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Background: Periodontitis is a complex disease, and bacterial factors play a crucial role in its initiation. The contributions of genetic and epigenetic factors to the pathogenesis of periodontal disease are increasingly recognized. Single-nucleotide polymorphisms (SNPs) in various molecules, including cytokines, are of particular interest due to their established involvement in numerous diseases. This study investigates the influence of SNPs in the IL-10 gene at positions −592 (rs1800872) C>A and −1082 (rs1800896) T>C (also referred to as 1082A>G) on the severity of periodontitis in a cohort of Bulgarian patients. Methods: In the recent study, both clinical and paraclinical methodologies were employed to comprehensively assess the periodontal status of the participants. The genotypic characterization of IL-10 polymorphisms was performed by PCR RFLP analysis. Statistical analyses, including principal component analysis (PCA), were executed utilizing IBM SPSS Statistics Version 21. Results: We have established a statistically significant association between the presence of at least one A-allele in the patients' genotype and the incidence of severe periodontitis (p = 0.047). Conclusions: IL-10 single-nucleotide polymorphisms (SNPs) could be effectively considered as biomarkers for the severity of periodontitis. [ABSTRACT FROM AUTHOR]

Eggplant, or brinjal, is one of the most consumed and important tropical solanaceous vegetable crops grown worldwide. Little leaf is a disease associated with the presence of phytoplasmas especially widespread in brinjal in India. To clarify the epidemiology of this disease, a verification of its transmission through seeds to seedlings and their progeny derived from symptomatic mother plants was performed. Brinjal seeds field-collected in the Dharwad district of Karnataka State, India, were sowed in a greenhouse under insect-proof conditions. DNA was extracted from seedlings and their progeny and from symptomatic plant samples collected in the field. The first- and second-generation seedlings obtained *under these conditions were tested at various time points after germination by amplification of the 16S rRNA gene of phytoplasmas. The amplicons obtained were subjected to restriction fragment length polymorphism (RFLP) analysis and sequencing for the identification of detected phytoplasmas. Ribosomal groups 16SrI, 16SrII, 16SrIII, 16SrV, 16SrVI, and 16SrXII were identified. Moreover, a number of fruits produced from the first-generation seedlings showed precocious seed germination, and the young seedlings resulted as phytoplasma-positive. The seed transmission of phytoplasmas in eggplants for two subsequent generations highlights the risk of additional sources of infection of the disease represented by asymptomatic and infected seedlings in the presence of insect vectors. The seed transmission could explain the continuous presence of epidemic outbreaks of phytoplasmas in brinjal cultivations in several cultivation areas. [ABSTRACT FROM AUTHOR]

Eggplant, or brinjal, is one of the most consumed and important tropical solanaceous vegetable crops grown worldwide. Little leaf is a disease associated with the presence of phytoplasmas especially widespread in brinjal in India. To clarify the epidemiology of this disease, a verification of its transmission through seeds to seedlings and their progeny derived from symptomatic mother plants was performed. Brinjal seeds field-collected in the Dharwad district of Karnataka State, India, were sowed in a greenhouse under insect-proof conditions. DNA was extracted from seedlings and their progeny and from symptomatic plant samples collected in the field. The first- and second-generation seedlings obtained *under these conditions were tested at various time points after germination by amplification of the 16S rRNA gene of phytoplasmas. The amplicons obtained were subjected to restriction fragment length polymorphism (RFLP) analysis and sequencing for the identification of detected phytoplasmas. Ribosomal groups 16SrI, 16SrII, 16SrIII, 16SrV, 16SrVI, and 16SrXII were identified. Moreover, a number of fruits produced from the first-generation seedlings showed precocious seed germination, and the young seedlings resulted as phytoplasma-positive. The seed transmission of phytoplasmas in eggplants for two subsequent generations highlights the risk of additional sources of infection of the disease represented by asymptomatic and infected seedlings in the presence of insect vectors. The seed transmission could explain the continuous presence of epidemic outbreaks of phytoplasmas in brinjal cultivations in several cultivation areas.

