LINE1 Research Articles

Inhibition of angiotensin II type 1 receptor by candesartan reduces tumor growth and ameliorates fibrosis in colorectal cancer.

Colorectal cancer (CRC) is an important cause of cancer-related mortality. Aberrant activation of the renin-angiotensin system (RAS) is reported to be associated with poor clinical outcomes in patients with CRC. This study was designed to explore the anti-tumor effects of the angiotensin receptor blocker Candesartan either alone or in combination with 5-FU in in vitro and in vivo models of CRC. The cytotoxic effects of Candesartan were assessed using the MTT assay in two colorectal cancer cell lines (CT-26 and SW-480). To investigate the potential regulatory role of Candesartan on tumor growth, apoptosis, and migration, the expression levels of Cyclin D1, Survivin, MMP3, MMP9, and E-cadherin mRNAs were evaluated. The oxidant/antioxidant balance was also examined by determining the levels of MDA, thiols, SOD, and CAT. We used a xenograft model of colon cancer to investigate the effects of Candesartan alone, or in combination with 5-FU, on tumor growth following histological staining (Hematoxylin & Eosin and Masson trichrome staining) and biochemical studies as well as gene expression analyses by RT-PCR and western blotting. Candesartan suppressed tumor cell proliferation and migration by modulating Cyclin D1, MMP3/9, and E-cadherin. Treatment with Candesartan either alone, or in combination with 5-FU decreased tumor size in the mouse model, and also increased the level of oxidative markers MDA and reduced CAT, SOD, and thiols. Histological evaluation showed that Candesartan increased tumor necrosis, reduced tumor density and attenuated collagen deposition reducing tumor fibrosis in tumor xenograft. Candesartan, an inhibitor of the RAS, when used in combination with 5-FU reduced tumor growth by inhibiting fibrosis and inducing ROS production, supporting further clinical studies on this therapeutic approach for treatment of CRC.

Application of inter-SINE amplified polymorphism (ISAP) markers for genotyping of Cucumis melo accessions and its transferability in Coleus spp

Abstract. Sormin SYM, Purwantoro A, Setiawan AB, Teo CH. 2021. Application of inter-SINE amplified polymorphism (ISAP) markers for genotyping of Cucumis melo accessions and its transferability in Coleus spp. Biodiversitas 22: 2918-2929. An unambiguous characterization of melon (Cucumis melo L.) accessions based on their morphological traits is often laborious and affected by environment when compared with molecular marker genotyping. Short interspersed nuclear elements (SINEs) are highly abundant non-autonomous and non-coding retrotransposons that are widely scattered over all chromosomes of eukaryotes. They can serve as a good molecular marker for routine genotyping in plant breeding and marker-assisted selection. This study aimed to apply inter-SINE amplified polymorphism (ISAP) markers for genotyping of Cucumis melo accessions and its transferability in Coleus spp. Twenty-one C. melo accessions, one C. metuliferus E. Mey. ex Naudin, and 15 accessions of Coleus spp. were used for ISAP marker development. A copy of cucumber-specific long interspersed nuclear element (LINE) and multiple copies of melon-specific SINE were identified and isolated. ISAP primers were designed from the highly conserved region of the SINEs and LINE. The melon and cucumber-specific ISAP markers showed a higher degree of polymorphism (87.5%-100%) than potato ISAP markers (60%-100%) in all the tested melon accessions. The unweighted pair group method with arithmetic average (UPGMA) dendrogram generated from polymorphic ISAP bands clearly distinguish the Cucumis melo accessions from their distantly related wild species C. metuliferus. The transferable nature of Cucumis ISAP marker system was demonstrated in Coleus species, where the marker differentiates the tested accessions.

A Birst of LINE‐1 and IAP Retrotransposon Expression in the Mouse Brain Following Acute Behavioral Stress

