Genetic polymorphism of white mistletoe (Viscum album L.) in Ukraine

Aim. The aim of the study was to establish genetic differences between V. album growing in different parts of Ukraine. Methods. White mistletoe samples collected in different regions of Ukraine were used in the study. The method of estimating the intron length polymorphism of β-tubulin genes was used. Amplified DNA fragments were fractionated by non-denaturing polyacrylamide gel electrophoresis and visualized by silver nitrate staining. Results. The genotypes of 91 white mistletoe plants were analyzed. DNA profiles of white mistletoe with a specific amplicons of β-tubulin gene introns were obtained, which allowed to differentiate the samples from each other. Fingerprinting data were used for cluster analysis and dendrogram construction. Conclusions. It was found that the analyzed mistletoe samples did not differ by geographical factor and were characterized by a low level of genetic diversity in the studied samples. Keywords: Viscum album L., intron length polymorphism, β-tubulin, genetic variability, Ukraine.

Cytogenetic Studies on Three Species of Genus Burmagomphus of Family Gomphidae (Odonata: Anisoptera) from India

Male germ cell chromosomes of Burmagomphus divaricatus, Burmagomphus pyramidalis and Burmagomphus sivalikensis of family Gomphidae have been investigated by using conventional staining, C-banding, silver nitrate staining and sequence specific staining. The species were collected from Punjab and Himachal Pradesh, India. All the species possess the chromosome number 2n = 23 which is the type number of the family. Terminal C bands and NOR’s are present at the autosomal bivalents and X chromosome is C positive and NOR rich in all the three species, while m bivalents show variation in distribution of C- heterochromatin and NOR’s. In the sequence specific staining, whole complement shows bright DAPI signals in B. divaricatus, bright CMA3 signals in B. pyramidalis and both DAPI and CMA3 signals in B. sivalikensis.

Postnatal Growth Restriction in Mice Alters Cardiac Protein Composition and Leads to Functional Impairment in Adulthood

Postnatal growth restriction (PGR) increases the risk for cardiovascular disease (CVD) in adulthood, yet there is minimal mechanistic rationale for the observed pathology. The purpose of this study was to identify proteomic differences in hearts of growth-restricted and unrestricted mice, and propose mechanisms related to impairment in adulthood. Friend leukemia virus B (FVB) mouse dams were fed a control (CON: 20% protein), or low-protein (LP: 8% protein) isocaloric diet 2 weeks before mating. LP dams produce 20% less milk, inducing growth restriction. At birth (postnatal; PN1), pups born to dams fed the CON diet were switched to LP dams (PGR group) or a different CON dam. At PN21, a sub-cohort of CON (n = 3 males; n = 3 females) and PGR (n = 3 males; n = 3 females) were euthanized and their proteome analyzed by two-dimensional differential in-gel electrophoresis (2D DIGE) and mass spectroscopy. Western blotting and silver nitrate staining confirmed 2D DIGE results. Littermates (CON: n = 4 males and n = 4 females; PGR: n = 4 males and n = 4 females) were weaned to the CON diet. At PN77, echocardiography measured cardiac function. At PN80, hearts were removed for western blotting to determine if differences persisted into adulthood. 2D DIGE and western blot confirmation indicated PGR had reductions in p57kip2, Titin (Ttn), and Collagen (Col). At PN77, PGR had impaired cardiac function as measured by echocardiography. At PN80, western blots of p57kip2 showed protein abundance recovered from PN21. PN80 silver staining of large molecular weight proteins (Ttn and Col) was reduced in PGR. PGR reduces cell cycle activity at PN21, which is recovered in adulthood. However, collagen fiber networks are altered into adulthood.

Evaluation of genetic diversity of honey bee in Ukraine analyzed by the SSR-markers

Aim. The aim of the work was to analyze current genetic structure of honey bee populations in Ukraine that belong to different subspecies: A. meliffera meliffera, A. meliffera carnica, A. meliffera macedonica using microsatellite markers. Methods. SSR-analysis was used for evaluation of the honey bee polymorphism. Amplified fragments were fractionated by electrophoresis in non-denaturing polyacrylamide gel. DNA bands were detected using silver nitrate staining. Results. The analysis of the sample of honey bees (workers and male-bees) collected from different regions of Ukraine was performed by using two SSR-markers (Ac011 and A007). In this sample reasonably high polymorphism was observed, especially for the SSR-marker A007. Conclusions. It was estimated that SSR-analysis method can be applied in molecular-genetic analysis of honey bees for evaluation of genetic diversity and cross-subspecies hybridization. Keywords: microsatellite markers, Apis meliffera, PIC (Polymorphism Information Content).

