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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1009-1019

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.

                               https://doi.org/10.2298/GENSR1903009D

Original scientific paper

INCIDENCE OF GENETIC CAUSES OF IDIOPATHIC MALE INFERTILITY

IN SERBIA – TEN YEARS’ EXPERIENCE OF SINGLE CENTRE

Bojana DOBRIC¹, Danijela RADIVOJEVIC¹, Tanja LALIC¹, Marijana MISKOVIC¹,

Sanja CIRKOVIC¹, Maja DJORDJEVIC^2,3, Marina DJURISIC¹

¹Laboratory of Medical Genetics, Mother and Child Health Care Institute of Serbia “Dr Vukan Cupic”, Belgrade, Serbia

²Metabolic Department, Mother and Child Health Care Institute of Serbia “Dr Vukan Cupic”, Belgrade, Serbia

³School of Medicine, University of Belgrade, Belgrade, Serbia

Abstract

dobric B., D. Radivojevic, T. Lalic, M. Miskovic, S. Cirkovic, M. Djordjevic, M. Djurisic (2019): Incidence of genetic causes of idiophatic male infertility in Serbia-ten years’ experience of single centre. - Genetika, Vol 51, No.3, 1009-1019.

The aim of the study was to estimate the type and the prevalence of chromosomal abnormalities and Y-chromosome microdeletions, analysed together for the first time in idiopathic infertile men in Serbia. During 10 years period among 823 couples with infertility problems, in 110 cases the cause of infertility was severe oligospermia or azoospermia in male partners. All of them underwent cytogenetic analysis, performed according to standard techniques. Testing for the presence of Y-chromosome microdeletions in AZF regions using multiplex PCR was done in all patients with normal karyotype (97) and in three cases with cytogenetically visible aberrations of Y chromosome, in order to specify the breakpoints. The overall prevalence of chromosomal abnormalities in the group of 110 infertile men was 11.82%. The most frequent aberration was Klinefelter syndrome (47, XXY), being found in 5.45%. Chromosomal aberrations were found in 13.89% in group of men with azoospermia, and in 7.89% in group of men with severe oligospermia. Among the infertile men with normal karyotype, the incidence of microdeletions of AZF regions was 7.22%. Two types of deletions were identified: AZFc and AZFbc, with frequencies of 6.19% and 1.03%, respectively. Y-chromosome microdeletions were found in 6.45% of azoospermic patients, and in 8.57% of severe oligospermia group of patients. Our findings demonstrate the presence of higher frequency of chromosome aberrations and Y-microdeletions in a group of infertile men with azoospermia/oligospermia in Serbia. Results confirmed importance of offering these tests as part of genetic counselling of infertile couples in our country.

Keywords: male infertility, azoospermia, oligospermia, karyotype, Y-chromosome microdeletions

Corresponding author: Bojana Dobric, Laboratory of Medical Genetics, Mother and Child Health Care Institute of Serbia “Dr Vukan Cupic”, Radoja Dakica str.6-8, 11070 Belgrade, Serbia,Tel: ++381 11 3108 273,Fax: ++381 11 3108 276,E-mail: bojana_dobric@yahoo.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1021-1030

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.630

https://doi.org/10.2298/GENSR1903021G

Original scientific paper

GENETIC VARIABILITY ANALYSIS IN Peganum harmala L. BY SCOT AND SRAP MOLECULAR MARKERS

Tayebeh GHOLAMI¹, Maryam PEYVANDI^1*, Hossein ABBASPOUR¹, Zahra NOORMOHAMMADI², Fariba SHARIFNIA¹

¹Department of Biology, Faculty of Biological Sciences, North-Tehran Branch, Islamic Azad University, Tehran, Iran.

²Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Abstract

Gholami T., M.Peyvandi, H. Abbaspour, Z. Noormohammadi, F. Sharifnia (2019): Genetic variability analysis in Peganum harmala L. by SCOT and SRAP molecular markers.- Genetika, Vol 51, No.3, 1021-1030.

Peganum harmala L. is a perennial herbaceous plant and it has long been used for medicinal purposes as herbicide due to the presence of harmine. P. harmala is a famous medicinal plant used in the Iranian traditional medicine, due to the antimicrobial compounds found in its seeds and roots. Population genetic study is an essential scientific approach for studying medicinal plants; as it produces data on genetic variability, genetic structure, and gene flow versus genetic fragmentation of these plants. We have no detailed information on genetic structure of this plant species in the country. Therefore, a through population genetic study was conducted in four geographical populations of these valuable plants by using start codon targeted (SCoT), and sequence related amplified polymorphic (SRAP) molecular markers. Both these molecular markers are highly reproducible and polymorphic molecular markers and are very efficient in genetic variation studies in plants. In present study, both multidimensional scaling (MDS) plot and analysis of molecular variance (AMOVA) based on both SCOT and SRAP molecular markers revealed genetic differentiation of the studied populations. This indicates that local populations may be genetically isolated and due to that gained specific genetic content. This is good news for genetic conservation of these medicinally important plants. We should continue for hunting new P. harmala wild populations in the country and study their genetic structure; in this way we can broaden P. harmala gene pools which can be utilized for future breeding studies.