Retrotransposons belong to mobile genetic elements that account for up to ~40% of the human genome and are propagated by being transcribed into RNA, then reverse-transcribed into DNA and inserted into a new site in the genome. Physiological regulation and possible effects of such retrotranspositions in cells of an adult organism are largely unknown. In the mouse genome LINE-1 (L1) and intracisternal A particle (IAP) are among functional and the most transpositionally active retrotransposons, capable of self-copying and insertion into DNA. Recent findings indicate that L1 is active in neuronal precursor cells during differentiation, and its de novo insertions can change expression of neuron-specific genes and cause somatic mosaicism in the adult brain. Furthermore, clinical studies have identified increased L1 expression and copy numbers in the brain of patients with psychiatric disorders. However, there has been limited understanding whether such retrotranspositions happen in the adult brain, when and how they occur, and what is their role in the CNS function. In our experiments we test the hypothesis that retrotranspositions can be activated in the brain by behavioral and particularly cognitive stimuli and that this process can influence adaptive neural plasticity. In previous experiments we showed an increase in L1 DNA copy number in hippocampus, frontal cortex and cerebellum of the mouse brain, but not in the muscular and testicular tissues after a prolonged stress. In this study we tested whether an acute stress can lead to L1 and IAP mRNA expression changes in the mouse brain. We used male C57Bl/6 mice at the age of 5 months. To assess the effect of acute stress, the animals were subjected to immobilization for 30 minutes or 2 hours. The frontal cortex and hippocampus were taken for qPCR studies. Compared to naive home cage controls, mice that underwent 30 min immobilization had significantly increased mRNA levels of L1 and IAP in frontal cortex and L1 in hippocampus. After 2 hours of immobilization IAP mRNA levels were significantly increased in both frontal cortex and hippocampus compare to control group. Therefore, L1 expression is up-regulated prior to IAP expression, perhaps, due to the differences in the transcription regulation of these two elements. Thus, we have shown that expression of both L1 and IAP is rapidly induced in several brain regions by behavioral stress. Since our previous data demonstrated a stress-induced increase of L1 and IAP DNA copy number in the same brain structures, together it points to a full retrotransposition event. We further suggest that reverse transcription involved in L1 and IAP retrotransposition by behavioral stimuli may also contribute to long-term plasticity and memory formation in the nervous system.

Distribusi Herpetofauna di Kawasan Hutan Lindung Gambut Sungai Buluh Tanjung Jabung Timur

Hutan Lindung Gambut (HLG) Sungai Buluh is one of the village forests in East Tanjung Jabung. Peat soil has high water content and is herpetofauna habitat. Herpetofauna acts as a prey and predator in the peat ecosystem. Some of them occupy the position of top predators on food networks in the peat ecosystem. The existence of this group of animals also acts as an environmental bioindicator which can be indicated by its species richness and individual count. This research was conducted on two habitat conditions (terrestrial and aquatic) with the aim of this study were the distribution and habitat characteristics of herpetofauna in the HLG Sungai Buluh. This study used the Visual Encounter Survey (VES) method which was combined by transect lines in each habitat. Sampling was done nocturnal and diurnal. The specimens obtained then preserved in 70% alcohol. The results showed total of 105 individual herpetofauna consisting of 4 Anura families (Bufonidae 1 species, Dicroglossidae 3 species, Rhacophoridae 3 species, and Ranidae 4 species) and 4 Reptile families (Agamidae 3 species, Scincidae 2 species, Gekkonidae 1 species, and Natricidae 1 species). Eleven species were found in the terrestrial habitat transect while eleven species were found in aquatic habitat transects. Furthermore, four species were found in both habitat transects. From these results it can be concluded that the HLG Sungai Buluh has a suitable habitat for herpetofauna. However, further research needs to be carried out covering the entire are of HLG Sungai Buluh

Tumor Long Interspersed Nucleotide Element-1 (LINE-1) Hypomethylation in Relation to Age of Colorectal Cancer Diagnosis and Prognosis.

