Scientific Studies 2012

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

Hana Polzerová, Josef Patzak, Marie Greplová, Lukáš Kreuz, Jaroslava Domkářová
Study of variability in population of somatic hybrids Solanum bulbocastanum (+) S. tuberosum (A)
Hana Polzerová, Marie Greplová, Jaroslava Domkářová
Preliminary experience with hybridization at Allium genera
Renata Švecová, Jiří Ptáček, Jaroslava Domkářová
The utilization of molecular markers at selected interspecific hybrids of Solanum species
Renata Švecová, Hana Polzerová, Jaroslava Domkářová
Petri dish test for determination of resistance to potato late blight (Phytophthora infestans) in wild potato species Solanum acaule
Miloš Faltus, Petr Dědič, Vendulka Horáčková
Efficiency of cryotherapy for virus elimination in potato
Petr Dědič, Tomáš Moravec, Noemi Čeřovská, Helena Plchová, Oldřich Navrátil
A possibility to elucidate transgenic resistance of potatoes for Potato Leafroll Virus (PLRV)
Jan Bárta, Veronika Bártová, Adéla Brabcová, Vendulka Horáčková
Electrophoretic profiles of tuber proteins and isozymes in cultivated and primitive potato species (Solanum spp.)
Jaroslava Domkářová, Bohumil Vokál, Václav Čermák
Dependence between tuber starch content and cooking quality in selected potato varieties
Jiří Diviš, Jan Kulík, Jan Bárta
The effect of leaf fertilizers at selected potato cultivars
Helena Kusá, Pavel Růžek, Pavel Kasal
Effect of mineral nitrogen fertilizers application into the ridge on potato yield and nitrogen efficiency
Petr Doležal, Ervín Hausvater, Jitka Dejmalová, Vladimíra Sedláková
Efficacy of chemical and biological products on Colorado potato beetle (Leptinotarsa decemlineata (Say, 1824)) in the Czech Republic in 2010–2012
Tomáš Litschmann, Petr Doležal, Ervín Hausvater
Sensitivity analysis of selected models for potato late blight forecast under conditions of Czech Republic

Study of variability in population of somatic hybrids Solanum bulbocastanum (+) S. tuberosum (A)

POLZEROVÁ, H. – PATZAK, J. – GREPLOVÁ, M. – KREUZ, L. – DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský, Havlíčkův Brod, 2012, 20: 9-20

Variability in forty somatic hybrids [S. bulbocastanum Dun. (2n=2x=24; cytoplasm type Wγ) + S. tuberosum (2n=2x=24; cytoplasm type Tßγ)] was assessed using in vitro morphology (general habitus, leaf shape and growth vigour), ploidy level, random amplified polymorphic DNA, microsatellites, mitochondrial DNA and chloroplast DNA analyses. Somatic hybrids with the sum of parental ploidy levels were not recorded in the expected percent. In contrast, hexaploid regenerants were more common. Some somatic hybrids were asymmetric. The influence of ploidy level and genome loss had no impact on in vitro morphology. Cytoplasm type influenced leaf shape only. Chloroplasts segregated in a random manner while cosegregation of chloroplasts was sporadic. However, the co-existence of parental types was recorded in all cases in the mitochondrial genome. Somatic hybrids with the cytoplasm type Tβγ showed mostly standard habitus close to in vitro S. tuberosum plants, whereas the hybrids with Wβγ cytoplasm showed branched or standard habitus. Variability among somatic hybrids can be successfully distinguished at the in vitro stage of the plant. This raises the possibility of preselection of prospective somatic hybrids for use in subsequent breeding programmes.

chloroplasts; cytoplasm-nuclear interaction; in vitro; mitochondria; somatic hybridization

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

Preliminary experience with hybridization at Allium genera

POLZEROVÁ, H. – GREPLOVÁ, M. – DOMKÁŘOVÁ, J.

There were two hybridization methods verified at four species of Allium genera (A. ursinum, A. cyathophorum, A. flavum). The first method was based on flower emasculation and immediate pollen application with little brush. In the second method, the castrated and fertile flowers were technically isolated together. The effectiveness and laboriousness of both hybridization methods were discussed. The possibility of self-pollination was tested.

