Zarema Kokaeva

Studying plant genome variation using molecular markers

The authors’ studies on the organization and variation of plant genome with the use of molecular markers are briefly reviewed with special emphasis on random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), sequence characterized amplified region (SCAR), and cleaved amplified polymorphic sequence (CAPS) markers detected with the use of polymerase chain reaction (PCR). These markers have been demonstrated to be promising for identifying cultivars and determining the purity of genetic strains of pea. Genetic relationships between strains, cultivars, and mutants of pea have been studied. The role of molecular markers in molecular genetic mapping and localizing the genes of commercially important characters of pea has been shown. The possibility of the use of molecular markers for studying somaclonal variation and detecting mutagenic factors in plants during long-term spaceflights is considered. The prospects of using DNA markers for understanding the organization and variability of higher plant genomes are discussed.The authors’ studies on the organization and variation of plant genome with the use of molecular markers are briefly reviewed with special emphasis on random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), sequence characterized amplified region (SCAR), and cleaved amplified polymorphic sequence (CAPS) markers detected with the use of polymerase chain reaction (PCR). These markers have been demonstrated to be promising for identifying cultivars and determining the purity of genetic strains of pea. Genetic relationships between strains, cultivars, and mutants of pea have been studied. The role of molecular markers in molecular genetic mapping and localizing the genes of commercially important characters of pea has been shown. The possibility of the use of molecular markers for studying somaclonal variation and detecting mutagenic factors in plants during long-term spaceflights is considered. The prospects of using DNA markers for understanding the organization and variability of higher plant genomes are discussed.

Identification and mapping of polymorphic RAPD markers of pea (Pisum sativum L.) genome

Various pea cultivars, lines, and mutants were studied by the RAPD method. Polymorphic fragments characteristic of certain pea genotypes and which can be used for identifying genotypes were detected. Inheritance of some polymorphic RAPD fragments was studied. Mendelian inheritance of these fragments was shown. By analyzing the data obtained in studies of RAPD polymorphism, genetic distances between different pea cultivars, lines, and mutants were calculated and a genealogic dendogram showing a varying extent of differences between RAPD patterns was constructed. Ten new RAPD markers linked to various pea genes were detected. Genetic distances between RAPD markers and genes to which they are linked were calculated, and the respective disposition of RAPD markers on chromosomes was established.

Effects of MTHFR gene polymorphism on the clinical and electrophysiological characteristics of migraine

BackgroundIt was previously shown that the MTHFR gene polymorphism correlated with an increased risk of migraine, particularly migraine with aura. The substitution of cytosine for thymine at the position 677 of the MTHFR gene leads to formation of the thermolabile form of the protein and development of hyperhomocysteinemia, which increases the probability of migraine. The purpose of this study was to determine whether the replacement of C677T in the gene MTHFR influenced any particular symptoms of the disease.MethodsWe have analyzed clinical and electrophysiological characteristics of 83 patients with migraine (migraine with aura (MA), 19 patients, and migraine without aura (MO), 64 patients, according to the ICHD-II (2003)) taking into account their genotypes of C677T variant of MTHFR.ResultsWe have shown that MA was significantly more prevalent among the T-allele carriers (37.2%), as compared to the СС genotype patients (0%), р < 0.0001. Patients with TT genotype were not only more likely to have accompanying symptoms (significant differences were found only for photophobia), but also more sensitive to migraine attack triggers. In RP-VEP test results we observed a trend that the T-allele carriers were presented with the decreased N75/P100 amplitudes and a positive habituation index, as compared to the СС genotype patients.ConclusionsThus, according to our data, the MTHFR genotypes are associated with several clinical and electrophysiological characteristics of migraine.

The study of the genetic effects in generation of pea plants cultivated during the whole cycle of ontogenesis on the board of RS ISS

Results of studies on growth and development of offspring of two genetically marked dwarf pea lines planted during the whole ontogenesis cycle in the Lada space greenhouse on board of Russian Segment of International Space Station (RS ISS) are presented. The offspring of M1 and M2 plants grown from seeds formed during space flight was examined under conditions of Earth-based cultivation. It had been shown that growth and developmental characteristics, frequency of chromosome aberrations in primary root meristem and level of molecular polymorphism revealed in individual plants via RAPD method show no significant differences between offspring of “space-grown” and control seeds.

