Bolitophilidae, three species of Diadocidiidae, two species of Ditomyiidae, 34 species of Keroplatidae and 202 species of Mycetophilidae. 230 and 188 species are recorded from Georgia and the whole of Transcaucasia for the first time, respectively. Three new species – Sciophilageorgei sp. nov., Leiakatae sp. nov. and Anatellametae sp. nov. – are described including detailed illustrations of the male terminalia. Photographs are provided for an additional 38 species to highlight a variability of their general facies. Combined with earlier published data, the number of fungus gnat species in Georgia is set at 246. The estimated diversity of fungus gnats in Georgia is calculated using non-parametric methods and discussed with respect to other Western Palaearctic regions.The fungus gnats of Georgia are studied based on 2682 specimens collected from 57 localities during 2011–2019. Altogether, 245 species are recorded including four species of  Diptera  species from Marelisi, NW of Borjomi. Later on, Transcaucasia, the area southwards from the Greater Caucasus Mountains that includes the countries of Georgia, Azerbaijan and Armenia, is considered one of the biodiversity hotspots of the world, with a remarkable number of endemic species . Howevererbaijan  and seveerbaijan . From Geerbaijan  prior toDuring the last decade, a considerable amount of fungus gnat material from Georgia has accumulated in the author’s possession. The aim of the current contribution is to provide results of the study based on that material along with summarising all available published information on Georgian fungus gnats.The material was collected from 2011 to 2019 using different methods in the course of 61 collecting events from 57 localities in Georgia , Tartu, Estonia;ZFMKZoological Research Museum Alexander Koenig, Bonn, Germany.IZBE which is not repeated in the species list for every specimen. However, the material collected by X. Mengual  in 2019 is divided between three institutions and the depository is specified in listed material sections below.The majority of the studied specimens are deposited in Bolitophilidae, three species of Diadocidiidae, two species of Ditomyiidae, 34 species of Keroplatidae and 202 species of Mycetophilidae including three species described as new to science. One additional species of Keroplatidae was included from the literature data . 230 and 188 species are recorded from Georgia and the whole Transcaucasia for the first time, respectively. In the species list, all available literature sources are cited for the species recorded earlier in Georgia and/or in Transcaucasia generally. The studied material is listed, using abbreviations of collecting events provided in Table http://www.sciaroidea.info/) except in two cases. Firsty, the subfamily Platyurinae of Keroplatidae is used according to Brachycampta Winnertz, 1863 is reinstated to the generic status from a subgenus of Allodia Winnertz, 1863 in accordance with a thorough study by Altogether, 2682 studied specimens were identified to 245 different species, viz. four species of ure data . MoreoveTaxon classificationAnimaliaDipteraMycetophilidaeB92821CC-8394-5DDE-8E2F-3199603BAFBChttp://zoobank.org/D9E0ED72-E487-480C-A89F-4E6DDD98C406Holotype. Male, Georgia, Kakheti, Lagotekhi NR, Matsimi river gorge, 41°47.75'N, 46°17.12'E, 350 m a.s.l., 17–27.v.2011, Malaise trap, leg. G. Japoshvili [see Table IZBE). Paratype. Male, same as holotype .Sciophilageorgei sp. nov. can be distinguished by combination of the characters of the male terminalia as follows: lateral branch of gonostylus ventrally with two apical spine-like setae, small internal branch of gonostylus with one spine-like seta, tergite 9 large with medially rising apical margin that bears two large and simple setae, parameres straight and long, extending over tergite 9 apically, aedeagus small, star-shaped.Male. Body length 2.7–2.8 mm (n = 2). Coloration. Head with vertex and frons dark brown, face and clypeus brown and mouthparts including palpus pale yellow. Scape and pedicel yellow. First three or four flagellomeres yellowish, rest of flagellomeres light brown. Scutum entirely dark brown, antepronotum and proepisternum yellowish, anepisternum, anepimeron and katepisternum light brown, laterotergite and mediotergite brown, scutellum brown. Thoracic setae all yellowish. Wing hyaline, all veins brown including radial veins somewhat darker. Halter with stem and knob pale yellow. All coxae, femora and tibiae yellow, tarsi yellow but seem darker because of dense brown setae. Tibial setae brown, spurs yellowish. Abdomen with tergites light brown, 1–3 tergites somewhat lighter, all sternites yellowish. Abdominal vestiture yellow. Terminalia brown. Head. Ocelli in a shallow triangular arrangement. Medial ocellus somewhat smaller than laterals. Frontal furrow complete. Clypeus subrounded, about as long as broad. Fourth flagellar segment about as long as wide, apical flagellar segment 2.25 times as long as wide basally. Flagellar segments with dense yellowish short setae. Thorax. Scutum covered with short setae, with marginal and prescutellar setae stronger. Antepronotum with 8–9 setae. Proepisternum with 6–7 setae. Anepisternum with 5–6 setae on upper part, katepisternum and anepimeron non-setose. Laterotergite with 7–9 setae on posterior half. Mediotergite with 10–15 setae on lower part. Metepisternum with setulae. Scutellum with setulae and marginal setae not arranged in pairs. Wing. Length 2.5–2.8 mm, length to width 2.4–2.7. Wing membrane uniformly covered with micro- and macrothichia. All veins setose, except sc-r, Rs, 2+3R. Costa reaches about one fifth from 4+5R to 1M. Sc ending on C before level of furcation of posterior fork. Sc-r located slightly before Rs. r-m about two times as long as m-stem. 4M basally very faint or shortly interrupted at base. Legs. Ratio of femur to tibia for fore, mid and hind legs: 0.83–0.93; 0.89–0.97; 0.84–0.92. Ratio of tibia to basitarsus for fore, mid and hind legs: 1.26; 1.42–1.65; 