Samples of various host plants from different locations in Serbia showing foliar symptoms resembling those caused by the cucumber mosaic virus (CMV) were collected. Samples were considered CMV-infected after detection by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Fifteen CMV isolates were selected for developing a reverse transcription-polymerase chain reaction (RT-PCR) restriction fragment length polymorphism (RFLP) protocol and a more detailed molecular characterization of all five genes. Based on in silico RFLP, the following restriction enzymes were selected: HindIII, SacII (1a gene), MluI (2a gene), StuI, SalI (2b gene), BaeI (movement protein (MP) gene of CMV), SfcI, and HaeIII (capsid protein (CP) gene), which were capable of distinguishing between subgroups of CMV based on the obtained characteristic restriction patterns. According to in situ RFLP, the predominant haplotype IA; IA, IA; IA, IA was determined in 14, while the distinct haplotype II; II, II; II, II was found in only one isolate. Sequence analyses of two selected Serbian CMV isolates with different restriction patterns and haplotype profiles confirmed the RFLP results, showing that isolates 674-11 and 137-13 belong to subgroups IA and II, respectively. Different restriction patterns after digestion of all five CMV genomic regions proved to be a simple way to investigate the natural population of CMV. This study provides insight into the genetic structure of the CMV population in Serbia. [ABSTRACT FROM AUTHOR]

Cowpea plants exhibiting virescence, phyllody and little leaf symptoms were observed in the fields of Zhanjiang City, Guangdong province of China in October 2020. Total DNA was extracted from symptomatic cowpea leaves. Using phytoplasma universal primer pairs (R16mF2/R16mR1, P1/P7) and a secA primer pair (secAfor1/secArev3), expected PCR fragments of approximately 1400, 1800 and 800 bp were amplified from all symptomatic cowpea leaves. MUSCLE analysis revealed that the representative 16S rRNA gene sequence of cowpea phyllody phytoplasma (CPP) shared 98.67% nucleotide identity with the reference strain WBDL of 'Ca. Phytoplasma aurantifolia' (GenBank accession number U15442); and 100% nucleotide identity with 16SrII-V phytoplasmas associated with 'Praxelis clematidea' phyllody (GenBank accession number KY568717); 'Desmodium triflorum' little leaf (GenBank accession number MT452308); and 'Ixeris chinensis', 'Emilia sonchifolia', and 'Desmodium ovalifolium' witches' broom (GenBank accession number MT416114, MT420682 and MK956144, respectively). Phylogenetic analyses based on the 16S rRNA and secA gene sequences revealed that the CPP strain clustered with the 16SrII-V subgroup and 16SrII group, respectively. Virtual restriction fragment length polymorphism analysis of the 16S rRNA sequence revealed that CPP was associated with the 16SrII-V subgroup. To our knowledge, this is the first report of the16SrII-V subgroup of phytoplasmas associated with cowpea phyllody disease. [ABSTRACT FROM AUTHOR]

