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Biotehnology
Issue № | Name | ||
№1 (2020) | Marker-associated selection by the Sh1 gene forthe creation the anthocyanincolored sugar maize line | Annotation Marker-associated selection by the Sh1 gene forthe creation the anthocyanincolored sugar maize line UDC 57.084:633.15 Grain Crops. 2020. 4 (1). 38–45.
Psolova A. O., Denysiuk K. V., Satarova T. M. SEinstitute of grain cropsof national academy of agrarian sciences, 14 Volodymyr Vernadskyi Str., Dnipro, 49027,ukrainе
Today, there is a trend in the world to create new maize genotypes with a controlled content of anthocyanins, carotenoids, proteins and carbohydrates in grains and use them as functional foods. A high content of anthocyanins is characterized by some maize varieties and hybrids of dental, scaly and flintly subspecies on the Ukraine territory, however, sugar maize with anthocyanin colored grains is not widespread. The aim of our work was to obtaining the later generations of plants from the crossing of yellow grains sugar maize with anthocyanin colored grains maize and identification from them the homozygous plants for marker phi033 of Sh1 gene to create a inbred of sugar maize with anthocyanin colored grains. The material of the study was the CE401 sugar maize inbred with yellow grains, the population Chornosteblova, which has a dental anthocyanin grains, the F1 hybrid (CE401хChornosteblova) and the self-pollination hybrid generations F1(CE401хChornosteblova). All grains of all F1 hybrid studied plants had a yellow color and a semi-dental structure (not wrinkled endosperm). Splitting in the F2–F3 generations had more compound display. Splitting by color and grain structure was observed. The anthocyanin colored grains with wrinkled endosperm typical for sugar maize were obtained in the F4 generation from self-pollination of the CE401хChornosteblova hybrid. Molecular genetic analysis was carried out in parallel. According to the results of the polymerase chain reaction, the molecular marker phi033 of the Sh1 gene is represented with the 236 bp allele or/and 310 bp allele in the CE maize sugar inbred. This marker is represented with the 264 bp allele in the Chornosteblova population with a dental structure of grains. Heterozygotes with the marker phi033 of the Sh1 gene with the 236 bp/264 bp and 310 bp/264 bp are occurred among plants of the F1(CE401хChornosteblova) hybrid. Alleles of parental forms and their various combinations are noted also in the self-pollination F4 ge-neration of the CE401хChornosteblova hybrid for the marker phi033 of the Sh1 gene. The allele of the only maternal sugar marker form with 236 bp was observed at 15 among 35 analyzed F4 plants that indicating homozygosity for this marker. Self-pollination and reproduction of such plants is recommended for obtaining the sugar maize inbred with anthocyanin colored grains. Key words: maize, polymerase chain reaction, geneSh1, phi033, anthocyanin colored grains.
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2 (2018) | The content of anthocyanins in stalks of different maize genotypes | Annotation The content of anthocyanins in stalks of different maize genotypes UDC 581.192:633.15 https://doi.org/10.31867/2523-4544/0028 Psolova A. O., Derkaсh K. V., Bielikov Ye. I., Satarova T. M. Grain Crops. 2018. 2 (2).218–225. SEinstitute of grain cropsof national academy of agrarian sciences, 14 Volodymyr Vernadskyi Str., Dnipro, 49027,ukrainе A large group of water-soluble pigments in the kingdom of plants are anthocyanins belonging to the class of flavonoids and providing a color from orange-red to black-violet. The presence of food products rich in antioxidants, including anthocyanins, reduces the risk of vascular diseases, the emergence of tumors, it has an anti-mutagenic effect. Anthocyanins are registered in the European Union as a food and dietary sup-plement in the section Е100-Е199 (dyes) under the number Е163. Due to the fact that antioxidants are often used as food additives in a balanced diet of animals and humans, the estimation of the content of anthocyanins in crops, especially in maize, is very relevant. The material for the study were the following maize (Zea mays L.) genotypes: the population Bilosnizhka which is a sweet corn with green stalk, flint maize inbred lines AiD121, IKS226, BS4626-6 / 1, BS37-10 and AI178-325 with a green stalk and a population of dent maize Chornosteblova with a dark purple stalk. The content of anthocyanins was determined by the modified differential spectrophotometry method by Lu. E. Rodriguez-Saona, R.E. Wrolstad and M. M. Giusti, R. E. Wrolstad Determination of the content of anthocyanins was carried out during the flowering period, and the eighth internode of the stalk was used for the analysis. The content of anthocyanins in the stalk of the studied maize genotypes varies in the range of 168,64 – 38362,23 mg/kg, that is, 0,02–3,83 % of the absolutely dry weight of the stalk. According to the total content of anthocyanins in the stalk, the studied samples were divided into genotypes with low (168,64 mg/kg – population Bilosnizhka), medium (338,89, 404,15, 425,93, 463,45 and 996,13 mg/kg, respectively in inbred lines AiD121, IKS226, BS4626-6/1, BS37-10 and АI178-3253) and super-high content of anthocyanins (38362,23 mg/kg – the population Chornosteblova). Іn the studied genotypes with a green stalk color, the total content of anthocyanins varied in the wide range from 168,64 mg/kg to 996,13 mg/kg, that is, it differed 5,91 times. Тhe