Purpose. Analysis of the allelic state of Ppd-1 genes, which control sensitivity to photoperiod, in varieties and lines of bread winter wheat, and comparison of the results obtained with field observations on the duration of periods before heading and flowering, whose originators were the Nosivska Breeding and Research Station of the V. M. Remeslo Myronivka Institute of Wheat National Academy of Agrarian Sciences of Ukraine and Poltava State Agrarian Academy of the Ministry of Education and Science of Ukraine. Methods. The following methods were used in the work: DNA extraction, allele-specific PCR, agarose and polyacrylamide gel electrophoresis, analysis of variance. Results. It was revealed that ‘Yuvivata 60’ variety has a recessive Ppd-1 genotype and belongs to the III haplotype by a combination of mutations in the structure of Ppd-D1 gene. Line ‘L41/95’ was heterogeneous by alleles of Ppd-D1 gene, which corresponded to the presence of haplotypes III and VII. All other tested cultivars and lines were characterized by alleles Ppd-A1b, Ppd-B1b and Ppd-D1a, and assigned to haplotype VII. According to the results of statistical data processing, the duration of the period from May, 1 to heading was the smallest for the variety ‘Donskaya polukarlikovaya’ in the conditions of both the Forest-Steppe and Polissia-Forest-Steppe regions of Ukraine, the longest – in the varieties ‘Yuvivata 60’, ‘Myronivska 61’ and ‘L41/95’. The differences between these groups were significant and amounted to 10 days. Conclusions. A breeding material with a high adaptive ability for growing conditions in Polissia-Forest-Steppe zone was studied by the allelic state of the Ppd-1 genes. A low level of polymorphism in the studied varieties and lines was revealed by the alleles of Ppd-1 genes [12.5% – Ppd-D1b (²²²), 12.5% – Ppd-D1à/b (²²²/VII), 75% – Ppd-D1a (VII)], that agrees with the hypothesis that breeders gave a greater preference for the photoperiod-insensitive wheat genotype under Ukrainian conditions. The genotypes with the dominant Ppd-D1a (VII) gene almost completely dominate in the south of Ukraine. At the same time, in northern latitudes, weather conditions negate the advantages of the genotypes with Ppd-D1a gene

Purpose. To create a multiplex system for identification glyphosate-tolerant sugar beet by using PCR. Methods. Molecular genetic analysis. Results. The article presents the results of studies to determine the parameters of the polymerase chain reaction (PCR) in order to develop a multiplex system for identification of the structural elements of the design of transgenic gene cp 4 epsps, which provides tole­rance to glyphosate. For amplicon target DNA sequences, the following values of temperature conditions of PCR were determined: step 1 (initial denaturation) 95 °C – 3 min; step 2 (specific reaction products accumulation): denaturation 95 °C – 45 s; hybridization of primers 55 °C – 50 s; elongation 72 °C – 1 min; number of cycles – 40; step 3 (final elongation) 72 °C – 6 min. A series of PCR were carried out for the purpose of selecting the optimal amount of DNA matrix for efficient estimate of transgenic sugar beet plants for the presence of specific sequences. Conclusions. To identify transgenic glyphosate-tolerant sugar beet, it is advisable to determine 35S promoter and gene cp 4 epsps in individual genotypes. It was found that during the selection of temperature parameters of multiplex reaction a 5 °C rise in primer hybridization temperature did not affect the identification of gene als that allowed to include specific primers for determination of this sequence as an internal control. Based on the results of test multiplex reactions, concentrations of dNTPs and Mg2+ ions were determined that allowed to exclude the possibility of non-specific fragments and false-negative results. The optimum amount of matrix DNA (100–150 ng) for an efficient estimate of transgenic sugar beet plants for the presence of specific sequences was determined. Obtained results allowed to develop a multiplex test system for identification of transgenic glyphosate-to­lerant sugar beet which can be used for simultaneous determination of the 35S promoter, cp 4 epsps gene and als gene as an internal reaction control.