Simple SummaryThe rising incidence of early-onset cancers diagnosed before 50 years in many body sites (including the colorectum) is a growing concern. Despite the many studies of early-onset colorectal cancer, the characteristics of colorectal cancer diagnosed at age 50 or slightly after (close to age 50) have not been adequately examined. Although epigenetic alterations are considered to play a role in early-onset colorectal cancer, the association of LINE-1 hypomethylation (that reflects global DNA hypomethylation) with the age of colorectal cancer diagnosis has not been thoroughly clarified. Using a database of 1356 colorectal cancers, we found that tumor LINE-1 hypomethylation was increasingly more common with decreasing age of colorectal cancer diagnosis and was associated with higher colorectal cancer-specific mortality. Our findings support the “age continuum” model that has substantial implications in research on cancers in not only the colorectum but also many other body sites.Evidence indicates the pathogenic role of epigenetic alterations in early-onset colorectal cancers diagnosed before age 50. However, features of colorectal cancers diagnosed at age 50–54 (hereafter referred to as “intermediate-onset”) remain less known. We hypothesized that tumor long interspersed nucleotide element-1 (LINE-1) hypomethylation might be increasingly more common with decreasing age of colorectal cancer diagnosis. In 1356 colorectal cancers, including 28 early-onset and 66 intermediate-onset cases, the tumor LINE-1 methylation level measured by bisulfite-PCR-pyrosequencing (scaled 0 to 100) showed a mean of 63.6 (standard deviation (SD) 10.1). The mean tumor LINE-1 methylation level decreased with decreasing age (mean 64.7 (SD 10.4) in age ≥70, 62.8 (SD 9.4) in age 55–69, 61.0 (SD 10.2) in age 50–54, and 58.9 (SD 12.0) in age <50; p < 0.0001). In linear regression analysis, the multivariable-adjusted β coefficient (95% confidence interval (CI)) (vs. age ≥70) was −1.38 (−2.47 to −0.30) for age 55–69, −2.82 (−5.29 to −0.34) for age 50–54, and −4.54 (−8.24 to −0.85) for age <50 (Ptrend = 0.0003). Multivariable-adjusted hazard ratios (95% CI) for LINE-1 methylation levels of ≤45, 45–55, and 55–65 (vs. >65) were 2.33 (1.49–3.64), 1.39 (1.05–1.85), and 1.29 (1.02–1.63), respectively (Ptrend = 0.0005). In conclusion, tumor LINE-1 hypomethylation is increasingly more common with decreasing age of colorectal cancer diagnosis, suggesting a role of global DNA hypomethylation in colorectal cancer arising in younger adults.

Hepatitis C virus infection restricts human LINE-1 retrotransposition in hepatoma cells.

LINE-1 (L1) retrotransposons are autonomous transposable elements that can affect gene expression and genome integrity. Potential consequences of exogenous viral infections for L1 activity have not been studied to date. Here, we report that hepatitis C virus (HCV) infection causes a significant increase of endogenous L1-encoded ORF1 protein (L1ORF1p) levels and translocation of L1ORF1p to HCV assembly sites at lipid droplets. HCV replication interferes with retrotransposition of engineered L1 reporter elements, which correlates with HCV RNA-induced formation of stress granules and can be partially rescued by knockdown of the stress granule protein G3BP1. Upon HCV infection, L1ORF1p localizes to stress granules, associates with HCV core in an RNA-dependent manner and translocates to lipid droplets. While HCV infection has a negative effect on L1 mobilization, L1ORF1p neither restricts nor promotes HCV infection. In summary, our data demonstrate that HCV infection causes an increase of endogenous L1 protein levels and that the observed restriction of retrotransposition of engineered L1 reporter elements is caused by sequestration of L1ORF1p in HCV-induced stress granules.

MET Exon 14 Skipping: A Case Study for the Detection of Genetic Variants in Cancer Driver Genes by Deep Learning.

Background: Disruption of alternative splicing (AS) is frequently observed in cancer and might represent an important signature for tumor progression and therapy. Exon skipping (ES) represents one of the most frequent AS events, and in non-small cell lung cancer (NSCLC) MET exon 14 skipping was shown to be targetable. Methods: We constructed neural networks (NN/CNN) specifically designed to detect MET exon 14 skipping events using RNAseq data. Furthermore, for discovery purposes we also developed a sparsely connected autoencoder to identify uncharacterized MET isoforms. Results: The neural networks had a Met exon 14 skipping detection rate greater than 94% when tested on a manually curated set of 690 TCGA bronchus and lung samples. When globally applied to 2605 TCGA samples, we observed that the majority of false positives was characterized by a blurry coverage of exon 14, but interestingly they share a common coverage peak in the second intron and we speculate that this event could be the transcription signature of a LINE1 (Long Interspersed Nuclear Element 1)-MET (Mesenchymal Epithelial Transition receptor tyrosine kinase) fusion. Conclusions: Taken together, our results indicate that neural networks can be an effective tool to provide a quick classification of pathological transcription events, and sparsely connected autoencoders could represent the basis for the development of an effective discovery tool.

The Transposable Element Environment of Human Genes Differs According to Their Duplication Status and Essentiality.

