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Використання EST-SSR маркерів для аналізу поліморфізму сортів салату посівного (Lactuca sativa L.) вітчизняної селекції | Использование EST-SSR маркеров для анализа полиморфизма сортов салата посевного (Lactuca sativa L.) отечественной селекции | Application of EST-SSR markers for analysis of polymorphism of lettuce varieties (Lactuca sativa L.) of domestic breeding
2020
Хареба, О. В. | Лещук, Н. В. | Стариченко, Є. М. | Шитікова, Ю. В. | Присяжнюк, Л. М.
Цель. Определение молекулярно-генетического полиморфизма сортов салата посевного отечественной селекции с помощью EST-SSR маркеров.Методы. Молекулярно-генетический анализ, статистические методы.Результаты. Представлены результаты изучения молекулярно-генетического полиморфизма 7 сортов салата посевного с помощью 7 EST-SSR маркеров. В результате анализа по исследуемым маркерам идентифицировано 37 аллелей, в среднем 5,29 аллели на локус. Наиболее полиморфным оказался маркер KSL-92, с помощью которого выявлено 7 аллелей, Polymorphism Information Content (РІС) – 0,98. Наименьшее значение РІС – 0,57 было у маркера KSL-37, с помощью которого идентифицировано наименьшее количество аллелей – 3. Для оценки генетического разнообразия сортов салата с помощью EST-SSR маркеров были определены генетические дистанции между сортами с применением меры близости Жаккара. Наиболее близкими оказались сорта, генетические дистанции между которыми составляли 0,17: ʻЗорепад’ и ʻМалахит’, ʻМалахит’ и ʻДублянский’, ʻДублянский’ и ʻСмуглянка’, ʻКрутянский’ и ʻСмуглянка’. Наиболее удаленным оказался сорт ʻСкарб’ со значениями генетических дистанций 0,00 по отношению к другим исследуемым сортам. Согласно рассчитанным генетическим дистанциям сорта ʻЗорепад’ и ʻМалахит’, ʻМалахит’ и ʻДублянский’, которые относятся к одной разновидности Lactuca sativa L. var. secalina, имели высокую степень генетической близости. Сорта ʻСкарб’ и ʻПогоныч’, которые относятся к разновидностям Lactuca sativa L. var. longifolia и Lactuca sativa L. var. angustana, соответственно, были генетически отдаленными, что подтверждают значения генетических дистанций. Значение индекса Шеннона внутри разновидности салата посевного по исследуемым EST-SSR маркерам составляло 0,61, между разновидностями – 0,96, среднее значение – 1,57.Выводы. Наиболее высокий уровень полиморфизма был отмечен по маркеру KSL-92, наименьшее количество аллелей (3 аллели) было идентифицировано с помощью маркера KSL-37. Сорта, генетические дистанции между которыми составляли 0,17, оказались наиболее близкими. Наиболее отдаленным сортом по исследованным EST-SSR маркерам оказался сорт ʻСкарб’. | Мета. Визначення молекулярно-генетичного поліморфізму сортів салату посівного вітчизняної селекції за EST-SSR маркерами.Методи. Молекулярно-генетичний аналіз, статистичні методи.Результати. Представлено результати вивчення молекулярно-генетичного поліморфізму 7 сортів салату посівного за 7-ма EST-SSR маркерами. За досліджуваними маркерами ідентифіковано 37 алелів, у середньому 5,29 алеля на локус. Найполіморфнішим виявився маркер KSL-92, за яким було виявлено 7 алелів, Polymorphism Information Content (РІС) – 0,98. Найменше значення РІС – 0,57 було у маркера KSL-37, за яким ідентифіковано найменшу кількість алелів – 3. Для оцінювання генетичного різноманіття сортів салату за EST-SSR маркерами було визначено генетичні дистанції між сортами із застосуванням міри