DNA Fingerprinting Studies Research Articles

Molecular and Morphological Profiling of Rice Cultivars Using Hypervariable Microsatellite Markers and DUS Descriptors

Identification and protection of varieties is one of the most important tasks in the plant breeding programme. As the number of new varieties increasing year by year, there is a need to protect and maintain the purity of the varieties. DNA fingerprinting studies allows us to identify the plant varieties with high precision as it is not influenced by environment and other factors. In this context, the present study has been taken up for morphological and molecular profiling of 20 rice genotypes using 39 DUS descriptors and SSR markers. The field experiment was conducted during the year Kharif 2021 and Kharif 2022 at Agricultural Research Station, Bapatla and molecular profiling studies were conducted at RARS, Lam, Guntur. Significant variations were observed for 16 morphological traits at different growth stages, which are relatively more informative in the identification and characterization of rice genotypes. A total of 45 polymorphic SSR markers were used to screen 20 rice genotypes, which produced 336 alleles. The number of alleles produced by these markers ranged from 3 (RM1048) to 15 (RM124) with an average number of alleles of 7.46 per marker. The PIC values of these markers ranged from 0.54 (RM1048) to 0.92 (RM124) with an average PIC value of 0.75. SSR based molecular profiles were developed using 12 highly polymorphic SSR markers viz., RM495, RM6933, RM489, RM6006, RM3351, RM510, RM418, RM3215, RM105, RM6364, RM144 and RM2972 for discrimination of each genotype as well as easy identification.

Evaluation of DNA extraction methods for molecular traceability in cold pressed, solvent extracted and refined groundnut oils

Groundnut oil (GNO)/ peanut oil is one of the agro-food products with great economic value and hence an attractive target for adulteration and mislabeling. Simple Sequence Repeats (SSR) are markers of choice for DNA fingerprinting studies as they exhibit high polymorphism due to variable number of repeats. Hence, this study was designed to evaluate and optimized a method for DNA isolation from groundnut oil and study the possibility of using the isolated DNA for molecular traceability using SSR markers. Four methods to isolate DNA from groundnut oil were evaluated. All the four methods were modified CTAB protocols, but differed in procedures forextraction, buffer compositions, amount of oil used and DNA carriers. For molecular traceability of oils, extraction and recovery of DNA from edible oil is a key step, especially in refined oils. A method that employed DNA enrichment prior to extraction with CTAB buffer yielded amplifiable DNA from cold pressed GNO, crude hexaneextracted GNO and refined GNO. The optimized method for isolation of DNA from groundnut oil is simple, efficient, less costly and reproducible when compared to chromatography and spectroscopy based techniques.

Molecular Analysis of Genetic Variability and Relationship among Pearl Millet (Pennisetum glaucum) Genotypes by Using SSR Markers

Pearl millet has a place with sort Pennisetum, and family Poaceae. Pearl millet is the most broadly developed grain crop in Asia and Africa representing close to half of the worldwide millet creation. In India, pearl millet is the fourth most broadly developed food crop after rice, wheat, and maize. We used molecular techniques to investigate the genetic diversity and relatedness of six genotypes viz:ICMV155, Dhanshakti, PC-612, Sampada, HHB-67, and HHB-197.the six genotypes was collected from National agriculture research project Aurangabad. The present study was conducted at the Department of Plant Biotechnology at K. K. Wagh College of Agricultural Biotechnology, Nashik.
 DNA was isolated by fixing a sample in alcohol without using liquid nitrogen; six genotypes were analyzed through SSR primers to determine the extent of molecular characterization. PCR amplification using 10 SSR primers generated a total of 111 no. of bands were scored corresponding an average of 11 bands per primer with 77 bands showing polymorphism (67%) and 34 bands showing monomorphism (30%). The PIC value ranged from 0.35 to 0.68 with an average of 0.4. Jaccard’s coefficient based on SSR analysis 0.38 to 0.90. The dendrogram wasconstructed using the UPGMA method. It has two main clusters Cluster-1 consisting of C1- HHB 67, V6-ICMV-155, C2- HHB 197 and C4- Sampada. Cluster-2 comprised C3-Dhanshakti and V5-PC 612 as an out group. C2- HHB 197 and C4- Sampada genotypes have the highest similarity coefficient 0.76. Among all the genotypes, C1- HHB 67 and V5- PC 612 was found most diverse as it separated from all other genotypes at a very low similarity coefficient of 0.6. The identified markers can prove useful for identification of diverse germplasm and future DNA fingerprinting studies.

