MicroRNAs (miRNAs) are small noncoding RNA molecules that play crucial post-transcriptional regulatory roles in plants, including development and stress-response signaling. However, information about their involvement in secondary metabolism is still limited. Murraya koenigii is a popular medicinal plant, better known as curry leaves, that possesses pharmaceutically active secondary metabolites. The present study utilized high-throughput sequencing technology to investigate the miRNA profile of M. koenigii and their association with secondary metabolite biosynthesis. A total of 343,505 unique reads with lengths ranging from 16 to 40 nt were obtained from the sequencing data, among which 142 miRNAs were identified as conserved and 7 as novel miRNAs. Moreover, 6078 corresponding potential target genes of M. koenigii miRNAs were recognized in this study. Interestingly, several conserved and novel miRNAs of M. koenigii were found to target key enzymes of the terpenoid backbone and the flavonoid biosynthesis pathways. Furthermore, to validate the sequencing results, the relative expression of eight randomly selected miRNAs was determined by qPCR. To the best of our knowledge, this is the first report of the M. koenigii miRNA profile that may provide useful information for further elucidation of the involvement of miRNAs in secondary metabolism. These findings might be crucial in the future to generate artificial-miRNA-based, genetically engineered M. koenigii plants for the overproduction of medicinally highly valuable secondary metabolites.

The pharmacological properties of plants lie in the content of secondary metabolites that are classified into different categories based on their biosynthesis, structures, and functions. MicroRNAs (miRNAs) are small non-coding RNA molecules that play crucial post-transcriptional regulatory roles in plants, including development and stress-response signaling; however, information about their involvement in secondary metabolism is still limited. Cumin is one of the most popular seeds from the plant Cuminum cyminum, with extensive applications in herbal medicine and cooking; nevertheless, no previous studies focus on the miRNA profile of cumin. In this study, the miRNA profile of C. cyminum and its association with the biosynthesis of secondary metabolites were determined using NGS technology. The sequencing data yielded 10,956,054 distinct reads with lengths ranging from 16 to 40 nt, of which 349 miRNAs were found to be conserved and 39 to be novel miRNAs. Moreover, this work identified 1959 potential target genes for C. cyminum miRNAs. It is interesting to note that several conserved and novel miRNAs have been found to specifically target important terpenoid backbone, flavonoid biosynthesis, and lipid/fatty acid pathways enzymes. We believe this investigation will aid in elucidating the implications of miRNAs in plant secondary metabolism.

Secondary metabolites are bioactive compounds believed to contribute to the pharmacological properties of plants. MicroRNAs (miRNAs) are small non-coding RNA molecules involved in post-transcriptional regulation and are thought to play an important role in regulating secondary metabolism biosynthesis. Nevertheless, the extent of miRNA involvement in secondary metabolism remains minimal. Nigella sativa (black cumin/black seed) is a popular medicinal and culinary plant known for its pharmaceutical properties; however, its genomic information is scarce. In this study, next-generation sequencing (NGS) technology was employed to obtain the miRNA profile of N. sativa, and their involvement in secondary metabolite biosynthesis was explored. A total of 25,139,003 unique reads ranging from 16 to 40 nucleotides were attained, out of which 240 conserved and 34 novel miRNAs were identified. Moreover, 6083 potential target genes were recognized in this study. Several conserved and novel black cumin miRNAs were found to target enzymes involved in the terpenoid, diterpenoid, phenylpropanoid, carotenoid, flavonoid, steroid, and ubiquinone biosynthetic pathways, among others, for example, beta-carotene 3-hydroxylase, gibberellin 3 beta-dioxygenase, trimethyltridecatetraene synthase, carboxylic ester hydrolases, acetyl-CoA C-acetyltransferase, isoprene synthase, peroxidase, shikimate O-hydroxycinnamoyltransferase, etc. Furthermore, sequencing data were validated through qPCR by checking the relative expression of eleven randomly selected conserved and novel miRNAs (nsa-miR164d, nsa-miR166a, nsa-miR167b, nsa-miR171a, nsa-miR390b, nsa-miR396, nsa-miR159a, nsa-miRN1, nsa-miRN29, nsa-miRN32, and nsa-miRN34) and their expression patterns were found to be corroborated with the sequencing data. We anticipate that this work will assist in clarifying the implications of miRNAs in plant secondary metabolism and aid in the generation of artificial miRNA-based strategies to overproduce highly valuable secondary metabolites from N. sativa.

