., Burza W., Smiech M.
Journal of Applied Genetics
|
1996
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vol. 37
|
issue 1
65-78
EN
Regeneration of cucumber from leaf explants resulted in a new species phenotype designated mosaic (msc).It is characterizes by two types of spots on teh leaves (zucchini-like and chlorophyllous) and has many altered morphological and physiological properties including slower growth, smaller organs, poorly germinating or empty seeds and smaller number of flowers per node.In msc plants the shape of the first leaf is always altered, and in about 76% of the flowers the crown is reduced and distorted to a varying degree.Chloroplasts of the zucchini-like sectors are filled with large starch grains, and some of the embryos die at various stages of development.The msc phenotype is ransmitted uniparentally only by the male plants and no segregation is observed in the F2 and subsequent generations.Possible mechanisms responsible for msc phenotype and its transmission are discussed.
4

Generation of selectable marker-free transgenic plants

81%
Zuzga S., Burza W., Malepszy S.
|
2003
|
issue 4
32-46
EN
Since the first reports of successful plant transformation appeared, there have been steady improvements of the transformation methods. Nowadays, transgenic plants without the incorporation of selection genes for antibiotic or herbicide resistance would be the only ones acceptable to the public, so elimination of these genes from transgenic crops prior to their field release and commercialization seems inevitable. Several strategies have been developed to generate marker-free transgenic plants, including: positive selection, alternative marker genes, co-transformation, site-specific recombination, transpozon-mediated approaches and intrachromosomal homologous recombination. The efficiency of these methods is various as comparing to the traditional marker assisted selection - higher in case of alternative procedures and lower in others.
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