Potential resource materials from Ohio plants

Previously, the Northern Regional Research Center (NRRC) has studied chemical and botanical features of about 800 plant species in a program to identify potential renewable sources of industrial raw materials. In this program, another 64 species from northwestern and southwestern Ohio were studied for the present report. Aboveground samples were quantitatively analyzed for moisture, ash, crude protein, “oil,” “polyphenol,” and “hydrocarbon.” Plant oils were examined for classes of constituents. Oils were saponified and analyzed for yields of organic acids and unsaponifiable matter. Hydrocarbons were examined for the presence of rubber, gutta, and waxes. Rubber and gutta were analyzed for weight-average molecular weight and molecular weight distribution. Data are presented for 8 species of the 64 that gave the higher chemical yields.Rhus typhina gave outstanding yields of oil (6.5%) and polyphenol (30.8%) (moisture- plus ash-free basis).Liatris aspera yielded substantial amounts of oil (4.7%) and polyphenol (22.7%), whereasCornus racemosa yielded a substantial amount of oil (4.4%) but a more typical yield of polyphenol (11.0%).Nuphar advena andEpilobium angustifolium had considerable amounts of polyphenol (16.2, 16.3%) but little oil.Nuphar advena contained the most crude protein (24.8%). Noteworthy amounts of hydrocarbon were extracted fromCalamagrostis canadensis (1.1%),Aster umbellatus (0.8%), andSolidago riddellii (0.7%). Polyisoprenes in the hydrocarbon fractions of the latter 3 species were identified as gutta for C. canadensis and rubber forA. umbellatus andS. riddellii. Botanical features of the 8 species are briefly discussed.     

Plant species evaluated for new crop potential

About 500 plant species from various regions of the United States have been screened previously at the USDA Northern Regional Research Center for their multipurpose, energy-producing potential. Most collections have been from the rich flora of central Illinois. For this report, 92 additional species were collected from southern Illinois and evaluated by criteria previously established at this Center. Plant samples were analyzed for “oil,” “polyphenol,” “hydrocarbon,” and protein. Oil fractions of selected species were analyzed for classes of lipid constituents and were saponified to determine yields of unsaponifiable matter and fatty acids. Hydrocarbon fractions of selected species were analyzed for rubber, gutta, and waxes. Average molecular weight and molecular weight distribution of rubber and gutta were determined. of the 92 species, complete analytical data are presented for 16 selected species. Substantial quantities of oil were obtained fromPhiladelphias coronarius (5.0%; dry, ash-free sample basis),Cacalia muhlenbergii (4.1%),Lindera benzoin (4.1 %), andKoelreuteria paniculata (4.0%). High yields of polyphenol were obtained fromAcer ginnala (33.1%),Cornus obliqua (20.8%), andSalix caprea (20.0%). Maximum yields of hydrocarbon and protein were fromElymus virginicus (0.6%) andLindera benzoin (11.1%), respectively. Data are discussed with respect to species previously analyzed at this Center.     

Xerophytic species evaluated for renewable energy resources

Previously, the USDA Northern Regional Research Center has examined 600 plant species in 88 families for their multipurpose, energy-producing potential. About three-fourths of these species have been from central and southern Illinois, but only about 2% have been from arid or semiarid regions of the United States. For this report, 100 species collected from Arizona were evaluated, bringing the total number of species evaluated at this Center to 700 in 96 families. Plant specimens were analyzed for yields of fractions referred to as “oil,” “polyphenol,” “hydrocarbon,” and protein and were examined for botanical characteristics. Oil and hydrocarbon fractions of selected species were partially characterized. Ten species gave high yields of oil and/or polyphenol. For example,Asclepias linaria yielded 8.7% oil (dry, ash-free sample basis) + 11.7% polyphenol (1.9% hydrocarbon).Rhus choriophylla yielded 7.0% oil + 20% polyphenol (0.4% hydrocarbon) andJuglans major yielded 7.0% oil + 9.4% polyphenol (0.2% hydrocarbon).Pittosporum tobira gave the highest yield of hydrocarbon (23%). Fourteen species contained at least 18% protein. In general, the percentages of species yielding substantial amounts of oil and/or polyphenol were considerably higher for the Arizona species than for those 600 species previously analyzed. Complete analytical data are presented for 38 species and are discussed in relationship to the 600 species previously reported.     

