The Crucifers family (Brassicaceae) includes a large number of economically important crops, particularly
Brassica rapa
, which is a widely used model plant for molecular genetic studies of oilseeds.
...B. rapa
is a highly polymorphic species that includes many of genetically distinct subspecies. Considering this fact, the intraspecific hybridization of
B. rapa
subspecies is considered a promising breeding approach aimed at increasing the genetic diversity of the crop. Previously, the authors have shown that one of such hybrids,
B. rapa
subsp.
oleifera
f.
biennis
× (subsp.
rapifera
×
pekinensis
), could be a valuable oil feedstock due to its increased productivity. However, obtaining hybrids and their subsequent breeding would require the involvement of various molecular marker systems. So far, the method of estimating the length polymorphism of the first (TBP) and second (cTBP) introns of β-tubulin has demonstrated its high accuracy and reliability in the identification (DNA-barcoding) of flowering plant taxonomic units at different levels. In the present study, the productivity of such hybrid oil tyfon (
B. rapa
subsp.
oleifera
f.
biennis
× (subsp.
rapifera
×
pekinensis
)) was evaluated and DNA-barcoding of different hybrid tyfon lines (
B. rapa
subsp.
oleifera
f.
biennis
× (subsp.
rapifera
×
pekinensis
)) and its parental
B. rapa
subspecies using the β-tubulin intron length polymorphism assessment approach was carried out. Based on the data of the molecular genetic analysis, which included the assessment of length polymorphism of the first and second introns of β-tubulin genes, we were able to confirm the origin of the oil tyfon hybrid (
B. rapa
subsp.
oleifera
f.
biennis
× (subsp.
rapifera
×
pekinensis
)) hybrid from Dutch leaf tyfon (
B. rapa
subsp.
rapifera
×
pekinensis
) and winter turnip (
B. rapa
subsp.
oleifera
) with high confidence. Along with that, it was possible to differentiate var.
glabra
and var.
laxa
accession of napa cabbage (
B. rapa
subsp.
pekinensis
) for the first time using combined TBP and cTBP analyses. A variation in the number of amplified regions of β-tubulin introns was noted in different genotypes; however, these differences did not appear to be a specific feature of a particular subspecies/hybrid. This suggests that
B. rapa
hybrids most likely do not differ in ploidy compared to their parental genotypes. In addition, it was shown that the mentioned oil tyfon hybrid lines of Ukrainian breeding show a significant level of morphological variation despite their common breeding pedigree.
Peach is one of the most important fruit crops, and its cultivation occupies the third largest area among all fruit crops grown in the temperate climate zone. Cultivation of this crop under less ...favorable climate conditions would require the creation of new resistant genotypes via intra- or interspecific hybridization, including crossing with almond. Efficient breeding of hybrids and their long-term selection will require a rapid and accurate method of DNA barcoding that would be able to distinguish closely related genotypes or to detect interspecific hybrids. One such approach is TBP-analysis, which is based on the evaluation of intron length polymorphism of β-tubulin genes. However, the correct interpretation of the results of such analysis should be based on data on the diversity of the β-tubulin gene panel in the genomes of the analyzed species. Thus, here we report on the successful whole-genome identification and on comprehensive analysis of the phylogeny and synteny of the β-tubulin genes of
P. persica
and
P. dulcis
, and the possibility is demonstrated to use such data of the genomic search for interpretation of data of TBP genotyping of intra- and interspecific hybrids of peach and almond species. In general, 11 β-tubulin genes were identified within the
P. persica
genome and 10 genes in the
P. dulcis
genome accounting for pseudogenes. Additionally, phylogenetic and synteny analyzes of the identified genes made it possible to identify the orthologues in the genomes of
A. thaliana
and
A. lyrata
as well as to designate identified β-tubulins to specific isotypes. Genotyping via the TBP-method allowed obtaining distinct molecular profiles for 11 investigated accessions, among which eight were intra- or interspecific hybrids. Based on the obtained results of genotyping, a cluster analysis was carried out, the results of which correlated well with the breeding history of the analyzed genotypes, which additionally confirmed the effectiveness and accuracy of the genotyping approach used.
Based on the analysis of the intron polymorphism of β-tubulin genes, the genetic variability of old Quercus robur L. trees from “Holosiivsky” NPP was investigated. The genotyping of 55 old Q. robur ...trees was carried out; 40 polymorphic and one monomorphic (about 880 bp) TBR fragments were found. High frequency (70–90%) of occurrence of fragments with an approximate molecular weight of 275, 490, 500, and 1110 bp was observed.The genetic polymorphism of old Q. robur trees was assessed as quite high: РІС is 0.22 – 0.39, the effective number of alleles per locus was 1.174–1.268. The Shannon information index was in the range of 0.204–0.269.The geographical differentiation of the genetic structure of centuries-old oak trees from “Holosiivsky” NPP was not pronounced. The share of inter-selection genetic variability (AMOVA) accounts for about 6% of genetic variability, and the geographic component – about 1%. Around 93% of genetic variability is concentrated on the individual level. Using the ТВР method, we found that Q. robur forest stands do not have a stabilized genetic and visible spatial structure, but at the same time they possess a sufficiently large genetic diversity.