1.33–1.37. Fore tibia with a spur 2.29–2.81 times of tibial maximum width. Mid tibia with anterior spur 3.08–3.15 times and posterior spur 3.42–3.69 times of tibial maximum width. Hind tibia with anterior spur 2.50–2.60 times and posterior spur 3.47–3.57 times of tibial maximum width. Terminalia  in recognition of his contribution to study of the insects’ diversity in Georgia and his invaluable help in collecting the fungus gnat material that underlies the current communication. He was also the collector of the type material of this species.Sciophila Meigen are known wordwide  and parameres .More than 190 species of wordwide ; the moswordwide  that proTaxon classificationAnimaliaDipteraMycetophilidae69C08F18-98A3-5FC4-B902-31C0E6026560http://zoobank.org/FDD299DF-4281-4DD0-9BAC-8B6050D98226Holotype. Male, Georgia, Shida-Kartli, W of Surami, 42°01.57'N, 43°29.88'E, 940 m a.s.l., 18.v.2013, sweeping, leg. O. Kurina [see Table IZBE). Paratype. Male, GEORGIA, Samegrelo-Zemo-Svanethi, near Ushguli, path to glacier, 42°56.62'N, 43°03.23'E, 2220 m a.s.l., 15–17.vi.2019, Malaise trap, leg. X. Mengual [see Table ZFMK)Leiakatae sp. nov. can be distinguished by the combination of characters as follows: thorax bicolored , wing tinged yellowish, with faint preapical brownish band, male terminalia with bipartite gonostylus .Male. Body length 6.7–6.9 mm (n = 2). Coloration. Head with vertex brown, frons yellow, face, clypeus and mouthparts including palpus pale yellow. Scape and pedicel pale yellow. First two flagellomeres yellowish, flagellomeres 3–14 brown. Thorax bicoloured: scutum yellow with three brown longitudinal stripes, which are posteriorly completely fused, lateral stripes begin at a distance of one third from anterior margin, medial stripe shortly split anteriorly, lateral parts of scutum yellow; antepronotum, proepisternum and anepisternum yellow, posterior margin of anepimeron light brown, katepisternum with lower half brown and upper half yellow, laterotergite brown with posterior half yellowish, mediotergite brown, scutellum basally yellowish, apically brown. Thoracic setae all yellow. Wing with yellowish tinge and preapical very faint transverse brownish band reaching to 2M, all veins yellowish including radial veins somewhat darker. Halter with stem and knob pale yellow. All coxae and femora yellow, except hind femur apically with narrow brown band, all tibiae, tarsi yellow but seem darker because of dense brown setae. Tibial setae brown, spurs yellowish. Abdomen with all tergites brown and sternites yellow. Abdominal vestiture yellow. Terminalia brown with gonocoxite medially and gonostylus anteriorly yellow. Head. Ocelli in a linear arrangement. Medial ocellus about twice smaller than laterals, which are separated from eye margins by less than their own diameter. Frontal furrow complete. Clypeus obovoid. Fourth flagellar segment about as long as wide, apical flagellar segment 2.5 times as long as wide basally. Flagellar segments with dense yellowish short setae. Thorax. Scutum densely covered with setae, with marginal and prescutellar setae stronger. Antepronotum with 6–7 strong and a number of weaker setae. Proepisternum with one very strong seta at anterior margin about 10 weaker setae. Anepisternum, katepisternum and anepimeron non-setose. Laterotergite with long fine setae on posterior half. Mediotergite non-setose. Scutellum with a row of marginal setae including two pairs remarkably stronger. Wing. Length 5.3–5.7 mm, length to width 2.3–2.8. All veins setose, except Sc, sc-r, Rs and extreme base of 1M. Sc ending on C at level of furcation of posterior fork. 4+5R 3.3 times as long as R1. r-m 1.47 times as long as m-stem. 1M and 2M apically convergent, apical third of both veins faint. 4M interrupted at base. Rs located distally of the anterior fork. Legs. Ratio of femur to tibia for fore, mid and hind legs: 1.16; 0.95; 0.86. Ratio of tibia to basitarsus for fore, mid and hind legs: 1.00; 1.66; 2.00. Fore tibia with a spur 2.95 times of tibial maximum width. Mid tibia with anterior spur 3.33 times and posterior spur 3.96 times of tibial maximum width. Hind tibia with anterior spur 3.33 times and posterior spur 4.58 times of tibial maximum width. Terminalia , an enthusiastic student of biology at the University of Tartu (Estonia). She participated in a trip to Georgia in 2017 that yielded several specimens of this study and she always insists we call her Kata.Leia Meigen species known worldwide including 33 in the Palaearctic region . Paratype. Male, same as holotype .Anatellametae sp. nov. is characterized by the presence of a strong posteroventral fringe on mid femora with a row of strong setae, absence of anterior spur on mid tibia, absence of setae on hind coxa basally. The new species is closest to A.atlanticiliata Chandler and Ribeiro but differs in characters of the male terminalia: ventral branch of the gonostylus about twice as long as the dorsal branch, dorsal branch of the gonostylus with long and slender medial prong, medial branch of the gonostylus slender and apically hooked.Male. Body length 2.7–2.9 mm (n = 2). Coloration. Head with vertex, frons, face and clypeus brown, mouthparts including palpus pale yellow. Scape, pedicel and base of first flagellomere yellow, rest of flagellum light brown. Thorax with scutum and lateral parts light brown. Thoracic setae yellowish to brown, with thicker setae darker than finer ones. Wing hyaline, unmarked with yellowish tinge. Halter with stem and knob pale yellow. Legs yellow, tarsi yellow but seem darker because of dense brown setae. All setae on legs brown, tibial spurs yellowish. Abdomen mainly brown with first two segments somewhat lighter. Abdominal vestiture brown. Terminalia light brown. Head. Ocelli two, very close to eye margins, with dark brown patches at anterior margin. Frontal furrow complete. Clypeus rectangular. Fourth flagellar segment about 2.5 times as long as wide, apical flagellar segment 2.5 times as long as wide basally. Flagellar