Search Results (12)

Six novel green extraction techniques were evaluated and optimized to extract pyrethrin from dried Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir./Sch.Bip.). This approach offers a promising natural alternative to conventional chemotherapeutics. Four methods are presented for the first time in this study: microwave-assisted extraction (MAE), high-voltage electric discharge (HVED) extraction, subcritical water extraction (SWE), and deep eutectic solvent (DES) extraction, together with supercritical CO₂ extraction (SC-CO₂) and ultrasound-assisted extraction (UAE), for pyrethrin extraction from Dalmatian pyrethrum. The study revealed that supercritical CO₂ extraction was the most effective method for extracting all six pyrethrins, yielding the highest total amount of 124.37 ng/mg. This approach offers a “natural” insecticide produced with a clean, environmentally friendly technology that can contribute to the development of sustainable and effective insecticide strategies that are in line with environmental safety and organic production standards. In addition, this research highlights the potential application of pyrethrins as antiparasitic agents, emphasizing their role in environmentally friendly and ecological practices. Full article

Tanacetum cinerariifolium, a perennial Asteraceae plant, is renowned for its ornamental value and natural insecticidal compounds, especially pyrethrins. These compounds, primarily stored in flower heads, are highly effective as insecticides with low toxicity to mammals, making them crucial for organic agriculture, along with the sesquiterpene (E)-β-farnesene (EβF), play critical roles in T. cinerariifolium defense mechanisms. However, the spatiotemporal patterns of these secondary metabolites in stems and flower heads, as well as their regulatory mechanisms, remain unclear. This study investigated the biosynthesis and regulation of pyrethrins and EβF across developmental stages (S1–S4) in flowers and stems using GC-MS and transcriptomics. Transcriptome analysis revealed that the expression of pyrethrin biosynthetic genes was not synchronized with pyrethrin accumulation. The main pyrethrin biosynthetic genes exhibited coordinated expression patterns, peaking during early flowering stages (S1–S2), while pyrethrin accumulation was primarily observed during mid-flower development. In contrast, the biosynthetic genes of EβF showed synchronized expression with EβF accumulation, with the highest activity observed in stems and early flowers. WGCNA identified jasmonic acid signaling, trichome differentiation, and terpene transport pathways as potentially associated with pyrethrin biosynthesis. Hub genes including MYC2 were identified as playing pivotal roles in regulating secondary metabolite biosynthesis. These findings provide new insights into the regulation and biosynthesis of pyrethrins and EβF, offering a foundation for optimizing bioactive compound production and advancing sustainable pest management strategies. Full article

Growing demands for environmentally safe and sustainable pest management have increased interest in biopesticides as alternatives to synthetic chemical pesticides. This review presents the current status of bioacaricides, defined as commercial biopesticide products based on microorganisms (microbial acaricides) and biologically active substances of microbial, plant or animal origin (biochemicals and semiochemicals) used in crop protection against spider mites (Tetranychidae) and other plant-feeding mites. The most important microbial bioacaricides are mycopesticides, which are products manufactured from living propagules of Beauveria bassiana s.l. and several other acaropathogenic fungi. Products based on avermectins and milbemycins, secondary metabolites of actinomycetes, are well-known examples of biochemicals of microbial origin. Among the biochemicals of plant origin, the most widely used to date have been the products based on pyrethrum—obtained from the Dalmatian daisy, Tanacetum cinerariifolium (Asteraceae)—and azadirachtin, obtained from the Indian neem tree, Azadirachta indica (Meliaceae). In recent years, products based on essential oils from aromatic plants belonging to the families Lamiaceae, Myrtaceae, Rutaceae and others have also gained increasing importance in the market. Special emphasis in this review is given to the compatibility of bioacaricides with predatory mites of the family Phytoseiidae as biological control agents used in the integrated management of plant-feeding mites. Full article