Purpose. Defining marker-trait associations of microsatellite markers with specific regions of the genome that control important agronomical traits in the investigated varieties originated in the Plant Breeding and Genetics Institute – National Center of Seed and Cultivar Investigations and entered into the State register of plant varieties suitable for dissemination in Ukraine during different years. Methods. Molecular genetic methods (extraction of genomic DNA, polymerase chain reactions (PCR), electrophoresis of amplification products in polyacrylamide gel), field methods (phenological observations of heading date and analysis of plant height, visual assessment of the colour and length of wheat ear and awns), statistical methods (evaluation of trait means by descriptive statistic instruments of EXCEL package, ANOVA method performed by GLM instrument from AGROBASE 21 package). Results. During four gro­wing years (2010/11, 2011/12, 2012/13, 2013/14), 47 bread winter wheat varieties were phenotypically measured and analyzed with 17 microsatellite loci. 35 marker-trait associations (MTA) for heading date, 39 for plant height, 33 for awn size, 20 for ear colour and 8 for ear size were found to be stable and significant during two–four different gro­wing years. Conclusions. Microsatellite markers that showed substantial and stable during different growing years associations with agronomical traits can be useful and suitable for marker-assisted selection (MAS) in Ukrainian wheat breeding programs.

Purpose. To review publications relating to the key point of the genotyping technology that is competitive allele-specific polymerase chain reaction (which is called now Kompetitive Allele Specific PCR, KASPTM) and its use in various genetic-breeding researching (a study of maize). Results. The essence of KASP-genotyping, its advantages are highlighted. The requirements for matrix DNA are presented, since the success of the KASP-analysis depends on its qua­lity and quantity. Examples of global projects of plant breeding for increasing crop yields using the KASP genoty­ping technology are given. The results of KASP genotyping and their introduction into breeding and seed production, in particular, for determining genetic identity, genetic purity, origin check, marker-assisted selection, etc. are presented using maize as an example. It is demonstrated how geno­mic selection according to KASP genotyping technology can lead to rapid genetic enhancement of drought resistance in maize. Comparison of the effectiveness of creating lines with certain traits (for example, combination of high grain yield and drought resistance) using traditional breeding approaches (phenotype selection) and molecular genetic methods (selection by markers) was proved that it takes four seasons (two years in case of greenhouses) in order to unlock the potential of the plant genotype using traditional self-pollination, test-crossing and definitions), while using markers, the population was enriched with target alleles during one season. At the same time, there was no need for a stress factor. Conclusions. KASP genotyping technology is a high-precision and effective tool for modern genetics and breeding, which is successfully used to study genetic diversity, genetic relationship, population structure, gene­tic identity, genetic purity, origin check, quantitative locus mapping, allele mapping, marker-assisted selection, marker-assisted breeding. It is expedient and timely to introduce KASP genotyping technology in our country to solve a wide range of modern genetics, breeding, seed production tasks.

Purpose. Literature review of DNA banks creation as the current strategy of plant genetic resources conservation. Results. The current state of plant genetic resources conservation was analyzed in the context of the threat of gene­tic erosion. The importance of DNA banks was shown which function is to store DNA samples and associated products and disseminate them for research purposes. The main DNA banks in the world were described, including the Republican DNA Bank of Human, Animals, Plants and Microorganisms at the Institute of Genetics and Cytology of the National Academy of Sciences of Belarus. Stages of DNA banking were considered: tissue sampling (usually from leaves), cell destruction, DNA extraction, DNA storage. Different methods of tissue sampling, extraction and DNA storage were compared. The need for Plant DNA Bank creation in Ukraine was highlighted. Conclusions. DNA collections is an important resource in the global effort to overcome the crisis in biodiversity, for managing world genetic resources and maximi­zing their potential.

This paper presents the results of molecular studies on testing and evaluation of multiplex test systems for the genetically modified organisms’ (GMOs) identification by PCR in real time. For the detection of genetic modifications in plants of the cabbage family (Brassicaceae) screening of major regulatory sequences is insufficient, as in genetic engineering manipulations 35S promoter – of DNA-containing cauliflower mosaic virus (CaMV) is used, which presence in the test material can lead to false positive results. In this work the feasibility of testing for the presence of viral CaMV’s DNA and analysis of screened gene sequences of interest for the usage during the analysis of rapeseed (Brassica napus L.) samples. It was established that the analysis of the presence/absence of a positive result for the CaMV 35S promoter is not giving complete answers to the absence of GMO in the sample. The appropriateness of GMO’s screening in rape seeds with NOS terminator and genes of interest.