Keywords: Genetic variability, Peganum harmala, SCoT, SRAP

Corresponding author: Maryam Peyvandi, Department of Biology, Faculty of Biological Sciences, North-Tehran Branch, Islamic Azad University, Tehran, Iran e-mail: m_peyvandi@iau-tnb.ac.ir; Phone: 098 9122899867

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp.1031-1038

© 2019 Serbian Genetics Society

S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.633

https://doi.org/10.2298/GENSR1903031H

Original scientific paper

MARKER AIDED SELECTION (MAS) FOR QUALITATIVE TRAITS IN SEGREGATING F₂ POPULATIONS OF RICE

A.HAJIAQATABAR¹, G. KIANI¹*, S. K. KAZEMITABAR¹, M. ALAVI²

¹Department of Plant Breeding, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

²Genetics and Agricultural Biotechnology Institute of Tabarestan

Abstract

Hajiaqatabar A., G. Kiani, S. K. Kazemitabar, M. Alavi (2019): Marker aided selection (MAS) for qualitative traits in segregating F₂ populations of rice.- Genetika, Vol 51, No.3, 1031-1038.

The selection and screening of desirable traits at early stages of plant development is possible with the advancement of marker assisted selection (MAS). Marker aided selection can be used for monitoring the presence or absence of genes in breeding populations and can be combined with conventional breeding approaches. In this study, crosses between high-yielding cultivar, Nemat, with 4 local aromatic varieties were made followed by phenotypic selection for desirable individual plants in F₂ populations. Then, MAS applied to genotypic selection of aromatic plants using allele specific amplification (ASA) marker for fragrance (fgr) and SSR marker RM190 linked with intermediate amylose content (AC), gel consistency (GC) and gelatinization temperature (GT) in these populations. Molecular screening based on RM190 and fragrance markers showed that 11 lines were homozygous aromatic and 32 lines had desirable cooking quality in segregating F₂ population. Results demonstrated that marker assisted selection in F₂ segregating populations reduced the time and cost effective for accelerating cultivar development in rice. Further analyses are underway on these plants on subsequent generations through pedigree breeding method. These plants promise to develop new aromatic rice lines through classical and molecular breeding in the near future in Iran.

Keywords: aroma, markers, quality, rice, screening

Corresponding author: G. Kiani, Department of Plant Breeding, Sari Agricultural Sciences and Natural Resources University, Sari, Iran, Email: ghkiani@gmail.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1039-1052

© 2019 Serbian Genetics Society

S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.

https://doi.org/10.2298/GENSR1903039I

Original scientific paper

SPECIFICITY AND SENSITIVITY OF THREE PCR-BASED METHODS FOR DETECTION OF Erwinia amylovora IN PURE CULTURE AND PLANT MATERIAL

Milan IVANOVIĆ^1*, Nemanja KUZMANOVIĆ², Katarina GAŠIĆ³, Anđelka PROKIĆ¹,

Nevena ZLATKOVIĆ¹, Aleksa OBRADOVIĆ¹

¹University of Belgrade, Faculty of Agriculture, Belgrade-Zemun, Serbia

²Institute for Epidemiology and Pathogen Diagnostics, Federal Research Centre for Cultivated Plants, Julius Kühn-Institut (JKI), Braunschweig, Germany

³Institute for Plant Protection and Environment, Belgrade, Serbia

Abstract

Ivanović M., N. Kuzmanović, K. Gašić, A. Prokić, N. Zlatković, A. Obradović  (2019): Specificity and sensitivity of three PCR-based methods for detection of Erwinia amylovora in pure culture and plant material.- Genetika, Vol 51, No.3, 1039-1052.

Three PCR methods, referred in this study as „conventional“, „nested“ and „chromosomal“ PCR and suggested for routine detection of Erwinia amylovora in pure culture and plant material, were evaluated according to their specificity and sensitivity. Specificity of PCR methods was analyzed by using 42 strains of E. amylovora, originating from different locations and plant species, with diverse PFGE profiles, representing distant populations of the pathogen. Sensitivity of PCR protocols in pure culture was studied by using nine different concentrations of E. amylovora in sterile ultrapure water as a template in PCR reactions. In order to study inhibitory effect of plant DNA and other inhibitors on sensitivity of the three PCR methods bacterial dilutions were mixed with plant macerate of pear, apple and quince prior to the PCR reaction. In specificity assays, tested PCR protocols were able to detect all E. amylovora strains regardless of the host of the strain, its origin or PFGE group, indicating primer specificity. On the other hand, sensitivity among tested methods varied, depending on bacterial concentration and selected plant material used in the PCR. When working with pure cultures nested PCR showed the greatest sensitivity by detecting 1.9 bacterial cells per PCR reaction, followed by detection limit of 9.5 cells per PCR reaction with conventional PCR and 1.9´10⁵ cells/PCR reaction with chromosomal PCR. In spiked samples plant inhibitors either did not affect or they decreased the sensitivity of the PCR reaction, depending on the protocol and/or type of plant macerate. In our experiments, inhibitors from pear and quince macerates did not affect sensitivity of nested PCR, while apple macerate reduced its sensitivity by a factor of 10. Conventional PCR protocol was able to detect 95 cells/PCR reaction in pear and apple macerate, but only 9.5´10³ cells/PCR in quince macerate. Greatest decrease in sensitivity of the PCR method was observed in spiked samples with chromosomal PCR since bacterial DNA was not detected in each of the spiked samples. Our research shows that all three PCR protocols are specific for detection of E. amylovora, but nested PCR proved to be most sensitive when working with pure cultures and plant material.

Keywords: molecular detection, conventional PCR, nested PCR, quantification, fire blight

Corresponding author: Milan Ivanović, University of Belgrade, Faculty of Agriculture, Nemanjina 6, 11080 Belgrade-Zemun, Serbia, Phone: 011 441 3301, e-mail: milanivanovic007@yahoo.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1053-1060

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.

https://doi.org/10.2298/GENSR1903053Z

Original scientific paper

PRP GENE POLYMORPHISM IN THE RED DEER (Cervus elaphus L.)