New sources of gutta-percha in Garrya flavescens and G. wrightii

Analyses of extracts from whole-plant samples revealed that Garrya flavescens and Garrya wrightii produced gutta (trans-1,4-polyisoprene) as the major hydrocarbon in yields of 1.2 and 0.4 %, respectively. The average MWs of gutta were 142000 for G. flavescens and 10000 for G. wrightii. These species also yielded moderate amounts of ‘oil’ and ‘polyphenol’. To our knowledge, these are the first species of Garryaceae reported to produce gutta.     

Renewable resources from wild sunflowers (Helianthus spp., asteraceae)

Twenty-eight taxa of Helianthus collected throughout the United States and grown in afield nursery were evaluated for yields of oil, polyphenol hydrocarbon, protein, and ash in above-ground biomass. Hydrocarbons were examined for the presence of rubber, gutta, and waxes. Rubber and gutta were analyzed for weight-average molecular weight (ìw) and molecular weight distribution (MWD). Helianthus ciliaris had the highest oil yield (3.7%) and was analyzed for yield of fatty acids and unsaponifiable matter. Most taxa had low polyphenol yields (<10%), with H. strumosus having the highest (13.9%). Helianthus salicifolius yielded the most hydrocarbon (1.6%) and H. ciliaris had the highest protein content (13.4%). Natural rubber was present in 13 species of wild sunflowers. Helianthus maximiliani had the lowest ìw (29.8 x 10³), while H. laevigatus had the highest (73.3 x 10³). The MWD of rubber from sunflowers were within the range of those for commercial rubbers. The lower molecular weight rubbers may have potential as plasticizing additives in commercial processing of synthetic polyisoprenes and as hydrocarbon feedstock for a synthetic petroleum industry.     

Laticiferous taxa as a source of energy and hydrocarbon

Twenty-nine laticiferous taxa of Apocynaceae, Asclepiadaceae, and Sapotaceae were screened for suitability as alternative sources of renewable energy, rubber, and phytochemicals and to select the most promising ones for large-scale cultivation. Of these,Allamanda violacea (14.9% protein, 13.8% polyphenol, 8.6% oil, 3.2% hydrocarbon),Catharanthus roseus (15.4% protein, 10.4% polyphenol, 11.5% oil, 1.9% hydrocarbon), andHolarrhena antidysenterica (14.2% protein, 16.4% polyphenol, 5.4% oil, 4.8% hydrocarbon) of Apocynaceae;Asclepias curassavica (19.3% protein, 6.5% polyphenol, 3.9% oil, 2.0% hydrocarbon), Calotropis gigantea (18.5% protein, 6.8% polyphenol, 7.0% oil, 2.8% hydrocarbon) of Asclepiadaceae;Mimusops elengi (11.3% protein, 9.7% polyphenol, 7.2% oil, 4.0% hydrocarbon) of Sapotaceae show promising potential for future petrochemical plantations; of all these taxa,Holarrhena antidysenterica yielded an unusually high percentage (4.8%) of hydrocarbon fraction followed byMimusops elengi (4.0%). NMR spectra confirmed the presence of cis-polyisoprene in all species studied exceptNerium indicum (white-flowered var.). These data indicate that the majority of the species under investigation may be considered for large-scale cultivation as an alternative source of rubber, intermediate energy, and other phytochemicals.     

Combined application of botanical formulations and biocontrol agents for the management of Fusarium oxysporum f. sp. cubense (Foc) causing Fusarium wilt in banana

Plant products along with biocontrol agents were tested against Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense (Foc). Of the 22 plant species tested, the leaf extract of Datura metel (10%) showed complete inhibition of the mycelial growth of Foc. Two botanical fungicides, Wanis 20 EC and Damet 50 EC along with selected PGPR strains with known biocontrol activity, Pseudomonas fluorescens 1, Pf1 and Bacillus subtilis, TRC 54 were tested individually and in combination for the management of Fusarium wilt under greenhouse and field conditions. Combined application of botanical formulation and biocontrol agents (Wanis 20 EC + Pf1 + TRC 54) reduced the wilt incidence significantly under greenhouse (64%) and field conditions (75%). Reduction in disease incidence was positively correlated with the induction of defense-related enzymes peroxidase (PO) and polyphenol oxidase (PPO). Three antifungal compounds (two glycosides and one ester) in D. metel were separated and identified using TLC, RP-HPLC (Reverse Phase-High Pressure Liquid Chromatography) and mass spectrometry. In this study it is clear that combined application of botanical formulations and biocontrol agents can be very effective in the management of Fusarium wilt of banana.     