A quick one-step fabrication of efficient SERS substrates by a modified approach based on a silver-mirror reaction (using Tollens’ reagent) is reported. Commercially available microscope slides or ...cover glass (coverslips) were used as-received, without special surface treatment. In contrast to the commonly used two-step process, the composition of the Tollens reagent was modified to use a single-step process. The obtained rather homogeneous films of densely packed nanoislands are promising for application as substrates for Surface-Enhanced Raman Scattering (SERS), as demonstrated by several different kinds of molecules as analytes. In particular, the achieved level of detection of a standard dye analyte, down to 10-14 M of Rhodamine 6G, is in the range of best values reported in the literature. Low concentrations of some biomolecules are also detected, such as lysozyme (10-4 M), adenine (10-4 M), and salicylic acid (10-5 M). For some analytes, stronger SERS was observed in the drop, and for others after the solvent was dried. The possible reasons for this effect are described. By applying thermal annealing in the inert gas atmosphere, the Ag film morphology can be partially converted into a coral-like 3D structure that may be advantageous for the localization of the analyte in the “hot spots” and allow additional spectral tunability of the plasmon resonance.
The biolistic transformation method was used for genetic improvement of three commercial cultivars of barley (Oksamytoviy, Vodogray, and Hetman). The plasmid pHLFTuBA containing target gene hLF ...encoding human lactoferrin under the control of the rice glutein B-1 promoter GluB-1 was used for transformation. The gene encoding mutant alfa-tubulin conferring resistance to trifluralin (dinitroaniline herbicide) was used as the selective marker. The screening of different trifluralin concentrations ranging from 0.1-30 μM was used for determination of selective concentration of the agent. Two transgenic barley lines of cultivars Oksamytoviy and Hetman's callus line were selected after 2-3 months of cultivation on 10 μM of trifluralin. To confirm stable integration of the transformed gene, the PCR analysis of leafs from regenerated plant after their adaptation on the ground was carried out. The 734 bp fragment of the target gene was amplified from both regenerated plants.
Tubulins play crucial roles in numerous fundamental processes of plant development. In flowering plants, tubulins are grouped into alpha-, beta- and gamma-subfamilies, while alpha- and beta-tubulins ...possess a large isotype diversity and gene number variations among different species. This circumstance leads to insufficient recognition of orthologous isotypes and significantly complicates extrapolation of obtained experimental results, and brings difficulties for the identification of particular tubulin isotype function. The aim of this research is to identify and characterize tubulins of an emerging biofuel crop Camelina sativa. We report comprehensive identification and characterization of tubulin gene family in C. sativa, including analyses of exon-intron organization, duplicated genes comparison, proper isotype designation, phylogenetic analysis, and expression patterns in different tissues. 17 alpha-, 34 beta- and 6 gamma-tubulin genes were identified and assigned to a particular isotype. Recognition of orthologous tubulin isotypes was cross-referred, involving data of phylogeny, synteny analyses and genes allocation on reconstructed genomic blocks of Ancestral Crucifer Karyotype. An investigation of expression patterns of tubulin homeologs revealed the predominant role of N.sup.6 (A) and N.sup.7 (B) subgenomes in tubulin expression at various developmental stages, contrarily to general the dominance of transcripts of H.sup.7 (C) subgenome. For the first time a complete set of tubulin gene family members was identified and characterized for allohexaploid C. sativa species. The study demonstrates the comprehensive approach of precise inferring gene orthology. The applied technique allowed not only identifying C. sativa tubulin orthologs in model Arabidopsis species and tracking tubulin gene evolution, but also uncovered that A. thaliana is missing orthologs for several particular isotypes of alpha- and beta-tubulins.
Using electrophoretic analysis of 11 enzyme systems, we studied the genetic structure and differentiation of eight natural populations of silver fir Abies alba Mill. in the Ukrainian Carpathians. Of ...24 isozyme loci identified, 66.8% proved to be polymorphic. The mean numbers of alleles and genotypes per locus in the populations were respectively 3.1 and 4.5. Each A. alba tree was on average heterozygous at 15.9% of genes. In six populations, the genotypic distribution for all of the loci examined corresponded to Hardy-Weinberg proportions. The populations studied had low levels of subdivision (F sub(ST) = 0.018; G sub(ST) = 0.019 ) and differentiation. Nei's genetic distances between the A. alba populations in the region ranged from 0.002 to 0.009, being on average 0.006.
—Camelina or false flax (
Camelina sativa
) is one of the most promising oilseed crops as a feedstock for biofuel production. It is thought that this species originated in the Eastern European ...region, where this species emerged due to polyploidization or crossing between wild representatives of the
Camelina
genus. As a result, camelina is characterized by a low level of genetic diversity, which imposes limitations on breeding and improvement of this crop. Although there are some studies discussing genetic polymorphism among the existing camelina cultivars, the assessments of Ukrainian genotypes are scarce, although this region belongs to the center of
C. sativa
’s origin. For the first time, an integrated assessment of genetic diversity among Ukrainian camelina breeding lines and cultivars, as well as analyzed the morphometric and yield parameters of their seed oil and its fatty acid composition, has been performed. Based on the results of chromatographic analysis, two camelina genotypes (FEORZhYaF-2 and FEORZhYaFD) with seed lipids to a greater extent composed of fatty acids with a short carbon chain (below C18) have been distinguished. Genetic distances between the studied genotypes have also been identified using ISSR, SSR, and ILP (actin and β-tubulin) markers. The obtained data were compared with the crop productivity and morphometric parameters, which allowed for differentiating the most promising pairs of phenotypes for further cross breeding. For example, crossing the FEORZhYaF-2 and FEORZhYaFD breeding lines with the cultivars Mirazh and Peremoha in different combinations may hypothetically provide the trait of heterosis in the first generation. This approach to the analysis of camelina breeding samples and genetic resources may become a powerful instrument for improving the given oil-bearing crop.
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