The Asteraceae family is a promising source of bioactive compounds, such as the famous Asteraceae plants Tanacetum cinerariifolium (pyrethrin) and Artemisia annua (artemisinin). As a result of our series of phytochemical studies of the subtropical plants, two novel sesquiterpenes, named crossoseamines A and B in this study (1 and 2, respectively), one undescribed coumarin-glucoside (3), and eighteen known compounds (4–21) were isolated from the aerial part of Crossostephium chinense (Asteraceae). The structures of isolated compounds were elucidated by spectroscopic methods, including 1D and 2D NMR experiments (¹H, ¹³C, DEPT, COSY, HSQC, HMBC, and NOESY), IR spectrum, circular dichroism spectrum (CD), and high-resolution electrospray ionization–mass spectrometry (HR-ESI–MS). All isolated compounds were evaluated for their cytotoxic activities against Leishmania major, Plasmodium falciparum, Trypanosoma brucei (gambiense and rhodesiense), and human lung cancer cell line A549 because of the high demand for the discovery of new drug leads to overcome the present side effects and emerging drug-resistant strains. As a result, the new compounds (1 and 2) showed significant activities against A549 (IC₅₀, 1: 3.3 ± 0.3; 2: 12.3 ± 1.0 μg/mL), L. major (IC₅₀, 1: 6.9 ± 0.6; 2: 24.9 ± 2.2 μg/mL), and P. falciparum (IC₅₀, 1: 12.1 ± 1.1; 2: 15.6 ± 1.2 μg/mL). Full article

Natural pyrethrins have high application value, and are widely used as a green pesticide in crop pest prevention and control. Pyrethrins are mainly extracted from the flower heads of Tanacetum cinerariifolium; however, the natural content is low. Therefore, it is essential to understand the regulatory mechanisms underlying the synthesis of pyrethrins through identification of key transcription factors. We identified a gene encoding a MYC2-like transcription factor named TcbHLH14 from T. cinerariifolium transcriptome, which is induced by methyl jasmonate. In the present study, we evaluated the regulatory effects and mechanisms of TcbHLH14 using expression analysis, a yeast one-hybrid assay, electrophoretic mobility shift assay, and overexpression/virus-induced gene silencing experiments. We found that TcbHLH14 can directly bind to the cis-elements of the pyrethrins synthesis genes TcAOC and TcGLIP to activate their expression. The transient overexpression of TcbHLH14 enhanced expression of the TcAOC and TcGLIP genes. Conversely, transient silencing of TcbHLH14 downregulated the expression of TcAOC and TcGLIP and reduced the content of pyrethrins. In summary, these results indicate that the potential application of TcbHLH14 in improving the germplasm resources and provide a new insight into the regulatory network of pyrethrins biosynthesis of T. cinerariifolium to further inform the development of engineering strategies for increasing pyrethrins contents. Full article

Pyrethrum (Tanacetum cinerariifolium) cultivation in Australia, which accounts for the majority of global production of natural insecticidal pyrethrins, is affected by a persistent yield decline which in part is caused by a complex of pathogens. Globisporangium and Pythium species were isolated from crown and roots of pyrethrum plants showing stunting and brown discoloration of crown tissue, and from soil adjacent to diseased plants from yield-decline-affected sites in Tasmania and Victoria, Australia. Ten known Globisporangium species (Globisporangium attrantheridium, G. erinaceum, G. intermedium, G. irregulare, G. macrosporum, G. recalcitrans, G. rostratifingens, G. sylvaticum, G. terrestris and G. ultimum var. ultimum), two new Globisporangium species (Globisporangium capense sp. nov. and Globisporangium commune sp. nov.) and three Pythium species (Pythium diclinum/lutarium, P. tracheiphilum and P. vanterpoolii) were identified through morphological studies and multigene phylogenetic analyses using ITS and Cox1 sequences. Globisporangium ultimum var. ultimum, G. sylvaticum, G. commune sp. nov. and G. irregulare were most abundant. Globisporangium attrantheridium, G. macrosporum and G. terrestris were reported for the first time in Australia. Seven Globisporangium species were pathogenic on both pyrethrum seeds (in vitro assays) and seedlings (glasshouse bioassays), while two Globisporangium species and three Pythium species only caused significant symptoms on pyrethrum seeds. Globisporangium irregulare and G. ultimum var. ultimum were the most aggressive species, causing pyrethrum seed rot, seedling damping-off and significant plant biomass reduction. This is the first report of Globisporangium and Pythium species causing disease in pyrethrum globally and suggests that oomycete species in the family Pythiaceae may have an important role in the yield decline of pyrethrum in Australia. Full article

