Havlíčkův Brod, Czech Republic
Havlíčkův Brod, Czech Republic

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Kodrik D.,Academy of Sciences of the Czech Republic | Kludkiewicz B.,Academy of Sciences of the Czech Republic | Navratil O.,Academy of Sciences of the Czech Republic | Skokova Habustova O.,Academy of Sciences of the Czech Republic | And 4 more authors.
Applied Biochemistry and Biotechnology | Year: 2013

Several recombinant derivatives of serine protease inhibitor called silk protease inhibitor 2 (SPI2), which is a silk component in Galleria mellonella (Lepidoptera, Insecta), were prepared in the expression vector Pichia pastoris. Both the native and the recombinant protease inhibitors were highly active against subtilisin and proteinase K. The synthetic SPI2 gene with Ala codon in the P1 position was fused with mGFP-5 to facilitate detection of the transgene and its protein product. A construct of the fusion gene with plant regulatory elements (promoter 35S and terminator OCS) was inserted into the binary vector pRD400. The final construct was introduced into Agrobacterium tumefaciens that was then used for genetic transformation of the potato variety Velox. The transgene expression was monitored with the aid of ELISA employing polyclonal antibody against natural SPI2. In vitro tests showed increased resistance to the late blight Phytophthora infestans in several transformed lines. No effect was seen on the growth, mortality, life span or reproduction of Spodoptera littoralis (Lepidoptera, Insecta) caterpillars, while feeding on transformed potato plants expressing the fusion protein, indicating that the transformed potatoes may be harmless to non-target organisms. © 2013 Springer Science+Business Media New York.


Mazakova J.,Czech University of Life Sciences | Zouhar M.,Czech University of Life Sciences | Rysanek P.,Czech University of Life Sciences | Taborsky V.,Czech University of Life Sciences | And 2 more authors.
Plant Protection Science | Year: 2010

A total of 195 isolates of Phytophthora infestans were collected from late blight-diseased potatoes grown in several localities in the Czech Republic during the years 2007-2008. The isolates were analysed for mating type using the pairing test, CAPS marker assays and PCR assays. Of the 195 tested isolates, 28% were of the Al mating type and 75% corresponded to the A2 mating type. Furthermore, oospores of P. infestans were microscopically detected in leaf samples from one locality.


Polzerova H.,Potato Research Institute Ltd. | Greplova M.,Potato Research Institute Ltd. | Frcek J.,Potato Research Institute Ltd.
Journal of Applied Botany and Food Quality | Year: 2011

The multiplication protocol of the medicinal plant Lepidium meyenii was established through axillary bud proliferation. Multiplication was initiated on modified MS supplemented with a combination of zeatin (2 μM) and gibberellic acid 3 (0.3 μM). The average number of proliferated shoots was 5.3 per segment. Shoots were rooted on modified MS medium with 9.8 μM of indole-3-butyric acid and 3 μM of a-naphthalene acetic acid. In vivo rooting was also successful and 94% regenerated plantlets were acclimatized.


Polzerova H.,Potato Research Institute Ltd. | Patzak J.,Hop Research Institute Co. | Greplova M.,Potato Research Institute Ltd.
Plant Cell, Tissue and Organ Culture | Year: 2011

Variability of 31 somatic hybrids of Solanum pinnatisectum Dun. with Solanum tuberosum L. for leaf morphology, plant vigor, resistance to Phytophthora infestans, ploidy level, and cytoplasm type was evaluated in vitro. The composition of these somatic hybrids was as follows: [S. pinnatisectum Dun. (2n = 2x = 24; cytoplasmic type Wγ) + S. tuberosum L. (2n = 4x = 48; cytoplasmic type Tß)]. Based on leaf morphology and plant growth vigor, plants were divided into three groups, including plants close to tbr parent with unlobed leaves, small plants with scarcely dissected leaves, and vigorous plants with asymmetrically and pinnately lobed leaves. Nine of the somatic hybrids were found to be highly resistant to P. infestans. Somatic hybrids were either tetraploid or hexaploid, with hexaploids being predominant. The cytoplasm of somatic hybrids was either Tßγ or Wßγ, with Tßγ being more common. Overall, in contrast to leaf morphology and growth vigor, level of resistance to P. infestans was not related to either ploidy level or type of cytoplasm. These findings demonstrate that early in vitro selection of promising hybrids can be useful in breeding programs. © 2010 Springer Science+Business Media B.V.