Мета. Класифікація вихідного матеріалу кукурудзи за ступенем холодостійкості, ідентифікація самозапилених ліній кукурудзи в лабораторних і польових умовах за холодостійкістю та основними господарсько-цінними показниками та їхнє генотипування на основі SSR маркерів. Методи. Польові та лабораторні методи, молекулярно-генетичний аналіз. Результати. У результаті досліджень на основі cold test проведено ранжування самозапилених ліній кукурудзи за рівнем холодостійкості. Встановлено, що на результати ранжування не впливає тип зерна, оскільки до найбільш холодостійких належать лінії з різним типом зерна. Визначено, що польова схожість ліній кукурудзи варіювала залежно від строку сівби та становила 32,1–87,8% за першого (6–6,5 °С) строку сівби, 41,8–88,5% – за другого (8–8,5 °С) та 51,1–90,0% – за третього (10–10,5 °С). Встановлено, що самозапилені лінії: HLG 1203, HLG 1238, Co 255, UCH 37 та FV 243, Q170, AK 135, F2, L155 та P165 мають найкращу регенеративну здатність та найвищу схожість за холодного пророщування насіння та її збереження відносно контролю. За показниками урожайності самозапилених ліній визначено, що вони по різному реагують на строки сівби. Визначений генетичний поліморфізм холодостійких ліній за 5 SSR маркерами. Відповідно до отриманих результатів встановлено наявність від 3 до 7 алелів. Індекс поліморфності локусу (РІС) склав 0,56–0,86. За трьома маркерами – bnlg1129, bnlg1782 та phi064 у досліджуваних ліній було виявлено внутрішньолінійний поліморфізм. Результати досліджень дозволили визначити 4 кластери, які відобразили ступінь генетичної близькості за досліджуваними маркерами. Встановлено, що лінії Ak 135 та Ak 153, які увійшли в один кластер є найбільш спорідненими, а найбільш віддаленими – Со225 та Q 170. Отримані дані свідчать, що досліджувані лінії кукурудзи сформували кластери відповідно до їхнього походження і деякі – відповідно до їхньої холодостійкості. Висновки. За результатами досліджень виділено 7 самозапилених ліній кукурудзи (Co 255, HLG 1203, HLG 1238, Q 170, UCH 37, Ak 135, FV 243), які є цінним вихідним матеріалом у селекції на холодостійкість.