Fatty acids and sterols of selected hyphochytriomycetes and chytridiomycetes

The fatty acids and sterols of eight Chytridiomycetes and two Hyphochytriomycetes, and fatty acids of the OomycetePythium gracile, were analyzed by gas-liquid chromatography. In addition to the fatty acids anticipated for fungi, the two Hyphochytriomycetes (Hyphochytrium catenoides andRhizidiomyces apophysatus) and four of the Chytridiomycetes (Catenaria anguillulae, Blastocladiella emersonii, Monoblepharella sp., andAllomyces macrogynus) contained arachidonic acid as a major fatty acid of the polar lipid fraction, and this fatty acid was detected as a minor component ofRhizophlyctis rosea andSpizellomyces punctatum. Eicosapentaenoic acid constituted 4.6% of the polar lipid fatty acids inMonoblepharella sp., and trace amounts were detected in several other species. Both the gamma (ω-6) and alpha (ω-3) isomers of linolenic acid were detected in all of the species analyzed. Cholesterol was the predominant (≥73%) sterol ofB. emersonii, R. rosea, A. macrogynus, andChytridium confervae, and a minor (<12%) component ofC. anguillulae, andH. catenoides. The major sterols of the other species included lanosterol (C. anguillulae, 45%), stigmasta-5,22-dien-3β-ol (H. catenoides, 51%), 24-ethyl-cholesterol (S. punctatum, 38%;H. catenoides, 17%;Monoblepharella sp., 70%; andR. apophysatus, 84%), 24-methyl-cholesterol (H. catenoides, 23%;R. apophysatus, 14%;S. punctatum, 53%), and 24-methylene cholesterol (Rhizophydium sphaerotheca, 51%). Neither ergosterol nor fucosterol was detected in any of the species studied.     

Breakdown of plant cell biomass by filamentous fungi

Three species of filamentous fungi, Botrytis cinerea, Sporotrichum thermophile and Trichoderma viride, have been selected to assess the potential of utilizing filamentous fungi to degrade plant cell biomass produced by mass cell culture techniques. All three fungal species grew comparatively well on plant cell biomass with no requirement for supplementary nutrients. Of the three species assessed B. cinerea demonstrated the most growth. This species also produced the greatest yield of d-glucose. However, when culture conditions were modified, yields of d-glucose were markedly reduced indicating that the combination of species and culture conditions must be thoroughly investigated to ensure maximum product yield. The growth of filamentous fungi on plant cells also markedly affected the nature of the resulting fungal-plant cell residue, increasing the levels of soluble carbohydrates and essential amino acids with the largest increase in these materials being promoted by B. cinerea.     

Forage quality of native pastures in a Mediterranean area

Three field sites in the Hyblean region of Sicily were studied for their forage quality and botanical composition. A total of 70 plant species were analyzed. Another 36 were rare and of insufficient quantity for analysis. Forty-three bulk samples combining all existing species were also prepared and analyzed. The chemical analyses included were dry matter, ash, neutral-detergent fiber (NDF), acid-detergent fiber (ADF), lignin, total nitrogen, nonprotein nitrogen (NPN), soluble nitrogen and degradable protein. Quality varied over the growing season declining with forage maturity. The nutritional quality was high overall. Nongrass nonlegume species dominated. The most abundant species of high quality was a Compositae, Calendula arvensis, which represented approximately 22.8% of the available forage.     

DNA fingerprinting—A useful tool in the taxonomy of apomictic plant groups

DNA fingerprinting has proved useful for the analysis of genotypic distribution and estimation of genetic relatedness in apomictic plant groups likeRubus, Amelanchier, Hieracium andTaraxacum. Speciation through interspectific hybridization has been demonstrated in one case involving apomicticRubus species. The application of molecular markers to progeny derived from experimental cross-pollinations has yielded information on levels of apomixis as well as on its inheritance and regulation in e.g.Rubus, Poa, Pennisetum andTripsacum.     