Molecular profiling of therapeutically important Alpinia and Kaempferia species using RAPD, ISSR, and SSR markers

Alpinia and Kaempferia species require precise identification and diversification to improve the accuracy in drug discovery and industrial demands. In this current study, four Alpinia species (Alpinia malaccensis, A. nigra, A. galanga and A. calcarata) and three Kaempferia species (Kaempferia galanga, K. rotunda and K. parishii) were selected for molecular profiling analysis. Three types of molecular markers (PCR based) such as RAPD, ISSR, and SSR were used to increase the accuracy of analysis. About 91 polymorphic loci were amplified in these 7 species using 9 RAPD primers out of 30. The resolving power (RP) was varied from 4.56 to 10.06 and the polymorphic information content (PIC) ranged between 0.8 and 0.93. Out of 40 ISSR primer 13 primers showed total 118 polymorphic loci. The maximum RP and PIC were 9.36 and 0.97 respectively. In case of SSR analysis, only 11 SSRs out of 35 showed 201 polymorphic loci. The RP were found between 3.12 and 17.86. The PIC in SSR primers were in between 0.78 and 0.97. This study indicated that unique banding pattern was observed in specific primers of each marker. D20, D18, and D8 primers were found as the best RAPD markers. (GA)8T, (GA)9T, and (GTGC)4 were the best ISSR markers which can produce distinct banding pattern. SSR2 and SSR14 have also successfully amplified in Alpinia and Kaempferia species with distinct banding pattern. Although SSR marker is more advance than other two but multiple markers based molecular profiling would have enough significance. This present study has enormous importance towards DNA fingerprinting study of different species of Alpinia and Kaempferia. The unique banding pattern in specific primer could be efficiently useful for species identification and future improvement of the taxa.

Effective bioremediation of soil from the Burgan oil field (Kuwait) using compost: A comprehensive hydrocarbon and DNA fingerprinting study

An innovative combination of metagenomic profiling of microbial communities and GC-MS & Pyrolysis-GC-MS fingerprinting methods were used to assess the biodegradation of contaminated soil from the Burgan oil field in Kuwait. The soil was treated with (sludge) compost in microcosms to evaluate the feasibility of this material for bioremediation purposes. The most favourable trial showed a > 80% decrease in TPH, thereby indicating strong potential for full-scale application using a cost-effective technology and thus in line with the principles of the circular economy. The microbial study showed that compost addition enhanced the organic matter and nutrient content of the soil. However, the microorganisms in the compost did not seem to play a relevant role in bioremediation, meaning that compost amendments serve as a biostimulation rather than a bioaugmentation approach. The chemical study of the distinct oil fractions revealed rapidly biodegraded compounds (alkanes, alkyl-aromatics, etc.) and others that were much more refractory (hopanes, benzohopanes, etc.). Of note, although heavy fractions are usually considered recalcitrant to biodegradation, we observed incipient degradation of the asphaltene fraction by means of double-shot thermodesorption and pyrolysis. Finally, chemical fingerprinting also revealed that the treated soil contained some of the compounds found in the compost, such as coprostanol, cholesterol, and plant sterols. This observation would support the use of these compounds as proxies to monitor the effects of compost and to adjust dosages in real-scale bioremediation treatments.

Phenotypical and Genotypical Comparison of Clostridium difficile Isolated from Clinical Samples: Homebrew DNA Fingerprinting versus Antibiotic Susceptibility Testing (AST) and Clostridial Toxin Genes.