Ervin ZECEVIC, Admir DOKSO, Alma RUSTEMPASIC and Muhamed BRKA

Faculty of Agriculture and Food Sciences University of Sarajevo

Abstract

Zecevic E., A. Dokso, A.Rustempasic and M. Brka (2019): PRP gene polymorphism in the red deer (Cervus elaphus L.).- Genetika, Vol 51, No.3, 1053-1060.

Transmissible spongiform encephalopathy (TSE) belongs to the group of contagious disease that affects the nervous system with fatal outcome. Chronic wasting disease belongs to this group and is characteristic for animals from the deer family. In this study polymorphisms in eleven codons located in exon 3 of the PrP gene has been investigated red deer (Cervus elaphus L.) population. Exon 3 regions, 628 bp long, were amplified by the PCR method from genomic DNA. Sequencing was performed by applying the Sanger dideoxi method. Results showed presence of eight different genotypes and eight different haplotypes. Susceptible genotypes were not found at a high frequency.

Keywords: PrP, transmissible spongiform encephalopathy, Deer (Cervus elaphus L.)

Corresponding author: Ervin Zecevic Faculty of Agriculture and Food Sciences University of Sarajevo, Bosnia and Herzegovina, Phone +387 61 171 176; E-mail: e.zecevic@ppf.unsa.ba

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1061-1073

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.

https://doi.org/10.2298/GENSR1903061I

Original scientific paper

Genetic, MOrphological and chemical characterisation of the GRAPE variety 'Probus' (Vitis vinifera L.)

Dragoslav Ivanišević¹, Mladen Kalajdžić¹, Gabriele Di Gaspero², Mato Drenjančević³, Nada Korać¹, Florian Schwander⁴, Ulrike Braun⁴, Goran Barać¹, Serena Foria^2,*

¹ University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia

² Istituto di Genomica Applicata, Udine, Italy

³ University of J.J. Strossmayer, Faculty of Agriculture, Osijek, Croatia

⁴ Julius Kühn-Instiut, Institute for Grapevine Breeding, Siebeldingen, Germany

* current affiliation: Dr. Schär R&D, AREA Science Park Trieste, Italy

⁵University of Sarajevo-Institute for Genetic Engineering and Biotechnology, Sarajevo, B&H

Abstract

Ivanišević D., M. Kalajdžić, G. Di Gaspero, M. Drenjančević, N. Korać, F. Schwander, U. Braun, G. Barać, S. Foria (2019): Genetic, morphological and chemical characterisation of the grape variety 'Probus' (Vitis vinifera L.).- Genetika, Vol 51, No.3, 1061-1073.

In the middle of the 20^th century, varieties of Vitis vinifera locally grown in the Balkans were crossed with varieties from Western Europe with the aim of improving wine quality. The variety 'Probus' originated from the crossing of 'Kadarka' and 'Cabernet Sauvignon' and was released in 1983. The acreage of 'Probus' has recently increased in Serbia, along with the reputation of its wine. A detailed characterisation of this variety is therefore desirable. Traits with high heritability such as phenology, leaf morphology, anthocyanin profile, and microsatellite DNA were compared between 'Probus' and its parents were compared. 'Probus' and 'Cabernet Sauvignon' showed a synchronized phenology and highly similar leaf shape. The anthocyanin composition of 'Probus' wines was more similar to 'Cabernet Sauvignon' than to 'Kadarka', due to high anthocyanin content and high relative abundance of tri-substituted monoglucosides and acetylated conjugates, which are usually associated with deeper and more durable colour during the ageing of red wines. For total anthocyanin content, 'Probus' performed even better than 'Cabernet Sauvignon' in the environmental conditions of Northern Serbia. 'Probus' has compact bunches and large berries typical of the high-yielding 'Kadarka'. DNA analysis confirmed the alleged parentage of 'Probus' and revealed the high heterozygosity of its genome, as result of the cross between distantly related parents. These results confirm that 'Probus' is a promising variety for the production of quality red wines in Southeastern Europe.

Keywords: ampelography, anthocyanins, grapevine microsatellite markers, Vitis vinifera, 'Probus'

Corresponding author: Dragoslav Ivanišević, University of Novi Sad, Faculty of Agriculture, Trg Dositeja Obradovića 8, 21000 Novi Sad, Serbia. Email: idragoslav@polj.uns.ac.rs, Phone: +381 21 485 3253

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1075-1087

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.630

https://doi.org/10.2298/GENSR1903075J

Original scientific paper

GENETIC ADVANCE AND REGRESSION ANALYSIS IN SUNFLOWER

Milan JOCKOVIĆ¹, Siniša JOCIĆ¹, Slaven PRODANOVIĆ², Sandra CVEJIĆ¹,

Jelena JOCKOVIĆ³, Aleksandra RADANOVIĆ¹, Bojan JOCKOVIĆ¹

¹Institute of Field and Vegetable Crops, Novi Sad, Serbia

²Agriculture Faculty, University of Belgrade, Zemun, Serbia

³Faculty of Sciences, University of Novi Sad, Serbia

Abstract

Jocković M., S. Jocić, S. Prodanović, S, Cvejić, J. Jocković, A. Radanović, B. Jocković (2019): Genetic advance and regression analysis in sunflower.- Genetika, Vol 51, No.3, 1075-1087.

