Identifying Terpenoid Biosynthesis Genes in Euphorbia maculata via Full-Length cDNA Sequencing.

The annual herb Euphorbia maculata L. produces anti-inflammatory and biologically active substances such as triterpenoids, tannins, and polyphenols, and it is used in traditional Chinese medicine. Of these bioactive compounds, terpenoids, also called isoprenoids, are major secondary metabolites in E. maculata. Full-length cDNA sequencing was carried out to characterize the transcripts of terpenoid biosynthesis reference genes and determine the copy numbers of their isoforms using PacBio SMRT sequencing technology. The Illumina short-read sequencing platform was also employed to identify differentially expressed genes (DEGs) in the secondary metabolite pathways from leaves, roots, and stems. PacBio generated 62 million polymerase reads, resulting in 81,433 high-quality reads. From these high-quality reads, we reconstructed a genome of 20,722 genes, in which 20,246 genes (97.8%) did not have paralogs. About 33% of the identified genes had two or more isoforms. DEG analysis revealed that the expression level differed among gene paralogs in the leaf, stem, and root. Whole sets of paralogs and isoforms were identified in the mevalonic acid (MVA), methylerythritol phosphate (MEP), and terpenoid biosynthesis pathways in the E. maculata L. The nucleotide information will be useful for identifying orthologous genes in other terpenoid-producing medicinal plants.

Association of Participation in Health Check-ups with Risk Factors for Cardiovascular Diseases.

BACKGROUND: We compared the risk factors for cardiovascular diseases (CVDs) among Koreans who did and did not participate in national periodic health check-ups, after adjustment for demographic factors, socioeconomic status, and lifestyle factors. METHODS: This cross-sectional study used data from the Korea National Health and Nutrition Examination Survey (KNHANES) from 2007 to 2018. Study subjects were classified as participants or non-participants in health check-ups, based on attendance at national periodic health check-ups during the previous two years. RESULTS: Comparison of participants and non-participants in health check-ups indicated statistically significant differences in age, gender, region, education level, monthly income, employment status, obesity, smoking, alcohol consumption, exercise, and marital status. After adjustment for demographic, socioeconomic factors, and health-related behaviors, woman non-participants were more likely to have metabolic syndrome, pre-hypertension, hypertension, prediabetes, and diabetes, and man non-participants were more likely to have pre-diabetes and diabetes. CONCLUSION: Subjects who participated in periodic health check-ups had fewer CVD-related risk factors than non-participants. Thus, health care providers should encourage non-participants to attend periodic health check-ups so that appropriate interventions can be implemented and decrease the risk for CVDs in these individuals.

Plastid Genomes of the Early Vascular Plant Genus Selaginella Have Unusual Direct Repeat Structures and Drastically Reduced Gene Numbers.

The early vascular plants in the genus Selaginella, which is the sole genus of the Selaginellaceae family, have an important place in evolutionary history, along with ferns, as such plants are valuable resources for deciphering plant evolution. In this study, we sequenced and assembled the plastid genome (plastome) sequences of two Selaginella tamariscina individuals, as well as Selaginella stauntoniana and Selaginella involvens. Unlike the inverted repeat (IR) structures typically found in plant plastomes, Selaginella species had direct repeat (DR) structures, which were confirmed by Oxford Nanopore long-read sequence assembly. Comparative analyses of 19 lycophytes, including two Huperzia and one Isoetes species, revealed unique phylogenetic relationships between Selaginella species and related lycophytes, reflected by structural rearrangements involving two rounds of large inversions that resulted in dynamic changes between IR and DR blocks in the plastome sequence. Furthermore, we present other uncommon characteristics, including a small genome size, drastic reductions in gene and intron numbers, a high GC content, and extensive RNA editing. Although the 16 Selaginella species examined may not fully represent the genus, our findings suggest that Selaginella plastomes have undergone unique evolutionary events yielding genomic features unparalleled in other lycophytes, ferns, or seed plants.

Biological effects of cyclosporin A on CD3-CD161+ and CD3+CD161+ lymphocytes.