Background Clostridium (Clostridioides) difficile is recognized as the major cause of healthcare antibiotic-associated diarrhea. We surveyed a molecular epidemiological correlation between the clinical isolates from two general hospitals in Iran through clustering toxigenic types and antibiotic susceptibility testing (AST) accuracy. Methods Study population included 460 diarrhoeic specimens from inpatients with a history of antibiotic therapy. All samples underwent enriched anaerobic culture, confirmed by detection of gluD gene with PCR. Toxin status and AST were assessed by the disk diffusion method (DDM) and minimal inhibitory concentrations (MICs) of metronidazole, vancomycin, and rifampin. C. difficile outbreak was analyzed through conventional PCR by tracing toxin genes and Homebrew pulsed-field gel electrophoresis (PFGE) for characterizing isolates within our healthcare systems. Results A total of 29 C. difficile strains were isolated by enriched anaerobic culture from the clinical samples. Among them, 22 (4.8%) toxigenic profiles yielded toxins A and B (tcdA, tcdB) and binary toxins (cdtA, cdtB). The minimum inhibitory concentration (MIC) was 18.1% and 9% for vancomycin and metronidazole, and all isolates were susceptible to rifampicin and its minimum inhibitory concentration was at <0.003 μg/mL. The most dominant toxigenic and antibiotic-resistant “pulsotype F” was detected through PFGE combined with multiple Clostridial toxigenic pattern and AST. Conclusions DNA fingerprinting studies represent a powerful tool in surveying hypervirulent C. difficile strains in clinical settings. Resistance to vancomycin and metronidazole, as first-line antibiotics, necessitate accomplishment of proper control strategies and also prescription of tigecycline as a more appropriate option.

Identification of unique alleles and assessment of genetic diversity of soybean genotypes using SSR markers and seed traits

Thirty polymorphic SSR markers and seven seed traits were used to determine the genetic diversity among twenty five soybean genotypes that differ with respect to seed coat color. A total of 133 alleles were detected and the number of alleles for each SSR locus varied from two to seven with an average of 4.0. Polymorphic information content varied from 0.81 to 0.07. Jaccard’s similarity coefficient grouped the genotypes into two major clusters. To test the goodness of fit of clusters to SSR markers, cophenetic correlation was estimated. Cophenetic value of 0.98 indicated a very good fit. A combination of SSR and seed traits data grouped the genotypes into two major clusters with grouping of all the brown seeded genotypes into one subcluster. A total of 21 unique alleles were identified in twenty five soybean genotypes which is a valuable resource for DNA fingerprinting studies. A set of nine SSR markers (Satt 600, Satt 463, Satt 371, Satt 193, Satt 538, Satt 126, Satt 286, Satt 281 and Satt 656) could differentiate all the soybean genotypes. Key WordsGenetic diversity, SSR markers, Soybean, unique alleles, seed traits

English

DNA fingerprinting is rapid, easy, and efficient method for discrimination, identification and characterization of various genotypes for protection of plant breeder’s rights (PBRs). Present study was designed for DNA fingerprinting and genetic diversity assessment of 25 GM cotton genotypes (possessing Cry1Ac gene) using 297 SSR markers through conventional PCR and Polyacrylamide gel electrophoresis. Out of 297 SSR markers, 25 markers were not amplified, 28 were monomorphic and 244 were polymorphic. A total of 1537 alleles were amplified among which 1294 (84.18%) were polymorphic. PIC value in our study ranged from 0.08 to 0.93 with an average of 0.73. Unique allelic pattern was observed for nineteen genotypes whereas six genotypes were identified using two-step identification methods. The UPGMA dendrogram divided the genotypes into two distinct clusters. Cluster I was comprised of 20 genotypes whereas cluster II was comprised of four genotypes. MNH-1020 did not obey any clustering and remained separated. The results of the structure analysis were complementary to cluster analysis and the population was divided into two subgroups. Our results evidenced narrow genetic base of the cotton genotypes cultivated in Punjab Pakistan due to use of common parents in the pedigree/parentage. Further, we proposed a core set of markers for future DNA fingerprinting and genetic diversity studies. The information generated in this study will be helpful in variety registration and subsequent protection under PBRs. Further our findings will be useful in selection of SSR markers for future studies which are focused on DNA fingerprinting and genetic diversity assessment. © 2021 Friends Science Publishers

Evaluation of DNA extraction methods for molecular traceability in cold pressed, solvent extracted and refined groundnut oils.

Groundnut oil (GNO)/peanut oil is one of the agro-food products with great economic value and hence an attractive target for adulteration and mislabeling. Simple Sequence Repeats (SSR) are markers of choice for DNA fingerprinting studies as they exhibit high polymorphism due to variable number of repeats. Hence, this study was designed to evaluate and optimize a method for DNA isolation from groundnut oil and study the possibility of using the isolated DNA for molecular traceability using SSR markers. Four methods to isolate DNA from groundnut oil were evaluated. All the four methods were modified CTAB protocols, but differed in procedures for extraction, buffer compositions, amount of oil used and DNA carriers. For molecular traceability of oils, extraction and recovery of DNA from edible oil is a key step, especially in refined oils. A method that employed DNA enrichment prior to extraction with CTAB buffer yielded amplifiable DNA from cold pressed GNO, crude hexane extracted GNO and refined GNO. The optimized method for isolation of DNA from groundnut oil is simple, efficient, less costly and reproducible when compared to chromatography and spectroscopy based techniques.