The knowledge about the magnitude and nature of variability that is present in a breeding population is an important prerequisite for designing efficient breeding programme in order to improve the yield potential of genotypes. The objective of this research was to evaluate heritability and genetic advance of important quantitative traits in new crosses of sunflower as well as to evaluate ratio of dominant and recessive genes in parental genotypes. The plant material selected for this research consisted of 6 sunflower genotypes, which according to literary data possess important characteristics for the production of sunflower. According to presented results there is significant variability of evaluated quantitative traits. Phenotypic variance was higher than genotypic demonstrating strong environment effect in expression of traits. The broad sense heritability was found very high for plant height (83.25%), high for 1000 seed weight (69.33%), moderate for seed yield/plant (46.53%) and head diameter (56.89%), while low for oil content (29.35%). Genetic advance expressed as a percentage of the mean ranged between 2.23% and 19.96%. Placement of array points displayed that the highest frequency of dominant genes for seed yield/plant, 1000 seed weight and head diameter was found in parental genotype Rodnik. Position of expected line of regression pointed over dominance in inheritance for seed yield/plant, oil content and head diameter, while for 1000 seed weight and plant height additive gene action played role in inheritance suggesting that selection in early generations for these traits will be effective. By testing the coefficients of regression interallelic interaction was not determined.

Key words: components of variance, heritability, inheritance, interallelic interaction

Corresponding author: Milan Jocković, Institute of Field and Vegetable Crops, Novi Sad, Serbia,e-mail: milan.jockovic@ifvcns.ns.ac.rs; jockovic@gmail.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp.1089-1101

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.630

https://doi.org/10.2298/GENSR1903089S

Original scientific paper

SOME PHYSICOCHEMICAL CHARACTERISTICS OF BLACK AND WHITE MULBERRY GENOTYPES FROM BOSNIA AND HERZEGOVINA

Azra SKENDER¹, Mirsad KURTOVIĆ², Sezai ERCISLI³, Dinko BEĆIRSPAHIĆ¹

¹Biotechnical Faculty of University Bihać, Bihać, Bosnia and Herzegovina

² Faculty of Agricultural and Food Sciences of University Sarajevo, Sarajevo, Bosnia and Herzegovina

³Department of Horticulture, Faculty of Agriculture, Ataturk University, Erzurum, Turkey

Abstract

Skender A., M. Kurtović, S. Ercisli, D. Bećirspahić (2019): Some physicochemical characteristics of black and white mulberry genotypes from Bosnia and Herzegovina.- Genetika, Vol 51, No.3, 1089-1101.

Recently, interest in fruit consumption and the practice of healthy eating is growing. Therefore, the focus of this research is to determine some physicochemical properties of mulberry fruits from the area of ​​northwest Bosnia, where significant amount genetic resources of mulberries are available. In total, six genotypes of black and eight genotypes of white mulberries were analyzed. From physical properties, the fruit mass, fruit height, fruit width and length were determined. Black mulberries are found richer than white mulberries for most of the searched parameters. The average water, soluble solids, total acidity, total sugar, ash, vitamin C, total phenol and total anthocyanin of black mulberry genotypes were 86.46%, 14.88%, 0.08%, 7.30%, 0.68%, 45.84 mg/100 g FW (fresh weight), 50.67 mg GAE/100 g FW and 562.42 cyanidin-3-glucoside mg/g. These values were 82.49%, 17.51%, 0.09%, 7.34%, 0.78%, 43.79 mg/100 g FW and 29.49 mg GAE/100 g FW for white mulberries. The obtained results can serve for future steps in the breeding of these species, and because of the high quality fruit characteristics, they can affect the conservation of these species due to the increasing genetic erosion of this species.

Keywords: Morus nigra, Morus alba, physical and chemical characteristics, mulberry

Corresponding author: Azra Skender, PhD. Biotecnical faculty, University of Bihać, Luke Marjanovića bb, 77 000 Bihać, Bosna i Hercegovina. Telephone number: ++ 387 37 228 059; Fax number: ++ 387 37 228 059; e-mail: skender.azra71@gmail.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1103-1112

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.630

https://doi.org/10.2298/GENSR1903103P

Original scientific paper

APPLICATION OF SSR MARKERS FOR ASSESSMENT OF GENETIC DIFFERENTIATION OF SILVER FIR (Abies alba Mill.) ORIGINATING FROM JAVOR MOUNTAIN

Vladan POPOVIĆ¹, Aleksandar LUČIĆ¹, Ljubinko RAKONJAC¹, Jelena MILOVANOVIĆ²,

Snežana MLADENOVIĆ DRINIĆ³, Danijela RISTIĆ³

¹Institute of Forestry, Belgrade

²Singidunum University, Belgrade

³Maize Research Institute, Zemun Polje, Belgrade

Abstract

Popović V., A. Lučić, Lj. Rakonjac, J. Milovanović, S. Mladenović Drinić, D. Ristić (2019): Application of SSR markers for assessment of genetic differentiation of silver fir (Abies alba Mill.) originating from Javor mountain.- Genetika, Vol 51, No.3, 1103-1112.

The process of plant breeding and conservation of gene pool among other things depends on the knowledge of the level of genetic differentiation. The aim of research in this paper was to determine the genetic differentiation of silver fir (Abies alba Mill.) populations of regular type and atypical genotypes with pyramidal crown that can be found on Javor mountain, at the site Ogorijevac. The genetic differentiation of silver fir was determined using SSR (Simple Sequence Repeat) markers. Nine SSR pairs of primers gave 29 alleles, while the average number of alleles was 3.2. The primer NFH15 gave the smallest number of alleles (two), while the primer SF78 gave the greatest number of alleles (five). Dice coefficient of the genetic similarity was used to obtain a dendrogram by UPMGA analysis using NTSYSpc statistical program. The genetic similarity recorded among the individuals P1 and P2 was the largest (0.89), while the populations VI and individual P2 showed the lowest similarity (0.61).