The Gly-Asp-Ser-Leu (GDSL) motif of esterase/lipase family proteins (GELPs) generally exhibit esterase activity, whereas transferase activity is markedly preferred in several GELPs, including the Tanacetum cinerariifolium GDSL lipase TciGLIP, which is responsible for the biosynthesis of the natural insecticide, pyrethrin I. This transferase activity is due to the substrate affinity regulated by the protein structure and these features are expected to be conserved in transferase activity-exhibiting GELPs (tr-GELPs). In this study, we identified two amino acid residues, [N/R]208 and D484, in GELP sequence alignments as candidate key residues for the transferase activity of tr-GELPs by two-entropy analysis. Molecular phylogenetic analysis demonstrated that each tr-GELP is located in the clusters for non-tr-GELPs, and most GELPs conserve at least one of the two residues. These results suggest that the two conserved residues are required for the acquisition of transferase activity in the GELP family. Furthermore, substrate docking analyses using ColabFold-generated structure models of both natives and each of the two amino acids-mutated TciGLIPs also revealed numerous docking models for the proper access of substrates to the active site, indicating crucial roles of these residues of TciGLIP in its transferase activity. This is the first report on essential residues in tr-GELPs for the transferase activity. Full article

Pyrethrins are a mixture of terpenes, with insecticidal properties, that accumulate in the aboveground parts of the pyrethrum (Tanacetum cinerariifolium). Numerous studies have been published on the positive role of MYB transcription factors (TFs) in terpenoid biosynthesis; however, the role of MYB TFs in pyrethrin biosynthesis remains unknown. Here, we report the isolation and characterization of a T. cinerariifolium MYB gene encoding a R3-MYB protein, TcMYB8, containing a large number of hormone-responsive elements in its promoter. The expression of the TcMYB8 gene showed a downward trend during the development stage of flowers and leaves, and was induced by methyl jasmonate (MeJA), salicylic acid (SA), and abscisic acid (ABA). Transient overexpression of TcMYB8 enhanced the expression of key enzyme-encoding genes, TcCHS and TcGLIP, and increased the content of pyrethrins. By contrast, transient silencing of TcMYB8 decreased pyrethrin contents and downregulated TcCHS and TcGLIP expression. Further analysis indicated that TcMYB8 directly binds to cis-elements in proTcCHS and proTcGLIP to activate their expression, thus regulating pyrethrin biosynthesis. Together, these results highlight the potential application of TcMYB8 for improving the T. cinerariifolium germplasm, and provide insight into the pyrethrin biosynthesis regulation network. Full article

Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevis.) Sch. Bip.) is an outcrossing plant species (2n = 18) endemic to the eastern Adriatic coast and source of the natural insecticide pyrethrin. Due to the high repeatability and large genome (1C-value = 9.58 pg) our previous attempts to develop microsatellite markers using the traditional method were unsuccessful. Now we have used Illumina paired-end whole genome sequencing and developed a specific procedure to obtain useful microsatellite markers. A total of 796,130,142 high-quality reads (approx. 12.5× coverage) were assembled into 6,909,675 contigs using two approaches (de novo assembly and joining of overlapped pair-end reads). A total of 31,380 contigs contained one or more microsatellite sequences, of which di-(59.7%) and trinucleotide (25.9%) repeats were the most abundant. Contigs containing microsatellites were filtered according to various criteria to achieve better yield of functional markers. After two rounds of testing, 17 microsatellite markers were developed and characterized in one natural population. Twelve loci were selected for preliminary genetic diversity analysis of three natural populations. Neighbor-joining tree, based on the proportion of shared alleles distances, grouped individuals into clusters according to population affiliation. The availability of codominant SSR markers will allow analysis of genetic diversity and structure of natural Dalmatian pyrethrum populations as well as identification of breeding lines and cultivars. Full article