Cyclosporin A (CSA) is a widely used drug to prevent the immune cell function. It is well known that CSA blocks transcription of cytokine genes in activated T cells. The connection between T cells and CSA has been well established. However, the effect of CSA on natural killer (NK) cells is not thoroughly understood. Therefore, in the present study, splenocytes and peripheral blood mononuclear cells (PBMCs) were treated with CSA in the presence of concanavalin A (Con A) or interleukin-2 (IL-2). CSA at higher concentrations induces apoptosis and inhibition of proliferation, while lower concentrations showed synergistically enhanced proliferation in splenocytes and PBMCs. Further, CSA favored the in vitro conversion of CD3+CD161+ cells. Splenocytes and PBMC were found to have synergistic proliferation with Con A, and PBMC exhibited significantly higher expression of NKp30, NKp44, and granzyme B along with enhanced cytotoxicity against K-562 cells in CSA-treated animals. Proliferation assay also showed that proliferation of CD161+ cells was higher in CSA-treated animals. Collectively, our results suggest that CSA differentially influences the population, function, and expression of the NK cell phenotype.

Ultravist Induces the Expression of MCP-1 and VCAM-1 in IL-4-Stimulated HUVECs.

The goal of the present study focused on the adverse reaction of contrast medium (CM) via the induction of inflammatory molecules in human umbilical vein endothelial cells (HUVECs). Ultravist-induced monocyte chemoattractant protein-1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) gene expression was markedly increased in interleukin-4 (IL-4)-pretreated HUVECs in a time- and dose-dependent manner and was paralleled by concomitant production of MCP-1 and VCAM-1 proteins. MCP-1 and VCAM-1 gene expression by Ultravist in combination with IL-4 was mediated by the c-Jun N-terminal kinases (JNK1/2) signaling pathway. IL-4-pretreated Ultravist-stimulated HUVECs showed greatly increased migration and adhesion of THP-1 cells. Cell migration was decreased by treatment of CCR2 antagonist, and cell adhesion was also decreased by VCAM-1 blocking antibody. Furthermore, when tested in vivo under similar conditions, MCP-1 protein was significantly increased in Ultravist combined with IL-4-injected mice. Taken together, our findings suggest that MCP-1 blocking may be crucial in preventing the endothelial dysfunction induced by contrast medium in patients with inflammatory disease and atherosclerosis.

Adversarially Learned Total Variability Embedding for Speaker Recognition with Random Digit Strings.

Over the recent years, various research has been conducted to investigate methods for verifying users with a short randomized pass-phrase due to the increasing demand for voice-based authentication systems. In this paper, we propose a novel technique for extracting an i-vector-like feature based on an adversarially learned inference (ALI) model which summarizes the variability within the Gaussian mixture model (GMM) distribution through a nonlinear process. Analogous to the previously proposed variational autoencoder (VAE)-based feature extractor, the proposed ALI-based model is trained to generate the GMM supervector according to the maximum likelihood criterion given the Baum-Welch statistics of the input utterance. However, to prevent the potential loss of information caused by the Kullback-Leibler divergence (KL divergence) regularization adopted in the VAE-based model training, the newly proposed ALI-based feature extractor exploits a joint discriminator to ensure that the generated latent variable and the GMM supervector are more realistic. The proposed framework is compared with the conventional i-vector and VAE-based methods using the TIDIGITS dataset. Experimental results show that the proposed method can represent the uncertainty caused by the short duration better than the VAE-based method. Furthermore, the proposed approach has shown great performance when applied in association with the standard i-vector framework.

Development of CACTA transposon derived SCAR markers and their use in population structure analysis in Zea mays.

Molecular marker technologies have proven to be an important breakthrough for genetic studies, construction of linkage maps and population genetics analysis. Transposable elements (TEs) constitute major fractions of repetitive sequences in plants and offer a wide range of possible areas to be explored as molecular markers. Sequence characterized amplified region (SCAR) marker development provides us with a simple and time saving alternative approach for marker development. We employed the CACTA-TD to develop SCARs and then integrated them into linkage map and used them for population structure and genetic diversity analysis of corn inbred population. A total of 108 dominant SCAR markers were designed out of which, 32 were successfully integrated in to the linkage map of maize RIL population and the remaining were added to a physical map for references to check the distribution throughout all chromosomes. Moreover, 76 polymorphic SCARs were used for diversity analysis of corn accessions being used in Korean corn breeding program. The overall average polymorphic information content (PIC) was 0.34, expected heterozygosity was 0.324 and Shannon's information index was 0.491 with a percentage of polymorphism of 98.67%. Further analysis by associating with desirable traits may also provide some accurate trait specific tagged SCAR markers. TE linked SCARs can provide an added level of polymorphism as well as improved discriminating ability and therefore can be useful in further breeding programs to develop high yielding germplasm.