Major Extractable Components in Asclepias linaria (Asclepiadaceae) and Ilex verticillata (Aquifoliaceae), Two Potential Hydrocarbon Crops

The major extractable components of two species identified as having high oil or polyphenol contents were characterized in detail.Asclepias linaria, a desert milkweed, contains 30.3% extractable material on a dry-weight basis, andIlex verticillata contains 41.5% extractable material on a dry-weight basis. Important components inA. linaria oil fractions are triterpene alcohols and esters, wax, and natural rubber; fatty acid triglycerides were nearly absent.Ilex verticillata oil fractions were predominantly triglycerides with some triterpene fatty acid esters. The more polar polyphenol fraction contained sugars and sugar esters of fatty acids and triterpene acids. The polyphenol fraction from these plants is better described as a saponin fraction. Because the crude saponin fraction represents 10.7% of the dry weight of A. linaria and 18.9% of the dry weight ofI. verticillata and because the saponin fractions showed good emulsifying properties, the refined extract of these plants might be used as a biodegradable surfactant.     

Correlation of diversity of rust fungi and their host plants with disturbance and conservation of vegetation in western Panama

The occurrence of rust fungi (Basidiomycota) infecting vascular plants on 14 sites in western Panama differing in altitude, climate, and vegetation type was analyzed. Three vegetation-types are investigated: (1) Disturbed sites where Asteraceae, Fabaceae, and Poaceae are most speciose, (2) slightly disturbed high canopy forests with sparse herbaceous undergrowth and no dominant plant family, and (3) relatively undisturbed forests with rich herbaceous undergrowth where Orchidaceae and ferns are most speciose. The majority of species of Pucciniales were found in disturbed sites on Asteraceae, Cyperaceae, Fabaceae, and Poaceae, whereas high canopy forests with sparse herbaceous layers yielded only few species of Pucciniales that were not common on any specific plant family. In undisturbed forests, a higher number of Pucciniales was found infecting mostly pteridophytes and Orchidaceae. Generally, undisturbed sites yielded a lower number of Pucciniales in relation to the number of plant species of which several species have a wide host range within the same site, and a low percentage of samples containing teliospores. Disturbed sites yielded a high number of Pucciniales in relation to the number of plant species, many rusts having a wide geographical distribution and a wide host range outside the sites, and a higher percentage of samples containing teliospores.     

Genetic Diversity in Old Portuguese Durum Wheat Cultivars Assessed by Retrotransposon-Based Markers

Retrotransposon-based markers, such as the inter-retrotransposon-amplified polymorphism (IRAP) and the retrotransposon microsatellite-amplified polymorphism (REMAP) are highly informative, multilocus, and reveal insertional polymorphisms among plant individuals. These markers have been used for evolutionary studies, genetic diversity assessment, DNA fingerprinting, genetic mapping linkage, and for the detection of genetic rearrangements induced by polyploidization. In this study, we used IRAP and REMAP markers to assess the genetic diversity among 51 old Portuguese durum wheat cultivars belonging to 27 botanical varieties and to define their genetic relationships. Five IRAP and four REMAP primer combinations were used. IRAP markers revealed 66.3% of polymorphism and an average of 18.4 bands per primer combination which ranged in size from 450 to 3,100?bp. The REMAP technique allowed the detection of 86.36% of inter-cultivar polymorphism and an average of 11 bands per primer combination. The molecular weight of the REMAP bands ranged from 250 to 2,750?bp. The durum wheat cultivars analyzed here belong to 27 botanical varieties of the subspecies Triticum turgidum subsp. turgidum L. [syn. T. turgidum] and Triticum turgidum L. subsp. durum [syn. T. durum] (Desf.) Husn.. Our results showed that the genetic variability assessed by both the IRAP and REMAP markers did not allow the clustering of the durum wheat cultivars according to their taxonomical criteria (subspecies or botanical variety) or homonymy. Nonetheless, these markers were useful for the assessment of genetic diversity at the individual level, for the definition of genetic relationships among cultivars, and for estimation of the genetic structure of the Old collection under analysis. The achieved data could be valuable for future experiments of DNA fingerprinting, genetic improvement, and germplasm conservation in wheat.     