The primary experiments in the intraspecific and interspecific hybridizations were beginning.

A. cyathophorum; A. flavum; A. ursinum; A. victorialis; fertility; germination capacity; hybridization

The utilization of molecular markers at selected interspecific hybrids of Solanum species

ŠVECOVÁ, R. – PTÁČEK, J. – DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 31-40

The selected wild Solanum species were chosen for crossing by means of traditional breeding methods. These wild species, maintained in genebank of Potato Research Institute in Havlíčkův Brod, are a good source of a resistance to various biotic and abiotic factors. 90 hybrids from 23 combinations, which were arisen by reciprocal crossing procedures (wild species x variety or dihaploid) and 19 parental genotypes were tested by means of PCR methods. We obtained the crossing materials with wild species S. acaule, S. berthaultii a S. demissum and with varieties Žofie, Red Anna, Westamyl, Romanze, Krumlov, Ornella and dihaploid genotype number 314. The hybridity of progeny after interspecific hybridization was visually checked by morphological traits and RAPD method. The hybridity was only found in hybrids from 4 crossing combinations. The hybrid materials will be conserved and used to next experiments.

crossing; wild species; RAPD method

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

Petri dish test for determination of resistance to potato late blight (Phytophthora infestans) in wild potato species Solanum acaule

ŠVECOVÁ, R. – POLZEROVÁ, H. – DOMKÁŘOVÁ, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 41-48

Wild potato species Solanum acaule has been long-term hold in the gene bank of Potato Research Institute Havlíčkův Brod. It is maintained in five genotypes under in vitro culture. All Solanum acaule genotypes were transferred from in vitro culture into greenhouse conditions, following 8 weeks samples of well-developed and healthy leaves were taken. Genotypes S. acaule 00026, S. acaule 00238, S. acaule 00027, S. acaule 00029 and S. acaule 00030 were used for Petri dish test to determine resistance to Phytophthora infestans (in Solanum genus). Statistical assessment was performed using analysis of variance (ANOVA) and Tukey’s test. Mean LGR values of evaluated S. acaule genotypes ranged between 0.68 and 1.66 and were significantly more resistant compared to control variety Valfi, reaching mean LGR value 5.45. In Tukey’s test genotypes S. acaule did not differ regarding LGR value. As a result, the wild species S. acaule was determined as medium resistant to potato late blight (Phytophthora infestans).

Solanum acaule; Phytophthora infestans; test; resistance

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

Efficiency of cryotherapy for virus elimination in potato

FALTUS, M. – DĚDIČ, P. – HORÁČKOVÁ, V.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 49-60

The efficacy of the cryotherapy method for elimination of three potato viruses was tested in 21 selected potato genotypes. Eight genotypes were infected with Potato Leaf Roll Virus (PLRV), four with Potato Virus Y (PVY) and nine with Potato Virus S (PVS). Before cryotherapy, the potato explants were acclimated osmotically with sucrose and subsequently air-dehydrated above silica-gel. Afterwards, the potato shoot tips were plunged into liquid nitrogen. After 60 minutes, the explants were removed from liquid nitrogen and planted on the regeneration medium. Survival and regeneration were evaluated after 2 and 8 weeks of cultivation, respectively. The presence of viral pathogens was evaluated preliminary in in vitro explants and subsequently (in the case of a negative result) in seedlings from the greenhouse by means of ELISA. It was demonstrated that cryotherapy was effective for elimination of pathogens PLRV and PVY, but ineffective for elimination of the pathogen PVS. Average virus elimination by the cryotherapy method was 66 % in genotypes infected with PLRV and 65 % in genotypes infected with PVY. No virus-free plants were obtained after cryotherapy of genotypes infected with the pathogen PVS.

cryopreservation; PLRV; PVS; PVY; recovery; Solanum tuberosum L.; virus

Corresponding author:
Ing. Miloš FALTUS, Ph.D., Crop Research Institute,
Drnovská 507, 161 06 Prague 6
phone: +420 233 022 362, fax: +420 233 022 286, e-mail: faltus@vurv.cz

A possibility to elucidate transgenic resistance of potatoes for Potato Leafroll Virus (PLRV)

