Scientific Studies 2014

ote: Papers marked with (A) are written in English

  • Marie Greplová, Hana Polzerová, Renata Švecová, Jaroslava Somkářová
    Involvement of somatic hybrids into sexual hybridization (A)
  • Petr Dědič, Noemi Čeřovská, Helena Plchová, Tomáš Moravec
    Recombinant antibodies and possibilities of their utilization for laboratory diagnosis of potato viruses
  • Jiří Ptáček, Jaroslav Matoušek, Renata Švecová, Jaroslava Domkářová
    The detection of Potato virus Y by means of DNA-RNA hybridization
  • Karel Hamouz, Jaromír Lachman, David Bečka, Matyáš Orsák, Jiří Cimr, Vladimír Pivec, Jaroslav Čepl
    Influence of selected factors on the content of chlorogenic acid and antioxidant activitiy of potato tubers with different flesh colour (A)

Involvement of somatic hybrids into sexual hybridization (A)

GREPLOVÁ, M. – POLZEROVÁ, H. – ŠVECOVÁ, R. – DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2014, 22: 9-34

Aim of this work was to evaluate cross ability of two somatic hybrids SH292 and SH1003 with high level of resistance to Phytophthora infestans (PI) originated from protoplast fusion 1EBN diploid Solanum pinnatisectum 8166 + 2EBN Solanum tuberosum dihaploid 299. The pollination of these hybrids was successful after application of pollen  tbr cvs: Barbora, Valfi, Flavie and Karin. More than four hundred flowers was pollinated which led to production of 19 berries, 343 seeds, 90 seedlings in crossing generation (CG)1. All seed germination averaged 21 %. Almost all genotypes bloomed and had good pollen fertility. 38 % of genotypes had long stolons. Repeated laboratory test showed again high level of resistance to PI at chosen genotypes. The genotypes tested in the field were moderately resistant in most cases and three genotypes had high level of field resistance. One hybrid had very good table quality but it was moderate resistant to PI. In CG2, more than one thousand flowers were pollinated. As result of that pollination, 27 berries, 1032 seeds and 418 seedlings were obtained. All seeds germination of CG2 reached in average 41 %.  One third of them flowered; half of them had good pollen productivity as well as fertility. No genotype had stolons. Chosen genotypes were again highly resistant to PI in laboratory test. Reciprocal crosses between somatic hybrids failed and the same situation was in reciprocal crosses of them with two hybrids of CG1. During the years 2009–2014, the laboratory test on Petri dish showed good predictive value for first selection of breeding materials.
This work demonstrated the possibility of somatic hybrids to be sexually crossed with Solanum tuberosum cultivars and also proved that the resistance to PI from wild Solanum pinnatisectum 8166 was passed to somatic hybrids SH292 and SH1003 and also to CG1 and CG2 progenies.

late blight resistance; Phytophthora infestans; progeny; sexual hybridization; Solanum pinnatisectum; S. tuberosum; somatic hybrids

Corresponding author:
Ing. Marie GREPLOVÁ, Ph.D., Potato Research Institute Havlíčkův Brod,
Dobrovského 2366, 580 01 Havlíčkův Brod, Czech Republic
phone: +420 569 466 239, e-mail: greplova@vubhb.cz

Recombinant antibodies and possibilities of their utilization for laboratory diagnosis of potato viruses

DĚDIČ, P.  –  ČEŘOVSKÁ, N. – PLCHOVÁ, H. – MORAVEC, T.
Vědecké práce  –  Výzkumný ústav bramborářský Havlíčkův Brod, 2014, 22: 35-44

The genes encoding the coat protein (CP) of Potato virus X and  Potato virus Y (PVYN-Wi), respectively, were isolated,  cloned into expression vectors and  expressed in Escherichia coli. Purified recombinant proteins of both viruses were used for rabbit immunization and obtained polyclonal antibodies were tested for the detection of both viruses in host plants  Nicotiana tabacum, N. benthamiana  and Solanum tuberosum. The antibodies of both viruses specifically detected the homolog antigens by Western blots as well as the whole spectra of tested virus isolates using indirect plate trapped antigen IPTA ELISA. The recombinant antibodies of both viruses did not react in double antibody sandwich (DAS) ELISA as coating  of plates, as well as the conjugations of both antibodies with enzyme alkalic phosphatase were unsuccessful. 

 ELISA modification; virus coat protein; potato virus X; potato virus Y; detectability

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

The detection of Potato virus Y by means of DNA - RNA hybridization

PTÁČEK, J. – MATOUŠEK, J. – ŠVECOVÁ, R. – DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2014, 22: 45-56

The detection methods of Potato Virus Y (PVY) by means of DNA-RNA hybridization were tested. Various methods of total RNA isolation were used. The specific PVY hybridization probe was prepared (based on the sequence of selected PVY isolate), the probe was labelled nonradioactively and radioactively, respectively. These probes were used for the PVY detection. The use of PVY hybridization probe (with all methods of probe labelling) for the virus detection was easy, sensitive and reliable. The achieved results were compared with the ELISA method from the point of view sensitivity and reliability.

potatoes; Potato Virus Y; DNA-RNA hybridization

Corresponding author:
RNDr. Jiří PTÁČEK, CSc., Potato Research Institute Havlíčkův Brod,
Dobrovského 2366, 58001 Havlíčkův Brod
phone: +420 569 466 244, e-mail: ptacek@vubhb.cz

Influence of selected factors on the content of chlorogenic acid and antioxidant activity of potato tubers with different flesh colour (A)

HAMOUZ, K. – LACHMAN, J. – BEČKA, D. – ORSÁK, M. – CIMR, J. – PIVEC, V. – ČEPL, J.
Vědecké práce – Výzkumný ústav bramborářský, Havlíčkův Brod, 2014, 22: 57-70

In precise field trials in the years 2010 and 2011 the effect of genotype and location in varieties with yellow, white, purple or red flesh on content of chlorogenic acid (CA) and in 2011 and 2012 the effect of organic cultivation was evaluated. The results show a statistically significant effect of genotype to CA content, which ranged from 74.0 mg/kg FM (Agria) to 824.6 mg/kg FM (Vitelotte). Demonstrable effect of flesh colour on CA content between varieties with coloured flesh was found (429.9 mg/kg FM) in comparison with varieties with yellow or white flesh (71.1 mg/kg FM). For varieties with coloured flesh ruled rather the genotype specific variety than the purple or red flesh colour. In terms of the influence of location higher CA levels were found on warm locations with frequent periods of drought in comparison with locations of high altitude climatic conditions which are favourable for potato cultivation. In the organically grown potatoes significantly higher levels of CA were found as compared with conventional variant. Linear correlation between antioxidant activity and content of tubers CA (r = 0.914, p £ 0.01) was found.

potato; variety; yellow, purple and red flesh; ecological farming; location; year

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

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