Цель. Классификация исходного материала кукурузы по степени холодоустойчивости, идентификация самоопыленных линий кукурузы в лабораторных и полевых условиях по холодостойкости и основным хозяйственно-ценными показателями и их генотипирование на основе SSR маркеров.Методы. Полевые и лабораторные методы, молекулярно-генетический анализ.Результаты. В результате исследований на основе cold test проведено ранжирование самоопыленных линий кукурузы в соответствии с уровнем их холодоустойчивости. Установлено, что на результат ранжирования не влияет тип зерна, поскольку к наиболее холодостойким принадлежат линии с различным типом зерна. Определено, что полевая всхожесть линий кукурузы варьировала в зависимости от срока посева и составила 32,1–87,8% при первом (6–6,5 °С) сроке посева, 41,8–88,5% – при втором (8–8,5 °С) и 51,1–90,0% – при третьем (10–10,5 °С). Установлено, что самоопылённые линии: HLG 1203, HLG 1238, Co 255, UCH 37 и FV 243, Q170, AK 135, F2, L155 и P165 имеют лучшую регенеративную способность и высокую всхожесть в условиях холодного проращивания семян и ее сохранение относительно контроля. По результатам полевых исследований выделены линии Co 255, HLG 1203, HLG 1238 и Q 170, в которых были отмечены высокие показатели процента сохранения всхожести по сравнению с контролем. По показателям урожайности самоопыленных линий кукурузы определено, что они по-разному реагируют на сроки посева. В результате ПЦР анализа 13 холодостойких линий определен генетический полиморфизм по 5 SSR маркерам. Согласно полученным результатам установлено наличие от 3 до 7 аллелей. Индекс полиморфности локуса (РІС) составил 0,56–0,86. Определено, что по трем маркерами bnlg1129, bnlg1782 и phi064 в исследуемых линиях был обнаружен внутрилинейный полиморфизм. Результаты исследований позволили определить четыре кластера, которые отражают степень генетической близости по исследуемым маркерам. Установлено, что линии Ak 135 и Ak 153, которые вошли в один кластер, являются наиболее родственными, а наиболее удаленными – Со225 и Q 170. Полученные данные свидетельствуют, что исследуемые линии кукурузы сформировали кластеры в соответствии с их происхождением и некоторые – в соответствии с их холодоустойчивостью.Выводы. По результатам исследований выделено 7 самоопыленных линий кукурузы (Co 255, HLG 1203, HLG 1238, Q 170, UCH 37, Ak 135, FV 243), которые являются ценными источниками при селекции на холодоустойчивость. | Purpose. Classification of the source material of maize by the rate of cold resistance, identification of self-pollinated maize lines in laboratory and field conditions by cold resistance and main economically valuable indicators, and their genotyping based on SSR markers.Methods. Field and laboratory methods, molecular genetic analysis.Results. As a result of studies based on the cold test, the ranking of self-pollinated lines of corn on 6 groups was carried out in accordance with the level of their cold resistance. It was revealed that the type of grain does not affect the ranging result, since lines with different types grain belong to the most cold-resistant ones. It was determined that field germination of maize lines varied depending on the sowing period and amounted to 32,1–87,8% at the first (6–6,5 °С), 41,8–88,5% at the second (8–8,5 °С) and 51.1–90.0% at the third (10–10,5 °С) sowing dates. It was determined that self-pollinated lines: HLG 1203, HLG 1238, Co 255, UCH 37 and FV 243, Q170, AK 135, F2, L155 and P165 have the best regenerative ability and high germination under conditions of cold germination of seeds and its maintaining relatively to control. Based on the results of field studies, the lines Co 255, HLG 1203, HLG 1238, and Q 170 were identified, in which the germination rate (in percent) was high compared to the control. Based on the assessment of the yield of self-pollinated maize lines, it was determined that they react differently to the sowing dates. As a result of PCR analysis of 13 cold-resistant lines, genetic polymorphism was determined by 5 SSR markers. According to obtained results, the presence of 3 to 7 alleles was revealed. PIC was 0.56–0.86. It was determined that, using the three markers bnlg1129, bnlg1782 and phi064, intralinear polymorphism was detected in the studied lines. The results of the studies allowed to obtain four clusters which reflect the degree of genetic proximity to the studied markers. It was found that lines the Ak 135 and Ak 153 entered the same cluster are the most related and the most distant lines are Co225 and Q 170. The obtained data indicate that the studied maize lines formed clusters according to their origin and some lines according to their cold resistance.Conclusions. According to the results of studies, 7 self-pollinated lines of corn (Co 255, HLG 1203, HLG 1238, Q 170, UCH 37, Ak 135, FV 243) were indentify. They are valuable sources for breeding to cold resistance. | Мета. Класифікація вихідного матеріалу кукурудзи за ступенем холодостійкості, ідентифікація самозапилених ліній кукурудзи в лабораторних і польових умовах за холодостійкістю та основними господарсько-цінними показниками та їхнє генотипування на основі SSR маркерів.Методи. Польові та лабораторні методи, молекулярно-генетичний аналіз.Результати. У результаті досліджень на основі cold test проведено ранжування самозапилених ліній кукурудзи за рівнем холодостійкості. Встановлено, що на результати  ранжування не впливає тип зерна, оскільки до найбільш холодостійких належать лінії з різним типом зерна. Визначено, що польова схожість ліній кукурудзи варіювала залежно від строку сівби та становила 32,1–87,8% за першого (6–6,5 °С) строку сівби, 41,8–88,5% – за другого (8–8,5 °С) та 51,1–90,0% – за третього (10–10,5 °С). Встановлено, що самозапилені лінії: HLG 1203, HLG 1238, Co 255, UCH 37 та FV 243, Q170, AK 135, F2, L155 та P165 мають найкращу регенеративну здатність та найвищу схожість за холодного пророщування насіння та її збереження відносно контролю. За показниками урожайності самозапилених ліній визначено, що вони по різному реагують на строки сівби. Визначений генетичний поліморфізм холодостійких ліній за 5 SSR маркерами. Відповідно до отриманих результатів встановлено наявність від 3 до 7 алелів. Індекс поліморфності локусу (РІС) склав 0,56–0,86. За трьома маркерами – bnlg1129, bnlg1782 та phi064 у досліджуваних ліній було виявлено внутрішньолінійний поліморфізм. Результати досліджень дозволили визначити 4 кластери, які відобразили ступінь генетичної близькості за досліджуваними маркерами. Встановлено, що лінії Ak 135 та Ak 153, які увійшли в один кластер є найбільш спорідненими, а найбільш віддаленими – Со225 та Q 170. Отримані дані свідчать, що досліджувані лінії кукурудзи сформували кластери відповідно до їхнього походження і деякі – відповідно до їхньої холодостійкості.Висновки. За результатами досліджень виділено 7 самозапилених ліній кукурудзи (Co 255, HLG 1203, HLG 1238, Q 170, UCH 37, Ak 135, FV 243), які є цінним вихідним матеріалом у селекції на холодостійкість.