Association of Blood Pressure with Blood Lead and Cadmium Levels in Korean Adolescents: Analysis of Data from the 2010-2016 Korean National Health and Nutrition Examination Survey.

BACKGROUND: We evaluated the association of blood pressure (BP) with blood levels of cadmium, lead, and cadmium and lead together (cadmium + lead) in a representative sample of adolescents from Korea. METHODS: We used 2010-2016 data from the Korean National Health and Nutrition Examination Survey. This cross-sectional study enrolled adolescents aged at 10-18 years-old who completed a health examination survey and had blood measurements of lead and cadmium. The association of adjusted mean differences in diastolic and systolic BP with doubling of blood lead and cadmium were estimated by regression of BP against log2-transformed blood metals and their quartiles after covariate adjustment. Adjusted odds ratio for prehypertension were calculated for log2-transformed blood levels of lead and cadmium and their quartiles. RESULTS: Our analysis of adolescents in Korea indicated that blood levels of lead and cadmium were not significantly associated with increased BP or risk of prehypertension. However, the cadmium + lead level was associated with prehypertension. Previous studies showed that blood levels of lead and cadmium were associated with increased BP and risk of hypertension in adult populations. We found no such association in Korean adolescents. CONCLUSION: We found that the cadmium + lead level was associated with prehypertension. The differences between adults and adolescents are because adolescents generally have lower levels of these blood metals or because adolescents only rarely have hypertension.

Construction of genetic linkage map and identification of QTLs related to agronomic traits in maize using DNA transposon-based markers.

Transposable elements (TEs), are a rich source for molecular marker development as they constitute a significant fraction of the eukaryotic genome and impact the overall genome structure. Here, we utilize Mutator-based transposon display (Mu-TD), and CACTA-derived sequence-characterized amplified regions (SCAR) anchored by simple sequence repeats and single nucleotide polymorphisms to locate quantitative trait loci (QTLs) linked to agriculturally important traits on a genetic map. Specifically, we studied recombinant inbred line populations derived from a cross between dent corn and waxy corn. The resulting linkage map included 259 Mu-anchored fragments, 34 SCARs, and 614 SSR markers distributed throughout the ten maize chromosomes. Linkage analysis revealed three SNP loci associated with kernel starch synthesis genes (sh2, su1, wx1) linked to either Mu-TD loci or SSR markers, which may be useful for maize breeding programs. In addition, we used QTL analysis to determine the chromosomal location of traits related to grain yield and kernel quality. We identified 24 QTLs associated with nine traits located on nine out of ten maize chromosomes. Among these, 13 QTLs involved Mu loci and two involved SCARs. This study demonstrates the potential use of DNA transposon-based markers to construct linkage maps and identify QTLs linked to agronomic traits.

A bioinformatics approach for identifying transgene insertion sites using whole genome sequencing data.

BACKGROUND: Genetically modified crops (GM crops) have been developed to improve the agricultural traits of modern crop cultivars. Safety assessments of GM crops are of paramount importance in research at developmental stages and before releasing transgenic plants into the marketplace. Sequencing technology is developing rapidly, with higher output and labor efficiencies, and will eventually replace existing methods for the molecular characterization of genetically modified organisms. METHODS: To detect the transgenic insertion locations in the three GM rice gnomes, Illumina sequencing reads are mapped and classified to the rice genome and plasmid sequence. The both mapped reads are classified to characterize the junction site between plant and transgene sequence by sequence alignment. RESULTS: Herein, we present a next generation sequencing (NGS)-based molecular characterization method, using transgenic rice plants SNU-Bt9-5, SNU-Bt9-30, and SNU-Bt9-109. Specifically, using bioinformatics tools, we detected the precise insertion locations and copy numbers of transfer DNA, genetic rearrangements, and the absence of backbone sequences, which were equivalent to results obtained from Southern blot analyses. CONCLUSION: NGS methods have been suggested as an effective means of characterizing and detecting transgenic insertion locations in genomes. Our results demonstrate the use of a combination of NGS technology and bioinformatics approaches that offers cost- and time-effective methods for assessing the safety of transgenic plants.

Environmental exposures to lead, mercury, and cadmium among South Korean teenagers (KNHANES 2010-2013): Body burden and risk factors.

