Scientific Studies 2011

Note: Papers marked with (A) are written in English.

  • Pavel Kasal, Pavel Růžek, Helena Kusá, Jaroslav Čepl
    Effective ways of mineral nitrogen fertilizer applications in potatoes and their effect on potato yield
  • Karel Hamouz, Jaromír Lachman, Kateřina Hejtmánková, Jiří Cimr, Milan Čížek
    Effect of flesh colour on the content of major antioxidants in potato tubers from organic cultivation (A)
  • Václav Krejzar, Iveta Pánková, Ervín Hausvater, Petr Doležal, Václav Kůdela
    Relative incidence of potato blackleg pathogens in the Czech Republic in 2010-2011
  • Renata Švecová, Jiří Ptáček, Jaroslava Domkářová
    The utilization of molecular markers at assessment of subcollection genotypes of Solanum wild species
  • Vendulka Horáčková, Oldřich Navrátil, Viktor Kopačka, Jaroslava Domkářová
    Evaluation of resistance to potato late blight in transformants with the transgene for SPI2 :: GFP fusion protein
  • Petr Dědič
    New rational procedures of potato virus laboratory diagnosis using Multiplex ELISA and Luminex-xMAP technology
  • Hana Polzerová, Marie Greplová, Jaroslava Domkářová
    Analysis of somatic hybrid performance in the field (A)
  • Marie Greplová, Hana Polzerová
    Experience with the axenic culture establishment and with hybridization at Allium genera

Effective ways of mineral nitrogen fertilizer applications in potatoes and their effect on potato yield

KASAL, P.  – RŮŽEK, P. –  KUSÁ, H. – ČEPL, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 9-18

In field trials during 2008 – 2011 various variants of mineral N fertilizer applications were evaluated in potatoes under conditions of de-stoning technology. Broadcast application of granulated ammonium sulphate prior to planting was compared to local application at potato planting. Furthermore, use of locally applied mineral nitrogen fertilizers containing nitrification and urease inhibitors was verified. In these variants the effect on potato yield was studied. For retention of more rain water, the shape of the ridge was modified, forming an infiltration groove on the top. Year had the highest effect on the results. Local application of ammonium sulphate positively affected tuber yield in all years compared to broadcast application, on average by 10 %. Potato yield was also positively affected by fertilizer containing nitrification inhibitor Alzon and urease inhibitor UREAstabil. Modification in the shape of the ridge, forming infiltration groove on the top, had a tendentious positive impact on potato yield in three out of four trial years.

potato fertilization; local application of nitrogen fertilizers; nitrification and urease inhibitor-containing fertilizers; nitrogen use; potato yield

Corresponding author:
Ing. Pavel KASAL, Ph.D., Potato Research Institute Havlíčkův Brod, Ltd., 
Dobrovského 2366, 580 01 Havlíčkův Brod,
phone: +420 569 489 209, fax: +420 5569 421 578, e-mail: kasal@vubhb.cz

Effect of flesh colour on the content of major antioxidants in potato tubers from organic cultivation (A)

HAMOUZ, K. – LACHMAN, J. – HEJTMÁNKOVÁ, K. – CIMR, J. – PAZDERŮ, K. – ČÍŽEK, M.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 19-26

In 2010 a field trial at the Experimental station of the Czech University of Life Sciences in Prague Uhříněves in terms of organic conditions with 6 blue- or red-fleshed potato varieties and with the yellow-fleshed control Agria variety was performed. After harvest, tubers were analyzed for the content of chlorogenic acid (ChA), ascorbic acid (AA) and total anthocyanins (TA). Flesh colour significantly influenced the ChA content, which was in the varieties with coloured flesh from 1.29 to 1.88 times higher in comparison with the yellow-fleshed Agria variety. Significant differences in the content of ChA were found among the different varieties with coloured flesh, a decrease of the variety with the lowest content to the highest content was 31.6 %. In contrary, AA content did not depend on the colour of the flesh, but the genotype of each variety, as the difference of AA contents between the Blaue Elise (176.7 mg/kg FM – blue-fleshed), Agria (170.6 mg/kg FM - yellow-fleshed) and Highland Burgundy Red (160.3 mg/kg FM – red-fleshed) varieties was inconclusive. TA content was demonstrably influenced by the genotype of the variety, the highest value reached the Blaue Elise variety (84.6 mg cyanidin/kg FM).

potato varieties; colour of flesh; antioxidants; chlorogenic acid; ascorbic acid; anthocyanins