Chromosome Dynamics Regulating Genomic Dispersion and Alteration of Nucleolus Organizer Regions (NORs)

The nucleolus organizer regions (NORs) demonstrate differences in genomic dispersion and transcriptional activity among all organisms. I postulate that such differences stem from distinct genomic structures and their interactions from chromosome observations using fluorescence in situ hybridization and silver nitrate staining methods. Examples in primates and Australian bulldog ants indicate that chromosomal features indeed play a significant role in determining the properties of NORs. In primates, rDNA arrays that are located on the short arm of acrocentrics frequently form reciprocal associations (“affinity”), but they lack such associations (“non-affinity”) with other repeat arrays—a binary molecular effect. These “rules” of affinity vs. non-affinity are extrapolated from the chromosomal configurations of meiotic prophase. In bulldog ants, genomic dispersions of rDNA loci expand much more widely following an increase in the number of acrocentric chromosomes formed by centric fission. Affinity appears to be a significantly greater force: associations likely form among rDNA and heterochromatin arrays of acrocentrics—thus, more acrocentrics bring about more rDNA loci. The specific interactions among NOR-related genome structures remain unclear and require further investigation. Here, I propose that there are limited and non-limited genomic dispersion systems that result from genomic affinity rules, inducing specific chromosomal configurations that are related to NORs.

Distribution of mistletoe (Viscum album L.), which parasitizes different woody plants species, in Kyiv and its genetic characteristics

Aim. The aim of the work was to study the species diversity of mistletoe host trees with the establishment of damage degrees and the study of the mistletoe genetic characteristics in Kiev. Methods. The β-tubulin first intron length polymorphism evaluating method (TBP) was used. Amplified fragments DNA were fractionated by electrophoresis in non-denaturing polyacrylamide gel. DNA bands were detected using silver nitrate staining. Results. The species diversity of Viscum album L. host-trees was studied in Kiev. Among them, plants of the genus Pinus and angiosperms that belong to 8 genera (Acer, Fraxinus, Acacia, Populus, Tilia, Salix, Malus, Sorbus), wherein 47% belong to the genus Acer. It was revealed that among infected trees, 30% had severe damage to mistletoe, 28% moderate and 42% weak. Based on the analysis of DNA marker polymorphisms, the molecular-genetic profiles of V. album, which grows on various species of host trees, were obtained and analyzed. Conclusions. The species diversity of V. album host-trees was studied and the degree of their infection was assessed in Kiev. Mistletoe, which grows on angiosperms, is characterized by a greater degree of genetic polymorphism than that which grows on gymnosperms. Keywords: Viscum album L., host-plant, introns of genes, length polymorphism, β-tubulin.

Intravarietal intron-length polymorphism of β-tubulin genes in belorussian landraces of Linum usitatissimum L.

Aim. The aim of the work was to evaluate the possibilities of using the β-tubulin intron length (TBP, Tubulin Based Polymorphism) for genetic differentiation of ancient flax varieties (landraces), plants that was historically formed in Belarus. Methods. The β-tubulin first intron length polymorphism evaluating method (TBP) was used. Amplified fragments (β-tubulin introns) were fractionated by electrophoresis in non-denaturing polyacrylamide gel. DNA bands were detected using silver nitrate staining. Results. The size of the amplified fragments varied from 400 bp to 1900 bp. It was found that 25 of 30 studied varieties (landraces) were genetically heterogeneous. The total number of allele phenotypes was 7, and the value of PIC (Polymorphism Information Content) varied from 0.0 to 0.72. Conclusions. The data obtained make it possible to recommend landraces as a source of genes for increasing the genetic diversity of the existing flax gene pool, and the TBP method can be applied both in molecular-phylogenetic analysis and in molecular selection of flax. Keywords: Linum usitatissimum, landrace, molecular markers, DNA fingerprinting, β-tubulin genes, intron polymorphism.

Short Communication: Genetic diversity of fumonisin producing Fusarium isolates from rice using PCR-RFLP of IGS-rDNA region

Nayak S, Dhua U, Chhotaray A, Samanta S, Sengupta C. 2018. Short Communication: Genetic diversity of fumonisin producing Fusarium isolates from rice using PCR-RFLP of IGS-rDNA region. Biodiversitas 19: 571-576. Fusarium verticillioides (Sacc.) and related species produce carcinogenic mycotoxin known as Fumonisins in several agricultural crops including rice. However, this principal food crop has been infected by genetically diverse Fusarium species. Odisha belongs to the coastal part of India and many popular rice varieties are in the food chain in this region. Many Fusarium species producing fumonisins have been found to be associated with these rice varieties. Hence, the genetic diversity of twenty eight Fumonisin producers and non producers of Fusarium pathogens in this region was carried out in the current study. The IGS regions of 28 Fusarium isolates (both fumonisin producing and non producing) were amplified and the PCR products were restriction digested with ECoRI and HhaI. The digested products were separated on PAGE and bands were visualized by Silver Nitrate Staining. The 28 isolates could be separated into 14 IGS haplotypes. The lowest similarity was detected to be of 33% between F40 and F47. A group containing 14 isolates represented the biggest haplotypes. The isolates in which the FUM gene had not been detected (fumonisin non producer) were in a separate group having 90% similarity with each other and placed consistently in separate branch from others. Presence of unique band for this group was observed at 1650bp where as absence of specific bands was observed at 380bp and 300bp. The result of this study indicated a high degree of genetic variation among 28 Fusarium isolates. PCR RFLP of IGS region was also found to be useful for diversity study in Fusarium.