Transposable elements (TEs) are major components of eukaryotic genomes and represent approximately 45% of the human genome. TEs can be important sources of novelty in genomes and there is increasing evidence that TEs contribute to the evolution of gene regulation in mammals. Gene duplication is an evolutionary mechanism that also provides new genetic material and opportunities to acquire new functions. To investigate how duplicated genes are maintained in genomes, here, we explored the TE environment of duplicated and singleton genes. We found that singleton genes have more short-interspersed nuclear elements and DNA transposons in their vicinity than duplicated genes, whereas long-interspersed nuclear elements and long-terminal repeat retrotransposons have accumulated more near duplicated genes. We also discovered that this result is highly associated with the degree of essentiality of the genes with an unexpected accumulation of short-interspersed nuclear elements and DNA transposons around the more-essential genes. Our results underline the importance of taking into account the TE environment of genes to better understand how duplicated genes are maintained in genomes.

EDUCATION FOR SUSTAINABLE DEVELOPMENT FROM THE PERSPECTIVE OF CHRISTIANITY: PEDAGOGIES AND PROSPECTS

With rising arguments on the fluid idea of education for sustainable development where an established framework is lacking, this review aims to look into the Christian bible for messages of sustainability and how these messages could enhance education for sustainable development in themes and pedagogies. This review examined only the old and new testaments of the bible common to most, if not all Christian denominations. It examined verses with clear connection to sustainability encompassing the environmental, social and economic aspects, particularly the environmental aspect. Discourse analysis of the verses comprising instructions, dialogues and lines of letters was conducted by delving into the historical and sociocultural contexts of the verses to exegete their sustainability implications. This review yielded ten verses with sustainability themes ranging from human responsibilities, ecological value, balance and conservation, responsible consumption and production, conservation of resources, traditional wisdom, equity, social welfare and sustainability to anthropogenic causes of environmental degradation. The main pedagogies for education for sustainable development implicated from this review are problem-based learning, experiential learning, exploratory learning, scenario- and simulation-based learning, reflective learning and deep learning, with specific references to the biblical sustainability themes. This review constitutes one of the very few literature probing sustainability and its pedagogies from Christianity’s perspective by qualitatively examining the most pertinent biblical verses. It contributes to the enrichment of sustainability education and pedagogies through religious wisdom. It is crucial in advancing sustainability education in religious studies. Article visualizations:

El mercado global de apuestas deportivas online: terreno fértil para fraudes y crímenes

Las nuevas tecnologías han modificado radicalmente la industria de las apuestas deportivas. En la actualidad, millones de personas apuestan en línea y en tiempo real, en una amplia gama de eventos que se desarrollan por todo el mundo. Este mercado global ha generado diversas oportunidades para el crimen y el fraude. En el panorama actual, el amaño de partidos ha evolucionado de ser un fenómeno histórico dentro del mundo de los deportes a un problema criminal global que involucra a organizaciones criminales transnacionales, a jugadores profesionales, a empresarios, árbitros, deportistas e intermediarios. A pesar de que la manipulación de competiciones se ha posicionado en la agenda pública como la mayor amenaza contra la sustentabilidad del deporte, la complejidad y extensión del nuevo mercado de apuestas generan áreas de riesgo para el delito que van más allá del simple amaño de partidos. El artículo tiene el objetivo pedagógico y práctico de explicar los cambios en el mercado y tipos de apuestas deportivas y describir las nuevas estructuras de oportunidad para el delito y el fraude que surgen del nuevo marco comercial. Todas estas cuestiones, poco estudiadas, abren una ventana de oportunidad para nuevas líneas de investigación multidisciplinar sobre deporte, criminalidad, economía y políticas públicas.

De novo transcriptome sequencing of triton shell Charonia lampas sauliae: Identification of genes related to neurotoxins and discovery of genetic markers.