The plant Tanacetum coccineum (painted daisy) is closely related to Tanacetum cinerariifolium (pyrethrum daisy). However, T. cinerariifolium produces large amounts of pyrethrins, a class of natural insecticides, whereas T. coccineum produces much smaller amounts of these compounds. Thus, comparative genomic analysis is expected to contribute a great deal to investigating the differences in biological defense systems, including pyrethrin biosynthesis. Here, we elucidated the 9.4 Gb draft genome of T. coccineum, consisting of 2,836,647 scaffolds and 103,680 genes. Comparative analyses of the draft genome of T. coccineum and that of T. cinerariifolium, generated in our previous study, revealed distinct features of T. coccineum genes. While the T. coccineum genome contains more numerous ribosome-inactivating protein (RIP)-encoding genes, the number of higher-toxicity type-II RIP-encoding genes is larger in T. cinerariifolium. Furthermore, the number of histidine kinases encoded by the T. coccineum genome is smaller than that of T. cinerariifolium, suggesting a biological correlation with pyrethrin biosynthesis. Moreover, the flanking regions of pyrethrin biosynthesis-related genes are also distinct between these two plants. These results provide clues to the elucidation of species-specific biodefense systems, including the regulatory mechanisms underlying pyrethrin production. Full article

Pyrethrum (Tanacetum cinerariifolium) is one of the most important industrial crops for the extraction of pyrethrins, which are natural insecticidal compounds. Progress in pyrethrum molecular breeding with the objective of increasing pyrethrin content has been slow for lack of a suitable gene transfer system. Regeneration recalcitrance is a crucial barrier to establishing a genetic transformation system in pyrethrum. Therefore, in this study, an Agrobacterium-mediated transformation system in pyrethrum was developed using shoot apical meristems from germinated seedlings. Factors affecting transformation efficiency were optimized. Optimal conditions included explants at the “no true leaf” stage with a half apical meristem, an Agrobacterium tumefaciens cell density of OD₆₀₀ = 0.5, two days of cocultivation, and the incorporation of 1.5 mg L⁻¹ 6-BA and 30 mg L⁻¹ kanamycin into the selection medium. Under the optimized conditions, two expression cassettes (proTcCHS-GUS and proRbcS-TcCHS) were successfully transformed into pyrethrum. Polymerase chain reaction (PCR), Southern blotting, reverse-transcription quantitative PCR (RT-qPCR), and histochemical staining confirmed the identity of proTcCHS-GUS transgenic plants. PCR and RT-qPCR analyses confirmed the identity of proRbcS-TcCHS transgenic plants. The transformation efficiency was 0.83% (5 transgenic lines/600 infected explants). The relative concentration of pyrethrins in proRbcS-TcCHS transformants (OX T0-1: 1.50% or OX T0-2: 1.24%) was higher than that in nontransformed plants (WT: 0.76%). Thus, the genetic transformation system overcame the low regeneration efficiency and integrated a foreign gene into the pyrethrum genome. The new system is a suitable and effective tool for creating high-yielding cultivars of pyrethrum. Full article

Pyrethrin is a specialized metabolite of Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip.), Asteraceae, known worldwide as an effective bioinsecticide. It consists of six active compounds: Pyrethrin I and II, cinerin I and II, and jasmolin I and II. Pyrethrin accumulates mainly in the flower heads and its content depends on numerous factors, such as the flower developmental stage. This study aims to investigate the accumulation patterns of six pyrethrin compounds in the flower heads of Dalmatian pyrethrum over the eight developmental stages (FS1 to FS8), and to make a comparison in six natural populations. Ultrasound assisted extraction was used to extract the pyrethrin, while qualitative and quantitative analysis was performed by High performance liquid chromatography. The accumulation patterns of different pyrethrin compounds were generally similar and also synchronous between different populations, while the pyrethrin I/pyrethrin II ratio showed irregular patterns. In all populations studied, the highest increase of all compounds was observed from FS1 to FS2. Their concentration continuously increased, reaching the highest values at FS4 stage (2–5 rows of open disc flowers), and generally decreased gradually towards FS6 or FS7 and then stagnated until the FS8 stage. Despite the very similar accumulation pattern of pyrethrin compounds in the different populations, the significant differences in their content suggest a different genetic background. Knowledge of the dynamics of pyrethrin compounds accumulation across flower development stages is valuable for determining the optimal harvest time of pyrethrum flower heads. Full article