Based on the cluster analysis it can be concluded that the studied populations and genotypes of silver fir with different types of crown are clearly differentiated. The basic insight into the level of the genetic diversity of the natural populations of silver fir with the various types of crown has been provided using selected SSR markers. The obtained results can be used for creating further strategy for the conservation of the available gene pool and the regeneration of silver fir forests in Serbia.

Keywords: Abies alba Mill., SSR (Simple Sequence Repeat) markers, population, differentiation

Corresponding author: Vladan Popović; Institute of Forestry; Kneza Višeslava 3; 11000 Belgrade; Serbia; Phone: 011-3553355; e-mail: vladanpop79@gmail.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp 1113-1126

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.

https://doi.org/10.2298/GENSR1903113S

Original scientific paper

COMET ASSAY AND CYTOGENETIC FINDINGS IN DIFFERENTIAL DIAGNOSIS OF FANCONI ANEMIA

Karolina SUNJOG*^1,2, Sanja ĆIRKOVIĆ³, Branka VUKOVIĆ-GAČIĆ¹, Marija GUĆ-ŠĆEKIĆ⁴,

Marijana MIŠKOVIĆ³, Dragana VUJIĆ⁵,⁶ and Dejan ŠKORIĆ⁷

¹University of Belgrade, Faculty of Biology, Chair of Microbiology, Center for Genotoxicology and Ecogenotoxicology, Belgrade, Serbia

²University of Belgrade, Institute for Multidisciplinary Research, Department of biology and inland waters protection, Belgrade, Serbia

³Mother and Child Health Care Institute of Serbia “Dr Vukan Čupić”, Laboratory for medical genetics, Belgrade, Serbia

⁴University of Belgrade, Faculty of Biology, Chair of Genetics and Evolution, Belgrade, Serbia

⁵University of Belgrade, Faculty of Medicine, Chair of Pediatrics, Belgrade, Serbia

⁶Mother and Child Health Care Institute of Serbia “Dr Vukan Čupić”, Department of Bone Marrow Transplantation with Laboratory for Cryobiology, Belgrade, Serbia

⁷University of Belgrade, University Children’s Hospital, Faculty of Medicine, Department of Hematology – Oncology, Belgrade, Serbia

Abstract

Sunjog K., S. Ćirković, B. Vuković-Gačić¹, M. Guć-Šćekić, M. Mišković, D. Vujić and D. Škorić (2019): Comet assay and cytogenetic findings in differential diagnosis of fanconi anemia.- Genetika, Vol 51, No.3, 1113-1126.

Fanconi anemia (FA) is a complex genetic disease with a variety of congenital and hematological symptoms, including the predisposition for cancer development. The main hallmark of FA cells, an increased chromosomal fragility, in the presence of the DNA-interstrand cross-linking chemicals, mitomycin C or diepoxybutane (DEB), makes the diagnosis of FA much easier. Cytogenetic method can detect the FA patients with highly elevated chromosomal breakage, but also some of the patients with borderline sensitivity to DEB no matter if they have FA or not. These particular circumstances lead us to introduce comet assay along with cytogenetic analysis, in order to determine DNA lesions and chromosomal fragility in untreated and DEB-treated lymphocytes of full blood from seven patients with clinical features of FA. Highly elevated DEB induced chromosomal sensitivity confirmed the diagnosis in five patients (FA group: 0.48-4.47 breaks/cell vs control group: 0.00-0.08 breaks/cell). Borderline DEB sensitivity (FA* group: 0.15-0.44 breaks/cell) was found in the remaining two patients. Results of the comet assay showed higher baseline and DEB-induced DNA damage values (Olive tail moment and tail intensity) in all five FA and one FA* patient, when compared to the control group. These findings could provide a new model of FA screening test algorithm, including comet assay as additional and very useful accurate tool, beside the DEB test, in differential diagnosis of FA.

Keywords: Fanconi anemia, genetic disease, comet assay, chromosomal fragility, diepoxybutane

Corresponding author: Karolina Sunjog, Center for Genotoxicology and Ecogenotoxicology,Chair of Microbiology Faculty of Biology University of Belgrade,11000 Belgrade,Serbia,Mob: +381 63 87 23 804,E-mail: sunjogkarolina@yahoo.com,E-mail: sunjogkarolina@imsi.rs

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Published in „GENETIKA“ Vol. 51, No.3(2019), pp. 1127-1138

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-ZemunSerbia

UDC 575.

https://doi.org/10.2298/GENSR1903127S

Original scientific paper

Spatial genetic analysis of roe deer from the Northern Serbian Province

of Vojvodina

Milomir STEFANOVIĆ^1*, Nikoleta KARAISKOU², Nevena VELIČKOVIĆ¹,

Milutin KOVAČEVIĆ³, Zoran RISTIĆ³, Mihajla DJAN¹

¹University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Novi Sad, Serbia

²Department of Genetics, Developmental and Molecular Biology, School of Biology, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece

³University of Novi Sad, Faculty of Sciences, Department of Geography, Tourism and Hotel Management, Novi Sad, Serbia

Abstract

Stefanović M., N. Karaiskou, N. Veličković, M. Kovačević, Z. Ristić, M. Djan (2019): Spatial genetic analysis of roe deer from the northern Serbian province of Vojvodina.- Genetika, Vol 51, No.3, 1127-1138.