Absence of strict monogamy in the Eurasian jackdaw, Coloeus monedula

Abstract The Eurasian Jackdaw is thought to be archetypically monogamous, but recent tagging research uncovered extra-pair copulations in the species. Here we examined extra-pair paternity (genetic monogamy) in Eurasian jackdaws breeding in the Judean Hills, Israel, at the global edge of the species range, using a set of highly polymorphic molecular microsatellites. We found roughly a sixth of nests sampled showed deviations from monogamy, more than previously found in DNA fingerprinting studies of jackdaws, suggesting a mixed mating strategy in this population. These findings support the trend of extra-pair paternity in avian species, even when social monogamy remains the rule, and highlight the importance of continued study of species throughout their geographical range.

Ethiopia’s transforming wheat landscape: tracking variety use through DNA fingerprinting

Ethiopia is the largest wheat producer in sub-Saharan Africa yet remains a net importer. Increasing domestic wheat production is a national priority. Improved varieties provide an important pathway to enhancing productivity and stability of production. Reliably tracking varietal use and dynamics is a challenge, and the value of conventional recall surveys is increasingly questioned. We report the first nationally representative, large-scale wheat DNA fingerprinting study undertaken in Ethiopia. Plot level comparison of DNA fingerprinting with farmer recall from nearly 4000 plots in the 2016/17 season indicates that only 28% of farmers correctly named wheat varieties grown. The DNA study reveals that new, rust resistant bread wheat varieties are now widely adopted. Germplasm originating from CGIAR centres has made a significant contribution. Corresponding productivity gains and economic benefits have been substantial, indicating high returns to investments in wheat improvement. The study provides an accurate assessment of wheat varietal status and sets a benchmark for national policy-makers and donors. In recent decades, the Ethiopian wheat landscape has transformed from local tetraploid varieties to widespread adoption of high yielding, rust resistant bread wheat. We demonstrate that DNA fingerprinting can be applied at scale and is likely to transform future crop varietal adoption studies.

Development of a core SNP panel for cacao (Theobroma cacao L.) identity analysis.

Single nucleotide polymorphisms (SNPs) are preferred markers for DNA fingerprinting and diversity studies in cacao (Theobroma cacao L.). Yet, a consensus SNP panel with a minimum number of SNPs for optimal identity analysis is unavailable for cacao. An initial set of 146 SNP panels of varying sizes were assembled based on heterozygosity, linkage disequilibrium (LD), linkage group (LG) distribution, major allele frequency, minor allele frequency (MiAF), polymorphism information content (PIC), and random distribution. These panels were assessed to determine their ability to distinguish among a training set of 155 accessions. The panels with the best separation ability were supplemented with additional SNPs to create 16 designer panels, which separated all 155 accessions. The 16 designer SNP panels were then assessed on a dataset of 1220 accessions coming from 10 ancestral groups. Increasing the number of SNPs generally yielded improved resolution of genetic identities with concomitant reduction of synonymous groups. The number and choice of SNPs were critical factors with LD, MiAF, and PIC being important selection attributes but an even LG distribution was unnecessary. A robust set of 96 SNPs is recommended as a minimal core SNP panel for cacao DNA fingerprinting to the international cacao community.

Elucidate Genetic Diversity and Population Structure of Bread Wheat (Triticum Aestivum L.) Cultivars Using IRAP and REMAP Markers