Lachman J.,Czech University of Life Sciences | Hamouz K.,Czech University of Life Sciences | Orsak M.,Czech University of Life Sciences | Pivec V.,Czech University of Life Sciences | And 4 more authors.
Food Chemistry | Year: 2012

The impact of selected factors - cultivar, storage, cooking and baking on the content of total anthocyanins (TAC) in coloured-flesh potato cultivars has been studied. TAC ranged from 248.5 to 2257.8 mg kg -1 dry matter (DM). TAC difference between cultivars was statistically significant. Cold storage (4°C) influenced TAC differentially. In the Violette and Highland Burgundy Red cultivars TAC increased by 18.5% and 12.1% respectively, and in the Valfi cultivar it decreased by 33.9%. Baking increased TAC 3.34 times whereas cooking in boiled water increased it 4.22 times. Correlation between antioxidant activity (AOA) and TAC (r 2 = 0.659) has been found. The Violette, Vitelotte and Highland Burgundy Red cultivars with the highest TAC showed high AOA and the Shetland Black cultivar and the cultivars Salad Blue and Blue Congo with a "marbled" texture showed the lowest TAC and AOA. Individual anthocyanidins are fingerprints of colour-fleshed potato cultivars. © 2011 Elsevier Ltd. All rights reserved.


Simkova D.,Potato Research Institute Ltd. | Lachman J.,Czech University of Life Sciences | Hamouz K.,Czech University of Life Sciences | Vokal B.,Potato Research Institute Ltd.
Food Chemistry | Year: 2013

In recent time the interest of industry increases particularly in processing and use of potato high amylopectin (AMP) starches. Therefore the plant breeders effort to obtain "waxy" potato cultivars with low amylose (AMS) content. In this four-year study sixteen potato cultivars grown on five experimental locations were evaluated on the percentage of AMS/AMP by enzymatic method, starch content by the underwater weight method, phosphorus (P) content in starch digests spectrophotometrically, and starch granule size determined by laser diffraction method. Between enzymatic and iodine-potassium iodide method good correlation has been revealed (r = 0.71). The correlation analysis between AMS and P levels showed a clear negative correlation. For all measured parameters (starch, AMS, P, starch granule size) significant impact of cultivar has been determined. Location and year have lower, but significant impact. No statistically significant effect of year on AMS has been found. The cultivar Amado distinguished with the highest AMP and P contents and the cultivar Westamyl showed all positive values interesting for growers and processors. © 2013 Elsevier Ltd.


Lachman J.,Czech University of Life Sciences | Hamouz K.,Czech University of Life Sciences | Musilova J.,Slovak University of Agriculture | Hejtmankova K.,Czech University of Life Sciences | And 5 more authors.
Food Chemistry | Year: 2013

The impact of peeling and three cooking treatments (boiling, baking and microwaving) on the content of selected phytochemicals in white-, yellow-, red- and purple-fleshed potatoes was investigated. Ascorbic acid and chlorogenic acid contents were determined by HPLC-DAD, total anthocyanin content by pH-differential spectrophotometry, glycoalkaloid, α-chaconine and α-solanine contents by HPLC-ESI/MS/MS. All cooking treatments reduced ascorbic and chlorogenic acid contents, total glycoalkaloids, α-chaconine and α-solanine with the exception of total anthocyanins. The losses of ascorbic and chlorogenic acids were minimised with boiling and total anthocyanin levels retained the highest. Boiling of peeled tubers decreased contents of total glycoalkaloids (α-chaconine and α-solanine) and appeared as the most favourable among the three tested methods. Moreover, due to higher initial levels, red- and purple-fleshed cultivars retained higher amounts of antioxidants (ascorbic acid, chlorogenic acid and total anthocyanin) after boiling and may be healthier as compared with white or yellow cultivars. © 2012 Elsevier Ltd. All rights reserved.