Charonia lampas sauliae (triton snails, triton shells or tritons; Mollusca, Caenogastropoda, Littorinimorpha, Ranellidae) is a marine species with a wide distribution. In Korea, this species is listed as vulnerable and is regionally protected as an endangered species. Here, we report the first comprehensive transcriptome dataset of C. lampas sauliae obtained using the Illumina HiSeq 2500 platform. In total, 97.68% of raw read sequences were processed as clean reads. Of the 577,478 contigs obtained, 146,026 sequences were predicted to contain coding regions. About 89.34% of all annotated unigene sequences showed homologous matches to protein sequences in PANM DB (Protostome database). Further, about one-third of the unigene sequences were annotated using the UniGene, Swiss-Prot, Clusters of Orthologous Groups (COG) and Gene Ontology (GO) databases. In total, 190 enzymes were predicted under key metabolic pathways under stood through Kyoto Encyclopedia of Genes and Genomes (KEGG) database annotation. Repetitive elements such as long terminal repeats (LTRs), short interspersed nuclear elements (SINEs), long interspersed nuclear elements (LINEs), and DNA elements were enriched in the unigene sequences. Among the identified transcripts were the channel proteins, some of which were blocked by tetrodotoxin, which is thought to be synthesized by symbiotic bacteria inhabiting the shells. In addition, conotoxin superfamily peptides, such as B-conotoxin, conotoxin superfamily T and alpha-conotoxin, were identified, which may have relevance to biomedical and evolutionary research. A transcriptome-wide search for polymorphic loci identified 21,568 simple sequence repeats (SSRs) in the unigene sequences. Most SSRs were dinucleotides, among which AC/GT was the dominant SSR type. The molecular and genetic resources revealed in this study could be utilized for investigations on the fitness of the species in the marine environment and sustainability in a changing habitat.

Diversity of short interspersed nuclear elements (SINEs) in lepidopteran insects and evidence of horizontal SINE transfer between baculovirus and lepidopteran hosts

BackgroundShort interspersed nuclear elements (SINEs) belong to non-long terminal repeat (non-LTR) retrotransposons, which can mobilize dependent on the help of counterpart long interspersed nuclear elements (LINEs). Although 234 SINEs have been identified so far, only 23 are from insect species (SINEbase: http://sines.eimb.ru/).ResultsHere, five SINEs were identified from the genome of Plutella xylostella, among which PxSE1, PxSE2 and PxSE3 were tRNA-derived SINEs, PxSE4 and PxSE5 were 5S RNA-derived SINEs. A total of 18 related SINEs were further identified in 13 lepidopteran insects and a baculovirus. The 3′-tail of PxSE5 shares highly identity with that of LINE retrotransposon, PxLINE1. The analysis of relative age distribution profiles revealed that PxSE1 is a relatively young retrotransposon in the genome of P. xylostella and was generated by recent explosive amplification. Integration pattern analysis showed that SINEs in P. xylostella prefer to insert into or accumulate in introns and regions 5 kb downstream of genes. In particular, the PxSE1-like element, SlNPVSE1, in Spodoptera litura nucleopolyhedrovirus II genome is highly identical to SfSE1 in Spodoptera frugiperda, SlittSE1 in Spodoptera littoralis, and SlituSE1 in Spodoptera litura, suggesting the occurrence of horizontal transfer.ConclusionsLepidopteran insect genomes harbor a diversity of SINEs. The retrotransposition activity and copy number of these SINEs varies considerably between host lineages and SINE lineages. Host-parasite interactions facilitate the horizontal transfer of SINE between baculovirus and its lepidopteran hosts.

Possibili direzioni di senso nella malattia oncologica: un’esperienza di liminalità, meaning making e progettazione esistenziale

The oncological disease experience is counted as a wound in the body and mind attributable to a traumatic experience that fragments and disorients the person’s biography. The neoplasia leaves marks and scars in both somatic and existential level. The illness experience suggests to patient to look for meaning that cannot be unheard. The literature associating the concept of liminality in oncological disease to understand the process of meaning making. The definition of new horizons of meaning, generated by crossing the limen, opens up a new self-awareness and worldview. The autopoietic dimension present in oncological disease reveals new scenarios of generativity and existential planning in the direction of self-realization in the cipher of the possible. The contribution presents the story of Anastasia, a 41-years-old nurse. A few months after her new marriage and close to a professional promotion, she receives the diagnosis of cholangiocarcinoma: a very aggressive, low-incidence, neoplasia, without specific therapeutic lines and with a negative prognosis. Anastasia’s words, thoughts and emotions, collected in the course of some testimonies in a university classroom, are analyzed and deciphered within the pedagogical paradigm, refering to the perspective of pedagogical problematicism and phenomenological pedagogy.

Battle lines drawn over plastics bill

Battle lines drawn over plastics bill

Global DNA hypermethylation in peripheral blood mononuclear cells and cardiovascular disease risk: a population-based propensity score-matched cohort study

BackgroundDNA methylation plays an important role in the pathogenesis and progression of cardiovascular disease (CVD) but the prospective association of DNA methylation with CVD has not been evaluated. Here, we...