Analysis of genetic diversity and population structure in bread wheat is an essential step in their conservation, utilization, and breeding. Retrotransposons are ubiquitous and abundant a throughout the plant genomes, therefore extensively used as ideal molecular markers for genetic variability, DNA fingerprinting and genetic mapping studies in plant species. In the current research, we used two retrotransposon-based marker systems, inter-retrotransposon amplified polymorphisms (IRAPs), and the retrotransposon-microsatellite amplified polymorphisms (REMAPs) markers to evaluate the genetic diversity and survey activity of long terminal repeat retrotransposon (LTR-retrotransposon) elements in a collection of 49 bread wheat (Triticum aestivum L.) cultivars that mainly bred in Iran. In general, 90 and 126 loci were amplified using 9 IRAP and 20 REMAP primers, respectively. Both techniques produced a satisfactory number of bands for cultivar analysis; however, the technique IRAP, particularly single primer Nikita generated a large number of bands, indicating the wide activity of Nikita family under various environmental conditions of bread wheat. The percentage of polymorphic loci (PPL) in the studied collection for IRAP and REMAP markers was 81.78 and 86.40%, respectively. A model-based Bayesian method, Principal coordinate analysis (PCoA) and cluster analysis using Minimum Evolution (ME) algorithm hinted of the existence of two groups. This grouping was in agreement with the growing season and conformed by the high within-group bootstrap value. These results demonstrated that these markers developed using transpositionally active retrotransposons (RTNs) are efficient and reliable markers in determining level of genetic diversity and population structure in bread wheat in breeding programs.

Comparative DNA Fingerprinting and Botanical Study of Certain Haworthia and Gasteria Species Growing in Egypt

Introduction: Haworthia Duval and Gasteria Duval are succulent plants, native to South Africa. They have been used worldwide in folk medicine. They are members of the Alooideae subfamily, which has been always viewed as a taxonomically confusing sub-family. The present study aims to differentiate between Haworthia limifolia Marloth, Gasteria carinata (Mill.) Duval and Gasteria minima Poelln. growing in Egypt. Methods: A comparative botanical and genetic investigation is presented. Macromorphological and micromorphological botanical features of the leaves and stems of the species under investigation using the entire specimens, transverse sections and isolated elements. Moreover, their DNA was extracted from the leaf samples and RAPD-PCR analysis was made using 10 primers. Results: The comparative botanical criteria of the examined leaves and stems were identified. Furthermore, the total number of amplified products produced by the ten primers was 98 fragments. Primers OPB-07, OPB-01 and OPB-02, recording high percentage of polymorphism (78.6, 70.6 and 57.1%, respectively), can be used to differentiate between the examined species. While primers OPB-01, OPB-04, OPB-07 and OPB-08, generating fragments with wide molecular size (17, 14, 14 and 11fragments, respectively), can be used for the identification of Haworthia limifolia, Gasteria carinata and Gasteria minima. Conclusion: The macromorphological, micromorphological and genetic identifying parameters presented in the current study revealed good crieteria to authenticate and differentiate between Haworthia limifolia Marloth, Gasteria carinata (Mill.) Duval and Gasteria minima Poelln.

SSR marker based DNA fingerprinting and diversity studies in mustard (Brassica juncea)

SSR marker based DNA fingerprinting and diversity studies in mustard <i>(Brassica juncea)</i>

Development of a set of SNP markers for population genetics studies of Ipe (Handroanthus sp.), a valuable tree genus from Latin America

A combination of restriction associated DNA sequencing (RADSeq) and low coverage MiSeq genome sequencing was used for the development of single nucleotide polymorphisms (SNP) and INDEL (insertion/deletions) genetic markers for Ipe (Handroanthus sp.). Of the 402 putative loci identified, 389 SNPs and INDELs (315 nuclear SPNs, six chloroplast INDELs, 15 chloroplast SNPs, 12 mitochondrial INDELs and 41 mitochondrial SNPs) were successfully genotyped at 93 individuals from Brazil, Bolivia and French Guiana using a MassARRAY® iPLEX™ platform. This set of markers will be invaluable for population genetics, phylogeography and DNA fingerprinting studies

Identification of common, unique and polymorphic microsatellites among 73 cyanobacterial genomes.

Microsatellites also known as Simple Sequence Repeats are short tandem repeats of 1-6 nucleotides. These repeats are found in coding as well as non-coding regions of both prokaryotic and eukaryotic genomes and play a significant role in the study of gene regulation, genetic mapping, DNA fingerprinting and evolutionary studies. The availability of 73 complete genome sequences of cyanobacteria enabled us to mine and statistically analyze microsatellites in these genomes. The cyanobacterial microsatellites identified through bioinformatics analysis were stored in a user-friendly database named CyanoSat, which is an efficient data representation and query system designed using ASP.net. The information in CyanoSat comprises of perfect, imperfect and compound microsatellites found in coding, non-coding and coding-non-coding regions. Moreover, it contains PCR primers with 200 nucleotides long flanking region. The mined cyanobacterial microsatellites can be freely accessed at www.compubio.in/CyanoSat/home.aspx. In addition to this 82 polymorphic, 13,866 unique and 2390 common microsatellites were also detected. These microsatellites will be useful in strain identification and genetic diversity studies of cyanobacteria.