Corresponding author: 
Prof. Ing. Karel HAMOUZ, CSc., Czech University of Life Sciences,
Kamýcká 129, 165 21 Prague 6 - Suchdol
phone: +420 224 382 548, fax: +420 224 382 535, e-mail: hamouz@af.czu.cz

Relative incidence of potato blackleg pathogens in the Czech Republic in 2010 – 2011

KREJZAR, V. –  PÁNKOVÁ, I. –  HAUSVATER, E. –  DOLEŽAL, P. –  KŮDELA, V.
Vědecké práce  – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 27-34 

Blackleg is a seed-borne disease of potato caused by different soft rot bacteria depending mainly on prevailing climate. In this study, we evaluate incidence of various soft rot bacteria in potato stems sampled from six localities in the Czech Republic in 2010 and 2011. We analysed microbiologically altogether 81 samples of potato stems, 52 samples in 2010 and 29 in 2011. Relative incidence of individual potato blackleg pathogen was expressed as percentage of the total number of identified pathogens in a particular year. Of a total 196 isolates obtained from symptomatic blackleg stems, Pectobacterium carotovorum was surprisingly prevailing as it was determined in 63 % and 58 % in 2010 and 2011 respectively. On the other hand, P. atrosepticum was determined only in 6 % and 16 % in 2010 and 2011 respectively. Dickeya chrysanthemi was determined in 16 % namely in samples collected in 2010. This is the first published report of D. chrysanthemi associated with potato blackleg in the Czech Republic. Pseudomonas viridiflava, P. putida and P. fluorescens figured among determined isolates in 2 – 3% in 2010 and in 0 – 20% in 2011.

blackleg; Dickeya chrysanthemi; Pectobacterium atrosepticum; P. carotovorum subsp. carotovorum; Pseudomonas fluorescens; P. putida; P. viridiflava; potatoes;

Corresponding author:
Ing. Václav KREJZAR, Ph.D., Crop Research Institute,
Drnovská 507, 161 06 Prague 6
phone: +420 233 022 427, fax: +420 233 311 592, e-mail: krejzar@vurv.cz

The utilization of molecular markers at assessment of subcollection genotypes of Solanum wild species

ŠVECOVÁ, R. – PTÁČEK, J. –  DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 35-42 

The subcollection „Solanum wild species“ is maintained under in vitro conditions in the gene bank of PRI Havlíčkův Brod (twenty-two tuber-forming, one tuberless). These wild species are the source of resistance to the diseases, pests and abiotic stresses.

Forty eight genotypes from seven wild species (S. acaule, S. berthaultii, S. bulbocastanum, S. brevidens, S. demissum, S. pinnatisectum and S. verrucosum) were tested by means of PCR. The plant material was transferred from in vitro conditions to the greenhouse and grown there. The DNA was isolated from these plants by means of GenElute Plant Genomic DNA Miniprep Kit (Sigma). DNA markers (PVY, late blight) were chosen and tested. The selected markers will be used for subsequent testing of diploid and tetraploid potato collections. The wild species bearing resistance markers will be used for crossing experiments.

wild species; Solanum; tested; DNA; markers

Corresponding author:
Ing. Renata ŠVECOVÁ, Potato Research Institute Havlíčkův Brod, Ltd.,
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 225, fax: +420 469 421 578, e-mail: svecova@vubhb.cz

Evaluation of resistance to potato late blight in transformants with the transgene for SPI2 :: GFP fusion protein

HORÁČKOVÁ, V. – NAVRÁTIL, O. – KOPAČKA, V.  –  DOMKÁŘOVÁ, J.
Vědecké práce - Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 43-52 

In the study results are elaborated concerning laboratory and field evaluation of resistance to potato late blight (Phytophthora infestans Mont. de Bary) in potato lines of the variety Velox, derived after transformation by gene for SPI 2 :: GFP (silk proteinase inhibitor 2) fusion protein from Galleria mellonella. A set of 21 modified Velox lines was tested under in vitro conditions using plant inoculation with P. infestans spore suspension. Based on plant infection extent, the material was divided into three resistance groups. The field challenge test of resistance was not treated by any fungicides during the whole vegetation. Planting was done in three years; determination was always initiated when first visual symptoms of infection occurred, on six dates in regular intervals and was expressed as infection percentage. Slightly positive results of resistance in transgenic clones were recorded; they are not necessary the expression of the gene, but they could be a result of random genetic changes only indirectly associated with transformation. Further verification of gene expression and subsequently stability of resistance level will be necessary.