Ecological and evolutionary processes responsible for genetic variation are affected by geographical distribution of samples, therefore combined analysis of genetic and spatial information can help to a better understanding of these processes. Mitochondrial DNA control region was sequenced in 103 individuals from 11 different localities crossing Northern Serbian Province of Vojvodina. Sixteen haplotypes were detected, with high haplotype diversity values (0.898 ± 0.012), that were clustered in two spatially and genetically differentiated groups named: East and West. The haplotype diversity was higher in East group, while nucleotide diversity and average number of nucleotide differences were higher in West group. Conservation plans and management strategies should always take into consideration genetic data that helps to understand spatial genetic patterns and/or effect of landscape features in dispersal between animal populations.

Keywords: Capreolus capreolus, genetic clustering, mtDNA, Serbia

Corresponding author: Milomir Stefanović, Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovića 2, 21000 Novi Sad, Serbia, e-mail: milomir.stefanovic@dbe.uns.ac.rs

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1139-1149

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.

https://doi.org/10.2298/GENSR1903139C

Original scientific paper

ASSOCIATION OF RHEUMATOID ARTHRITIS ACCORDING TO THE DEGREE

OF GENETIC HOMOZYGOSITY AND GENDER: PILOT STUDY

Suzana CVJETICANIN¹, Milan TERZIC², Dejan NIKOLIC^2,3*

¹Institute for Human Genetics, Faculty of Medicine, University of Belgrade, Belgrade, Serbia

²Faculty of Medicine, University of Belgrade, Belgrade, Serbia

³Department of Physical Medicine and Rehabilitation, University Children’s Hospital, Belgrade, Serbia

Abstract

Cvjeticanin S., M.Terzic, D. Nikolic (2019): Association of rheumatoid arthritis according to the degree of genetic homozygosity and gender: pilot study.- Genetika, Vol 51, No.3, 1139-1149.

Rheumatoid arthritis (RA) is a chronic synovial inflammatory autoimmune disease with multifactorial origin. With epigenetic and genetic mechanisms playing a role in the development of RA, the aim of our study was to evaluate the anthropogenetic variability in tested individuals that were diagnosed with rheumatoid arthritis, and the possible influence of gender in expression of illness. 100 patients with rheumatoid arthritis (RA) and 100 healthy control individuals were evaluated. For the estimation of the degree of recessive homozygosity, the homozygously recessive characteristics (HRC) test was performed testing 20 HRCs. There was a significant difference in the individual variations of 20 HRCs between the individuals of the control group and patients with RA (SX²=135.191; p<0.001). The mean values of the tested HRCs significantly differed between individuals of the control group and RA group (MV±SD_{Control group}-5.97±2.02, MV±SD_{RA group}-7.34±2.00, p=<0.001). There was a decrease in variability in the RA group versus the control group (V_{RA group}=27.19%; V_{Control group}=33.79%).There was significant difference in the frequencies of HRCs between those with and without RA in males (p<0.023) and in females (p<0.001). Our findings pointed to the higher degree of recessive homozygosity along with decreased variability in RA patients compared to a healthy control group. Therefore, it may be assumed that different genes in different proportions have certain influence in the processes responsible for RA susceptibility and its different degrees of expression.

Key words: gender, genetic homozygosity, rheumatoid arthritis, variability

Corresponding author: Dejan Nikolic, Department of Physical Medicine and Rehabilitation, University Children’s Hospital, Belgrade, Serbia, Tirsova 10, Belgrade, Serbia,  email: denikol27@gmail.com

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Published in „GENETIKA“ Vol. 51, No. 3(2019), pp. 1151-1164

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

 UDC 575.630

                              https://doi.org/10.2298/GENSR1903151M

Original scientific paper

STABILITY ANALYSIS OF TUBER YIELD USING UNBALANCED DATA FROM POTATO VARIETY TRIALS

Zlatko MIJIĆ¹, Vinko.KOZUMPLIK², Hrvoje ŠARČEVIĆ², Vladimir MEGLIĆ³, Ivan VARNICA⁴ and Tihomir ČUPIĆ⁵

¹ State Inspektorate, Sector of inspection in Agriculture, Beli Manastir, Croatia

² University of  Zagreb, Facultyof Agriculture, Department of Plant Breeding, Genetics and Biometrics, Zagreb, Croatia

³ Agricultural Institute of Slovenia, Crop Science Department, Ljubljana, Slovenia

⁴ Croatian Agency for Agriculture and Food, Centar for Seed and Seedling, Osijek, Croatia

⁵ Agricultural Institute Osijek, Department for Breeding and Genetic of Forage Crop, Osijek, Croatia

Abstract

Mijić Z., V. Kozumplik, H. Šarčević, V. Meglić, I. Varnica, T. Čupić (2019): Stability analysis of tuber yield using unbalanced data from potato variety trials.- Genetika, Vol 51, No.3, 1151-1164.

Potatoes are grown in a wide region and in different environments unlike other crops, so it is known as one of the most spread world crops. In many agriculture areas around the world new genotypes are included in many multi-locational experiments at a large number of locations and years to determine the level and the stability of yields. However, there are a lot of problems with this traditional approach mostly due to the large number of genotypes that require consistent checking of every genotype at numerous locations and years. Multi-locational experiments require a lot of time and financial resources, and the possibility of error is greatly increased. In the present study we used the REML/BLUP method to predict tuber yield in an unbalanced set consisting of 54 potato genotypes investigated over twelve years at three locations. The data were subjected to stability analysis using the AMMI model based on estimated values. The analysis of variance showed that the greatest effect can be attributed to the environment (E), then to their interaction (G×E) and least to the genotype. The first two multiplicative interaction components explained 85.4% of the interaction sum of squares. AMMI analysis enabled the identification of stable and productive genotypes as well as genotypes adapted to specific environments and clearly separated the three locations as mega-environments. The genotype selection index (GSI), due to its nature of combining the assessments of stability measure and yield rank, provided more useful information for selection and recommendation. The results of this study indicate the superiority of the prediction model in comparison with the traditional multi-location experiment methods allowing the creation of recommended list of potato varieties based on the analysis of unbalanced data sets.