Absorption,translocation, and metabolism of14C-clomazone in soybean (Glycine max) and threeAmaranthus weed species

The patterns of clomazone (2-[(2-chlorophenyl) methyl-4,4-dimethyl-3-isoxazolidinone) absorption, translocation, and metabolism and their contribution to the plant selectivity of this herbicide were studied in tolerant soybean [Glycine max (L.) Merr.] andAmaranthus hybridus and in susceptibleA. retroflexus andA. lividus. Differential root absorption appeared to play a significant role in the differential response of these four plant species to clomazone. Absorption of root-applied¹⁴C-clomazone was greater by the two sensitiveAmaranthus weeds than by the tolerant soybean andA. hybri-dus. Following application of¹⁴C-clomazone to roots, most of the absorbed radioactivity was translocated to the leaves of all four species. Approximately 50% of the absorbed¹⁴C-clomazone was metabolized by all four plant species as early as 12 h after treatment. Thin layer Chromatographic (TLC) analysis of plant tissue extracts from all four species revealed the formation of two major metabolites of clomazone. These unidentified metabolites had Rf values of 0.4 and 0.8, respectively, in a butanol∶acetic acid∶water (12∶3∶5, vol/vol/vol) developing system. The Rf value of unaltered clomazone in this system was 0.95. Differential metabolism or differential rate of metabolism of clomazone was not observed in this study and did not seem to account for the tolerance of soybean andA. hybridus or the suceptibility ofA. retroflexus andA. lividus to this herbicide.     

DNA barcoding in Atlantic Forest plants: What is the best marker for Sapotaceae species identification?

The Atlantic Forest is a phytogeographic domain with a high rate of endemism and large species diversity. The Sapotaceae is a botanical family for which species identification in the Atlantic Forest is difficult. An approach that facilitates species identification in the Sapotaceae is urgently needed because this family includes threatened species and valuable timber species. In this context, DNA barcoding could provide an important tool for identifying species in the Atlantic Forest. In this work, we evaluated four plant barcode markers (matK, rbcL, trnH-psbA and the nuclear ribosomal internal transcribed spacer region - ITS) in 80 samples from 26 species of Sapotaceae that occur in the Atlantic Forest. ITS yielded the highest average interspecific distance (0.122), followed by trnH-psbA (0.019), matK (0.008) and rbcL (0.002). For species discrimination, ITS provided the best results, followed by matK, trnH-psbA and rbcL. Furthermore, the combined analysis of two, three or four markers did not result in higher rates of discrimination than obtained with ITS alone. These results indicate that the ITS region is the best option for molecular identification of Sapotaceae species from the Atlantic Forest.     

Germinating seeds or bulbils in 87 of 113 tested Arctic species indicate potential for ex situ seed bank storage

Arctic plant species are expected to lose range due to climate change. One approach to preserve the genetic and species diversity for the future is to store propagules in seed vaults. However, germinability of seeds is assumed to be low for Arctic species. We evaluated ex situ storage potential of 113 of the 161 native angiosperms of Svalbard by studying seed ripening and germination. Seeds or bulbils were collected, and germinability was tested after one winter of storage in the Svalbard Global Seed Vault. Twenty-six of the species did not produce ripe propagules, 8 produced bulbils, and 79 produced seeds. Bulbils sprouted to high percentages. Seeds of 10 species did not germinate, 22 had low germination (<20 %), 34 had germination of 21–70 %, and 13 had high germination percentages (>70 %). More than 70 % of the species belonging to Asteraceae, Brassicaceae, Caryophyllaceae, Juncaceae, Rosaceae, and Saxifragaceae germinated. Cold tolerant, common species had higher germination percentages than relatively thermophilous, rare species. Germination percentages were six times higher than observed in 1969 (n = 51) and 0.7 times that observed in 2008 (n = 22), indicating that recent climate warming improves germination in the Arctic. While in situ conservation is of vital importance, ex situ conservation in seed banks is a potential complementary conservation strategy for the majority of Arctic vascular plant species. For species that did not germinate, other methods for ex situ conservation should be sought, for example, growing in botanical gardens.     