segments with dense whitish setae about one third of segments’ width. Thorax. Scutum covered with setae, with marginal and prescutellar setae stronger. Antepronotum with 2 strong and 10–15 weaker setae. Proepisternum with two strong and 2–3 weaker setae. Anepisternum, katepisternum and anepimeron non-setose. Laterotergite with about 10 setae on upper half. Mediotergite non-setose. Scutellum with about 10 setae on upper surface. Wing. Length 2.39–2.70 mm, length to width 2.75–2.90. C, R, 1R, 4+5R setose, all other veins non-setose. C produced halfway between 4+5R and 1M. r-m about as long as m-stem. Posterior fork at the level of anterior fork or slightly before. CuA slightly sinuous. Legs. Ratio of femur to tibia for fore, mid and hind legs: 1.08–1.17; 0.97–1.00; 0.65–0.90. Ratio of tibia to basitarsus for fore, mid and hind legs: 0.96–1.00; 1.21–1.28; 1.41–1.77. Fore tibia with a spur 2.00 times of tibial maximum width. Mid tibia with anterior spur absent and posterior spur 2.27–2.40 times of tibial maximum width. Hind tibia with anterior spur 2.71–3.33 times and posterior spur 3.93–4.66 times of tibial maximum width. Strong posteroventral fringe of mid femora with row of strong setae. Hind coxa without basal setae. Terminalia , a keen naturalist who also participated in a trip to Georgia in 2017.Anatella Winnertz species known in the Holarctic region  ventral branch of the gonostylus twice as long as dorsal branch (only somewhat longer in A.atlanticiliata), (2) dorsal branch of the gonostylus with medial prong long, anchored to lateral prong basally , and (3) medial branch of gonostylus slender, apically hooked .There are about 50 gion cf. , the vasline cf. : fig. 27Taxon classificationAnimaliaDipteraBolitophilidae1.760DD7E7-5BF5-52A2-9515-0D090CDCB1BD2♀♀, SJ-7; 4♀♀, SJ-8. Total: 6♀♀.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraBolitophilidae2.585C6050-A66B-5B25-92E7-11A9C3C68BB61♀, MM-1. Total: 1♀.Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraBolitophilidae3.Meigen, 181836A13E06-BFD7-577F-80EB-E6664FDB2E5A1♂, MM-12. Total: 1♂.Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraBolitophilidae4.Edwards, 1941E764C905-F852-599E-8352-2F912BC50FD71♂, SJ-9. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraDiadocidiidae5.Mik, 187482387C38-2B44-5221-8C79-939284E382E41♀, SJ-1; 1♂, SK-1. Total: 1♂ 1♀.Shida Kartli, Samtskhe-Javakheti.Western Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraDiadocidiidae6.8C209075-9BBA-5FDF-90AC-22FEBB07089EZFMK); 2♂♂ 1♀, I-6; 2♂♂, I-9; 1♂, I-11; 1♂, A-1; 1♂, SJ-4; 1♂, SJ-7; 1♂, SJ-8; 4♂♂, SK-1. Total: 14♂♂ 1♀.1♂, SZS-3 ; 2♂♂, SK-1; 2♂♂, SJ-1 (ZFMK); 1♂, SJ-2 (IUTG); 1♂ 1♀, SJ-4; 2♂♂, SJ-7; 4♂♂ 3♀♀, SJ-9. Total: 13♂♂ 4♀♀.1♂, SZS-3 2DB9C662-7DE3-589A-8C0D-E962AAC24EAC2♂♂, I-9. Total: 2♂♂.Imereti.Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraDitomyiidae9.BC071DFE-4E2F-5EED-AE26-506DC6BEBC29ZFMK, 2♂♂ IUTG, 2♂♂ IZBE); 1♂, A-1; 1♂ 1♀, A-3; 1♀, I-3 (18.v–1.vi.2013); 1♂, I-6; 1♂ 1♀, I-9; 1♂, I-10; 2♂♂, I-11; 1♂, I-12; 4♂♂, SJ-1 ; 2♂♂, MM-12. Total: 20♂♂ 3♀♀.6♂♂, SZS-3 . Total: 1♂.1♂, SZS-3 5296B60A-9F13-5F7A-982F-2A72AFA5AC301♂, I-4. Total: 1♂.Imereti.Western Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraKeroplatidae21.Dalman, 1818EF3C443B-C58B-5F26-BDBD-63FEC08B75222♂♂, I-6. Total: 2♂♂.Imereti.Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraKeroplatidae22.95212B05-5ABD-5448-9D72-3FF4B76F9C05Adjara.Holarctic.I.semirufa. The other European species have the thorax yellow to orange with or without longitudinal stripes; also, see the next species and discussion by Taxon classificationAnimaliaDipteraKeroplatidae23.C50699F7-6CAE-5CAA-BCBE-D065A50339DD1♂, A-1; 1♂, A-5; 1♀, A-7; 1♂ 1♀, I-6. Total: 3♂♂ 2♀♀.Adjara, Imereti.Europe.I.semirufa), (2) wing with a distinct subapical band (anteriorly infuscated in I.semirufa) and (3) male terminalia with medial tooth of the gonostylys larger than the lateral one . I.czernyi is a rare European species known from the Mediterranean region and Slovakia . Total: 1♂.1♂, SZS-3 D977DD28-F5E3-5DAE-A828-7572B2648E6E1♂, K-2; 1♀, K-3. Total: 1♂ 1♀.Kakheti.Europe.Taxon classificationAnimaliaDipteraKeroplatidae27.5F94DBF1-DB7F-56F6-BE0D-11F82486C6831♂, SZS-1. Total: 1♂.Samegrelo-Zemo Svanethi.Europe.Taxon classificationAnimaliaDipteraKeroplatidae28.998EEC29-5151-5D99-B54D-1D832FCA78E5ZFMK, 3♂♂ IUTG, 2♂♂ IZBE); 1♂, A-3; 1♂, SJ-1 (ZFMK); 1♂, MM-7. Total: 10♂♂.7♂♂, SZS-3; 4♂♂, K-4; 1♂, K-6. Total: 171♂♂ 4♀♀.1♂, I-10 (holotype); 1♂, I-9 (paratype); 10♂♂, I-6 (paratypes). Additional material. 12♂♂ 4♀♀, A-1; 34♂♂, A-3; 7♂♂, A-5; 76♂♂, A-7; 1♂, A-8; 5♂♂, I-6; 12♂♂, I-11; 1♂, I-12; 5♂♂, SJ-1; 1♂, SJ-2 . Total: 36♂♂.1♂, I-5 (holotype). Additional material. 35♂♂, SZS-3 D65F51C5-DEAD-5EBD-87DA-F2440FE5794D16♂♂, SJ-4. Total: 16♂♂.Samtskhe-Javakheti.Europe.P.perpusilla species-group includes at least seven closely related species in Europe, separable only by small details of male terminalia  and the aedeagal sheath is interrupted medially on the ventral side . However, the Georgian specimens are slightly different from P.perpusilla as figured by P.alpicola. The material was compared to that of P.alpicola and P.oracula from North Italy 0A8E2391-907C-5DF5-90C8-3CE5112A615CIUTG, 1♂ IZBE); 1♂ 1♀, SJ-2 (ZFMK); 1♂, SJ-3; 4♂♂, MM-11. Total: 13♂♂ 1♀.2♂♂, A-5; 1♂, I-6; 2♂♂, I-11; 2♂♂, SJ-1 26C2DB5E-AFB6-5062-8F03-BF8B8116EF0FIZBE); 3♂♂, SJ-1 ; 1♂ 2♀♀, SJ-2 . Total: 5♂♂ 2♀♀.1♂, SZS-3 45B70A57-CAB3-5F1F-B32B-9ED35B4B091AZFMK, 1♂ IZBE). Total: 2♂♂.2♂♂, SZS-3 1D8C5EF7-B16B-5F1F-A7BE-04702F696BAE1♂, A-7. Total: 1♂.Adjara.Holarctic, extending to the Oriental region.Taxon classificationAnimaliaDipteraMycetophilidae37.Landrock, 192514E7A313-3C96-5BEB-9520-83526042BD61ZFMK). Total: 1♂.1♂, SZS-3 4895B3D3-A7C8-5875-BAE3-1E05BAB923CA1♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Holarctic, extending to the Oriental region.Taxon classificationAnimaliaDipteraMycetophilidae39.8C34CE3C-2248-5CBA-A57D-E38838550EF46♂♂, A-5; 1♂, A-7; 4♂♂, I-6; 2♂♂, SJ-3; 29♂♂, SJ-4; 1♂, SJ-9; 1♂, MM-7; 2♂♂, MM-14; 1♂, K-6. Total: 47♂♂.Adjara, Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi, Kakheti.Western Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae40.61EA199A-500A-50EF-A2CE-B6E8DA2FAB5FZFMK). Total: 1♂.1♂, SZS-3 25281D7A-AC5B-56FE-B95A-D5D9176686443♂♂, I-6; 1♂, I-17; 1♂, SJ-3; 2♂♂, MM-8; 1♂, MM-12. Total: 8♂♂.Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae42.3DD22F87-1AD5-5CF6-B3D3-9F8D60BF3F9A1♂, SJ-4. Total: 1♂.Samtskhe-Javakheti.Palaearctic, extending to the Oriental region.Taxon classificationAnimaliaDipteraMycetophilidae43.FF7DE2A9-8F7C-5B38-932A-8D557EE1C8591♂, A-7; 1♂, SJ-4. Total: 2♂♂.Adjara, Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae44.D682E1BD-A518-560A-9905-F5C975A5ACBDZFMK). Total: 1♂.1♂, SZS-4 2D79CC59-9BE6-55AF-94A5-22E3279E918CZFMK); 1♂, I-11. Total: 2♂♂.1♂, SZS-4 98985B94-602E-51D3-BD00-92B2B19634B91♂, SK-1; 1♂, SJ-8. Total: 2♂♂.Shida Kartli, Samtskhe-Javakheti.Palaearctic, extending to the Oriental region.Taxon classificationAnimaliaDipteraMycetophilidae47.495D18DF-70D2-56A7-90FC-74782D2A95C33♂♂, K-5; 1♂, K-6. Total: 4♂♂.Kakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae48.0DBC55C6-A12C-523A-8FF8-0E4C938638285♂♂, K-6. Total: 5♂♂.Kakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae49.Okada, 1939DF25961E-A463-5F46-B932-AE0FB8F592EC1♂, A-7; 2♂♂, I-6. Total: 3♂♂.Adjara, Imereti.Palaearctic.The species description from Hokkaido (Japan) by Okada was supplemented by a black and white figure of the general habitus including wing venation and pattern  are provTaxon classificationAnimaliaDipteraMycetophilidae50.DB8D5FAC-9D5B-5039-9277-3B1A2E399ECF1♀, I-6; 1♂ 1♀, I-14. Total: 1♂ 2♀♀.Imereti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae51.Winnertz, 1863D29AA442-C278-5CC1-89B2-E191581F4D131♂, I-6. Total: 1♂.Imereti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae52.Zaitzev, 19845789E4C1-C0D8-570E-9E8A-E845F3F1DF04ZFMK); 1♂, I-6. Total: 2♂♂.1♂, SZS-3 17ED99F2-7CD9-5C55-BDC4-214A370D0AEAZFMK, 1♂ IUTG, 1♂ IZBE); 2♂♂, A-3; 2♂♂, A-7; 3♂♂ 1♀, K-4; 1♂, K-5; 1♂, K-6. Total: 13♂♂ 1♀.4♂♂, SZS-3 7531BC30-B90C-53EB-81C4-7E94FA870802ZFMK). Total: 1♂.1♂, SJ-2 ; 3♂♂, A-1 (paratypes); 3♂♂ 3♀♀, A-3 (paratypes); 1♂, A-7 (paratype). Total: 8♂♂ 3♀♀.Adjara.Georgia.The species was recently described from material collected from Mtirala and Kintrishi National Parks in Adjara Taxon classificationAnimaliaDipteraMycetophilidae56.D08B9435-C761-558A-AD7B-DB940A759FB81♀, SJ-3. Total: 1♀.Samtskhe-Javakheti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae57.Strobl, 1898C384700E-8732-5365-BD58-A2FC230E3D731♂, I-11. Total: 1♂.Imereti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae58.0DF81BD0-A92E-5D8A-A2E0-6C46C67179812♂♂, I-6. Total: 2♂♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae59.225E1194-548C-54A0-9A92-AA678EFB7B943♂♂, I-6; 1♂, SK-1. Total: 4♂♂.Imereti, Shida Kartli.Holarctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae60.Zaitzev, 1994A290BF96-DECB-5791-A4AC-E60754CCEF34IZBE); 2♀♀, SZS-4 ; 3♂♂ 4♀♀, A-3; 1♂ 2♀♀, A-7; 2♂♂ 10♀♀, I-6; 1♂ 1♀, I-9; 2♂♂ 2♀♀, SK-1; 1♂, SJ-1 (IZBE); 4♂♂ 1♀, SJ-2 ; 2♂♂ 1♀, SJ-3; 1♂, KK-1. Total: 17♂♂ 24♀♀.1♀, SZS-3 , Georgia.Taxon classificationAnimaliaDipteraMycetophilidae61.Zaitzev, 198422356499-8476-5106-907F-83E6C85D0A451♂, MM-8; 1♂, K-4. Total: 2♂♂.Mtskhetha-Mthianethi, Kakheti.Caucasia.Known only from type locality in Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae62.Zetterstedt, 18521FE13894-88B6-5E0C-B321-27E001B5ACEC1♂, A-5. Total: 1♂.Adjara.Europe, with scattered distribution .Taxon classificationAnimaliaDipteraMycetophilidae63.Curtis, 1837CF4627B6-5F85-5D2E-8D9D-20C077CAE4CC1♂, I-6. Total: 1♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae64.6EE3AE2C-3471-5F13-BA0B-7E5D8502FBDCSee in species description above.Kakheti.Georgia.Taxon classificationAnimaliaDipteraMycetophilidae65.3134B590-2BCE-59CA-9FFC-A0CE93C7E6C0ZFMK); 1♂, SJ-1 (IZBE); 1♂, SJ-2 (IUTG). Total: 3♂♂.1♂, SZS-3 ; 1♂, SJ-2 (ZFMK). Total: 2♂♂.1♂, SZS-3 BF30F735-4D5E-55EA-9F95-A911BDF665A91♂, I-6. Total: 1♂.Imereti.Western Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae69.Zetterstedt, 1852633FA627-564F-5F1A-8B20-8A2A9D41B542ZFMK). Total: 2♂♂.1♂, SZS-2; 1♂, SZS-4 . Total: 4♂♂.4♂♂, SZS-4 48EC3D3A-5480-50A9-B8A9-4C5B5F93DCD95♂♂, A-1; 3♂♂, A-3. Total: 8♂♂.Adjara.Europe.The Georgian specimens have the ventral lobe of the gonostylus with a blunt small spine apically that is absent in studied specimens from Estonia and Sweden as well as in published figures e.g. . OtherwiTaxon classificationAnimaliaDipteraMycetophilidae72.Dziedzicki, 18855FA5CCF4-3BCF-50EB-84F6-82CBE46B2E42ZFMK, 1♂ IZBE). Total: 2♂♂.2♂♂, SZS-4 . Total: 1♂.1♂, SZS-4 ; 1♂, SJ-1 (IZBE); 1♂, K-6. Total: 12♂♂.10♂♂, SZS-3 . Total: 3♂♂.3♂♂, SZS-4 63D861C9-92F3-5D36-A993-E389101C3CB94♂♂, SJ-8; 5♂♂, SJ-9. Total: 9♂♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae77.64F04AF0-AEC2-5E3B-9C90-CEC84EC8EE90ZFMK); 8♂♂ 1♀, KK-1. Total: 9♂♂ 1♀.1♂, SZS-3 E27A7B78-1470-5913-B3E0-83E3673B132CZFMK); 1♀, I-6. Total: 1♂ 1♀.1♂, SZS-3 . Total: 1♂.1♂, SJ-2 ; 5♂♂, SZS-4; 2♂♂, SZS-3; 1♂, SJ-1. Total: 9♂♂ 712B5F01-42A6-54CB-8030-F90A70CD27C7ZFMK); 1♂, K-6. Total: 2♂♂.1♂, SJ-2 C73BA377-2968-5916-A3E4-A6752FDDF1A9ZFMK, 1♂ IZBE); 1♂, I-10; 1♂, SK-1. Total: 4♂♂.2♂♂, SZS-3 30D79A86-22F0-59D4-B417-D4ECBA9A9221ZFMK); 2♂♂, I-6. Total: 3♂♂.1♂, SZS-3 949C8852-9A16-5FDC-BEE7-2ECAC1C4987D1♂, A-1; 1♂, I-6. Total: 2♂♂.Adjara, Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae87.Meigen, 1818D9B33DD0-CD21-536A-A4B5-73B1EE00A3C3104♂♂, A-1; 54♂♂ 5♀♀, A-2; 70♂♂, A-3; 21♂♂ 2♀♀, A-4; 6♂♂ 4♀♀, A-5; 3♂♂, A-6; 173♂♂ 64♀♀, A-7; 14♂♂, I-1; 7♂♂, I-3 (18.v–1.vi.2013); 1♂, I-4; 138♂♂ 7♀♀, I-6; 25♂♂, I-7; 2♂♂ 3♀♀, I-9; 1♀, I-10; 8♂♂, I-11; 15♂♂, I-13; 2♂♂, I-16; 37♂♂, I-17; 1♂, MM-9. Total: 683♂♂ 86♀♀.Adjara, Imereti, Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae88.CA169BC7-C957-55D8-A597-66A52AB46E0AZFMK, 2♂♂ 1♀ IUTG, 2♂♂ IZBE); 1♂, K-4; 1♂, K-5. Total: 8♂♂ 2♀♀.6♂♂ 2♀♀, SZS-3 .A widely