Purpose. To develop a conceptually new method for determination of varietal purity (typicality), hybridity, steri­lity of seed lots. Methods of molecular biology (genomic DNA extraction, PCR with SSR markers application, capillary electrophoresis), genetic, statistical, mathematical analysis. Results. New method for determining the varietal qualities of seed lot was developed that consists of the following steps: simultaneous DNA extraction from a representative sample of aggregated seeds; PCR and further analysis of the amplification products by determination of the qualitative and quantitative composition of SSR-markers’ alleles; calculation of values of varietal seed lot quality using experimentally derived allele ratios. Conclusions. The developed method for determining varietal qualities of seed lots allows to reduce significantly the consumption of materials, time and labor during the analysis. Consistent qualification and quantification of alleles in the total sample of a seed lot is a conceptually new approach to establish varietal purity (typicality), hybridity, sterility.

Purpose. The analysis of allelic status of such key genes of carotenogenesis as gene of lycopene-ε-cyclase (lcyε) and gene of β-carotene hydroxylase (crtRB1) for DNA-markers in domestic maize lines and their hybrids. Methods. DNA isolation, PCR, gel electrophoresis. Results. Allelic status of key genes of carotenogenesis was investigated in eight maize inbred lines and their single crosses. Molecular genetic polymorphism in the studied sample of maize lines and hyb­rids has been detected in gene of β-carotene hydroxylase for marker crtRB1-3’TE. For this gene, codominant character of inheritance of alleles of parental lines in single crosses was confirmed. For markers of gene of lycopene-ε-cyclase lcyε-3’INDL and lcyε-SNP216, polymorphism in the group of investigated lines and hybrids has not been identified, genotypes included only one variant of alleles for each marker. For lines ‘DK253ZSZM’, ‘DK633/266zS,zM’, ‘DK366zS,zM’ and hybrids ‘DK296S×DK253ZSZM’, ‘DK272S×DK633/266zS,zM’ and ‘DK231S×DK366zS,zM’, the decrease of the activi­ty of β-carotene hydroxylase owing to the mutation of gene crtRB1 under the influence of transposone element at the 3’-end, the inhibition of β-carotene transition into β-cryptoxanthin can be expected, that allows to predict β-carotene accumulation in grain. Conclusions. The study of allelic status of carotenegenesis gene of lycopene-ε-cyclase in maize showed no polymorphism for markers lcyε-3’INDL and lcyε-SNP216 in eight inbred lines and their single crosses, along with this, for marker lcyε-3’INDL in genomes of all studied samples the allele was identified to be favorable for the accumulation of β-carotene. For marker crtRB1-3’TE of gene of β-carotene hydroxylase, the studied breeding material was polymorphic. Allele of crtRB1 being favorable for the accumulation of β-carotene was identified in lines ‘DK253ZSZM’, ‘DK633/266zS,zM’, ‘DK366zS,zM’ and hybrids ‘DK296S×DK253ZSZM’, ‘DK272S×DK633/266zS,zM’ and ‘DK231S×DK366zS,zM’. Single crosses inherit maternal and paternal alleles of gene of β-carotene hydroxylase codominantly.