A survey assessed cucumber phyllody disease incidence in Chikkaballapura, Bengaluru Rural, Bengaluru Urban, and Bagalkote districts of Karnataka, India. The average disease incidence ranged from 8 to 18 %. A total of 9 samples showing phyllody and little leaf symptoms were collected and association of phytoplasma was confirmed through 16S rRNA gene amplification, using universal primer pair P1/P7, and followed by nested PCR using primer R16F2n/R16R2. Further, characterization through multilocus sequence analysis targeting secY and rpl22 genes confirmed all nine strains as part of the 16SrI group ('Candidatus Phytoplasma asteris'). Additionally, four weed species (Lecusa aspera, Parthenium hysterophorus, Acaranthus sp., and Amaranthus viridis) found in cucumber fields tested positive for phytoplasma (16SrI) disease. In-silico RFLP analysis of the amplified F2n/R2 region of the 16S rRNA gene indicated that two phytoplasma strains (CuPP1 and CuPP3) from Chikkaballapura belonged to subgroup X (16SrI-X), one phytoplasma strain (CuPP2) belonged to subgroup B (16SrI–B). While the remaining strains from Bangalore Urban, Bengaluru Rural, and Bagalkote districts belonged to subgroup B (16SrI–B). A loop-mediated isothermal amplification (LAMP) detection protocol was also developed for 'Ca P. asteris', targeting the phytoplasma 16S ribosomal DNA. Comparison of assay sensitives demonstrated that LAMP technique is highly efficient and could detect the presence of phytoplasma up to 50 fg of template DNA. LAMP-based detection offers superior ease of use, cost-effectiveness, and rapid results. To the best of our knowledge, this is the first report of phytoplasma causing phyllody disease in cucumber crop in India. • Infected cucumber plants showed little leaf, phyllody, leaf proliferation, and shortened internodes. • ' Ca. Phytoplasma asteris' strain (16SrI) detected in cucumber phyllody symptoms via multi-locus gene sequence analysis (MLST). • A Loop‐mediated isothermal amplification (LAMP) assay was developed for detecting phytoplasma. • The present study reveals genetic diversity and impact of phyllody phytoplasma on cucumbers. • This is the first report of ' Ca. Phytoplasma asteris' (16SrI) causing cucumber phyllody in India. [ABSTRACT FROM AUTHOR]

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Abstract Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3βgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3β genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp-3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3βgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3β genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.

The review of literature presents analysis of the current state of molecular biological methods for research of Mycobacterium leprae. The characteristics and feasibility of application of the methods in use for diagnostics and epidemiological monitoring of the leprosy pathogen are discussed, as well as identification of possible genetic determinants of antibiotic resistance. The present knowledge is provided of genetic characteristics of mycobacteria of the above type, which enable to lay the foundations for development of methods for leprosy diagnostics, as well as also for their significant improvement.

The phenotypic and genotypic features of soybean nodule bacteria with different growth rates common in soils of Ukraine have been studied. The obtained results suggest high heterogeneity of soybean microsymbionts. It has been established that all studied strains are able to form active symbiotic relationships with soybean. When interacting with Chinese cowpea, intensive-growing strains form an ineffective symbiosis with a low nitrogen fixation level (phenotype Nod+Fix-), while slow-growing strains form complete nitrogen fixing systems (phenotype Nod+Fix+). Upon cultivation in the mannitol yeast medium, intensive-growing strain B. japonicum KC19 produces a significant amount of auxins and cytokinins, while the slow-growing strain B. japonicum KC23 is significantly superior by the amount of gibberellins. According to the organization of the genome defined in REP-PCR, studied strains of soybean rhizobia belong to two genetic groups, which corresponds to the division of strains into groups by the growth rate (slow- and intensive-growing). Based on the RFLP analysis of rpoB gene, using restriction enzymes MspI, HaeIII and NdeII, intensive-growing strains were combined into one rpoB type, while slow-growing strains belong to 2-3 rpoB types. [ABSTRACT FROM AUTHOR]