Investigation of the Organic Solvent Resistance Mechanisms in Rhodococcus and Lysinibacillus Using Several Molecular Forensic Tools

Rhodococcus erythropolis IBBPo5 (KM405337) and Lysinibacillus xylanilyticus IBBPo7 (KM405338) showed good tolerance to 5% (v/v) organic solvents (alkanes, aromatics). However, 5% (v/v) alkanes (i.e., cyclohexane, n-hexane, n-decane) were less toxic for R. erythropolis IBBPo5 and L. xylanilyticus IBBPo7 cells, compared with 5% (v/v) aromatics (i.e., toluene, o-xylene, ethylbenzene). The high organic solvent tolerance of these two Gram-positive bacteria could be due to the presence in their genome of some catabolic (alkB1, todM, xylM) and trehalose-6-phosphate synthase (otsA1) genes. R. erythropolis IBBPo5 possesses three catabolic genes (i.e., alkB1, todM, xylM), while L. xylanilyticus IBBPo7 possesses only one catabolic gene (i.e., alkB1). Numerous adaptations involved in organic solvents tolerance were depicted in R. erythropolis IBBPo5 and L. xylanilyticus IBBPo7 cells exposed to 5% (v/v) alkanes and aromatics (i.e., changes in the cell growth rate, membrane permeability, cell morphology, biosurfactant emulsification index, carotenoids profile, DNA fingerprinting). Therefore, microbiological, biochemical, and DNA fingerprinting studies of the bacteria isolated from polluted environments could provide valuable information that may complement or supplement other forensic investigations.

Genetic diversity and DNA fingerprinting in jute (Corchorus spp.) based on SSR markers

Genetic diversity analysis and DNA finger printing are very useful in breeding programs, seed conservation and management. Jute (Corchorus spp.) is the second most important natural fiber crop after cotton. DNA fingerprinting studies in jute using SSR markers are limited. In this study, 58 jute accessions, including two control varieties (Huangma 179 and Kuanyechangguo) from the official variety registry in China were evaluated with 28 pairs of SSR primers. A total of 184 polymorphic loci were identified. Each primer detected 3 to 15 polymorphic loci, with an average of 6.6. The 58 jute accessions were DNA-fingerprinted with 67 SSR markers from the 28 primer pairs. These markers differentiated the 58 jute accessions from one another, with CoSSR305-120 and CoSSR174-195 differentiating Huangma 179 and Kuanyechangguo, respectively. NTSYS-pc2.10 software was used to analyze the genetic diversity in the 58 jute accessions. Their genetic similarity coefficients ranged from 0.520 to 0.910 with an average of 0.749, indicating relatively great genetic diversity among them. The 58 jute accessions were divided into four groups with the coefficient 0.710 used as a value for classification, consistent with their species and pedigrees. All these results may be useful both for protection of intellectual property rights of jute accessions and for jute improvement.

Precision assessment of some supervised and unsupervised algorithms for genotype discrimination in the genus Pisum using SSR molecular data.

For the first time, prediction accuracies of some supervised and unsupervised algorithms were evaluated in an SSR-based DNA fingerprinting study of a pea collection containing 20 cultivars and 57 wild samples. In general, according to the 10 attribute weighting models, the SSR alleles of PEAPHTAP-2 and PSBLOX13.2-1 were the two most important attributes to generate discrimination among eight different species and subspecies of genus Pisum. In addition, K-Medoids unsupervised clustering run on Chi squared dataset exhibited the best prediction accuracy (83.12%), while the lowest accuracy (25.97%) gained as K-Means model ran on FCdb database. Irrespective of some fluctuations, the overall accuracies of tree induction models were significantly high for many algorithms, and the attributes PSBLOX13.2-3 and PEAPHTAP could successfully detach Pisum fulvum accessions and cultivars from the others when two selected decision trees were taken into account. Meanwhile, the other used supervised algorithms exhibited overall reliable accuracies, even though in some rare cases, they gave us low amounts of accuracies. Our results, altogether, demonstrate promising applications of both supervised and unsupervised algorithms to provide suitable data mining tools regarding accurate fingerprinting of different species and subspecies of genus Pisum, as a fundamental priority task in breeding programs of the crop.