MicroRNAs (miRNAs) are a class of non-coding RNAs about 20-24 nucleotides long. They play an important role in the gene regulation at the post-transcriptional level. They affect the plant genome response to environmental stress. The miRNA-based molecular markers is type of functional markers reported in very few plants. However, the information connected to the evaluation of genotypes by this type of markers within a single species is missing. Considering the stability, polymorphism, functionality and transferability potential of miRNA-based markers, the research was conducted to apply selected types of them (miR156b, miR408a and the combined type of miR156b/miR408a) for the genotyping analysis of eight flax genotypes of different origin together with the morphology analyses. A total of 145 miRNA loci were identified, of which 19 were unique. The highest numbers of miRNA loci (57) and unique fragments (9) as well as the highest percentage of polymorphism and the extent of polymerase chain reaction (PCR) amplification of miRNA fragments have been observed with the combination of miR156b-F and miR408-F markers. By means of the miRNA markers has been recorded the unique profile of the miRNA loci for individual accessions. The morphology study has shown that the genotypes are the same in the expression of selected morphological traits despite the different use and different places of origin. However, we have identified an interface between some of morphological traits and miRNA-based markers for genotyping the genetic resources of flax. By mutually linking these two types of markers, we were able to determine unique genotypes of flax.
Background. Structuring and phenotyping genetic diversity is an important aspect of the work with breeding sources and materials. In the Introduction, the authors pointed out the role of N.I. Vavilovs scientific foresight in defining the topical trend in researching the genetic diversity of a crop, particularly the analysis of its biochemical composition. As the target of their research, the authors chose biochemical characters identifiable in the process of metabolomic analysis conducted by means of gas chromatography with mass spectrometry. Materials and methods. The object was the grain of naked and covered forms of common oat (Avena sativa L.) from the collection held by the Oat, Rye and Barley Genetic Resources Department of VIR. The analysis of oil fatty acid content and metabolomic research were performed using the method of gas-liquid chromatography with mass spectrometry on the chromatograph Agilent 6850 (USA). Results. The obtained metabolomic spectra which reflected the metabolomic status of genotypes of various ecogeographic origin were compared among themselves using statistical (principal component) analysis methods. The results of the comparison are discussed by referring to the most important groups of metabolites significant for forming the traits of resistance to stressors as well as the characters related to food qualities of grain products. Special attention has been paid to biologically active compounds determining the functional value of the products for human nutrition: the sum of phenolics in covered forms is five times higher than that in naked ones and the content of glycine amino acid in covered forms is five times higher than in naked grain, with a similar proportion in the content of organic acids, sugars, etc. Conclusion. Differences between metabolomic profiles of naked and covered forms have been detected and statistically verified. Accessions with the most optimal nutritional composition have been identified for food purposes and for the development of resistance to biotic and abiotic environmental stresses.
This review is devoted to the description of chemical peculiarities of industrial oil crops cultivated (or prospective for cultivation) in Russia, which are stored in the VIR collection. Different crops have similar fatty acids biosynthesis pathways, but each species has its own individualities in the chemical composition of the oil and its genetic control. The diversity of oil crop chemical composition opens the possibility of its multipurpose utilization practically in all industrial segments. Sunflower, rapeseed, flax, mustard, camelina and safflower are cultivated in Russia as oil crops. Castor beans, perilla, lallemantia and noog are not cultivated on an industrial scale, but have original oil properties and are prospective for future cultivation. Hemp and poppy seeds contain oil valuable for food, but they are not widespread. Cotton and peanut oils are prospective for industrial purposes when early, already created varieties of these crops will be cultivated in Russia. Oil properties depend on the ratio of its basic fatty acids: saturated (stearic, palmitic) and unsaturated (oleic, linoleic, linolenic). As a rule, lauric, myristic and palmitoleic acids are determined in minor quantities. The oil of Brassicaceae crops also includes arachidic, eicosenoic, eicosadienoic, behenic, erucic and lignoceric acids. Fatty acids accumulation is influenced by growing conditions, though it has strict genetic control.
MicroRNAs represent small non-coding RNAs that play important role in regulating gene expression under various biotic and abiotic stresses as well as in different developmental stages of plants. They are involved in wide variety of biological and metabolic pathways of plants. The research was focused on the potential of selected miRNA-based molecular markers (miR156 and miR168) to map the polymorphism level of flax genome in the selected developmental stages (flower bud -floweringboll development) as a reflection of the activity of specific miRNAs in these flax organs and tissues. The miRNA polymorphism was evaluated on 8 flax genotypes by miRNA-based molecular marker assays and data were supported by the morphology measurements on buds, flowers, petals and developing bolls. Extent of PCR amplification of miRNA fragments ranged from 40 bp to 200 bp (miR168a-F/miR-R primer pair) or 35 bp to 120 bp (miR156b-F/miR-R primer pair), respectively. MiRNAbased primers amplified in total 196, respectively 158 miRNA loci. Based on the results can be concluded that the representation of miR168 loci have increased according to developmental stages ascending (stage of bud, flower and boll). In overall miRNA156b loci profile in all three analyzed developmental stages is possible to observe the increase of miRNA loci amplification almost in all genotypes. Taking into account the genetic background of genotypes, based on the peak analysis profile of amplified miRNA loci, implemented by GeneTools software, was possible to identified unique loci in linseed and intermediate genotypes in individual developmental stages. Have been found out that only the genotype Bolley Golden differ from other genotypes in both analysis (molecular and morphological) if the
Background. Linseed solin varieties were created for nutrition, but the effect of oil fatty acid (FA) composition on other characters is not clear. Materials and methods. Using 6 inbreeding generations from 26 heterogeneous flax accessions were generated 19 high (HL), 7 medium (ML) and 14 low linolenic (LL) lines. For each lines contents of 5 basic FA: palmitic, stearic, oleic (OLE), linoleic (LIO) and linolenic (LIN); the ratio LIO/LIN, oil iodine number, vegetative period (VP) phases and plants size were evaluated. Development of CAPS marker for LuFAD3A gene was performed using idtdna.com. Sequencing of LIN genes sites was done in the Centre MCT SPBGU and Eurogen. Results. ANOVA showed significant differences HL, ML and LL groups for PAL, OLE, LIO, LIN, LIO/LIN, IOD. Considerable decrease of LIN, causes asymmetric changes in FA ratio and correlations between them and other traits. Factor analysis revealed the influence of two factors. The first one divided lines according to their LIN level and characters associated with it, the second one according to the VP and OLE. LIN synthesis is controlled by two complementary genes LuFAD3A and LuFAD3B. Sequencing of LuFAD3A gene 1 exon of 6 lines revealed a mutation (G255 A255), resulting in formation of stop codon. Developed developed CAPS-marker confirmed the homozygosity of hybrids between LL (gc-391) and HL lines (gc-65, 109, 121). Descendants of hybrid between gc-109 and gc-391 ripened 8-10 days earlier than gc-391. CAPS markers of LuFAD3B gene revealed differences between HL, ML, LL lines. Sequencing of this gene first exon and the beginning of the second one in 3 lines (1HL, 2LL) showed that this method reveals a mutation in the second restriction site, located in the 2 exon (C6 T6), and causing the replacement Hys Tyr. Conclusion. Lines from GC have wide variability of FA and other agronomic characters, combination of which will expand the cultivation of solin.
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