Keywords: Potato, yield, unbalance data set, AMMI, Genotype Selection Index

Corresponding author: Tihomir Čupić, Agricultural Institute Osijek, Department for Breeding and Genetic of Forage Crop, Južno predgrađe 17, Osijek, Croatia; Phone : ++ 385 31 515 543; E-mail: Tihomir.Cupic@poljinos.hr

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Published in „GENETIKA“ Vol. 51 No. 3(2019), pp. 1165-1174

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.633.11

https://doi.org/10.2298/GENSR1903165M

Orginal scientific  paper

VARIATION IN NITROGEN USE EFFICIENCY OF WINTER WHEAT

Milan MIROSAVLJEVIĆ^1*, Vladimir AĆIN¹, Vladimir SABADOŠ², Danijela DOROTIĆ ²

¹Institute of Field and Vegetable Crops, Novi Sad, Serbia

²Agricultural Extension Service, Sombor, Serbia

Abstract

Mirosavljević M., V. Aćin, V. Sabadoš, D. Dorotić (2019): Variation in nitrogen use efficiency of winter wheat.- Genetika, Vol 51, No.3, 1165-1174.

Optimization of nitrogen fertilization to specific cultivar requirements is a major objective for improvement of trade-offs between grain yield, environmental sustainability and maximum profitable production. The aim of this study was to assess the effects of nitrogen fertilization on grain yield, and nitrogen use efficiency of modern wheat cultivars in different growing seasons. The trials with eight winter wheat cultivars and seven top-dressing nitrogen treatments were carried out in three successive growing seasons under rain-fed conditions of the southern Pannonian plain. The results from our study showed a significant variation in grain yield and nitrogen use efficiency among winter wheat cultivars when grown under different environmental and soil nitrogen level conditions. On average, grain yield ranged from 4961 to 6375 kg ha^-1 among winter wheat cultivars. Increase of N soil level resulted in significant grain yield increase and nitrogen use efficiency decrease compared to the control. In 2015/16, 2016/17, and 2017/18 growing seasons grain yield of winter wheat cultivars reached plateau at 156, 175, and 128 nitrogen soil level, respectively. Significant influence of cultivar by nitrogen fertilization interaction indicated that it is necessary to adjust nitrogen fertilization to each cultivar. Moreover, notable weather variability between different growing seasons is a major limiting factor for optimal nitrogen fertilizer application in winter wheat production under conditions of the Pannonian plain.

Keywords: cultivar, grain yield, growing season, NUE, Triticum aestivum L.

Corresponding author: Milan Mirosavljević, Institute of Field and Vegetable Crops, Maksima Gorkog 30, 21000 Novi Sad; Phone: 0214898220; Fax: 021 4898222; E-mail: milan.mirosavljevic@nsseme.com

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Published in „GENETIKA“ Vol. 51, No.3(2019), pp. 1175-1184

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.630

https://doi.org/10.2298/GENSR1903175N

Orginal scientific  paper

VARIABILITY OF NEEDLE MORPHO-ANATOMY OF NATURAL Pinus heldreichii POPULATIONS FROM SCARDO-PINDIC MOUNTAINS

Biljana NIKOLIĆ^1*, Zorica MITIĆ², Srdjan BOJOVIĆ³, Vlado MATEVSKI⁴,  Zoran KRIVOŠEJ⁵,

Petar D. MARIN⁶

^1*Institute of Forestry, Belgrade, Serbia

²University of Niš, Faculty of Sciences and Mathematics, Department of Biology and Ecology, Niš, Serbia,

³University of Belgrade, Institute for Biological Research “Siniša Stanković” Belgrade, Serbia

⁴University “Ss. Kiril and Metodij”and Macedonian Academy of Sciences and Arts, Institute of Biology, Faculty of Natural Sciences and Mathematics, Skopje, Republic of North Macedonia.

⁵University of Priština, Department of Biology, Faculty of Natural Sciences, Kosovska Mitrovica, Serbia.

⁶University of Belgrade, Faculty of Biology, Institute of Botany and Botanical Garden “Jevremovac“, Belgrade, Serbia

Abstract

Nikolić B., Z. Mitić, S. Bojović, V. Matevski,  Z. Krivošej  and P. D. Marin (2019): Variability of needle morpho-anatomy of natural Pinus heldreichii populations from Scardo-Pindic mountains.- Genetika, Vol 51, No.3, 1175-1184.

Eight morpho-anatomical properties of two-year-old needles of Pinus heldreichii (Bosnian pine) from the Scardo-Pindic mountain massif in Serbia (Kosovo, Mt. Ošljak) and North Macedonia (Mt. Galičica) were investigated. All measured characteristics, except for needle length, were inspected on mid-needle cross-section. Cross-sections were obtained with razorblade, while measurements were performed with Leica-Gallen III light microscope. The mean values of the analyzed characters were as follows: 5.91 cm (needle length), 1.35 mm (needle width), 0.85 mm (needle thickness), 25.05 μm (cuticle + epidermis thickness), 69.90 μm (height of hypodermal cells), 21.76 μm (resin duct diameter), 3.4 (number of hypodermis layers), and 3.6 (number of resin ducts). P. heldreichii needles also had 2-5 hypodermis layers and 0-12 resin ducts. The highest variation was in the number of resin ducts (CV=27%). In comparison with previously investigated needles from the Dinaric mountains, P. heldreichii needles from the Scardo-Pindic massif were shorter and had a thicker layer of hypodermis. PCA and CA visualize partial segregation of P. heldreichii populations between the two mountain massifs.