Galphimia spp. is popularly used in Mexican traditional medicine. Some populations of Galphimia exert anxiolytic and sedative effects due to the presence of the modified triterpenoids galphimines. However, the galphimine synthesis pathway has not yet been elucidated. Hence, in this study, a comparative transcriptome analysis between two contrasting populations of Galphimia spp., a galphimine-producer, and a non-galphimine-producer, is performed using RNA-Seq in the Illumina Next Seq 550 platform to identify putative candidates genes that encode enzymes of this metabolic pathway. Transcriptome functional annotation was performed using the Blast2GO in levels of gene ontology. For differential expression analysis, edgeR, pheatmap, and Genie3 library were used. To validate transcriptome data, qPCR was conducted. In producer and non-producer plants of both populations of Galphimia spp., most of the transcripts were grouped in the Molecular Function level of gene ontology. A total of 680 differentially expressed transcripts between producer and non-producer plants were detected. In galphimine-producer plants, a larger number of highly expressed transcripts related to acyclic and polycyclic terpene synthesis were identified. As putative candidate genes involved in the galphimine synthesis pathway, P450 family members and enzymes with kinase activity were identified.

El objetivo fue determinar si una solución salina ozonizada (SSO3) incrementa los neutrófilos polimorfonucleares (NPMN) endometriales (Exp 1) y desafiar el efecto preventivo de SSO3 sobre la endometritis subclínica (ESC) (Exp 2). En el Exp 1 se utilizaron 38 vacas Holstein primíparas. Se incluyeron vacas con (CAM) y sin antecedentes de metritis postparto (SAM); posteriormente se repartieron en subgrupos control (CTRL, solución salina) o SSO3 (6.7 ± 0.3 ppm). A los 55 días postparto se aplicó 50 ml de CTRL o SSO3 intrauterinamente y a las 48 h se registró la cantidad de NPMN por citología endometrial. En el Exp 2 se utilizaron 26 vacas Holstein primíparas SAM. Las vacas se repartieron aleatoriamente en CTRL o SSO3. Se administraron dos dosis de 50 ml con intervalo de 7 días (primera aplicación 11.3 ± 0.4 días postparto). Al día 30 postparto se diagnosticó ESC (≥6% NPMN). Las vacas CAM tuvieron mayor número de NPMN endometriales comparado con vacas SAM (13.9 ± 6.2 vs 1.0±0.46, P<0.05). El grupo CAM-CTRL presentó mayor número de NPMN que el SAM-CTRL (17.0 ± 9.6 vs 0.1 ± 0.1, P0.05) y SAM-CTRL y SAM-SSO3 (0.1 ± 0.1 y 1.8 ± 4.8, P>0.05) no fueron diferentes. Se observó una tendencia estadística (P=0.09) de menor porcentaje de ESC en grupo CTRL comparado con SSO3 (15.4 y 46.2 %, respectivamente). En conclusión, la SSO3 por vía transcervical no incrementa el número de NPMN endometriales y el tratamiento preventivo con SSO3 aplicado a vacas SAM no disminuyó la ESC. Los resultados sugieren un posible efecto antiinflamatorio del tratamiento solución salina ionizada.

Plants have been indispensable for human civilization, for example, in medicine, where their secondary metabolites play a fundamental role. MicroRNAs (miRNAs), short non-coding riboregulators, are essential in the control of gene expression and offer potential therapeutic and bioengineering applications. Fenugreek (Trigonella foenum-graecum) has been a staple in Asian and African cuisines for centuries and has been shown to have significant medicinal and cosmetic benefits thanks to its high concentration of secondary metabolites; however, little is known about its miRNA profile. In this study, next-generation sequencing (NGS) technology was used to explore the miRNA landscape of T. foenum-graecum and its relationship with secondary metabolite biosynthesis. The analysis revealed 23,951,014 distinct reads, ranging from 16 to 40 nucleotides, identifying 287 conserved and 92 novel miRNAs. Furthermore, 10,313 potential target genes for fenugreek miRNAs were identified. Conserved and novel miRNAs were found targeting enzymes crucial for the biosynthesis pathways of terpenoids, diterpenoids, ubiquinones, carotenoids and flavonoids. These findings open the panorama for the use of transgenics mediated by miRNAs and the production of plant secondary metabolites, in addition to promoting industry applications at a commercial and medicinal level by improving the bioactivity and economic value of botanical resources such as fenugreek.