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1

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

100%

Malepszy S., Burza W., Smiech M.

Journal of Applied Genetics

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1996

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vol. 37

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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.

2

Growth kinetics of embriogenic and nonembriogenic suspension cultures of cucumber (Cucumis sativus L.) in stirred bioreactor

100%

Kroliczak P., Grajek W., Tomkowiak M.

Biotechnologia

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1995

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issue 3

152-166

EN

Embryogenic and nonembriogenic suspension cultures of cucumber were cultivated in a 5-L stirred bioreactor.Significant differences between kinetic characteristics of these cell lines (growth rate, tissue morphology, sugar and nitrogen metbolism) are presented. Nonembryogenic culture demonstrated faster growth rate anr greater nutrient uptake.Mechanical sterss in stirred bioreactor under experimental conditions was negligible.

3

The selection of mosaic (MSC) phenotype after passage of cucumber (Cucumis sativus L.) through cell culture ? a method to obtain plant mitochondrial mutants

100%

Bartoszewski G., Havey M., Ziolkowska A., Dlugosz M., Malepszy S.

Journal of Applied Genetics

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2007

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vol. 48

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issue 3

1-9

EN

Mosaic (MSC) mutants of cucumber (Cucumis sativus L.) appear after passage through cell cultures. The MSC phenotype shows paternal transmission and is associated with mitochondrial DNA rearrangements. This review describes the origins and phenotypes of independently produced MSC mutants of cucumber, including current knowledge on their mitochondrial DNA rearrangements, and similarities of MSC with other plant mitochondrial mutants. Finally we propose that passage of cucumber through cell culture can be used as a unique and efficient method to generate mitochondrial mutants of a higher plant in a highly homozygous nuclear background.

4

Cucumber transformation methods - the review

100%

Yin Z., Bartoszewski G., Szwacka M., Malepszy S.

Biotechnologia

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2005

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issue 1

95-113

EN

Several aspects of cucumber transformation including the ways of transgene introduction, factors influencing the transformation efficiency and the fate of the introduced genes were reviewed. Various transgenes have been introduced into the cucumber genome mostly via the Agrobacterium-mediated transformation. The frequency of Agrobacterium-mediated transformation ranged from 0.8 to 10% and was influenced by the selection agent, the regeneration efficiency, activation of vir genes expression, the explant size, bacteria cell density, the length of exposure and the co-cultivation period. The transgenes were integrated mostly as single copy in the Agrobacterium-mediated transformation and as multiple copies in direct transformation. Variable levels of the transgene expression were observed. The transmission of the transgenes as well as the transgenic phenotype follow the Mendelian, and rarely non-Mendelian, ratio. The production of marker-free transgenic cucumber and use of an alternative transformation method are recommended.

5

Sequential fluorescent staining with CMA and DAPI for somatic chromosome identification in cucumber (Cucumis sativus L.)

100%

Plader W., Hoshi Y., Malepszy S.

Journal of Applied Genetics

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1998

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vol. 39

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issue 3

249-258

EN

Each of the seven chromosomes in cucumber (Cucumis sativus L.) was identified using sequential staining with Chromomycin A3 (CMA) and 4-6-diamidino-2-phenylindole (DAPI) as DNA base-specific fluorescent dyes. The present method using enzymatic digestion produced a high level of well-spread early-metaphase chromosome complements. After CMA and DAPI staining, reproducible fluorescence bands were obtained in mitotic prometaphase chromosomes. The CMA staining method made it possible to characterize whole chromosomes from prometaphase to mid-metaphase. Chromosome 1 had the largest and widest CMA-positive (CMA+) band from the proximal region to the interstitial region on the long arm in prometaphase. A large gap separating of the short arm from the long arm was always observed in chromosome 2 during prometaphase. The banding pattern of the short arm was similar to that of the long arm in chromosome 2. Chromosomes 1 and 2 in early metaphase had sharp and large CMA-positive and DAPI-negative (CMA+DAPI-) bands at the pericentromeric regions. In early metaphase, chromosome 3 was characterized by having a narrow CMA+DAPI- band on the pericentromeric region of the short arm. Chromosomes 4 and 5 showed similar chromosome length and had a large CMA+ band at the distal region of the long arm. Chromosome 4 did not show any clear band in the short arm, while chromosome 5 showed a telomeric CMA+ band at the short arm and a clear CMA+DAPI- band at the pericentromeric region. Chromosome 6 had a CMA+ band at the distal region and a weak CMA+ band at the proximal region in each of the arms. Chromosome 7 had an evident CMA+ band in the long arm and a CMA+DAPI- band in the pericentromeric region.

6

Tissue-culture-responsive and autotetraploidy-responsive changes in metabolic profiles of cucumber (Cucumis sativus L.)

100%

Filipecki M., Yin Z., Wisniewska A., Smiech M., Malinowski R., Malepszy S.

Journal of Applied Genetics

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2006

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vol. 47

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issue 1

17-21

EN

Somaclonal variation commonly occurs during in vitro plant regeneration and may introduce unintended changes in numerous plant characters. In order to assess the range of tissue-culture-responsive changes on the biochemical level, the metabolic profiles of diploid and tetraploid cucumber R1 plants regenerated from leaf-derived callus were determined. Gas chromatography and mass spectrometry were used for monitoring of 48 metabolites and many significant changes were found in metabolic profiles of these plants as compared to a seed-derived control. Most of the changes were common to diploids and tetraploids and were effects of tissue culture. However, tetraploids showed quantitative changes in 14 metabolites, as compared to regenerated diploids. These changes include increases in serine, glucose-6P, fructose-6P, oleic acid and shikimic acid levels. Basing on this study we conclude that the variation in metabolic profiles does not correlate directly with the range of genome changes in tetraploids.

