RESUMEN
The application of natural products for pest control is important in modern farming. In the present study, Artemisia santonicum L. and Artemisia lerchiana Weber essential oil and exudate profiles were determined, and their potential as inhibitors of seed germination, acetylcholinesterase, and phytopathogenic mycelium growth were evaluated. Essential oils (EO) were obtained via hydrodistillation and exudates (AE) by washing aerial parts of the species with acetone. EO and AE's composition was identified using GC/MS. Eucalyptol (1,8-cineole) and camphor were found to be the main components of A. lerchiana EO, while ß-pinene, trans-pinocarveol, α-pinene, α-terpineol, and spathulenol were established as major compounds of A. santonicum EO. Strong inhibition on Lolium perenne seed germination was found at 2 µL/mL and 5 mg/mL using aqueous solutions of EO and AE, respectively. An inhibitory effect on acetylcholinesterase was established, with an IC50 value of 64.42 and 14.60 µg/mL for EO and 0.961, >1 mg/mL for the AE of A. lerchiana and A. santonicum, respectively. The low inhibition on the mycelium growth of studied phytopathogenic fungi was established by applying 2 µL of EO and 15 µL of 100 mg/mL of AE, with the exception of A. lerchiana AE against Botrytis cinerea. These results show that the studied EO and AE exhibited strong phytotoxic and AChE inhibitory activities, providing new data for these species.
RESUMEN
The genus Phytophthora comprises many economically and ecologically important plant pathogens. Hybrid species have previously been identified in at least six of the 12 phylogenetic clades. These hybrids can potentially infect a wider host range and display enhanced vigour compared to their progenitors. Phytophthora hybrids therefore pose a serious threat to agriculture as well as to natural ecosystems. Early and correct identification of hybrids is therefore essential for adequate plant protection but this is hampered by the limitations of morphological and traditional molecular methods. Identification of hybrids is also important in evolutionary studies as the positioning of hybrids in a phylogenetic tree can lead to suboptimal topologies. To improve the identification of hybrids we have combined genotyping-by-sequencing (GBS) and genome size estimation on a genus-wide collection of 614 Phytophthora isolates. Analyses based on locus- and allele counts and especially on the combination of species-specific loci and genome size estimations allowed us to confirm and characterize 27 previously described hybrid species and discover 16 new hybrid species. Our method was also valuable for species identification at an unprecedented resolution and further allowed correct naming of misidentified isolates. We used both a concatenation- and a coalescent-based phylogenomic method to construct a reliable phylogeny using the GBS data of 140 non-hybrid Phytophthora isolates. Hybrid species were subsequently connected to their progenitors in this phylogenetic tree. In this study we demonstrate the application of two validated techniques (GBS and flow cytometry) for relatively low cost but high resolution identification of hybrids and their phylogenetic relations.
RESUMEN
During an investigation of rivers in Bulgaria, an isolate of Phytophthora (RLKam2016/61c) was recovered and initially identified as Phytophthora sansomeana based on internal transcribed spacer region (ITS) sequence data. However, the sequencing of the mitochondrial cytochrome c oxidase subunit I (cox1) gene revealed high similarity to Phytophthora sp. kelmania, and sequencing of other nuclear regions (ß-tubulin [Btub] and translation elongation factor 1-alpha [tef1]) revealed a significant number of polymorphisms, indicating a possible hybridization event. Additional cloning and sequencing of the nuclear ITS and Btub regions showed the presence of two distinct groups of alleles, one of which was highly similar to P. sansomeana, whereas the other was similar to a species complex that includes Phytophthora sp. kelmania. Therefore, the new hybrid was named Phytophthora × sansomeana. It is characterized by fast growth on V8 juice agar (V8A) and carrot agar (CA), moderate aerial mycelium with radiate pattern of the colonies and relatively slower growth rate on malt extract agar (MEA) and potato dextrose agar (PDA), and petaloid to rosaceous pattern of the colonies with fluffy aerial mycelium. The optimum growth temperature for P. × sansomeana was at 25 C, with an average growth rate of 9 mm per day. Abundant sporangium formation of the isolate in spring water was observed, but the hybrid was sterile in culture. Pathogenicity analyses of the hybrid were conducted in comparison with the most closely related subclade 8a species from our collection, P. pseudocryptogea. The inhibition effect on the root growth of young seedlings of two legumes, common pea and vetch, as well as on cuttings of the ornamental plant coleus induced by both phytopathogens was significant. No effect of either the new hybrid or P. pseudocryptogea on the growth of maize seedlings was observed.