близькості Жаккара. Найближчими виявились сорти з генетичними дистанціями 0,17: ‘Зорепад’ і ‘Малахіт’, ‘Малахіт’ і ‘Дублянський’, ‘Дублянський’ і ‘Смуглянка’, ‘Крутянський’ і ‘Смуглянка’. Найвіддаленішим виявився сорт ‘Скарб’ із значеннями генетичних дистанцій 0,00 щодо інших досліджуваних сортів. За розрахованими генетичними дистанціями сорти ‘Зорепад’ і ‘Малахіт’, ‘Малахіт’ і ‘Дублянський’, які належать до одного різновиду Lactuca sativa L. var. secalina, мали високий ступінь генетичної близькості. Сорти ‘Скарб’ і ‘Погонич’, які належать до різновидів L. sativa L. var. longifolia та var. angustana, відповідно, були генетично віддаленими, що підтвердили значення генетичних дистанцій. Значення індексу Шеннона всередині різновиду салату посівного за досліджуваними EST-SSR маркерами становило 0,61, між різновидами – 0,96, середнє значення – 1,57.Висновки. Найвищий рівень поліморфізму було зауважено за маркером KSL-92, найменшу кількість алелів (3 алеля) було ідентифіковано за маркером KSL-37. Сорти, генетичні дистанції між якими становили 0,17, виявились найближчими. Найвіддаленішим сортом за досліджуваними EST-SSR маркерами виявився сорт ‘Скарб’. | Purpose. Determination of molecular genetic polymorphism of lettuce cultivars of Ukrainian breeding by EST-SSR markers. Methods. Molecular genetic analysis, statistical methods. Results. The results of molecular genetic polymorphism study of 7 lettuce cultivars by 7 EST-SSR markers are presented. As a result of the analysis by studied markers, 37 alleles were detected with an average 5.29 alleles per locus. The KSL-92 marker, which identified 7 alleles, proved to be the most polymorphic (PIC – Polymorphism Information Content 0.98). The lowest PIC value (0.57) was noted for the KSL-37 marker by which 3 alleles were identified. For assessing the genetic diversity of lettuce cultivars by EST-SSR markers, genetic distances between cultivars were determined based on Jaccard’s similarity coefficient. It was determined that the most similar cultivars with genetic distances 0.17 were ‘Zorepad’ and ‘Malakhit’, ‘Malakhit’ and ‘Dublianskyi’, ‘Dublianskyi’ and ‘Smuhlianka’, ‘Krutianskyi’ and ‘Smuhlianka’. The most distant cultivar was ‘Skarb’ with a genetic distance 0.00 compared to other studied cultivars. According to the calculated genetic distances, the cultivars ‘Zorepad’ and ‘Malakhit’, ‘Malakhit’ and Dublianskyi’, which belong to the same variety Lactuca sativa L. var. secalina have a strong genetic similarity. The cultivars‘Skarb’ and ‘Pohonych’, which belong to the varieties Lactuca sativa L. var. longifolia and Lactuca sativa L. var. angustana, respectively, were genetically distant, what was confirmed by genetic distances values. It was determined that the Shannon index within the lettuce variety by 7 EST-SSR markers is 0.61, between varieties – 0.96, the average value is 1.57. Conclusions. According to the results of studies of 7 lettuce cultivars, it was found that the highest polymorphism was determined by the KSL-92 marker, the minimum of alleles number (3 alleles) was identified by KSL-37 marker. Based on calculated genetic distances, it was noticed that lettuce cultivars, genetic distances between which are 0.17, were the most similar. The most distant cultivar based on 7 EST-SSR markers was ‘Skarb’ cultivar.
Show more [+] Less [-]Inheritance of a sign of apricot color of ray flowers of sunflower (Helianthus annuus L.)