7

The selection of mosaic (MSC) phenotype after passage of cucumber (Cucumis sativus L.) through cell culture ? a method to obtain plant mitochondrial mutants

100%

Bartoszewski G., Havey M. J., Ziolkowska A., Dlugosz M., Malepszy S.

Journal of Applied Genetics

|

2007

|

vol. 48

|

issue 1

1-9

EN

Mosaic (MSC) mutants of cucumber (Cucumis sativus L.) appear after passage through cell cultures. The MSC phenotype shows paternal transmission and is associated with mitochondrial DNA rearrangements. This review describes the origins and phenotypes of independently produced MSC mutants of cucumber, including current knowledge on their mitochondrial DNA rearrangements, and similarities of MSC with other plant mitochondrial mutants. Finally we propose that passage of cucumber through cell culture can be used as a unique and efficient method to generate mitochondrial mutants of a higher plant in a highly homozygous nuclear background.

8

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

100%

Burza W., Wochniak P., Wroblewski T., Malepszy S.

Journal of Applied Genetics

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1995

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vol. 36

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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.

9

Marker proteins of somatic embryogenesis i cucumber (Cucumis sativus L.)

100%

Filipecki M. K., Przybecki Z.

Genetica Polonica

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1994

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vol. 35

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issue 1-2

1-9

10

Selected aspects of genetic transformation in cucumber, tomato, and sweet pepper

88%

Bartoszewski G., Szwacka M., Czarny M., Niemirowicz-Szczytt K., Malepszy S.

Biotechnologia

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2010

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issue 4

146-154

EN

Plant transformation is a technology widely used in gene functional analysis and crop improvement. In this article we have attempted to sum up the studies on plant transformation carried out by the Department of Plant Genetics, Breeding and Biotechnology, Warsaw University of Life Sciences, pointing out to recent developments in this field. Efficient Agrobacterium-based transformation protocols for cucumber and tomato were established and applied. Several traits, including fruit taste (thaumatin gene), chilling tolerance (pGT::DHN24), parthenocarpy (DefH9::iaaM), and virus resistance (TSWV nucleoprotein gene), were modified. Transgenic cucumber lines expressing mitochondrially targeted GFP protein were developed. Sensory evaluation of fruit traits and of unintended effects of cucumber expressing thaumatin gene was made. Cucumber and tomato transformation was also applied with the aim to carry out gene functional analysis. Having introduced overexpression, silencing, and promoter gene constructs, we were able to obtain several transgenic tomato lines. Attempts have been made to set up an efficient method of sweet pepper transformation.

11

Expression pattern of the pre-prothaumatin II gene under the control of the CaMV 35S promoter in transgenic cucumber (Cucumis sativus L.) flower buds and fruits

75%

Szwacka M., Siedlecka E., Zawirska-Wojtasiak R., Wisniewski L., Malepszy S.

Journal of Applied Genetics

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2009

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vol. 50

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issue 1

9-16

EN

Thaumatin II is an extremely sweet-tasting protein produced by fruits of the West African shrub Thaumatococcus daniellii Benth, so it can be used in biotechnology to improve the tastes of various plant products. This study is concerned with the spatial and temporal aspects of expression of the 35S-pre-prothaumatin II chimeric gene in flower buds and fruits of transgenic cucumber (Cucumis sativus L.) line 225. The activity of the 35S promoter in organs of line 225 was compared with its activity in 2 other transgenic lines. The accumulation of recombinant thaumatin varied spatially in flower bud tissues of transgenic lines. We found that these differences in the spatial accumulation of transgenic protein concerned the ovary of female buds and the perianth of male buds. In contrast to flower parts, recombinant thaumatin was found in nearly all parts of the young fruit from the transgenic plants. The pre-prothaumatin II gene expression was detected at a very early developmental stage in male buds, and its pattern was rather conserved as the buds aged. The expression of the transgene was also detected in vascular tissues of examined organs but was undetectable in pollen grains, in agreement with the generally held view that the CaMV 35S promoter is virtually silent in pollen. Immunocytochemical analyses of sections of control organs revealed endogenous homolog(s) of thaumatin when using polyclonal antisera, but not when using monoclonal antibodies for recombinant thaumatin detection in transgenic cucumber.

Refine search results

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

Growth kinetics of embriogenic and nonembriogenic suspension cultures of cucumber (Cucumis sativus L.) in stirred bioreactor

The selection of mosaic (MSC) phenotype after passage of cucumber (Cucumis sativus L.) through cell culture ? a method to obtain plant mitochondrial mutants

Cucumber transformation methods - the review

Sequential fluorescent staining with CMA and DAPI for somatic chromosome identification in cucumber (Cucumis sativus L.)

Tissue-culture-responsive and autotetraploidy-responsive changes in metabolic profiles of cucumber (Cucumis sativus L.)

The selection of mosaic (MSC) phenotype after passage of cucumber (Cucumis sativus L.) through cell culture ? a method to obtain plant mitochondrial mutants

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

Marker proteins of somatic embryogenesis i cucumber (Cucumis sativus L.)

Selected aspects of genetic transformation in cucumber, tomato, and sweet pepper

Expression pattern of the pre-prothaumatin II gene under the control of the CaMV 35S promoter in transgenic cucumber (Cucumis sativus L.) flower buds and fruits