distributed Palaearctic species that has expanded its range in Europe during recent decades and is locally common also in anthropogenic environments B10E0BD9-4088-50F6-B9F8-82D15A116032IZBE); 1♂, SK-1; 1♂, SJ-2 (ZFMK); 1♂, SJ-9; 1♂, MM-7. Total: 5♂♂.1♂, SZS-3 1C32D3A3-0325-5C23-98AB-752DBF5317A01♂ 1♀, I-6; 1♂, K-6. Total: 2♂♂ 1♀.Imereti, Kakheti.Western Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae93.237FE660-B57B-5DEB-91F9-5495E0480CE1See in species description above.Samegrelo-Zemo-Svanethi, Shida-Kartli.Georgia.Taxon classificationAnimaliaDipteraMycetophilidae94.Edwards, 1925F846E2BF-2A57-5FAD-AD1B-E67C802CA9C71♂, I-6. Total: 1♂.Imereti.Europe, with scattered distribution.Taxon classificationAnimaliaDipteraMycetophilidae95.Lehmann, 1822BDED0737-CDA0-5D3B-A7E3-F00BA38C75953♂♂, I-10; 1♂, SJ-8. Total: 4♂♂.Imereti, Samtskhe-Javakheti.Holarctic, extending to the Oriental region.Taxon classificationAnimaliaDipteraMycetophilidae96.Strobl, 1893CC3BD89D-C0FC-5ACB-B0F2-998DB7ACA30AZFMK, 1♂ IZBE); 1♂, K-4; 1♀, K-5. Total: 2♂♂ 2♀♀.1♂ 1♀, SJ-2 .According to the recent molecular study by Taxon classificationAnimaliaDipteraMycetophilidae97.Lundström, 19132229D9F3-DFB4-5203-B11C-71248796DFC21♂, I-6. Total: 1♂.Imereti.Europe.A rare species, recorded from Central and Northern Europe with the south-eastern record on the Crimean Peninsula . The curExechiiniTribe Taxon classificationAnimaliaDipteraMycetophilidae98.A2A998A9-4689-5EFA-A56A-A19241D0FFD4ZFMK, 1♂ IUTG, 1♂ IZBE); 7♂♂, SJ-3; 1♂, SJ-8; 1♂, MM-2; 1♂, MM-11; 1♂, MM-12. Total: 15♂♂.4♂♂, SZS-4 A3D5387A-CEDA-5A08-AACD-FA64096B4E891♂, SJ-9; 1♂, MM-12; 1♂, MM-14; 1♂, K-1. Total: 4♂♂.Samtskhe-Javakheti, Mtskhetha-Mthianethi, Kakheti.Holarctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae100.Edwards, 19210E9B6530-E974-52E7-908C-6CDBDCEEE3F41♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae101.23C27DB0-0CDB-5FD4-9F28-F0435E5D2E4A1♂, A-7; 2♂♂, MM-2; 1♂, MM-8. Total: 4♂♂.Adjara, Mtskhetha-Mthianethi.Holarctic.Listed to occur in Transcaucasia without further details .Taxon classificationAnimaliaDipteraMycetophilidae102.Zaitzev, 1982D06F43DF-EE77-52B0-8841-08173AC2FDEFZFMK); 1♂, SJ-9. Total: 2♂♂.1♂, SZS-4 094C77D4-DCE1-5795-9268-B233EBDAA6652♂♂, SJ-8; 1♂, MM-12. Total: 3♂♂.Samtskhe-Javakheti, Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae104.Dziedzicki, 19238DF71C57-33C3-5F69-A9CD-F7C591336DADZFMK); 3♂♂, SZS-4 . Total: 4♂♂.1♂, SZS-3 AD712DBD-B0A1-5985-ACB7-7F565E61E8D61♂, MM-14. Total: 1♂.Mtskhetha-Mthianethi.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae108.B49B6A48-FAF6-5DA0-A101-EB77014367DB1♂, MM-2. Total: 1♂.Mtskhetha-Mthianethi.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae109.8F4B8BFF-06B7-56B7-9B14-DFC2F0C9F72F4♂♂, I-6; 1♂, I-10; 1♂, SJ-3; 1♂, MM-5; 1♂, MM-8. Total: 8♂♂.Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae110.42E51E02-4040-5C87-805E-0FE15C17B64F1♂, MM-2. Total: 1♂.Mtskhetha-Mthianethi.Holarctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae111.Edwards, 1925102425D3-7B65-57A3-B9D0-02DEF8F20A8E1♂, I-6. Total: 1♂.Imereti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae112.BE4FEB63-41D6-5F74-A9F1-C99B13C4297B5♂♂, I-6. Total: 5♂♂.Imereti.Holarctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae113.Laštovka & Matile, 1974118594DB-2746-5235-A04D-7486D1780615ZFMK). Total: 1♂.1♂, SZS-4 1888F37D-94BF-5C07-B033-6CFCEDE5AAA5ZFMK); 1♂, A-8. Total: 2♂♂.1♂, SZS-4 94B62DE6-BD86-5E5A-A6E3-FCB214A0DE06ZFMK). Total: 1♂.1♂, SZS-4 741831C8-147F-5443-8E44-CD19CF4F05EC1♂, I-6; 1♂, SJ-8. Total: 2♂♂.Imereti, Samtskhe-Javakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae118.1D2704EF-F6CC-5F56-94AE-F9BE10D0516FZFMK, 3♂♂ IUTG, 2♂♂ IZBE); 1♂, I-1; 2♂♂, I-6; 1♂, SK-1; 1♂, SJ-6; 4♂♂, SJ-7; 1♂, SJ-9; 1♂, MM-2; 1♂, MM-14; 1♂, K-1. Total: 21♂♂.8♂♂, SZS-4 91DF01BD-E92C-51D2-8E25-8AFA85835F841♂, I-11, 2♂♂, SJ-7; 2♂♂, SJ-8; 1♂, SJ-9; 1♂, MM-14. Total: 7♂♂.Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Western Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae120.3A698037-EB4B-5661-8460-9460DF2F6BF02♂♂, I-6; 1♂, I-11; 1♂, SJ-8; 1♂, SJ-9; 1♂, MM-5; 2♂♂, MM-12; 1♂, MM-13. Total: 9♂♂.Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Palaearctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae121.0AA820C9-1F8C-552B-82D9-03E41124CA31ZFMK); 1♂, SZS-4 (IZBE); 1♂, SJ-7; 4♂♂, SJ-8; 1♂, SJ-9; 3♂♂, KK-1; 1♂, MM-2; 1♂, MM-8; 1♂, MM-14; 1♂, K-2. Total: 15♂♂.1♂, SZS-3 0D62428F-A3A5-569D-9457-0F6C0C6E7DC1IZBE); 5♂♂, SZS-4 ; 1♂, A-5; 1♂, A-7; 1♂, I-6; 1♂, SJ-2; 1♂ (IZBE), KK-1; 1♂ 2♀♀, MM-12; 1♂, K-6. Total: 13♂♂ 2♀♀.1♂, SZS-3 . Total: 2♂♂.2♂♂, SJ-2 ; 1♂, SJ-7; 1♂, KK-1. Total: 4♂♂.Imereti, Samtskhe-Javakheti, Kvemo Kartli.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae127.Laštovka & Matile, 1974501FCD44-1E4B-503A-B137-326732685DF8ZFMK, 1♂ IZBE). Total: 2♂♂.2♂♂, SJ-2 49F49B66-89CD-5DA2-9461-D85699A863A22♂♂, I-3 (18.v–1.vi.2013 and 5–19.x.2013); 1♂, SJ-12. Total: 3♂♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae129.Edwards, 192524192B6E-4153-5B0F-BD08-1BB35EF5D9BF1♂, I-14; 1♂, K-5. Total: 2♂♂.Imereti, Kakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae130.Edwards, 1925360CB74B-D21E-57CD-9E2E-02A06751DF5DIZBE); 45♂♂, SZS-4 ; 2♂♂, I-6; 2♂♂, K-4; 1♂, K-6. Total: 53♂♂.3♂♂, SZS-3 A71DB4AC-2C11-5165-AC25-44BCBE77EB9F4♂♂, A-5; 3♂♂, A-7; 1♂, I-9; 1♂, I-10; 1♂, I-11; 1♂, I-12; 1♂, MM-8; 1♂, MM-14; 1♂, K-2. Total: 14♂♂.Adjara, Imereti, Mtskhetha-Mthianethi, Kakheti.Holarctic.In Transcaucasia recorded from Azerbaijan .Taxon classificationAnimaliaDipteraMycetophilidae132.Lundström, 1916626B5C75-0A96-54CC-B0CE-008D92114B841♂, A-7. Total: 1♂.Adjara.Europe.Taxon classificationAnimaliaDipteraMycetophilidae133.19A91249-D44A-518A-9296-3948FED127951♂, SJ-7; 2♂♂, MM-12; 1♂, MM-13; 3♂♂, MM-14. Total: 7♂♂.Samtskhe-Javakheti, Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae134.90573DFF-C175-5755-A3A3-5885415B8DCA2♂♂, I-6; 1♂, SJ-3; 2♂♂, MM-2; 1♂, MM-8; 2♂♂, MM-12. Total: 8♂♂.Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae135.Johannsen, 19124313126A-4BD6-5D00-BD0B-A8C03AF7ADCF2♂♂, K-4. Total: 2♂♂.Kakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae136.Caspers, 19845D451BCF-0935-567E-8CC1-C0410060BA901♂, A-3; 1♂, I-3 (24.viii–7.ix.2013); 1♂, SJ-4; 1♂, MM-12; 1♂, K-5. Total: 5♂♂.Adjara, Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi, Kakheti.EuropeTaxon classificationAnimaliaDipteraMycetophilidae137.7C6A136E-A13F-5BD5-94BE-0BDE6566B4C21♂, A-5. Total: 1♂.Adjara.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae138.DC45407F-05C5-5889-8FB9-2080F4E812F31♂, I-12. Total: ♂.Imereti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae139.7B45499B-1357-5C76-83C4-27E546DD0E4C1♂ 1♀, MM-1. Total: 1♂ 1♀.Mtskhetha-Mthianethi.Europe.Taxon classificationAnimaliaDipteraMycetophilidae140.Laštovka & Matile, 197489ACE343-4B65-56D1-BC12-532866737E485♂♂, MM-2. Total: 5♂♂.Mtskhetha-Mthianethi.Palaearctic.In Transcaucasia recorded from Armenia .Taxon classificationAnimaliaDipteraMycetophilidae141.4F73EF82-103B-5EF8-BC26-2B9AC667EE231♂, MM-8; 2♂♂, MM-11. Total: 3♂♂.Mtskhetha-Mthianethi.Europe.Taxon classificationAnimaliaDipteraMycetophilidae142.31C34051-D7C7-56A5-A8CE-8402001B612D2♂♂, SJ-8; 1♂, SJ-9. Total: 3♂♂.Samtskhe-Javakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae143.Kjærandsen, 200960FA453D-A68E-560D-9FBC-6A980A27C1BA1♂, I-10. Total: 1♂.Imereti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae144.Winnertz, 18630C322548-4A24-5C17-9EF0-89D6D753F5171♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae145.51E1C0F0-6F0F-5F32-B31B-2F4BF1E406901♂, I-9. Total: 1♂.Imereti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae146.D1CEF64C-BE60-5867-8248-BE4BD5732CB01♂, A-5; 1♂, SK-1; 1♂, SJ-3; 5♂♂, SJ-8; 1♂, MM-2; 8♂♂, MM-8; 5♂♂, MM-11; 4♂♂, MM-12. Total: 26♂♂.Adjara, Shida Kartli, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae147.5EC4A53E-FB93-58BD-A179-BA75D53378FE1♂, A-1. Total: 1♂.Adjara.Europe.Taxon classificationAnimaliaDipteraMycetophilidae148.AC653FDF-F281-5499-BD1A-B2FF408491861♂, SJ-7. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae149.Strobl, 1895C879A0BE-D234-5323-8E6F-AEBAD1FC4B711♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae150.E83C84B8-F512-52E6-B493-AEFC2C45691BZFMK). Total: 1♂.1♂, SZS-3 447934FC-6655-505B-AA40-4CDE3F7E998D1♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae154.282B4262-4F7D-5F0E-91BE-A193474E650B1♂, K-6. Total: 1♂.Kakheti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae155.Chandler, 1981ABC06099-8A31-5F57-ADDF-DF21CA2850401♂, K-5. Total: 1♂.Kakheti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae156.0848D148-1392-52A1-8E33-F6613DEE3D5C1♂, K-4; 7♂♂ 4♀♀, K-5. Total: 8♂♂ 4♀♀.Kakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae157.Matile, 197768336123-D4DB-583D-B420-E55E30FBD6071♂, A-1; 1♂ 1♀, I-6; 1♂, MM-14; 1♂; K-2. Total: 4♂♂ 1♀.Adjara, Imereti, Mtskhetha-Mthianethi, Kakheti.Western Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae158.Dziedzicki, 18890892226D-9E66-5B5D-8652-1733EBC06E801♂, I-6; 1♂, K-4. Total: 2♂♂.Imereti, Kakheti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae159.Mik, 188428F22DB6-5FAA-5B9F-9A8A-485262233C451♂, SJ-8; 1♂, MM-13. Total: 2♂♂.Samtskhe-Javakheti, Mtskhetha-Mthianethi.Europe.Taxon classificationAnimaliaDipteraMycetophilidae160.Laffoon, 1965ACD09D52-0029-5E65-AFD0-F735B50AACABZFMK); 3♂♂, SJ-4; 2♂♂, SJ-9; 2♂♂, MM-8; 2♂♂, MM-11; 1♂, KK-1. Total: 50♂♂.1♂, A-3; 35♂♂, I-6; 1♂, I-9; 1♂, I-14; 1♂, SK-1; 1♂, SJ-1 ; 1♂, SJ-4; 1♂, SJ-8; 1♂, SJ-9. Total: 4♂♂.1♂, SZS-4 AF14881B-2BE0-546C-98F2-1BA77DDEADE9ZFMK). Total: 1♂.1♂, SZS-4 ; 1♂, I-14; 2♂♂, K-4. Total: 5♂♂.2♂♂, SZS-3 489DC62F-B1AC-5F09-985F-4C8F8C88A7AAIZBE); 19♂♂ 10♀♀, SZS-4 ; 1♂, I-3 (29.vi–13.vii.2013); 5♂♂ 2♀♀, I-6; 1♀, I-9; 1♂, I-10; 1♀, SK-1; 2♂♂, SJ-1 (IZBE); 2♀♀, SJ-3; 1♂, SJ-4; 2♂♂ 2♀♀, SJ-7; 4♂♂ 4♀♀, SJ-8; 2♂♂ 4♀♀, SJ-9; 1♂, SJ-10; 2♂♂, MM-3; 1♀, MM-6; 2♂♂ 7♀♀, MM-7; 1♂ 1♀, MM-8; 2♂♂ 2♀♀, MM-11; 1♀, MM-12; 1♂, MM-14; 1♂ 2♀♀, KK-1; 1♂, K-4. Total: 54♂♂ 43♀♀.6♂♂ 3♀♀, SZS-3 .Taxon classificationAnimaliaDipteraMycetophilidae169.Dziedzicki, 1884DC328F65-6563-5066-B5EA-95BFF7C03AFD4♂♂, SJ-8; 1♂, MM-2. Total: 5♂♂.Samtskhe-Javakheti, Mtskhetha-Mthianethi.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae170.Lundström, 19116FB7A634-74E7-508E-B98E-40BDBF81CEF4ZFMK, 1♂ IUTG, 1♂ IZBE); 1♂, I-3 (1–15.vi.2013); 3♂♂, I-6; 2♂♂, I-9; 1♂, I-12; 2♂♂, SK-1; 6♂♂, SJ-8; 5♂♂, SJ-9; 1♂, KK-1; 3♂♂, MM-8; 1♂, MM-13; 7♂♂, MM-14; 3♂♂, K-4; 1♂, K-5. Total: 40♂♂.4♂♂, SZS-4 ; 1♂, SZS-4 (IUTG). Total: 3♂♂.2♂♂, SZS-3 ; 1♂, MM-11; 3♂♂, MM-14. Total: 5♂♂.1♂, SZS-4 ; 1♂, SJ-6; 1♂, KK-1; 3♂♂, K-4. Total: 6♂♂.1♂, SZS-3 ; 1♂, SK-1; 1♂, SJ-2 (IZBE); 1♂, SJ-5; 3♂♂, SJ-8; 1♂, SJ-9. Total: 13♂♂.6♂♂, SZS-4 ; 2♂♂, A-1; 1♂, A-3; 1♂, A-7; 10♂♂, I-6; 1♂, I-10; 1♂, SK-1; 1♂, SJ-1 (IZBE); 1♂, KK-1; 1♂, MM-7; 1♂, MM-8; 4♂♂, MM-11; 1♂, MM-14; 1♂, K-4; 1♂, K-5; 1♂, K-6. Total: 33♂♂.5♂♂, SZS-3 . Total: 1♂.1♂, SZS-4 ; 1♂, I-10; 1♂, MM-4. Total: 3♂♂.1♂, SZS-4 ;1♂, I-6; 1♂, SJ-8; 1♂, SJ-9; 57♂♂, MM-13; 1♂, MM-14. Total: 67♂♂.6♂♂, SZS-3 ; 2♂♂, A-1; 2♂♂, A-3; 3♂♂, I-6; 1♂, I-9; 1♂, I-10; 1♂, MM-11; 4♂♂, MM-14; 1♂, K-2. Total: 16♂♂.1♂, SZS-4 ; 2♂♂, I-6. Total: 3♂♂.1♂, SZS-4 ; 3♂♂, SZS-4 ; 2♂♂, I-6; 1♂, SJ-4; 4♂♂, SJ-8; 1♂, SJ-9; 6♂♂, KK-1; 1♂, MM-11. Total: 19♂♂.1♂, SZS-3 ; 42♂♂, SZS-4 ; 1♂, I-10; 1♂, SJ-1 (IZBE); 1♂, SJ-2 (ZFMK). Total: 49♂♂.4♂♂, SZS-3 BD23232A-7516-5A32-883F-C4954A2F24059♂♂, SJ-8; 4♂♂, SJ-9. Total: 13♂♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae202.Zaitzev, 199869C3EFEE-8D2C-5945-A872-F526D23FF6161♂, SJ-9. Total: 1♂.Samtskhe-Javakheti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae203.Zaitzev, 199968D3862B-E35D-5A93-AAEC-DD76050A7DE01♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae204.29B8AD76-6C64-5F7D-A53C-A34D4823B2811♂, I-10. Total: 1♂.Imereti.Europe.Taxon classificationAnimaliaDipteraMycetophilidae205.Staeger, 18402AB9613F-06D8-52B9-8DBD-8430B6414A35ZFMK); 5♂♂, MM-13; 12♂♂, MM-14; 1♂, K-2. Total: 19♂♂.1♂, SZS-3 . Total: 1♂.1♂, SZS-4 . Total: 6♂♂.3♂♂, A-1; 