Objective. To study phenotypic and genotypic features of nodule bacteria - microsymbionts of leguminous plants of the Glycine, Vigna and Lupinus genera. Methods. Serological affinity of 12 rhizobia strains was determined in agglutination reactions. Host specificity of strains was studied in vegetation experiments. RFLP analysis of rpoB gene was performed using MspI, HaeIII and NdeII restrictases. Sequencing of the 16S-23S rDNA intergenic spacer of rhizobia was performed on ABI 3130 Genetic Analyzer. Results. It was established that studied rhizobia strains differ significantly in serological properties and belong to 7 serogroups: KB11, M8, 1967, 46, B1, B2 and 367a. Microsymbionts of soybean, cowpea and mung bean form the group of cross inoculation, however, are unable to infect lupine. Alternatively, lupine rhizobia B. lupini 367a and Bradyrhizobium sp. 4, enter into symbiotic interactions with lupine, however, they do not nodulate soybean. B. japonicum 631 strain is capable of symbiosis with leguminous plants of different tribes: Phaseoleae and Lupineae. Based on RFLP analysis of rpoB gene, rhizobia were grouped into four clusters with following microsymbionts: I - cowpea, II - mung bean and soybean, III - slow-growing soybean nodule bacteria, IV - lupine rhizobia and intensive-growing soybean rhizobia. 16S-23S rDNA sequencing confirmed pertinence of soybean rhizobia to the B. japonicum species (USDA 4, USDA 6, USDA 123 genetic groups). This species also included mung bean microsymbionts (USDA 4 group). Bradyrhizobium sp. 11 isolate was obtained for the first time from cowpea nodules, and it was identified as a new B. diazoefficiens species (USDA 110 group). ITS regions of B. lupini 367 and Bradyrhizobium sp. 4 rhizobia were found to be identical. B. japonicum 631 strain has a 100% similarity with B. lupini 367a strain according to 16S-23S rDNA, and with high degree of probability can be included to B. lupini species. Conclusions. Serological heterogeneity of rhizobia from root nodules of plants of the Glycine, Vigna and Lupinus genera has been demonstrated. It was established that microsymbionts of soybean, cowpea and mung bean belong to one group of cross inoculation. 16S-23S rDNA sequencing allowed to classify the studied strains as different genetic groups and identify them as B. japonicum, B. diazoefficiens and B. lupini. The serological grouping of nodule bacteria was found to coincide with the genetic (rpoB gene and ITS region) grouping, and their host specificity was related to species affiliation. [ABSTRACT FROM AUTHOR]

Spoilage caused by yeasts is a constant, widespread problem in the beverage industry that can result in major economic losses. Fruit juices provide an environment that allows the proliferation of yeast. Some factories in South Africa are not equipped with laboratory facilities to identify spoilage yeasts and outsourcing becomes a prolonged process which obstructs corrective action planning. This study aimed to establish yeast diversity and apply a rapid method for preliminary identification of spoilage yeasts associated with a small-scale fruit juice bottling factory. Yeast population in the factory was determined by isolation from the production environment, process equipment and spoiled products. PCR-RFLP analysis targeting the 5.8S-ITS region and D1/D2 sequencing was used for identification. A total of 207 yeasts belonging to 10 different genera (Candida, Lodderomyces, Wickerhamomyces, Yarrowia, Zygosaccharomyces, Zygoascus, Cryptococcus, Filobasidium, Rhodotorula/Cystobasidium and Trichosporon) were isolated and identified from the production environment and processing equipment. Candida intermedia, C. parapsilosis and Lodderomyces elongisporus were widely distributed in the factory. Zygosaccharomyces bailii, Z. bisporus, Zygoascus hellenicus and Saccharomyces cerevisiae were isolated from the spoiled products. The data provided a yeast control panel that was used successfully to identify unknown yeasts in spoiled products from this factory using polymerase chain reaction-restriction length polymorphism (PCR-RFLP) comparative analysis. [ABSTRACT FROM AUTHOR]

The present study was investigating the clinical pictures, prevalence, as well as the ecological conditions associated with Pseudomonas anguilliseptica outbreaks in four cultured seabream, Sparus aurata farms at different localities in Egypt during winter of 2016. The phenotypic and genotypic patterns of Pseudomonas isolates were investigated. The existence of intraspecific heterogeneity among different isolates was analyzed using Restriction Fragment Length Polymorphism (RFLP) technique. Attempts on disease control using antibiogram or dietary supplement were also considered. To achieve these goals, various commercial antibiotic discs were analyzed against Ps. anguilliseptica isolates using the disc diffusion method. Additionally, the impact of one-month dietary incorporation with 3% garlic extract or 0.5% potassium diformate on S. aurata viability and response for prolonged bathing treatment with florfenicol was evaluated following challenge with the virulent strain of Ps. anguilliseptica . Most of the naturally infected fish displayed spiral-swimming behavior with no obvious external lesions. The prevalence of infections in the four investigated farms (F1, F2, F3, and F4) were 44.9, 69.04, 67.72, and 83.4%, respectively. Water analysis revealed a significant variation in total hardness, pH, dissolved oxygen (D.O), ammonia and salinity among different localities. All isolates were rather uniform in most of the biochemical characteristics and were identical on the basis of RFLP analysis. The analyses of PAF-PAR gene pointed out specific amplification bands of 439 bp length. The antibiogram revealed a potential activity of florfenicol, ciprofloxacin, nitrofurantoin, and oxytetracycline against all isolates. Experimentally challenged fish fed on garlic extract or potassium diformate presented lower mortality and better therapeutic response to florfenicol than those fed on a normal basal diet. In conclusion, Ps. anguilliseptica is a prevalent pathogen among cultured seabream where dietary inclusion of 3% garlic extract or 0.5% potassium diformate seemed to improve seabream health status and subsequently, increase the efficacy of the treatment with the selective antibiotic. [ABSTRACT FROM AUTHOR]

