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1

Plant sweetness-inducing proteins and taste-modyfying proteins

100%
Szwacka M., Malepszy S.
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issue 1
64-82
EN
This work reviews the plant proteins which stimulate the sweet receptor in humans and primates. Identification of the sweet site of thaumatin is a promising target. Thaumatin as a sweetener and flavor enhancer has been used in a wide range of foods. Some of recombinant sweet proteins have been expressed in microbes below economically viable levels. Significance of high-quality production of sweet proteins in higher plant is discussed. Thaumatin and thaumatin-related family have been characterized.
2

Differences in the inheritance stability of kanamycin resistance between transgenic cucumbers (Cucumis sativus L.) containing two constructs

81%
Yin Z., Szwacka M., Malinowski R., Malepszy S.
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issue 3
307-314
EN
A phenotypic segregation of kanamycin resistance conferred by nosnptII was investigated in two groups of transgenic cucumber lines one containing the PR-2duidA-nosnptII construct and the other with the thaumatinII-nosnptII construct to the third and fifth generation, respectively. Expression of the nptII gene was more stable and predictable in the progeny of the PR-2d lines than that of thaumatin lines. In T2 and T3 generations representing seven PR-2d primary transformants, 78% of the hemizygous progenies exhibited a segregation ratio consistent with Mendelian inheritance; non-Mendelian ratio occurred in the remaining 22%. Mendelian segregation was observed in 46% of the hemizygous progenies derived from 11 thaumatin primary transformants. The segregation ratio for two and three independent loci appeared in single PR-2d and thaumatin lines, respectively.
3

Cucumber transformation methods - the review

81%
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.
4

Transformation of cucumber Cucumis sativus L.

81%
Szwacka M., Burza W., Palucha A., Malepszy S.
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EN
Different transformation methods were investigated. However a lot of fertile plants were obtained only in the case of A. tumefaciens and thaumatin with kanamycin as a selective marker. Small differences in the plant morphology and development were observed in T0 generation. Most of the plants showed the presence of the transgene but the level of mRNA was different.
5

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

71%
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.
6

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

71%
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.
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