INTRODUCTION: Limited information is available on the association of age and sex with blood concentrations of heavy metals in teenagers. In addition, factors such as a shared family environment may have an association. We analyzed data from the Korean National Health and Nutrition Examination Survey (KNHANES, 2010-2013) to determine whether blood levels of heavy metals differ by risk factors such as age, sex, and shared family environment in a representative sample of teenagers. METHODS: This study used data obtained in the KNHANES 2010-2013, which had a rolling sampling design that involved a complex, stratified, multistage, probability-cluster survey of a representative sample of the non-institutionalized civilian population in South Korea. Our cross-sectional analysis was restricted to teenagers and their parents who completed the health examination survey, and for whom blood measurements of cadmium, lead, and mercury were available. The final analytical sample consisted of 1585 teenagers, and 376 fathers and 399 mothers who provided measurements of blood heavy metal concentrations. RESULTS: Male teenagers had greater blood levels of lead and mercury, but sex had no association with blood cadmium level. There were age-related increases in blood cadmium, but blood lead decreased with age, and age had little association with blood mercury. The concentrations of cadmium and mercury declined from 2010 to 2013. The blood concentrations of lead, cadmium, and mercury in teenagers were positively associated with the levels in their parents after adjustment for covariates. CONCLUSION: Our results show that blood heavy metal concentrations differ by risk factors such as age, sex, and shared family environment in teenagers.

Carbohydrate Intake Exhibited a Positive Association with the Risk of Metabolic Syndrome in Both Semi-Quantitative Food Frequency Questionnaires and 24-Hour Recall in Women.

We compared the usual nutrient intake in both the semi-quantitative food frequency questionnaire (SQFFQ) and 24-hour recall methods and determined the association between metabolic syndrome (MetS) risk and nutrient intake calculated by both methods in Korea National Health and Nutrition Examination Survey (KNHANES; 2012-2014) data. Adjusted odds ratios for MetS were calculated according to the intake of macronutrients, measured by the 2 methods in 10,286 adults, while controlling for covariates associated with MetS. Fat and carbohydrate intake (energy percent) calculated by 24-hour recall and SQFFQ was significantly different between the MetS and non-MetS groups, particularly in women. The differences in other nutrient intakes determined by both methods were mainly non-significant. The correlation coefficients between the 2 methods were about 0.4 for most nutrients except total vitamin A and iron (Fe). Energy intake according to gender and MetS presence was similar between the 2 methods. Carbohydrate intake exhibited a positive association with the MetS risk, while fat intake showed a negative association in both methods. The association exhibited a gender interaction with carbohydrate and fat intake calculated by 24-hour recall: women exhibited a significant association. However, for the SQFFQ a gender interaction was evident only for carbohydrate intake. In diet quality index of SQFFQ the adequacy of vegetables and total fat intake was higher in the non-MetS than the MetS. In conclusion, the MetS prevalence exhibited a positive association with carbohydrate intake only in women, as assessed by 24-hour recall and SQFFQ. The SQFFQ can be used to assess the association between usual food intake and MetS risk in large population studies.

Decision-directed speech power spectral density matrix estimation for multichannel speech enhancement.

In this letter, a multichannel decision-directed approach to estimate the speech power spectral density (PSD) matrix for multichannel speech enhancement is proposed. There have been attempts to build multichannel speech enhancement filters which depend only on the speech and noise PSD matrices, for which the accurate estimate of the clean speech PSD matrix is crucial for a successful noise reduction. In contrast to the maximum likelihood estimator which has been applied conventionally, the proposed decision-directed method is capable of tracking the time-varying speech characteristics more robustly and improves the noise reduction performance under various noise environments.

High carbohydrate diets are positively associated with the risk of metabolic syndrome irrespective to fatty acid composition in women: the KNHANES 2007-2014.

We evaluated that carbohydrates and fatty acids intake modifies the prevalence of metabolic syndrome (MetS) and we also determined gender-nutrient interaction in 38,766 adults in KNHANES (2007-2014). Carbohydrate intake was positively associated, and fat intake inversely associated, with the incidence of MetS. The association exhibited a gender interaction with the macronutrient intake; this association was significant in females. Furthermore, saturated fatty acid (SFA), monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) intakes were inversely associated with MetS risk and only females showed the positive association. Both n-3 and n-6 fatty acids intake showed inverse associations with MetS risk, similar to PUFA intake. Among the MetS components, serum triglyceride levels and blood pressure had significant inverse associations with fatty acid intake irrespective of fatty acid types and exhibited a gender interaction. In conclusions, high carbohydrate intakes (≥74.2 En%) may increase the MetS risk and moderate fat intakes (≥20.7 En%), irrespective of fat types, may decrease it. These associations were significant only in women.