potatoes; transformants; proteinase inhibitors; Phytophthora infestans Mont de Bary; resistance tests

Corresponding author:
Ing. Vendulka HORÁČKOVÁ, CSc., Potato Research Institute Havlíčkův Brod, Ltd.,
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 220, fax: +420 569 421 578, e-mail: horackova@vubhb.cz

New rational procedures of potato virus laboratory diagnosis using Multiplex ELISA and Luminex-xMAP technology

DĚDIČ, P.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 53-62 

The basic variant of ELISA (Double-Antibody Sandwich - DAS) was evaluated as to detectability of particular virus using monovalent antibodies as well as using artificial antibody combinations (APAs). The results proved satisfactory efficiency up to six antibody mixtures and reveal the possibility of their applications for objectives where the necessity of virus differentiation is not essential. The practical applicability of the bi- and trivalent combinations is recommended.

In the preliminary experiments with new diagnostic method Luminex xMAP based on the fluorescent detection of labelled paramagnetic beds, its reasonable sensitivity was proved, as well as usability for simultaneous detection and differentiation of higher numbers of viruses. The new method might be useful for complex diagnosis at pre-basic breeding materials and for quarantine purposes.

DAS ELISA; complex diagnosis; APAs; differentiation and sensitivity of detection; utilization

Corresponding author:
Ing. Petr DĚDIČ, CSc., Potato Research Institute Havlíčkův Brod, Ltd.,
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 232, e-mail: dedic@vubhb.cz

Analysis of somatic hybrid performance in the field (A)

POLZEROVÁ, H.  –  GREPLOVÁ, M. – DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 63-74 

The somatic hybrid plants (SH) originated from protoplast fusions between diploid S. pinnatisectum 8166 (pnt 8166, 2n=2x=24) and S. tuberosum cv. Bintje (tbr cv. Bintje). The agronomic traits analysed, included: the development of plants during vegetation, incidence of viruses, tuber yield, number, morphology and general appearance. Differences in terms of sprouting time were significant between somatic hybrids and their parents. Hybrid plants were mostly robust and like their parents, initiated flowering. Pollen production was poor in tbr cv. Bintje and hybrid genotypes. Pollen fertility of S. pinnatisectum was 39 %. Only the genotype SH 316 set berries spontaneously. No significant difference in the set of tubers between somatic hybrids and tbr parent was found. The yield of somatic hybrids was significantly lower than for tbr parent. Postharvest ELISA test showed prolific incidence of viruses PVY and PVS in most tested hybrid materials. The taste test confirmed that all selected genotypes were edible.

Solanum tuberosum; Solanum pinnatisectum; somatic hybrid

Corresponding author:
Mgr. Hana POLZEROVÁ, Potato Research Institute Havlíčkův Brod, Ltd.
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 241, fax: +420 569 421 578,  e-mail: h.polzerova@vubhb.cz

Experience with the axenic culture establishment and with hybridization at Allium genera

GREPLOVÁ, M. – POLZEROVÁ, H.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2011, 19: 75-83 

The genotypes A. aflatunense, A. cyathophorum, A. fistulosum, A. flavum, A. schoenoprasum, A. ursinum and A. victorialis were used in the following experiments. The seeds were sown in both in vitro and in vivo conditions. The germination was low; no seeds germinated in A. aflatunense, A. cyathophorum or A. victorialis. In A. flavum, the germination was 49 % in greenhouse and 9 % under in vitro conditions. Three genotypes were only sown in vitro: A. fistulosum, A. ursinum and A. schoenoprasum with a germination of 87 %, 0 % and 35 % respectively. Only a percentage of germinated seeds produced whole plants. A few seedlings of A. flavum were successfully transferred to in vitro conditions. The surface of the plant materials was sterilised using 2% solution Chloramin BM (Bochemie) followed by 70% ethanol. Contamination was less than 3.6 %.The technique of flower castration, pollen separation and pollen viability testing was verified. The pollen viability was 86 – 97 % according to genotype. In total, 491 flowers were pollinated and 330 seeds were obtained.

A. aflatunense; A. cyathophorum; A. fistulosum; A. flavum; A. schoenoprasum; A. ursinum; A. victorialis; fertility; germination; crossing

Corresponding author:
Ing. Marie GREPLOVÁ, Potato Research Institute Havlíčkův Brod, Ltd.
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 241, fax: +420 569 421 578, e-mail: greplova@vubhb.cz

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