enhanced dark violet coloration of the stalk of plants of the Chornosteblova population is associated with a significant increase in the total content of anthocyanins in comparison with the green-stalk genotypes (38,5 to 227,5 times) due to the contribution of all species and forms of anthocyanins, from the forms of delphinidin (12,55 %) to forms of peonidin (27,31 %). Thus, studies have shown that the total content of anthocyanins in maize stalk of the Chornosteblova population with a dark violet color is 3,83 % of the dry weight of the stalk and it is more or less uniformly represented by all kinds of anthocyanins. The obtained results testify to the prospect of using maize stalks of the Chornosteblova population as a source of antioxidants in the production of the biological additive E163 –"anthocyanins". Key words: zea mays L.,antioxidants, anthocyanins,stalks, genotype. | |
№ 2 (2017) | Morphobiological features of maize transformated plants | Annotation Morphobiological features of maize transformated plants
UDC57.084:633.15 Derkach K. V.1*, Abraimova O. Ye.1, Cherhel V. Yu.1, Dzyubetskyi B. V.1, Morgun B. V.2, Nitovska I. O.2, Satarova T. M.1 Grain Crops, 2017, 1 (2), 232–241. 1SEinstitute of grain crops of national academy of agrarian sciences, 14 Volodymyra Vernadskyi Str., Dnipro, ukrainе, 49027,*e-mail: kvderkach@gmail.com 2Institute of Cell Biology and Genetic Engineering NASU, 148, akademika Zabolotnogo Str., Kyiv, ukrainе, 03680, ** e-mail: molgen@icbge.org.ua Key words: maize, resistance to herbicides, phosphinothricin, transformated plants, gene bar. The aim of the work was to characterize the morphobiological features of maize genetically transformated plants in generations Т0–Т6. The tasks of the study were to investigate seed germination, the survival of young seedlings after the Basta herbicide treatment, the duration of the periods between seedlings emergence and tassel/ear flowering, plant height. The maize hybrid PLS61хДК633266 which parent lines refered to PLS61 germplasm (PLS61) and Lancaster germplasm (ДК633266) was used as the initial research material. Genetic transformation was carried out via biolistic treatment of 10-20-days calli, which had been formed on scutellums of immature embryos. The control calli were not undergone biolistic treatment. Plasmid vector pAHC25 containing the gene of β-glucuronidase (uidA) from Escherichia coli and the gene of phosphinothricinacetyltransferase (bar) from Streptomyces hygroscopicus, both under the control of the promoter of the maize ubiquitin gene, was used for transformation. Initially the T0 plant regenerated from callus tissue of PLS61хДК633266, which had been biolistically treated with the plasmid vector, was pollinated with pollen of ДК633266 (self-pollination of the transformated plant was impossible owing to pollen sterility). The presence of bar gene in the T0 plant used for further studies has been confirmed by the polymerase chain reaction method. As a result, 40 seeds with T1 embryos of genotype: [T0(PLS61хДК633266)хДК633266] were received. T1 seeds partionally were sown into the soil in vegetation vessels. The seeds obtained from T1 plants via self-pollination had the genotype T2: [T0(PLS61хДК633266)хДК633266]1sp, where 1sp - the number of self-pollination (2079 pcs.). The seeds of the subsequent generations on plants Т2–Т6 were received only by self-pollination. The genotypes of embryos of these seeds and plants which were formed after their germination were, respectively, T3-6: [T0(PLS61хДК633266)хДК633266]2-5sp. 25 and more plants per generation were sown for cultivation of T2–T6. Five or more plants were self-pollinated in each generation. Morphological evaluation of plants in generations T0–T6 after bar-transformation allowed to characterize transformed plants on post-herbicide survival and basic growth parameters and to obtain seeds after self-pollination. It was found that 100% untransformated maize plants died within a week after treatment with Basta. Plants-descendants of biolistic transformed ones have reduced seed germination both in T2 and T3 generations. By average phenological parameters and height transformed plants, not treated with herbicide, in T2 do not differ significantly from untransformed untreated ones. In T3 generation they revealed an increase in the duration of interphase periods, a break in flowering male and female inflorescences as well as growth inhibition, which leads to the decrease of plant height. But generally, the shock reaction quickly passes in T2 and T3 progenies. The plants look like well developed, female and male inflorescences normally enter the flowering phases, form pollen and silk, produce seeds. There was a greater variation in morphological characteristics between plants within families T2 and T3 than in control variants, which suggests the existence of plants with various degrees of resistance to the herbicide. It can be explained with the presence or absence of bar gene in generations from self-pollination of the original regenerants obtained directly from biolistically treated callus tissue. The survival of T6 plants after herbicide treatment was 100%. The average values of the height of transformated plants without and after herbicidal treatment in T6 did not differ significantly. This fact indicates a successful screening on the selective phone in previous generations and the probable accumulation of transgene in plant genome in five cycles of homozygosity via self-pollination.
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