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KASP<sup>TM</sup> genotyping technology and its use in genetic-breeding programs (a study of maize)
2017
Н. Е. Волкова | В. М. Соколов
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.
Afficher plus [+] Moins [-]Assessment of stability and plasticity of new hybrids of maize (Zea mays L.) under the conditions of Polissia and Steppe zones of Ukraine
2016
Присяжнюк, Л. М | Шовгун, О. О | Король, Л. В | Коровко, І. І
Purpose. To select promising high productive maize hybrids of middle-early maturity group in terms of stability and plasticity of main economic characters. Methods. Field study, laboratory test, analytical procedure and statistical evaluation. Results. 14 maize hybrids recorded in the State Register of Plant Varieties Suitable for Dissemination in Ukraine in 2015 were studied for plasticity and stability of such traits as productivity, protein and starch content. Intensive highly-plastic hybrid ‘SI Tiptop’ was selected among the studied ones for productivity trait that can respond properly to changes of growing conditions. It was defined that for the starch content such hybrids as ‘SI Tiptop’, ‘SI Enigma’, ‘SI Arioso’, ‘Svich 38’, ‘Svich 35’, ‘HU 8653’, ‘Zdobutok’ and ‘SI Contrakt’ belonged to the intensive type and combined rather high values and the stability of the studied trait under variable conditions. The following hybrids as ‘NS 2642’, ‘DK S3016’, ‘Svich 38’, ‘NS 2632’ were qualified as intensive for protein content and appeared to be highly-plastic but stability values of this trait were low. ‘Svich 38’ hybrid was intensive simultaneously for two traits such as protein and starch content and showed rather high values of plasticity. ‘SI Tiptop’, ‘SI Enigma’ and ‘Svich 35’ were defined as hybrids of extensive type that provided stable protein content in adverse cultivation conditions. Conclusions. On the condition that intensive crop growing technologies should be used, for obtaining stable yields it is advisable to sow only highly-plastic hybrids that can adapt to unfavorable environmental factors, including ‘SI Tiptop’ – for productivity trait, ‘Zdobutok’ and ‘SI Kontrakt’ – for starch content, ‘MAC 24N‘, ‘NA 2642‘ and ‘Danubio’ – for protein content.
Afficher plus [+] Moins [-]KASP<sup>TM</sup> genotyping technology and its use in genetic-breeding programs (a study of maize)
2017
Волкова, Н. Е | Sokolov, V. M.
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.
Afficher plus [+] Moins [-]Bioinformatic analysis of maize gene encoding starch branching enzyme SBEIIb
2016
Сліщук, Г. І | Жернаков, Т. Ю | Волкова, Н. Е
Purpose. Investigation of maize ae1 gene polymorphism by bioinformatic methods. Methods. Global and local alignment of the nucleotide and amino acid sequences, in silico translation and transcription, translates modeling, primers design, phylogenetic analysis. Results. 255 nucleotide sequences of maize аe1 gene, 500 amino acid sequences of homology translates of maize ae1 gene (SBEIIb enzyme homologs) and 100 mRNA expressed from the maize ae1 gene were analyzed to establish phylogenetic relationships. Polymorphism of maize ae1 gene different regions was investigated by bioinformatic methods. Modeling of the maize enzyme SBEIIb was performed. Conclusions. According to the results of amino acid sequences of SBEIIb enzyme homologs alignment, it was found that ae1 gene orthologs are present only in monocots, paralogs – in monocots, dicots, and other taxa, including algae and animals. Based on the results of alignment of plants mRNA from which enzyme SBEIIb is translated, maize ae1 gene orthologs and the nearest paralogs encoding starch branching enzymes with chloroplast localization were defined; this suggests a possible origin of ae1 gene due to duplication of the gene encoding the 1,4-alpha-glucan-branching enzyme 2 with chloroplast or amyloplast localization. In the maize ae1 gene structure, regions were found that include polymorphic sites not defined previously. For the polymorphic sites design primers were developed that allowed to differentiate the maize lines. It was determined that the detection of polymorphism in theory can influence the enzyme function and, as a result, change the concentration of amylopectin in maize grain.
Afficher plus [+] Moins [-]Allelic status of key genes of carotenogenesis on DNA-markers in maize lines and their hybrids
2016
Гончаров, Ю. О | Сатарова, Т. М | Дзюбецький, Б. В | Черчель, В. Ю
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.
Afficher plus [+] Moins [-]Ukrainian cereals and oilseeds trade
2016
Духницький, Б. В | Сіліфонов, Т. В
Purpose. To analyze peculiarities of Ukrainian cereals and oilseeds trade, the situation on the world market, and determine future prospects of its development. Methods. Analysis and synthesis, comparative evaluation, graphic procedure. Results. The role and place of Ukraine in a total grain supply to the world market was determined. Ukraine is a world’s top ten grain producer. Among domestic agricultural products, cereals, oilseeds and sunflower oil are in the highest demand in the world. In recent years, our state has reinforced its status as one of the leading exporters of cereals. The commodity pattern of cereals and oilseeds export was analyzed with specifying most in-demand positions and the main countries purchasing these products. According to the results of 2015, Ukraine obtained the highest foreign currency revenue from export of corn, wheat and barley (in grain structure) including soybeans and rapeseed (among oil crops). Key domestic and multinational operators are the main exporters of cereals and oilseeds in Ukraine and still hold their leading position. It was found a significant excess of import price of seeds as compared with export price of crops grown in Ukraine. Assortment of maize and sunflower seeds offered by major companies-producers in Ukraine was studied. Main trends of the world grain market development are considered. Conclusions. It was established that Ukraine is one of the major exporters of cereals and oilseeds. However, volatility of their prices significantly affects the export revenue that was decreasing even with increasing export quantities in kind. The dependence of domestic grain industry development on high-quality imported seed of maize and sunflower hybrids was recorded. It is expected that in the years to come Ukraine will maintain its strong positions in the world’s grain market.