Battle lines drawn over plastics bill

Battle lines drawn over plastics bill

Responses of the antioxidative enzymes of few Sub1 rice (Oryza sativa) genotypes to prolonged submergence stress in Odisha

The present experiment was conducted in Department of Plant physiology, OUAT, Bhubaneswar during kharif 2017 and kharif 2019 to screen out the NILs rice genotypes for submergence adaptation traits under coastal regions of Odisha. The potential involvement of activated oxygen species by submergence stress was studied in twenty rice genotypes; thirteen sub1 NILs, two tolerant checks with six susceptible checks. These rice genotypes were subjected to 17 days of complete submergence. Under 17 days of complete submergence and after the submergence, the genotypes IR-85086-Sub 33 -3-2-1 and IR-88760-Sub 93-3-3 showed lower lipid Peroxidation in terms of malondialdehyde (MDA) level and also showed lower levels of ACC Oxidase activity (AAO) and presented higher activities of antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT), Peroxidase (POX) when compared to the susceptible checks. The levels of SOD activity indicated that detoxification of O2 - to H2O2 was maintained at a stable level throughout the submergence stress until up to 17 days in tolerant genotypes. These findings suggested that tolerance to submergence stress in rice might be proven by increased the capacity of antioxidative system. In addition, SOD and CAT activity has much higher affinity for scavenging H2O2 than POX. The present study evaluated thirteen pairs of Sub1 near-isogenic lines (NILs) together with FR13A and other check genotypes in pot culture conditions to assess the survival and growth processes occurring during submergence and recovery that are associated with Sub1.

Biochemical performance of some Sub1 rice (Oryza sativa) genotypes under submergence stress in Odisha

Recurring floods in Asia cause poor crop establishment. Yields decline drastically when plants are completely submerged for a few days. Traditional rice genotypes predominate because they have acquired moderate tolerance to flooding but they carry the penalty of inherently lower grain yields. Genotypes with tolerance to complete submergence were recently developed in the background of popular genotypes by transferring the submergence tolerance gene SUBMERGENCE1 (Sub1) from the highly tolerant Indian landrace FR13A. The present study evaluated thirteen pairs of Sub1 near-isogenic lines (NILs) together with FR13A and other check genotypes in pot culture conditions to assess the survival and growth processes occurring during submergence and recovery that are associated with Sub1. The present experiment was conducted in Department of Plant physiology, OUAT, Bhubaneswar during kharif 2017 and kharif 2019 to screen out the NILs rice genotypes for submergence adaptation traits under coastal regions of Odisha. The present study indicated that among the twenty genotypes, IR-85086-Sub 33-3-2-1(4.87 t ha-1), IR-88760-Sub 93-3-3 (4.7 t ha-1) and Swarna Sub -1 (4.57 t ha-1) contributed highest yield under submergence.

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress

Characterizing Repeats in Two Whole-Genome Amplification Methods in the Reniform Nematode Genome.

One of the major problems in the U.S. and global cotton production is the damage caused by the reniform nematode, Rotylenchulus reniformis. Amplification of DNA from single nematodes for further molecular analysis can be challenging sometimes. In this research, two whole-genome amplification (WGA) methods were evaluated for their efficiencies in DNA amplification from a single reniform nematode. The WGA was carried out using both REPLI-g Mini and Midi kits, and the GenomePlex single cell whole-genome amplification kit. Sequence analysis produced 4 Mb and 12 Mb of genomic sequences for the reniform nematode using REPLI-g and SIGMA libraries. These sequences were assembled into 28,784 and 24,508 contigs, respectively, for REPLI-g and SIGMA libraries. The highest repeats in both libraries were of low complexity, and the lowest for the REPLI-g library were for satellites and for the SIGMA library, RTE/BOV-B. The same kind of repeats were observed for both libraries; however, the SIGMA library had four other repeat elements (Penelope (long interspersed nucleotide element (LINE)), RTE/BOV-B (LINE), PiggyBac, and Mirage/P-element/Transib), which were not seen in the REPLI-g library. DNA transposons were also found in both libraries. Both reniform nematode 18S rRNA variants (RN_VAR1 and RN_VAR2) could easily be identified in both libraries. This research has therefore demonstrated the ability of using both WGA methods, in amplification of gDNA isolated from single reniform nematodes.