2019
Ведмедєва, К. В
Purpose. To reveal the nature of the inheritance of apricot color of the ray flowers of the sunflower and the type of interaction of genes causing different colors. Methods. Field experiment, genetic analysis. The statistical validity of the results was evaluated using Pearson’s criterion. Results. We conducted crosses of the ‘KG13’ line as the source of the sign of apricot color with sunflower lines that had yellow, orange and lemon colors of the ray flowers. In the first generation, from crossing the ‘KG13’ line with five lines, which had a yellow color, only a yellow color of ray flowers was observed. In the second generation, a 3 : 1 split was observed: three-quarters with yellow flowers and one with apricot flowers. Line ‘KG13’ was crossed with three lines (‘HA298’, ‘SL2966’, ‘LD72/3’), which had an orange color of flowers. In the first generation, orange flowers were observed; in the second generation, splitting was recorded: three-quarters of offsprings with orange-colored flowers and one-quarter with apricot flowers. The line ‘KG13’ was crossed with ‘KG107’ and ‘ZL678’, which had lemon-colored flowers. The resulting plants of the first generation had a yellow coloration of ray flowers. In the second generation, five classes of plants by coloration of ray flowers were obtained: yellow, orange, apricot, lemon, lemon-apricot in the ratio 6 : 4 : 3 : 2 : 1. According to these data, the genes of lemon and apricot color have a complementary effect, the homozygous state of orange allele is epistatic to the recessive homozygote of the lemon-colored gene. The ‘KG108’ line with a combination of genes responsible for apricot and light yellow color has its own light apricot color and in crossings with a yellow colored line in the second generation gives splitting in the ratio 9 : 3 : 3 : 1. Conclusions. It was revealed that the apricot color of the ray flowers of the sunflower line ‘KG13’ is due to the homozygous state of the allele of the same gene whose second allele causes an orange color in the lines ‘NA298’, ‘SL2966’ and ‘LD72/3’. The complementary action of alleles responsible for apricot and lemon, as well as apricot and light yellow coloration of ray flowers was determined. A case of epistasis of homozygotes along the allele controlling the orange color over the recessive homozygote of the gene, which is controlled by the lemon color in the crossing combination ‘ZL678’ / ‘KG13’, was revealed.
Show more [+] Less [-]Gnome 2 as a donor for winter rye (Secale cereale L.) short stem
2013
Скорик, В. В
A stabilizing (directed) selection has created a donor of short stem for winter rye (Secale cereale L.), plant height of which ranged from 50 to 60 cm. The plant height kept symmetry of its distribution curve and the frequency accumulation in central classes (positive excess). For the first time a symbolic designation to new short-stem related Hl-2Hl-2 allele and the donor name (Gnome 2) were proposed. 28 years of stabilizing selection showed that 57% of overall genetic variability of plant height resulted from adaptive genes available for directed selection by phenotype, and 43% from dominant and epistatic factors that predetermines the expression heterosis effect. Gnome 2 donor proved to have genetic additive correlation between the pants height and number of flows per ear, ear length, weight of seeds per plant , 100 seeds weight per plant; to have reverse correlation with ear density seeds weight per ear. The height of original parent components have displayed direct additive correlation with number of flowers per ear and reverse with the ear density. The additive correlation component directly exposes «genuine» impact of parental plants on the expression of the characteristics indicated among the offspring Productive bushing of parental plants, seed weight per plant directly, and seed size (100 seeds weight) indirectly, respectively, influence the height of offspring pants. The reverse additive correlation between the parents height and 100 seeds weight in the offspring is caused by pleiotropic effect of the genes impact thus enabling to combine the desirable characteristics in one genotype. Productive bushing is by 54% due to the impact of general genetic factors among the above, in particular, 30% due to that additive, 24 due to non-additive factors. The concept of genetic improvements for productive bushing of the Gnome 2 rye implies utilization of additive effect through the directed selection, as well as application of breeding techniques for controlling the effect of heterosis caused by the genes of dominant and epistatic impact. The selection paradigm requires simultaneous genotypes selection with immediate examination of the selection results by offspring while in parallel to develop inbred lines, combining these afterwards evaluating general and specific combining ability by productive bushing. It is also to be noted that the productive bushing essentially depends on the environmental conditions, which significantly corrects the implementation of productivity potent, thereby the issue of agronomical conditions aimed at extending the expression of characteristic in question remains.
Show more [+] Less [-]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.
Show more [+] Less [-]Gnome 2 as a Donor of Short Stem Characteristic for Winter Rye (Secale cereale L.)
2011
Скорик, В. В
Stabilizing (centripetal) breeding produced donor of short stem characteristic for winter rye (Secale cereale L.) with height of plants ranging from cm 50 to 60. Distribution curve of plant's height remained symmetric with cummulation of frequencies at central classes (leptokurtic). For the first time, designation for new allele of short-stem gene HI-2HI-2 and donor name Gnome 2 was suggested. Gnome 2 was found to have direct genetic correlation of plant height with number of flowers in the ear, length of the ear, weight of 100 seeds from the plant, as well as reverse correlation with ear density and weight of kernels from the ear.