Navratil O.,Academy of Sciences of the Czech Republic | Bucher P.,Potato Research Institute Ltd | Vacek J.,Potato Research Institute Ltd
Czech Journal of Genetics and Plant Breeding | Year: 2012

Cold-stored potato tubers gradually accumulate reducing sugars. A proposed reason is a cold-induced blocking of glycolysis. The introduction and expression of the bacterial gene Lbpfk coding for cold-tolerant phosphofructokinase might counteract this effect. We have recently introduced this gene into several Czech potato cultivars. The obtained transgenic lines were then tested for three years in field trials. In 17 transgenic lines derived from two of the cultivars we have investigated the accumulation of reducing sugars during two and four months of cold storage. Although in all transgenic lines the sugar content still increased between the 2 nd and the 4 th month of cold-storage, the level of reducing sugars was in all transgenic lines after both two and four months of cold storage considerably lower than in the original cultivars. The extent of sugar accumulation was also influenced by the parental genotype. No significant differences in sugar accumulation were observed between the transgenic lines from the same parent.


Hejtmankova K.,Czech University of Life Sciences | Kotikova Z.,Czech University of Life Sciences | Hamouz K.,Czech University of Life Sciences | Pivec V.,Czech University of Life Sciences | And 2 more authors.
Journal of Food Composition and Analysis | Year: 2013

The objective of this two-year study was to determine the main carotenoids in 15 varieties of potatoes (Solanum tuberosum L.) with different flesh colour - white, yellow, red and purple and two varieties of Solanum phureja. Content of total anthocyanins in pigmented potatoes was also determined and compared with the carotenoid level. Potatoes were cultivated in two localities with different soil-climatic conditions. Content of carotenoids and anthocyanins was affected by variety and locality. Moreover, carotenoid content was influenced by the growing year. Content of total carotenoids ranged from 0.779 to 13.3mg/kg dry matter (DM). The main carotenoid was lutein in all varieties (54-93%) furthermore violaxanthin, neoxanthin, zeaxanthin and β-carotene were identified in most of the analysed samples. The highest carotenoid content was found in S. phureja varieties. The pigmented varieties Blaue Anneliese, Violetta, Olivia (all purple) and H.B. Red (red) showed comparable carotenoid level with the yellow-fleshed variety Agria; other pigmented varieties contained similar carotenoid levels as white-fleshed varieties. Content of total anthocyanins ranged from 210 to 2419mg/kg DM and correspond to flesh colour intensity. A weak correlation between carotenoids and anthocyanins was found (r2=0.33). Thus some coloured varieties could be a good source of both studied phytochemicals. © 2013 Elsevier Inc.


Greplova M.,Potato Research Institute Ltd. | Polzerova H.,Potato Research Institute Ltd. | Ptacek J.,Potato Research Institute Ltd. | Domkarova J.,Potato Research Institute Ltd.
Acta Horticulturae | Year: 2015

Somatic hybridization is a technique for alternative breeding to increase genetic variability. The aim of this work was to establish protoplast culture of Allium ursinum. The initial genotypes originated from three localities. The homogeneity of these populations was tested using RAPD markers. These analyses showed some differences among tested genotypes. According to preliminary experiments, Allium ursinum protoplasts seemed to be recalcitrant for protoplast cultures. The influence of plant grow regulators, sugar type and electric field (AC, DC and combination) was tested. With some modifications, the protoplasts were isolated and affected by electric field according to Greplova et al. (2008). The application of indirect current seemed to be useful to stimulate cell division. Sugar type in culture medium appeared to be very important; the combination of glucose with sucrose proved to be the best. Putrescine showed to be crucial for cell viability and regeneration. © 2015 ISHS.

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