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Item EFFICIENCY OF MICROSATELLITE MARKERS IN GENOTYPING OF OROBANCHE CUMANA POPULATIONS(Iasi University of Life Sciences, 2021-10-21) Mutu, Ana; Clapco, Steliana; Duca, MariaMicrosatellites or simple sequence repeats (SSRs) markers are widely dispersed across the genome, have a codominant and multiallelic nature, high variability and informativeness, and a specific chromosomal location. For this reason, they are playing a significant role in different fundamental and applicable fields, including genetic studies at the individual, population, species levels, and molecular breeding programs. In this study, the utility of 15 microsatellite markers in discrimination of 33 Orobanche cumana (broomrape) populations from different geographical locations was assessed: Republic of Moldova (11 populations), Romania (1), Bulgaria (4), Serbia (7), Turkey (7) and China (3). The PCR amplification products obtained showed differences by number and size depending on the markers used (3 and 16 alleles). A total of 110 alleles with fragment sizes ranged from 76-343 bp were determined. The evaluation of the genetic polymorphism of SSR markers was performed by the effective number of alleles per locus (Ne) with an average of 5.31, Polymorphic Information Content index (PIC) – 0.75, Nei's genetic diversity index (H) – 0.78 and, Resolving Power index (Rp) – 5.79. High values of all indices revealed the significant informative capacity and effectiveness of markers studied. At the same time, most markers showed PIC values higher than 0.5, indicating a high polymorphism in O. cumana populations. Seven microsatellite markers (Ocum-052, Ocum-059, Ocum-074, Ocum-081, Ocum-087, Ocum-196, Ocum-197) were selected based on the analyzed statistical parameters, suggesting that these markers can efficiently measure genetic diversity in broomrape. Based on the Rp index and PIC values of these 15 SSR markers, genetic diversity of Turkish (Rp: 4.774; PIC: 0,722) and Moldavian (Rp: 4.394; PIC: 0,716) Orobanche cumana populations was higher than in other populations. However, the studied microsatellite (SSR) markers system characterized very well the genetic structure of all O. cumana populations included in this study. Selected markers could eventually be useful for breeders and sunflower seed producers to improve their control strategies for this parasitic plant.Item DISCRIMINATING ABILITY OF ISSR MARKERS IN THE REVEALING OF THE GENETIC POLYMORPHISM IN TURKISH BROOMRAPE POPULATIONS(Trakya University Publisher, 2021-08-29) Duca, Maria; Bivol, InaIn the present study, the analysis of the informativity level and discriminatory capacity for 14 ISSR-markers to identify polymorphism of seven Orobanche cumana populations from different geographical regions of Turkey was performed. ISSR markers utility was assessed by calculating of different parameters such as the polymorphism information content (PIC), observed and effective number of alleles (Na and Ne), Nei's and Shannon diversity indexes (H and I), number and percentage of polymorphic loci, resolving power (Rp), Simpson's coefficient (Hj). Out of 207 amplicons generated by microsatellites primers, 195 were found polymorphic (94.29%) and 12 were monomorphic with size range from 341 to 5353 bp. The average number of amplified bands was 14.79 fragments per primer. The PIC values for 14 primers ranged from 0.18 ((GATA)4) to 0.41 ((AG)8YA) with the mean value 0.34 Eight primers ((AG)8YA, (CT)8TC, BC807, BC841, (CA)6AC, (CTC)4RC, (CAG)5, BC835) were considered the most effective. The mean I value was 0.52 and ranged from 0.32 ((GATA)4) to 0.6 ((AG)8YA). The (AG)8YA, (CT)8TC, BC807, BC841, (CTC)4RC, (CA)6AC and (CAG)5 primers had the highest I values. The Rp ranged from 5.9 for primer (CA)6RG to 22.59 for primer (AG)8YA. Nine of the ISSR primers ((AG)8YA, (CTC)4RC, BC810, BC807, BC841, BC835, (CAG)5, BC857 and (CT)8TC) possessed high Rp values (22.59, 19.12, 16.83, 16.54, 16.49, 14.59, 13.32, 12.88 and 11.37, respectively) and were able to distinguish all 41genotypes. The higher discrimination potential calculating by the Simpson’s coefficient had primer BC810 (0.33) and the lower (CA)6RG (0.01), the mean value for all primers was 0.18. According to ISSR data analysis, the high proportion of the genetic variability (94.29%) was due to interpopulation variation and 45.82% of the diversity by cause of intrapopulation variations. In conclusion, in this comparative study of the discrimination capacity of 14 ISSR markers using to broomrape population genetic analysis, the tri- ((CTC)4RC, (CAG)5) and dinucleotide ((AG)8YA, BC807, BC841, (CT)8TC) primers proved more informative than tetranucleotide repeats. Moreover, these primers revealed the highest values for all analyzed polymorphism information indexes of genetic diversity (number of amplicons per primer from 15 to 28, level of polymorphism more than 88.89%, PIC≥0.36, I≥0.53, Hj≥0.11 and Rp≥11.37) for Turkish broomrape populations. The analysis of ISSR-markers showed that this type of marker is applicable for the assessment of intra- and interpopulation genetic polymorphism for species O. cumana.Item EFICIENȚA UNOR MARCHERI MOLECULARI ÎN DISCRIMINAREA POPULAȚIILOR DE LUPOAIE ORIGINARE DIN CHINA(Tipografia „Print Caro”, 2021) Duca, Maria; Mutu, Ana; Bivol, Ina; Clapco, StelianaIn this study, the effectiveness of different types of molecular markers in assessing genetic diversity of populations of O. cumana from China was determined. ISSR and SSR markers detected different levels of genetic variability among and within broomrape populations. SSR markers analysis showed high level of genetic variation within the populations as revealed by high average values of Nei's gene diversity (H=0,75) and Shannon's information index (I=1,44), while genotyping with ISSR markers showed greater ability to discriminate genotypes according to Resolving power (Rp=7,24). Thus, the combined use of ISSR and SSR markers allowed the detection of higher polymorphism than either set of marker alone.