Show more [+] Less [-]Gnom 3 as a Donor for Ultra Short- Stem Trait of Winter Rye
2011
Скорик, В. В
The article reflects the progress of genetic decrease of Rye F 3k- 10029/ Saratovske (Саратовське) 4 height by means of the shortest stem plants selection during the period from 1974 to 2010. 37 years selection of the shortest- stern genotypes decreased the plants height from 119.33 cm to 22.57cm. Targeted selection into minus direction decreased the plants height in 5,29 times on the background of the dominant HI expression. In average, the height of plants in the course of 27 breeding cycles were decreasing by 2.69 cm, but that was not going gradually. A new donor Gnom 3 had been created for ultra short-stem trait of the Winter Rye, with the marking of alleles HI-3HI-3. Relative influence on the minus selection efficiency has been established by height of plants for the selection differential (38%) and coefficient of inheritance in narrow sense (14,56%). Realized efficiency of selection in decrease of winter rye height in 72,08% of cases corresponded to predicted hit ration of the breeding. Analyzes of genetic and statistical parameters and correlation clusters of 11 utilitarian average characteristics of ultra short- stem rye Gnom 3 for the period of 1974 to 2010 has been performed.
Show more [+] Less [-]Allelic composition of puroindolinium genes and confectionery properties of flour of soft winter wheat samples
2020
Леонов, О. Ю | Шарипіна, Я. Ю | Усова, З. В | Суворова, К. Ю | Сахно, Т. В
Purpose. Identification of soft winter wheat varieties and lines from the Plant Production Institute nd. a. V. Ya. Yuryev, NAAS by allelic state of Pina–D1 and Pinb-D1 genes for targeted use in the breeding for high confectionery properties of flour. Methods. Identification of the Pina-D1 and Pinb-D1 genes allelic state was performed by polymerase chain reaction (PCR) using allele-specific primer pairs. Confectionery properties of flour were evaluated by determining the quality indicators: the water absorption capacity (WAC) of the flour, trial baking of cookies and evaluation of its quality. Results. According to the results of PCR analysis, 9 samples had an allelic composition of puroindoline genes (Pina-D1a and Pinb-D1a) characteristic for soft-grained varieties. Flour of the lines 'L137-26-0-2', 'L137-26-0-3' had the best confectionery properties, it had a WAC value less than 55%, cookies diameter 85 mm, cookies height 10 mm, estimation of a surface of cookies 7– 9 points, what meets the requirements for soft-grained wheat. 76% of the samples belonged to hard-grained varieties and had the corresponding alleles of the Pina-D1 or Pinb-D1 genes. In the studied sample, Pina-D1 gene is represented by 2 alleles: Pina-D1a and Pina-D1b. 27 samples had Pina-D1a allele, which also allows them to be used in breeding programs for grain quality when crossed with soft samples, 4 ones had Pina-D1b allele. As to Pinb-D1 gene, all hard grain samples had Pinb-D1b allele, and the 'Erythrospermum S 424-1 / 14' line was heterogeneous for Pinb-D1a / Pinb-D1b. The flour of these samples had typical for hard wheat quality indicators: WAC 68% and more, cookie diameter of 60–72 mm, cookie height of 13–15 mm, the surface evaluation of 1–4 points. Conclusions. The studies allowed to differentiate the breeding material and transfer a soft winter wheat cultivar of a confectionery use 'L137-26-0-3' ('Mazurok') which has an allelic structure of puroindolins genes (Pina-D1a and Pinb-D1a) characteristic for soft-grained varieties and high confectionery flour properties for qualification examination.
Show more [+] Less [-]Method for determination of varietal purity (typicality), hybridity, sterility of seed lots based on the establishment of the quantitative ratio of alleles of DNA markers
2016
Вдовиченко, Ж. В | Спиридонов, В. Г | Хомутовська, С. В | Парій, М. Ф
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.
Show more [+] Less [-]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.
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