2♂♂, A-7; 1♂, SJ-2 F77F38C2-E1BC-5C21-80E6-2D90D26220301♂, SJ-8; 1♂, SJ-9; 1♂, MM-12. Total: 3♂♂.Samtskhe-Javakheti, Mtskhetha-Mthianethi.Holarctic.In Transcaucasia recorded from Armenia .Taxon classificationAnimaliaDipteraMycetophilidae210.72FD0674-EA83-5E11-B8C2-5811E31D493C2♂♂, I-6. Total: 2♂♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae211.Dziedzicki, 18890DAC051C-2CF8-5F92-9A7F-18F84423308A1♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae212.FF0C737F-E055-5213-BC20-1E80FD73C7DC2♂♂, MM-2; 1♂, MM-8. Total: 3♂♂.Mtskhetha-Mthianethi.Holarctic.Listed to occur in Transcaucasia without further details .Taxon classificationAnimaliaDipteraMycetophilidae213.Winnertz, 186338D49FE8-D199-5D32-A78B-97CC0F7989C71♂, A-7; 1♂, SJ-4; 2♂♂, SJ-8; 2♂♂, SJ-9. Total: 6♂♂.Adjara, Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae214.Winnertz, 1863BAD7C9FD-3106-5F5D-ACF5-959E6306EFD2ZFMK); 8♂♂, I-6. Total: 9♂♂.1♂, SZS-4 C2464E28-4DA3-5FD0-A233-962B20CBA2322♂♂, I-6; 1♂, I-9; 1♂, I-10. Total: 4♂♂.Imereti.Palaearctic.Listed to occur in Transcaucasia without further details .Taxon classificationAnimaliaDipteraMycetophilidae216.Dziedzicki, 1889D9887017-89FA-5254-8022-6B772244F2841♂, SJ-8. Total: 1♂.Samtskhe-Javakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae217.Winnertz, 186353D73012-7EE7-598C-AD68-791855EA8D621♂, I-6. Total: 1♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae218.Dziedzicki, 1889334D7AFA-1FFA-5334-94C0-96AD497F9AC66♂♂, MM-12. Total: 6♂♂.Mtskhetha-Mthianethi.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae219.Winnertz, 18632BBFDCF1-AEBB-5E4B-B4F4-909F69F5933A9♂♂, I-6; 1♂, I-9; 4♂♂, I-10; 1♂, SJ-9; 8♂♂, MM-8; 1♂, MM-9. Total: 24♂♂.Imereti, Samtskhe-Javakheti, Mtskhetha-Mthianethi.Palaearctic.Listed to occur in Transcaucasia without further details .Taxon classificationAnimaliaDipteraMycetophilidae220.Winnertz, 1863E5C7B49E-CF57-56B8-B83B-702EEC76D8CCZFMK, 1♂ IZBE); 1♂, I-10; 1♂, MM-12. Total: 4♂♂.2♂♂, SZS-4 23A0F64A-3891-5000-92D1-5437E8E489B31♂, I-3 (13–27.vi.2013); 2♂♂, SJ-4; 1♂, K-4. Total: 4♂♂.Imereti, Samtskhe-Javakheti, Kakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae223.DD780163-2E5C-5AA7-A262-6A09799373281♂, K-2. Total: 1♂.Kakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae224.Chandler, 1991C200A2C5-524A-5A4F-8775-F72927C9AAEB18♂♂, MM-13; 18♂♂, MM-14; 8♂♂, K-2. Total: 44♂♂.Mtskhetha-Mthianethi; Kakheti.Western Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae225.Bechev, 19973ED2F776-BB4D-5845-90D9-EB00AA6DC7BB1♂, MM-5. Total: 1♂.Mtskhetha-Mthianethi.Europe.Taxon classificationAnimaliaDipteraMycetophilidae226.Edwards, 1925DBDAFE0F-73A7-53A5-B9E6-741DCC2CE330IZBE); 6♂♂, I-6; 3♂♂, I-9; 2♂♂, I-14; 1♂, SJ-2 (ZFMK); 1♂, KK-1; 10♂♂, MM-13; 14♂♂, MM-14. Total: 38♂♂.1♂, SZS-3 87654F15-0479-5360-ABF6-AD98069690532♂♂, A-3; 1♂, MM-13; 14♂♂, MM-14; 7♂♂, K-2. Total: 24♂♂.Adjara, Mtskhetha-Mthianethi, Kakheti.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae230.Edwards, 192589B4B4D0-243E-5505-907E-2A3C5725206EZFMK, 1♂ IUTG, 1♂ IZBE); 2♂♂, MM-12. Total: 6♂♂.1♂, SK-1; 3♂♂, SJ-2 DEAEFA39-2B5C-5EEA-89F4-8DAB9D2D8D611♂, I-6. Total: 1♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae233.Lundström, 1913D5A7B4C6-076C-5583-AC0B-12A352038A112♂♂, I-6; 1♂, KK-1; 1♂, MM-12. Total: 4♂♂.Imereti, Kvemo Kartli, Mtskhetha-Mthianethi.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae234.Lundström, 19116636485D-1F0D-5DF4-A574-291CDA56AC1B2♂♂, A-7. Total: 2♂♂.Adjara.Holarctic.Listed to occur in Transcaucasia without further details .Taxon classificationAnimaliaDipteraMycetophilidae235.Gagne, 198113B50CBB-AAE4-57FF-99FF-08B8A6A649DC1♂, I-6. Total: 1♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae236.van der Wulp, 1881690238D5-2DA9-5A02-B0D0-1C2C09E81FBA1♂, I-17. Total: 1♂.Imereti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae237.Zaitzev & Menzel, 1996EDFF1E14-CE1C-5D9D-9F0E-264979A5CF004♂♂, I-6; 1♂, SK-1. Total: 5♂♂.Imereti, Shida Kartli.Palaearctic.Taxon classificationAnimaliaDipteraMycetophilidae238.Lundström, 1909B7F405DA-A8A0-5447-85D1-466748A9FCD41♂, I-10. Total: 1♂.Imereti.Northern Europe.Kjæranden and Søli (2020) recently reinstated the species and provided detailed figures of the male terminalia of the allied species.Taxon classificationAnimaliaDipteraMycetophilidae239.F32B02E2-F1B1-527E-852B-9A19571484199♂♂, I-6; 1♂, SJ-4; 1♂, SJ-8; 2♂♂, K-4; 2♂♂, K-5. Total: 15♂♂.Imereti, Samtskhe-Javakheti, Kakheti.Holarctic.Taxon classificationAnimaliaDipteraMycetophilidae240.Loew, 186983BDCAF8-3701-5FF1-88FD-E4E9205138DBZFMK); 2♂♂, SJ-8; 1♂, SJ-9. Total: 4♂♂.1♂, SZS-4 1BCCDD70-5B78-5C06-9869-EDF88C2054BEIZBE); 5♂♂, SZS-4 ; 1♂, SJ-4; 2♂♂, MM-12; 1♂, MM-13; 3♂♂, MM-14. Total: 13♂♂.1♂, SZS-3 ; 1♂, SJ-4; 1♂, SJ-7; 1♂, KK-1; 5♂♂, K-4. Total: 9♂♂.1♂, SJ-2 210A89BA-8F63-5F49-BDBF-C7E3973EABEEZFMK); 1♂, I-6; 1♂, SJ-3; 1♂, SJ-5. Total: 4♂♂.1♂, SZS-4 . Total: 10♂♂.10♂♂, SZS-4 5A1DC03A-FFC5-5FD2-BF50-FC164B06B610ZFMK); 1♂, A-1; 2♂♂, SJ-9. Total: 5♂♂.1♂, SZS-2; 1♂, SZS-4  of fungus gnats are the result of a preliminary survey, while further sweeping studies will probably increase that number considerably . As expected, the majority of the recorded species are widely distributed in the Palaearctic or Holarctic regions , while 22% of species are restricted to Europe and 7% to the Western Palaearctic . Taking into account the mountainous landscape, high diversity of habitats, microclimates in Georgia and that several regions were not covered by sampling of the current study  followed by Orfeliageorgica (175 specimens from 14 samples). In the European boreal and temperate regions, the most abundant species belong frequently to the subfamily Mycetophilinae and/or to the genera Boletina Staeger and Mycomya Rondani. Several of the recorded species considerably increase the knowledge of their distribution, the most remarkable of them include: Neoempheriabrevilineata (earlier from Japan only), Clastobasisloici (earlier from Europe and Japan), Lusitanoneurachandleri (earlier from the Mediterranean Islands), Zygomyiasetosa (earlier with scattered distribution in Europe), Manotaunifurcata (earlier from Europe only).Surprisingly, the most abundant species was From the material underlining this study, four new species have been described earlier , three n