Aim of study: This study aims to provide basic information about physiological characteristics of isolates of Lactarius deliciosus (L.) Gray, Russula sanguinaria (Schumach.) Rauschert, Suillus collinitus (Fr) Kuntze, Suillus granulatus (L.) Rousell, Tricholoma batchii Gulden and Tricholoma imbricatum (Fr.) Kumm.Area of study: The isolates are obtained from Pinus heldreichii H. Christ forest in the south-eastern part of Montenegro.Material and methods: The isolates were molecularly characterised by internal transcribed spacer (ITS) sequencing and restriction fragment length polymorphism (RFLP) analysis. The effects of different temperatures (20, 22, 25°C), pHs (4, 4.5, 5.2, 5.8, 6.5, 7.5), and carbon (glucose, sucrose, dextrin, arabinose, xylose and starch) and nitrogen (NH4+, NO3- and protein) sources on their growth were examined under laboratory conditions.Main results: The studied factors established significant differences in the development of isolates. Isolates of R. sanguinaria, L. deliciosus and both Suillus, were characterised by faster growth at 22°C, while Tricholoma isolates grew faster at 25°C. S. granulatus, S. collinitus and T. imbticatum isolates grew well at lower pH values (4 - 5.2), while L. deliciosus, R. sanguinaria and T. bachii exhibited faster growth at pHs between 5.8 and 6.5. The examined isolates were able to utilize various carbohydrates as carbon sources. The biggest mycelial growth was characterised for sucrose, then glucose, dextrin, arabinose, starch and xylose. They grew on all examined nitrogen sources, while the biggest mycelia growth was achieved on ammonium, followed by nitrate and protein. Those characteristics varied amongst the species.Research highlights: Information about physiological characteristics of Tricholoma, Lactarius, Russula, as well as Suillus, are sparse. Hence, the data obtained in this study could contribute to the understanding of their function in ecosystems.

Phytoplasmas are cell wall-less microorganisms associated with plant diseases worldwide. Many important food, vegetable and fruits crops as well as ornamental plants can be severely affected by these pathogens, with significant economic impacts. Phytoplasma diseases of ornamentals have been described worldwide in a wide range of plant genera, and 11 different 16Sr groups have been identified. In Italy, many ornamental plant species belonging to several botanical families have been found to be infected by phytoplasmas, classified into the ribosomal groups 16SrI, 16SrII, 16SrV and 16SrXII. During a survey carried out in commercial gardens in Rome, some marguerite daisy (Argyranthemum frutescens) plants showing symptoms of phytoplasma-like disease, were collected and submitted to molecular analyses. Cloning and sequencing of the portion of the 16S rRNA gene followed by BLAST analysis, real and virtual restriction fragment length polymorphism anlaysis with AluI and RsaI, allowed assignment of the detected phytoplasma to the 16SrIX-C group (Picris echioides yellows, PEY).