Keywords: Bosnian pine, needle morphology, needle anatomy, PCA, Cluster Analysis

Corresponding author: Biljana Nikolić, ^*Institute of Forestry, Kneza Višeslava 3, 11000 Belgrade, Serbia, Corresponding author, E-mail: smikitis@gmail.com

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Published in „GENETIKA“Vol. 51, No.3(2019), pp. 1185-1196

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

UDC 575.630

https://doi.org/10.2298/GENSR1903185G

Orginal scientific  paper

VARIATION OF SPIKE INDEX OF CEREAL AND INDEX OF PEA PODS

IN MONOCROPS AND INTERCROPS SYSTEM OF CULTIVATION

Milosav GRČAK¹, Dragan GRČAK², Radivoje JEVTIĆ², Mirjana LALOŠEVIĆ²,

Danijela KONDIĆ³, Jelica ŽIVIĆ⁴, Desimir KNEŽEVIĆ¹

¹University of Priština, Faculty of Agriculture, Kosovska Mitrovica – Lešak, Lešak, Serbia

²Institute of Field and Vegetable Crops, Novi Sad, Serbia

³University of Banja Luka, Faculty of Agriculture Banjaluka, Banjaluka, Republika Srpska,

Bosnia & Herzegovina

⁴College of Agriculture and Food Technology, Prokuplje, Serbia

Abstract

Grčak M., D. Grčak, R. Jevtić, M. Lalošević, D. Kondić, J. Živić, D. Knežević (2019): Variation of spike index of cereal and index of pea pods in monocrops and intercrops system of cultivation.- Genetika, Vol 51, No.3, 1185-1196.

Parameters of plant productive organs (spike, pod etc.) are influenced by genotype of plant species and scientific technology farming measures as well environmental conditions. The aim of this study was to establish variability of spike harvest index for wheat, triticale, rye, oat and pea pods index, influenced by monocrops and intercrops system of cultivation. Four cereal species: wheat, triticale, rye and oat and one legume (pea) were included in investigation that carried out on field experimental conditions during one vegetation season. Each species were sown in monocrops and in intercrops wheat + pea, triticale + pea, rye + pea and oat + pea. Harvest index of spike variate between 65.2% (triticale intercrops) and 86.5% (oat solo). The values of spike weight and seed weight was different among the cereal species and higher in intercrops than in monocrop system of cultivation. However, only for rye in mixture with pea, harvest index (78.8%) was higher than in monocrops of rye (77.0%). Harvest index of pod variate in ratio 50.3% (pea solo) and 69.5% (in intercrops pea + rye). The values of pod weight and seed weight pod^-1 was different and higher in intercrops than in monocrops system of cultivation. Harvest pod index of peas was significantly higher in mixture with each small grains species genotypes than in pea’s monocrops. On the base of result we can conclude that intercropping cereals + peas, have positive effect on spike and pod characteristics which values were higher in intercrops than in monocrops.

Keywords: cereals, harvest index, intercropping, pea, pod, spike

Corresponding author: Desimir Knežević,University of Priština, Faculty of Agriculture, Kosovska Mitrovica – Lešak, Kopaonička bb, 38219 Lešak, Kosovo and Metohia, Serbia, deskoa@ptt.rs

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Published in „GENETIKA“Vol. 51, No.3(2019), pp. 1197-1225

© 2019 Serbian Genetics Society
S. Bajića 1, 11185 Belgrade-Zemun

Serbia

                                                                                              UDC 575.

https://doi.org/10.2298/GENSR1903197S

Orginal scientific  paper

FANCA AND CONTRIBUTION OF STUDIES FROM ASIAN POPULATIONS TO THE UNDERSTANDING OF FANCA MEDIATED FANCONI ANEMIA

Muhammad SHAHID and Sabika FIRASAT^*

Department of Animal Sciences, Faculty of Biological Sciences, Quaid-i-Azam University Islamabad, 45320 Pakistan.

Abstract

Shahid M. and S. Firasat (2019): Fanca and contribution of studies from Asian populations to the understanding of Fanca mediated Fanconi anemia.- Genetika, Vol 51, No.3, 1197-1225.

Fanconi anemia (FA) is a recessive disorder known to cause hematological and several congenital deformities in affected individuals worldwide. Out of 22 known FA causative genes, mutations in Fanconi Anemia Complementation Group A (FANCA) accounts for 60%-70% of FA cases. FANCA is a multi-functional protein essential for genome integrity. Although many physiological roles of FANCA have been delineated but exact etiopathomechanism of FANCA in FA phenotype is yet to be elucidated. FANCA is a hypermutable and highly polymorphic gene therefore identification and interpretation of mutations implicated in recessively inherited FA can fill gaps in existing knowledge of molecular mechanisms. This review is divided into two sections. The first section described known functions of FANCA important for genomic integrity perpetuation. The second part summarized all mutations of FANCA gene reported in FA patients from Asian populations on the basis of literature published till March 2019. It provides an overview of strategies used for these mutations identification, mutation hot spots for specific Asian populations and necessitates the need of extensive global as well as regional molecular genetics research efforts mainly in less explored inbred Asian countries to formulate diagnostic and targeted therapeutic measures.

Keywords: Asian population, Fanconi anemia, FANCA, mutations

Corresponding author: Sabika Firasat, Department of Animal Sciences, Faculty of Biological Sciences, Quaid-i-Azam University Islamabad, 45320 Pakistan, Email: sabika.firasat@qau.edu.pk

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