Aronia melanocarpa (Black Chokeberry) Reduces Ethanol-Induced Gastric Damage via Regulation of HSP-70, NF-κB, and MCP-1 Signaling.

Aronia melanocarpa (Michx.) Ell. belongs to the Rosaceae family. The purpose of this study is to explore the gastroprotective effect of the Aronia melanocarpa hydro-alcoholic extract (AMHAE) against ethanol-induced gastric ulcer in a rat model. Different concentrations (50, 100, and 200 mg/kg) of AMHAE, or 30 mg/kg of omeprazole, significantly inhibited the gastric injury formation. The ethanol-induced ulcer group showed significant increases of malondialdehyde (MDA), myeloperoxidase (MPO), tumor necrosis factor (TNF)-α, nuclear factor-kappaB p65 (NF-κB p65), and monocyte chemoattractant protein (MCP)-1, and decreased activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-px), and interleukin (IL)-4. However, AMHAE (200 mg/kg) pretreatment significantly reversed the altered pathophysiological levels of these biomolecules to near normal stages. The gastroprotective activity of AMHAE was abolished by pretreatment with l-NAME, naloxone, capsazepine, and indomethacin, demonstrating the participation of nitric oxide (NO), opioids, TRPV (vanilloid receptor-related transient receptor potential), and prostaglandins in AMHAE-assisted gastroprotection against ethanol-induced gastric injuries. This gastroprotective effect of AMHAE might be due to the downregulation of TNF-α-based NF-κB, MCP-1 signaling and strong antioxidant properties.

OPHTHALMIC EXAMINATION IN COMMON KESTRELS (FALCO TINNUCULUS) FROM SOUTH KOREA.

The purpose of this study was to establish reference ophthalmic findings in the common kestrel (Falco tinnunculus). Twenty healthy adult kestrels were included in this study. Ophthalmic examinations included slit lamp biomicroscopy, fundus exam, Schirmer tear test, conjunctival bacterial culture and isolation, corneal touch threshold, tonometry, and corneal diameter measurement. Mean tear production was 7.4 ± 3.27 mm/min, and mean intraocular pressure measured via applanation tonometry was 10.5 ± 3.15 mm Hg. In addition, the mean corneal touch threshold was 29.8 ± 20.1 mm, and the mean corneal diameter was 9.8 ± 1.1 mm. Of the 25 conjunctival swabs, 23 (92%) yielded bacterial growth. Most of these bacteria were gram positive (69.6%); the most predominant genus was Staphylococcus. This study presents reference values for ophthalmic examinations in common kestrels.

Ribosomal DNA locus variation and REMAP analysis of the diploid and triploid complexes of Lilium lancifolium.

Lilium lancifolium Thunb. (2n = 2x = 24) is a cytologically conspicuous species with both diploids and triploids in nature. Cytological and molecular genetic analyses were carried out in both diploids and triploids that were collected from 55 geographical locations in Korea, Japan, and China. While the 5S rRNA gene loci were located at duplicated loci on the long arm of chromosome 2, the 45S rRNA gene loci were present in chromosomes 1, 2, 4, 6, 7, and 11. While the loci on chromosomes 1 and 7 were constant, the loci on chromosomes 2, 4, 6, 7, and 11 were variable in some plants so that the L. lancifolium accessions were grouped into 7 cytotypes in diploids and 12 cytotypes in triploids. REMAP marker analysis revealed that the diploids were classified into seven clusters, and the triploids were classified into a large cluster. Geographic, cytological, and genetic differentiations were not related in both the diploid and triploid accessions of L. lancifolium. Thus, current genetic variations occurred prior to the geographic differentiation in both diploids and triploids, and the 45S rDNA cytotype variations occurred after geographic differentiation in the current habitats of L. lancifolium.

LTR-retrotransposons and inter-retrotransposon amplified polymorphism (IRAP) analysis in Lilium species.