Phytoplasmas are cell wall-less microorganisms associated with plant diseases worldwide. Many important food, vegetable and fruits crops as well as ornamental plants can be severely affected by these pathogens, with significant economic impacts. Phytoplasma diseases of ornamentals have been described worldwide in a wide range of plant genera, and 11 different 16Sr groups have been identified. In Italy, many ornamental plant species belonging to several botanical families have been found to be infected by phytoplasmas, classified into the ribosomal groups 16SrI, 16SrII, 16SrV and 16SrXII. During a survey carried out in commercial gardens in Rome, some marguerite daisy (Argyranthemum frutescens) plants showing symptoms of phytoplasma-like disease, were collected and submitted to molecular analyses. Cloning and sequencing of the portion of the 16S rRNA gene followed by BLAST analysis, real and virtual restriction fragment length polymorphism anlaysis with AluI and RsaI, allowed assignment of the detected phytoplasma to the 16SrIX-C group (Picris echioides yellows, PEY). [ABSTRACT FROM AUTHOR]

Abstract: Gaucher disease is an autosomal recessive lysosomal storage disorder resulting from deficient glucocerebrosidase activity. There have been nearly 300 mutations described to date. Novel mutations can potentially provide insight into the biochemical basis of the disease. Two novel mutations are described in two Type 1 Gaucher patients with N370S compound heterozygosity; a point mutation that causes an amino acid substitution at cysteine residue 23 for tryptophan, and a second point mutation within the splicing element at the 3′ end of intron 7. Both mutations were identified by PCR amplification and sequence analysis of patient glucocerebrosidase genomic DNA. Restriction fragment length polymorphism analysis was established for both novel mutations for efficient identification in future patients. Past literature suggests that mutations affecting cysteine residues involved in disulfide bridges, as well as mutations affecting splicing patterns of the glucocerebrosidase transcript, are detrimental to enzyme activity. However, compound heterozygosity with N370S, a mild mutation, will lead to a mild phenotype. The cases reported here support these past findings. [Copyright &y& Elsevier]

Nucleotide sequences in internal transcribed spacer (ITS)-1 region derived from dried nori products produced in Japan, China, and the Republic of Korea were compared. Thalli contained in the Japanese products were genetically homogenous, and their nucleotide sequences in ITS-1 were identical to those of the reference strains of Pyropia yezoensis f . narawaensis. In Chinese products, the thalli were related to P. yezoensis strain Minomiasakusa. In contrast, the thalli in the Korean products were genetically heterogeneous, and several different P. yezoensis strains and other Pyropia spp. were used for dried nori products. In some thalli produced in both China and Korea, the DNA sequences of the ITS-1 region were identical with that of Japan, suggesting that the cultivar strains might have been transplanted from Japan to China in recent years. The 432-bp-long nucleotide sequences in the ITS-1 region of thalli derived from Japanese origin were cleaved to two restriction fragments at 154 and 278 bp by cleavage of PCR-amplified products using MspI. Conversely, almost all of the corresponding sequences derived from China and Korea were lacking MspI or other restriction patterns, except for nori products from some areas that cultivate a closely related strain to the Japanese cultivar. [ABSTRACT FROM AUTHOR]

Restriction fragment length polymorphism tools is an R application which supports a complete workflow of polymerase chain reaction-restriction fragment length polymorphism ( PCR- RFLP), dealing with the problems which accompany analysis when PCR- RFLP is used in diversity studies. Large numbers of different RFLP samples obtained from multiple electrophoresis runs might lead to limitations or misidentifications due to the need for band matching in most existing software applications. Due to the common problem of variation in the density of bands (i.e. distances between bands or visual intensity) in the electropherograms, it is desirable to have options for handling samples with uncertain or faint bands. As a further step in the workflow, scientists often use DNA sequencing to identify individual genotypes, so that the use of specific software to combine these tasks might be helpful. With this background, we here present an application that supports a complete workflow, starting with the analysis of single species samples by PCR- RFLP, to PCR- RFLP genotype identification based on a reference data set and DNA sequencing followed by similarity analysis. RFLPtools is a freely available, platform-independent application which provides analysis functions for DNA fragment molecular weights (e.g. by RFLP analysis), including similarity calculations without the need for band matching. As it is written for the statistical software R, other statistical analyses might also be easily applied. [ABSTRACT FROM AUTHOR]