DĚDIČ, P. – MORAVEC, T. – ČEŘOVSKÁ, N. – PLCHOVÁ, H. – NAVRÁTIL, O.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 61-70

1725 bp long fragment of virus replicase gene ORF 2b was amplified by PCR from PLRV of previously sequenced Oděskij isolate. The amplicon was subcloned into pJET 1.2 blunt plasmid (Fermentas) and used for the construction of expression cassette further containing enhanced CaMV35S promotor and OCS terminator. The binary plasmid with the expression cassette was mobilized into Agrobacterium strain C58. Transgenic plants were prepared from two susceptible varieties Orbit a Red Anna which are also well suited for in vitro regeneration. In total we prepared and tested 8 transgenic lines of Orbit background (Orsvin 1-8) and 3 lines of Red Anna (Rasvin 4, 6, 11). Te resistance was assessed using artificial virus inoculation by grafting in the greenhouse. The plants were inoculated in three different time points of maturity and also the virus load was assayed by ELISA in three different intervals post inoculation. From positive virus titers it is evident that the tested transgenic lines do not show extreme resistance. However, further testing was focused to partial resistance components such as differences in infection resistance, virus multiplication and translocation. Substantial differences in the virus load and its multiplication and translocation between the transgenic lines of both varieties and their non-transgenic controls were observed especially in the second inoculation term. Both virus concentration in plant sap and symptom expression was assessed in greenhouse secondary infected plants. Nontransgenic Orbit plants showed significantly higher absorbance levels than all tested transgenic Orsvin lines (with the exception of line 7) in both assessed periods. It can be speculated that the transgenic plants only allowed the virus to multiply to lower levels. Significant differences were also observed in the expression of visual symptoms at the beginning of vegetation. Lower virus load in secondary infected Rasvin plants was apparent particularly in lines 6 and 11, the secondary inoculated transformants of line 11 were visually effectively symptomless. The results indicate that the majority of tested transgenic lines showed distinct level of resistance.

viral replicase; transgenic line; inoculation by grafting; virus translocation and concentration

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

Electrophoretic profiles of tuber proteins and isozymes in cultivated and primitive potato species (Solanum spp.)

BÁRTA, J. – BÁRTOVÁ, V. – BRABCOVÁ, A. – HORÁČKOVÁ, V.
Vědecké práce – Výzkumný ústav bramborářský, Havlíčkův Brod, 2012, 20: 71-78

Contribution was focused on evaluation of similarity relations among selected genotypes of cultivated and wild Solanum species using electrophoretic profiles of tuber proteins, esterase and peroxidase isozymes. In total, 20 genotypes from 15 Solanum species were evaluated (10 wild species, 5 genotypes of 4 non-traditional cultivated species and 5 cultivars of S. tuberosum). All of the three systems show resolution of the individual genotypes at least on the level of 4% dissimilarity (with only exception of two S. tuberosum cultivars Russet Burbank and Westamyl remaining non-resoluted in esterase profile) and that is why could be used for the wild, primitive and cultivated species characterization. All of the three chosen characterization system are also able to contribute to study of relationships between the wild, primitive and cultivated Solanum species; however for the detailed study these systems are not sufficient.

similarity; potato; electrophoresis; proteins; esterases; peroxidases

Corresponding author:
doc. Ing. Jan BÁRTA, Ph.D., University of South Bohemia, Studentská 13, 370 05 České Budějovice
phone: +420 387 772 432, e-mail: barta@zf.jcu.cz

Dependence between tuber starch content and cooking quality in selected potato varieties

DOMKÁŘOVÁ, J. – VOKÁL, B. – ČERMÁK, V.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 79-90