LTR-retrotransposons are ubiquitous and highly abundant in plant genomes. Moreover, LTR-retrotransposons can often cause genome obesity in plants. Although Lilium species have been known carrying large genomes among flowering plants, reports on the LTR-retrotransposons in Lilium species are rather limited. We isolated a novel Ty3/gypsy-like retrotransposon, LIRE-del, and two Ty1/copia-like retrotransposons, a LIRE-del and an unclassified, from a fosmid clone of Lilium longiflorum. Decayed internal ORF sequences indicated that they were non-autonomous elements. IRAP protocol was developed based on the LTR sequences of the isolated LTR-retrotransposons. Fourteen primer combinations showed clear distinctive PCR amplification bands that were highly informative in the analysis of species relationship among Lilium species. The phylogenetic relationship based on the IRAP profile revealed some discordant with phylogenetic studies based on the ITS sequences of 45S ribosomal gene and matK gene variations in a few species. Thus, the phylogenetic relationship among Lilium species may need to be re-evaluated with other tools such as cross compatibility and selectively neutral genetic markers.

Associations between blood mercury levels and subclinical changes in liver enzymes among South Korean general adults: analysis of 2008-2012 Korean national health and nutrition examination survey data.

INTRODUCTION: We herein used data from the Korean National Health and Nutritional Examination Survey (KNHANES) 2008-2012 to examine the associations between blood mercury levels and subclinical changes of liver function in a representative sample of the adult Korean population. METHODS: This study was based on data obtained from KNHANES, in which a rolling sampling design was used to perform a complex, stratified, multistage probability cluster survey of a representative sample of the non-institutionalized civilian population in South Korea. The associations between subclinical hepatic changes and blood mercury levels were assessed after adjustment for various demographic and lifestyle factors. RESULTS: Multivariate linear regression analyses revealed that each doubling of blood mercury increased serum aspartate transaminase (AST) by 0.676U/L and serum alanine transaminase (ALT) by 1.067U/L. The mean differences (95% CI) in serum AST and ALT between the lowest and highest quartiles were statistically significant at 1.249 (0.263-2.235)U/L and 2.248 (0.648-3.848), respectively. Logistic regression analysis showed that the odd ratios for having serum AST and ALT levels above the median were statistically significant in both the models according to the increase of blood mercury. The risks of having serum AST and ALT levels higher than the median among subjects in 4th quartile of blood mercury were 1.524 and 1.947, respectively. DISCUSSION: The present findings show that subclinical changes of liver function are associated with blood mercury levels. This is the first study to show an association between blood mercury levels and mild liver dysfunction, as a possible proxy measure of nonalcoholic fatty liver disease (NAFLD), in Asian population.

O- and N-Methyltransferases in benzylisoquinoline alkaloid producing plants.

BACKGROUND: Secondary metabolites such as benzylisoquinoline alkaloids (BIA) have attracted considerable attention because of their pharmacological properties and potential therapeutic applications. Methyltransferases (MTs) can add methyl groups to alkaloid molecules, altering their physicochemical properties and bioactivity, stability, solubility, and recognition by other cellular components. Five types of O-methyltransferases and two types of N-methyltransferases are involved in BIA biosynthesis. OBJECTIVE: Since MTs may be the source for the discovery and development of novel biomedical, agricultural, and industrial compounds, we performed extensive molecular and phylogenetic analyses of O- and N-methyltransferases in BIA-producing plants. METHODS: MTs involved in BIA biosynthesis were isolated from transcriptomes of Berberis koreana and Caulophyllum robustum. We also mined the methyltransferases of Coptis japonica, Papaver somniferum, and Nelumbo nucifera from the National Center for Biotechnology Information protein database. Then, we analyzed the functional motifs and phylogenetic analysis. RESULT: We mined 42 O-methyltransferases and 8 N-methyltransferases from the five BIA-producing plants. Functional motifs for S-adenosyl-L-methionine-dependent methyltransferases were retained in most methyltransferases, except for the three O-methyltransferases from N. nucifera. Phylogenetic analysis revealed that the methyltransferases were grouped into four clades, I, II, III and IV. The clustering patterns in the phylogenetic analysis suggested a monophyletic origin of methyltransferases and gene duplication within species. The coexistence of different O-methyltransferases in the deep branch subclade might support some cases of substrate promiscuity. CONCLUSIONS: Methyltransferases may be a source for the discovery and development of novel biomedical, agricultural, and industrial compounds. Our results contribute to further understanding of their structure and reaction mechanisms, which will require future functional studies.