Two new inbred lines, T589 (medium β-tocopherol content) and T2100 (high γ-tocopherol content), recently developed in CSIC, Cordoba, Spain, have been crossed to known tph1 and tph2 mutations which possessed the same phenotypes and which were obtained at VNIIMK, Krasnodar, Russia. Genetic identification of these recessive mutations with TLC profiles showed the new medium β-tocopherol mutation to be allelic to tph1 and the new high γ-tocopherol mutation to be allelic to tph2.
Near-infrared spectroscopy (NIRS) was used to estimate biochemical traits in turnip rape seeds. The purpose of the research was to develop graduating models for an IR-analyzer MATRIX-I for determination of oil content and contents of oleic, linoleic, and linolenic fatty acids in oil in unbroken seeds of turnip rape. The research was conducted in the biochemistry laboratory of V.S. Pustovoit All-Russian Research Institute of Oil Crops, Krasnodar. An object of the research was turnip rape seeds cultivated in 2019–2021 in the central zone of the Krasnodar region and in the southern forest-steppe of the Western Siberia. Oil content was determined on a NMR-analyzer AMV 1006М (Russia). Contents of oleic, linoleic, and linolenic fatty acids in oil were calculated by a method of percentage normalization of chromatographic analysis on a gas chromatograph “Chromatek-Kristall 5000” on a capillary column SolGelWax 30 m × 0.25 mm × 0.25 µM with automatic dripper of liquid samples. We obtained the best results on quality of the graduating models (root mean square error of prediction, R determination, and a meaning of a residual deviation of prediction for a rank reflected on a diagram) on determination of oil content (RMSEP = 0.25%, R2 = 98.5, and RPD = 8.2), contents of oleic (RMSEP = 0.59%, R2 = 98.9, and RPD = 9.7), linolic (RMSEP = 0.66%, R2 = 95.4, and RPD = 4.7), and linolenic (RMSEP = 0.37%, R2 = 98.0, and RPD = 7.1) fatty acids. The methodic “Turnip rape 51” for mass studying based on the developed graduating models for determination of oil content, contents of oleic, linoleic, and linolenic fatty acids in unbroken turnip rape seeds for IR-spectrometry in a middle sample (9–20 g) in cuvette with diameter 51 mm was obtained in a program OPUS LAB. This method allows conducting express-estimation of more than 100 samples of turnip rape seeds per an operating shift.
Influences of genetic background, ontogenesis and environment on tocopherol mutations in sunflower were estimated in this study. The content of α/β-tocopherols varied from 40/60 to 60/40% in seeds of different inbred lines containing the tph1 gene. The lines containing the tph2 gene ranged widely in α/γ-tocopherols, from 0/100 to 80/20%. A double mutation showed variability in different inbred lines in α/β/γ/δ-tocopherol contents from maximum expressivity of 0/0/60/40 to minimum 40/25/25/10%, due to incomplete expressivity of tph2. Seed maturation from 10 to 38 DAF influenced tocopherol composition in both normal and mutant genotypes by increasing the α-tocopherol content. The content varied from 81 to 97% in a normal genotype, from 33 to 50% in tph1 mutation and from 0 to 6% in tph2 mutation. Tocopherol mutations were shown to express their phenotype in different parts of a plant. All roots, hypocotyls, leaves, pollen and callus from the seeds, hypocotyls and leaves had normal, tph1, tph2 and double mutation tocopherol profiles depending on the genotype. The only exception was the absence of tph1 expressivity in the green tissue of the leaves. The experiment with day/night temperatures varying during seed development from 20/18 to 30/26°C showed an increased α-tocopherol content from 39 to 48% in tph1 mutation. Both a normal genotype (about 97% of α-tocopherol) and tph2 mutation (about 98% of γ-tocopherol) were constant in these two temperature regimes. Genetic background was the main factor that influenced the expressivity of tph2 in sunflower.
We used near-infrared reflectance spectroscopy (NIRS) to assess biochemical parameters in whole oil flax seeds, regardless of differences in seed coat color of the samples. At the first stage of work, the set the task to develop calibration models for the MATRIX-I IR analyzer to determine the oil and moisture content in flax seeds. The carried out the research in the laboratory of biochemistry on brown and yellow seed samples of oil flax, grown in 2015-2020 in various agro-ecological conditions of the Russian Federation. We determined the oil content on an AMV 1006M NMR analyzer in accordance with the GOST 8.597- 2010 measurement procedure; we assessed the moisture content by the standard method of GOST 10856- 96. We used the results of determination of the oil and moisture content of the seeds of test lot in accordance with the accuracy indicator of the calibration of GOST 32749-2014 to verify the reliability of the developed models. We received the best indicators of the quality of calibration models (root-mean-square prediction error, coefficient of determination and the value of the residual deviation of prediction for the rank displayed on the graph) by determining the oil content (RMSEP = 0.27 %, R2 = 99.2 and RPD = 11.2) and moisture content (RMSEP = 0.06 %, R2 = 99.9 and RPD = 39). In the OPUS LAB program we developed the “Flax 51” method for mass analysis based on the developed calibration models for the determination of oil and moisture content in whole oil flax seeds (9-20 g) in a sample cell with a diameter of 51 mm. It enables the quick carrying out a preliminary assessment of the breeding material at a high speed – more than 120 samples in 7 hours without seed destruction.
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