Based on results of four-year (2008–2011) field trials involving 71 potato varieties the dependence between potato tuber starch content and selected characteristics of cooked potatoes was evaluated. Simultaneously, an assessment of the dependence between tuber starch content and skin smoothness of raw potatoes was performed. Consistency, texture, mealiness, moisture, taste defects, after cooking blackening, quality stability and cooking type were determined on a nine-point scale in cooked potatoes. The dependence was evaluated for the whole studied set of varieties, according to earliness and cooking type. The results indicate that tuber starch content on average relatively uniformly increased with increasing growing period duration; however, growing period duration was not decisive for starch content in several varieties. The dependence between starch content and other studied characteristics using correlation coefficient was recorded for all studied parameters except for cooking type during evaluation of the whole range of ware potato varieties. In case of mealiness (r = 0.679**), texture (r = 0.618**), after cooking blackening (r = 0.347**) and taste defects (r = 0,296*) direct dependence was detected. For other parameters, i.e. moisture (r = -0.601**), quality stability (r = -0.327**), consistency (r = -0.268*), incl. skin smoothness (r = -0.456**) opposite tendency was found. The tendency determined for the whole set was not always valid for groups of varieties with various growing period duration and/or distinct cooking type. The fact that no dependence was confirmed between tuber starch content and cooking type demonstrated that cooking type and/or expected level of cooking quality could not be directly determined from the starch content. The results indicated that level of individual evaluated parameters affecting cooking quality is especially related to characteristics of individual varieties. The effect of weather course, soil and growing conditions must be also taken into account, since it could be expressed in a change of declared cooking type in individual years.

Solanum tuberosum varieties; starch content; cooking quality; cooking type

Corresponding author:
Ing. Jaroslava DOMKÁŘOVÁ, Ph.D., MBA, LL.M., Potato Research Institute Havlíčkův Brod, Ltd.
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 227, e-mail: domkarova@vubhb.cz

The effect of leaf fertilizers at selected potato cultivars

DIVIŠ, J. – KULÍK, J. – BÁRTA, J.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 91-100

The field experiment was conducted on locality with altitude of 380 m in 2011. The effect of urea and leaf fertilizers Lignohumate and Ligno SUPER NPK on potato tuber yield characteristics was evaluated for potato cultivars of different maturity class – cultivar Magda (very-early maturing), Adéla (early maturing), Laura (semi-early maturing), Satina (semi-late maturing) and Bionta (late maturing). The evaluated yield factors were: total tuber yield, portion of marketable tuber yield (tubers under 40 mm), average weight of potato tubers under 40 mm and starch content in potato tubers. All of the evaluated cultivars reacted on urea application by yield increase. Increased yield of tubers after application of Lignohumate was recorded for cultivar Adéla, Laura and Bionta. Application of leaf fertilizer Ligno SUPER NPK increased tuber yield for the same cultivars. Differences between variants of leaf fertilizer applications on tuber yield were statistically insignificant. Increased portion of marketable tubers in the yield was reached after application of urea for cultivars Adéla, Satina and Bionta; application of leaf fertilizer Lignohumate increased yield of the marketable tubers in the case of cultivar Adéla, Laura and Bionta and finally the application of Ligno SUPER NPK caused an increase of marketable tuber yield for cultivars Magda, Adéla and Bionta. Effect of foliar fertilizers on increased proportion of tubers over 40 mm was not statistically significant. The average weight of potato tubers under 40 mm increased after application of urea in the case of cultivars Magda, Adéla and Laura; application of leaf fertilizer Lignohumate increased the average weight of potato tubers under 40 mm for cultivars Adéla, Laura and Satina; application of Ligno SUPER NPK caused accrual of this parameters in the case of cultivars Magda, Laura and Satina. The highest content of starch in potato tubers was determined for all of the leaf fertilizers only in the case of cultivar Laura. The other cultivars showed the higher starch content in control variants than in the variants of leaf fertilizer application. Differences in starch content were not statistically significant. The obtained results demonstrated that application of leaf fertilizer beyond all doubt belongs to potato production technology.

potato; leaf fertilizer; yield; tuber weight; starch content

Corresponding author:
doc. Ing. Jiří DIVIŠ, CSc., University of South Bohemia,
Studentská 13, 370 05 České Budějovice
phone: +420 387 772 444, e-mail: divis@zf.jcu.cz

Effect of mineral nitrogen fertilizers application into the ridge on potato yield and nitrogen efficiency

KUSÁ, H. – RŮŽEK, P. – KASAL, P.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 101-112