Variation in small-scale spatial heterogeneity of soil properties and vegetation with different land use in semiarid grassland ecosystem

Soil properties (i.e. soil organic carbon, SOC; soil organic nitrogen, SON; and soil C/N ratio) and vegetation in a semiarid grassland of Inner Mongolia, northern China, were studied with the method of geostatistical analysis. We examined the spatial heterogeneity of soil and plants, and possible impacts of land use on their heterogeneity and on the relationship between soil resources and plant richness. Land use affected small scale spatial heterogeneity in plants and soil. SOC, SON and C/N ratio displayed autocorrelation over a range of ～2 m under most circumstances on sites where livestock grazing had been excluded. The uncontrolled grazing site (UG, i.e. unregulated grazing by excessive livestock) displayed an increased range of spatial autocorrelation and the total amount of variability in soil nitrogen over the other land use types. Plant life forms and plant species exhibited spatial autocorrelation over a range of about 2 m on the grazing exclusion (GE) and mowed (MW) sites, while pattern of spatial autocorrelation for several less common species on the UG site were difficult to predict. Plant species richness was positively related with spatial heterogeneity of SOC, SON and C/N on both GE and MW sites, and with only SOC heterogeneity on the UG site. These suggest that spatial soil heterogeneity plays an active role in maintaining plant species richness. However, we call for caution in generalization of the control of spatial soil heterogeneity over plant richness when multiple modes of disturbances are present, as we found in this study that higher total amount of variation in soil nitrogen and C/N ratio on the over-grazed UG site did not lead to increased plant species richness, and that land use had apparent effects on the patterns of spatial heterogeneity in both vegetation and soil.     

Purification and some properties of polyphenol oxidase from the yam tubers, Dioscorea bulbifera

In a comparison of the polyphenol oxidase activity of various species of yam tubers the greatest enzyme activity was found in D. bulbifera. The enzyme was purified from acetone powder extracts of this plant. Ammonium sulphate fractionation, followed by ion exchange chromatography and gel filtration gave 22-fold purification. The final product gave a single band on polyacrylamide disc gel electrophoresis. The purified enzyme showed activity towards catechol, pyrogallol and dl-β-3,4-dihydroxyphenylalanine (dl-DOPA) and had a MW 115000 ± 2000. It was characterized by response to various inhibitors. β-Mercaptoethanol, dithioerythritol, l-cysteine, sodium metabisulphite and KCN inhibited strongly.     

Salicaceae-Feeding Leaf-Mining Insects in Siberia: Distribution,Trophic Specialization,and Pest Status

This paper provides an overview of the leaf-mining insect community feeding on willows (Salix spp.) and poplars (Populus spp.) in Siberia. According to published data and our own observations, 50 leaf-mining insect species (i.e., 24 species of Lepidoptera, 15 Coleoptera, 6 Diptera, and 5 Hymenoptera) feed on those two plant genera in Siberia. Using an integrative approach combining field work, morphological and DNA barcoding analyses, we identified 32 leaf-mining insect species from 14 regions across Siberia (i.e. 64% of all leaf-mining species known on Salicaceae in this part of Russia). Among them, 26 species most often found in parks and botanical gardens, represented new faunistic records for several poorly explored regions of Siberia. We have more than doubled the list of Salicaceae-feeding leaf-mining insects in Tomsk oblast, Altai krai, and the Republic of Tuva, and for the first time provided data on leaf-miners for the Khanty–Mansi Autonomous Okrug. The micromoth Phyllocnistis gracilistylella (Gracillariidae), recently described from Japan, was found on a new host plant (Salix caprea) in the south of Krasnoyarsk krai, is new for Russia. Eight leafmining insect species (i.e., five gracillariids: Phyllocnistis labyrinthella, Ph. unipunctella, Phyllonorycter apparella, Ph. sagitella, and Ph. populifoliella; two beetles: Zeugophora scutellaris and Isochnus sequensi; and one sawfly: Heterarthrus ochropoda) can outbreak on poplars, most often in urban plantations, botanical gardens, and plant nurseries in Siberia, and can also affect natural stands. Forty-five species of 50 leaf-mining insects known to feed on willow and poplar in Siberia also occur in Central and Eastern Europe. The remaining five species (Phyllocnistis gracilistylella, Phyllonorycter sibirica, Heterarthrus fasciatus, Tachyerges dauricus, and Isochnus arcticus) are recorded in Asia only. Species richness of the family Gracillariidae, the most diverse on Salicaceae in Siberia, displays 80% similarity to that in the European part of Russia and 71% to the Russian Far East. We discuss the faunal similarity of these regions and highlight the importance of applying an integrative approach combining ecological, morphological analyses, and DNA barcoding to explore and characterize the insect fauna of poorly studied regions of Asian part of Russia.