Afficher plus [+] Moins [-]Assessment of combining ability for grain yield of self-pollinated S5 maize (Zea mays L.) families of mixed germplasm
2016
Гайдаш, О. Л
Purpose. Assessment and selection of self-pollinated S5 maize (Zea mays L.) families with high combining ability and wide adaptive capacity obtained on the basis of specially produced hybrids by crossing elite lines of various genetic plasms with the best main agronomic characters. Methods. Field studies, mathematico-statistical evaluation. Results. An amplitude of grain yield ranging from 5.96–10.96 t/ha (x = 8.44 t/ha) in 2013 to 2.67–7.59 t/ha (x = 5.08 t/ha) in 2014 was determined in the course of study of the testcrosses of self-pollinated S5 families. It was found that different response of genotypes of the studied testcrosses to the year conditions significantly affected the average yield level, which decreased in the stressful 2014 by 3.4 t/ha as compared to 2013. The results of the assessment based on the general and specific combining ability of new parent material of mixed germplasm were shown. A significant variability of the estimates of GCA (general combining ability) effects depending on the year conditions was observed in the course of study. A marked difference in the estimates of GCA effects based on the grain yield was revealed. They were persistently high in 19% of the best self-pollinated families in both years, 14% of the families had persistently low estimates of GCA effects, 67% changed their value depending on the genotype and year conditions. Conclusions. 17 best self-pollinated families with persistently high estimates of GCA, 6 families featuring high tolerance to drought and 4 families with persistently high variances of SCA were selected. The selected families will be used as a parent material in selection programs aimed to create new high performance hybrids.
Afficher plus [+] Moins [-]Assessment of combining ability for grain yield of self-pollinated S5 maize (Zea mays L.) families of mixed germplasm
2016
О. Л. Гайдаш
Purpose. Assessment and selection of self-pollinated S5 maize (Zea mays L.) families with high combining ability and wide adaptive capacity obtained on the basis of specially produced hybrids by crossing elite lines of various genetic plasms with the best main agronomic characters. Methods. Field studies, mathematico-statistical evaluation. Results. An amplitude of grain yield ranging from 5.96–10.96 t/ha (x = 8.44 t/ha) in 2013 to 2.67–7.59 t/ha (x = 5.08 t/ha) in 2014 was determined in the course of study of the testcrosses of self-pollinated S5 families. It was found that different response of genotypes of the studied testcrosses to the year conditions significantly affected the average yield level, which decreased in the stressful 2014 by 3.4 t/ha as compared to 2013. The results of the assessment based on the general and specific combining ability of new parent material of mixed germplasm were shown. A significant variability of the estimates of GCA (general combining ability) effects depending on the year conditions was observed in the course of study. A marked difference in the estimates of GCA effects based on the grain yield was revealed. They were persistently high in 19% of the best self-pollinated families in both years, 14% of the families had persistently low estimates of GCA effects, 67% changed their value depending on the genotype and year conditions. Conclusions. 17 best self-pollinated families with persistently high estimates of GCA, 6 families featuring high tolerance to drought and 4 families with persistently high variances of SCA were selected. The selected families will be used as a parent material in selection programs aimed to create new high performance hybrids.
Afficher plus [+] Moins [-]Allelic status of key genes of carotenogenesis on DNA-markers in maize lines and their hybrids
2016
Ю. О. Гончаров | Т. М. Сатарова | Б. В. Дзюбецький | В. Ю. Черчель
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.
Afficher plus [+] Moins [-]Bioinformatic analysis of maize gene encoding starch branching enzyme SBEIIb.
2016
Г. І. Сліщук | Т. Ю. Жернаков | Н. Е. Волкова
Purpose. Investigation of maize ae1 gene polymorphism by bioinformatic methods. Methods. Global and local alignment of the nucleotide and amino acid sequences, in silico translation and transcription, translates modeling, primers design, phylogenetic analysis. Results. 255 nucleotide sequences of maize аe1 gene, 500 amino acid sequences of homology translates of maize ae1 gene (SBEIIb enzyme homologs) and 100 mRNA expressed from the maize ae1 gene were analyzed to establish phylogenetic relationships. Polymorphism of maize ae1 gene different regions was investigated by bioinformatic methods. Modeling of the maize enzyme SBEIIb was performed. Conclusions. According to the results of amino acid sequences of SBEIIb enzyme homologs alignment, it was found that ae1 gene orthologs are present only in monocots, paralogs – in monocots, dicots, and other taxa, including algae and animals. Based on the results of alignment of plants mRNA from which enzyme SBEIIb is translated, maize ae1 gene orthologs and the nearest paralogs encoding starch branching enzymes with chloroplast localization were defined; this suggests a possible origin of ae1 gene due to duplication of the gene encoding the 1,4-alpha-glucan-branching enzyme 2 with chloroplast or amyloplast localization. In the maize ae1 gene structure, regions were found that include polymorphic sites not defined previously. For the polymorphic sites design primers were developed that allowed to differentiate the maize lines. It was determined that the detection of polymorphism in theory can influence the enzyme function and, as a result, change the concentration of amylopectin in maize grain.
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