RAPD Analysis of Maize Somaclones

The genetic difference between maize line A188 and A188-derived somaclones was assessed via analysis of randomly amplified polymorphic DNA (RAPD). In total, 17 decanucleotide primers used each allowed amplification of 2–17 fragments ranging 200–2000 bp. The RAPD patterns did not differ between individual plants of line A188, which demonstrated again its high genetic homogeneity. The difference between the initial line and the somaclones was high, ranging 64–74%. On evidence of the genetic divergence, the somaclones formed two clusters. The distribution of somaclones between these clusters was consistent with their origin.

Platelet-Activating Factor-Acetylhydrolase Gene (PLA2G7) Expression in Children with a History of Food Anaphylaxis

Background: One of key functions in the pathogenesis of anaphylaxis is held by platelet-activating factor (PAF). However, no research on detection of transcriptional activity of PAF-acetylhydrolase gene (PLA2G7) in patients with anaphylaxis has been held so far. The aim of this study is to evaluate transcriptional activity (expression at mRNA level) of PLA2G7 gene (plateletactivating factor acetylhydrolase) in patients with a history of anaphylaxis. Methods: 97 children were enrolled in the study: 27 children with a history of food anaphylaxis (group 1); 35 children with atopic dermatitis (group 2); 35 children without allergic diseases (group 3). Group 3 children were tested with Phadiatop and Phadiatop Infant (“Phadia 100”; “Phadia AB”, Sweden): 26 children were IgE-negative, 9 children were IgE-positive. RNA was extracted from blood samples with Trizol RNA Prep kits (“Isogen”, Russia). cDNA production was performed with MMLV RT kit (“Evrogen”, Russia). Real-time PCR was held with CFX-96 device (“Bio-Rad”, USA) and qPCRmix-HS PCR mix (“Evrogen”, Russia). Oligonucleotide primers and probes were synthesized at “DNKSintez” (Russia). Results: In children with a history of food anaphylaxis statistically significant decrease in PLA2G7 gene expression was detected (p<0.05) in compare with children free of allergic diseases. Transcriptional activity of PLA2G7 was significantly lower in children with severe anaphylaxis and in children with moderate anaphylaxis in compare with mild anaphylaxis children (p<0.05). In moderate/severe anaphylaxis children PLA2G7 expression was more than three times decreased in compare with atopic dermatitis children (p<0.05). In children with anaphylaxis characterized by clinical symptoms of cardiovascular manifestations the PLA2G7 expression level was significantly reduced in compare with anaphylaxis children free of cardiovascular symptoms (p<0.05). Conclusion: Reduction of blood cells PLA2G7 mRNA level in children with a history of food anaphylaxis may be considered to be a biomarker of severe anaphylaxis.

Production of a SCAR marker in pea Pisum sativum L. using RAPD analysis

A polymorphic 750-bp fragment, RAPD marker, specific to particular pea genotypes (line L-111 and the Nord cultivar) was identified. Using this RAPD marker, SCAR was obtained. SCAR inheritance in the first and second generations was studied and its dominant character was shown.

EHMTI-0233. Clinical characteristics of migraine patients with dopaminergic gene polymorphisms RS1611115

The dopaminergic system plays a major role in migraine. Dopamine beta hydroxylase (DBH) is responsible for maintaining dopamine-to-norepinephrine ratio implicated in migraine pathophysiology. We aimed to look for association of polymorphisms in dopaminergic genes in genetic susceptibility to migraine in Russian population. In the present study DBH polymorphisms rs1611115 was selected. The aim of this study was to determine whether the polymorphisms rs1611115 in DBH gene influenced any particular symptoms of the disease.

EHMTI-0208. Positive association between -1021C/T polymorphism of dopamine-b-hydroxylase gene and level substance dependence in medication-overuse headache

Results The genotype frequencies of -1021C/T polymorphism DBH gene did not differ between the MOH and other groups. In contrast, we found that the presence of the -1021T allele was significantly associated with the monthly drug consumption and LDQ total score in the MOH group. We also found an interaction between the presence of DBH -1021T genotype and efficiency serotonin-norepinephrine reuptake inhibitors in patient with MOH. Conclusions

Brain-derived neurotrophic factor gene (BDNF) polymorphism among Moscow citizens

Recent studies showed that brain-derived neurotrophic factor (BDNF) can participate in pathogenesis of various CNS disorders, being connected with proliferation, differentiation, and survival of neurons. In present study, analysis of occurrence rate was performed for three single nucleotide polymorphisms (SNPs) located in BDNF gene (rs6267 (A/G) allele A—0.265; rs2049046 (A/T) allele A—0.407; rs11030107 (A/G) allele A—0.872) in randomized selection of Moscow citizens. Linkage disequilibrium of rs6165 and rs2049046 loci was shown. Differences in allele frequencies in studied selection and populations of other regions were discovered.