The effect of band application of various mineral fertilizers to potato cultivated in technology with stone separation has been studied in Valečov in 2008-2011. ¹⁵N-labelled dissolved fertilizers (ammonium sulphate = SA, urea with nitrification inhibitor = MOn, urea with urease inhibitor = MOu) were applied in band side of tubers at planting. The soakage groove was made on top of the ridge (treatments with MOn and MOu) for the retention of more rainwater, improving moisture conditions within the ridge and increasing N use efficiency from applied fertilizers. The obtained results were affected by year strongly. The highest tuber yields and nitrogen uptakes in mineral fertilized treatments (73 t.ha^-1 and 207 kg N.ha^-1) were reached in humid year 2011, the lowest ones (41 t.ha^-1 a 161 kg N.ha^-1) in 2008. The efficiency of mineral fertilizer nitrogen was from 42 % (in 2008) to 70 % (in 2011). The significant effect of soakage groove was observed only in more arid year 2008: nitrogen efficiency elevated from 40 to 48 % and from 33 to 44 % for MO and MOu respectively.

band application of mineral fertilizers; ¹⁵N; nitrification inhibitor; urease inhibitor; nitrogen efficiency; tuber yield

Corresponding author:
Ing. Helena KUSÁ, Ph.D., Crop Research Institute,
Drnovská 507, 166 06 Prague 6
phone: +420 233 022 266, e-mail: kusa@vurv.cz

Efficacy of chemical and biological products on Colorado potato beetle (Leptinotarsa decemlineata (Say, 1824)) in the Czech Republic in 2010–2012

DOLEŽAL, P. – HAUSVATER, E. – DEJMALOVÁ, J. – SEDLÁKOVÁ, V.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 113-132

The trials performed in 2010–2012 indicated high efficacy of insecticides from the group of neonicotinoids (Actara 25 WG, Biscaya 240 OD, Mospilan 20 SP, Monceren G) and also a product based on spinosad (SpinTor). Insecticide efficacy was near or equal to 100 %. In these years any reduced efficacy of Mospilan 20 SP that was recorded in the years 2007– 2009 was not confirmed in the trial locality (southern Moravia, Žabčice). In 2010 reduced efficacy of Monceren G was determined due to strong longer lasting field wetting, when shortly after planting washing off and dilution of dressing agent on seed tubers occurred. Botanical insecticide Neem Azal T/S (a.i. azadirachtin A) showed gradually increasing efficacy and confirmed reliable mechanism of acting. For Novodor FC based on Bacillus thuringiensis var. tenebrionis gradual reduction of its effect was recorded and in 2012 deactivation of the insecticide probably due to intensive solar radiation was found.

Colorado potato beetle; insecticide efficacy; neonicotinoids; azadirachtin; Bacillus thuringiensis var. tenebrionis; resistant population

Corresponding author:
Ing. Petr DOLEŽAL, Ph.D., Potato Research Institute Havlíčkův Brod, Ltd.
Dobrovského 2366, 580 01 Havlíčkův Brod
phone: +420 569 466 240, e-mail: dolezal@vubhb.cz

Sensitivity analysis of selected models for potato late blight forecast under conditions of Czech Republic

LITSCHMANN, T. – DOLEŽAL, P. – HAUSVATER, E.
Vědecké práce – Výzkumný ústav bramborářský Havlíčkův Brod, 2012, 20: 133-142

The study describes performed sensitivity analysis of two models for potato late blight forecast, based on data from 2011 and 2012 for the locality Lípa near Havlíčkův Brod considering potential inaccuracy in air humidity measurement. It is shown that in case of negative prognosis model a change in measured humidity values is expressed as a continuous systematic error, where the size of deviation in predicted data for value 150 reaches 1-2 days for 1% humidity change. In NoBlight model lesser changes are found in case of longer moisture periods (year 2011) and they are especially expressed at negative deviations in measured humidity; however, in situations, when shorter periods with high air humidity prevail (year 2012), the differences are more pronounced, they are again higher in case, when a sensor measures lower values than actual ones. Therefore it is necessary to perform a verification control of air humidity sensors in automated meteorological stations before the start of every season using of a simple calibration agent.

potato late blight; forecasting methods; air humidity

Corresponding author:
RNDr. Tomáš LITSCHMANN, Ph.D., AMET, Velké Bílovice
Žižkovská 1230, 691 02 Velké Bílovice
phone: +420 519 346 252, e-mail: amet@email.cz