Monothecabuxifolia methanolic extract and its various fractions were assessed against Macrophominaphaseolina, a soil-borne fungal pathogen of more than 500 vegetal species as well as rare and emerging opportunistic human pathogen. Different concentrations of methanolic extract (3.125 to 200 mg mL−1) inhibited fungal biomass by 39–45%. Isolated n-hexane, chloroform and ethyl acetate fractions suppressed fungal biomass by 32–52%, 29–50% and 29–35%, respectively. Triterpenes lupeol and lupeol acetate  were isolated from n-hexane while betulin, β-sitosterol, β-amyrin, oleanolic acid (3–6) were isolated from chloroform fraction. Vanillic acid, protocatechuic acid, kaempferol and quercetin (7–10) were isolated from the ethyl acetate fraction and identified using various spectroscopic techniques namely mass spectroscopy and NMR. Antifungal activity of different concentrations (0.0312 to 2 mg mL−1) of the isolated compounds was evaluated and compared with the activity of a broad spectrum fungicide mancozeb. Different concentrations of mencozeb reduced fungal biomass by 83–85%. Among the isolated compounds lupeol acetate (2) was found the highest antifungal against M.phaseolina followed by betulin (3), vanillic acid (7), protocatechuic acid (8), β-amyrin (5) and oleanolic acid (6) resulting in 79–81%, 77–79%, 74–79%, 67–72%, 68–71% and 68–71%, respectively. Rest of the compounds also showed considerable antifungal activity and reduced M.phaseolina biomass by 41–64%.Antifungal activity of  Macrophominaphaseolina is an important soil-borne plant pathogen that causes diseases over 500 plant species including economically important crops such as legumes, sunflower, cotton, sorghum and vegetables. Generally it causes charcoal rot disease in various crops; however, it also causes other diseases such as seedling and stem blight, damping off and wilt1. It has vast distribution in tropical and subtropical countries, however, its exposure to human may cause infection in immunosuppressed patients. Several strategies are being adopted to control fungal plant pathogens by synthetic fungicides. No doubt, these fungicides are effective in controlling plant diseases but they also pose severe hazards to human health and cause environmental pollution by accumulation in soil and water2. It necessitates alternative environmental friendly strategies for management of phytopathogens. Many recent studies have shown that crude plant extracts as well as purified compounds isolated from various plant species are very effective in the control of fungal plant pathogens5. Studies have shown that plant extracts of Chenopodium spp., Sennaoccidentalis and Cirsiumarvense can control growth of M.phaseolina and charcoal rot of mungbean10. From Azadirachta and Mango leaves, three flavonoids (–)-epi-catechin, (−)-epicatechin-3-O-β-glucopyranoside and 6-(phydroxybenzyl)taxifolin-7-O-β-d-glucoside were isolated and found effective against M.phaseolina12.Monothecabuxifolia  A. DC. is monotypic genus of the family Sapotacea, grows mainly in Malakand Dir district, Pakistan. M.buxifolia is reported for wide-range of pharmacological activities including cough, wound healing, headache, analgesic, antipyretic, antiseptic and urinary tract infections13. The antibacterial activity of crude ethanolic extract and fractions of M.buxifolia using agar well diffusion technique confirmed their antibacterial potential14. Javed et al.15 isolated and identified five triterpenes from the bioactive fractions of aerial parts of M.buxifolia and displayed potent cytotoxic activities in vitro. The other plants of family Sapotaceae are widely explored and reported for significant antimycotic activities in various experimental models, but there is not only a single report on antifungal activity and isolation of antifungal compounds from M.buxifolia16. Therefore, the present investigation was conducted to explore antifungal potential of M.buxifolia fractions and isolated compounds against M.phaseolina, a highly problematic phytopathogen for which there is not any registered fungicide up to now.m/z (relative intensity): 426(52) [M]+, 411(20), 393 (8), 218(42), 207(80), 189(100), 139(74). EIMS showed fragment peaks at m/z 411 [M+-CH3], 218 [M+-C14H20], 207(80) which are characteristic signals for pentacyclic triterpenes having isopropenyl group. HREI-MS m/z: 426.3820 . The 1H-NMR spectrum of 24 showed seven singlet for methyl protons at δ 0.74, 0.81, 0.85, 0.89, 0.91, 0.97 and 1.06 signals, one for each of seven methyl protons. A pair of multiplets at δ 4.20 and 4.21, each for H-29, identified the presence of terminal isopropenyl group, characteristic of lupane series of triterpenoids. 13C NMR spectrum showed 30 signals for the triterpenoid lupane skeleton, including C-3 at δ 79.2 bonded to the hydroxyl group. Compound 1 was identified as Lupeol19.EI-MS + 468.3967 correlated with C32H52O2 . EI-MS showed fragments 427 [M+—41], 408 [M+-AcOH], 249 [M+-C16H27] and 189 [M+-C16H27-AcOH]. 1H-NMR spectrum of 2 recorded seven tertiary methyl group at δ 0.76, 0.81, 0.83, 0.86, 0.92, 1.01, 1.31 and methyl acetate at 2.32. A pair of broad singlets at δ 4.47 and 4.44 showed presence of exomethylene group. A double doublet at δ 4.03, J = 9.8, 4.3 Hz was due to C-3 proton, bonded to acetoxy group. The coupling constant along with chemical shift describes the absolute configuration of acetoxy group, β and equatorial at C-3. The recorded data of 2 is comparable with the data reported for lupeol acetate in the literature20.HREI-MS showed [M]+ at 442.3819 deduced for molecular formula C30H50O2 depicted six degrees of unsaturation. 1H-NMR spectrum showed δ at 3.25  was attributed due to H-3 germinal to _OH proton. Two broad singlets appeared at δ 4.46 and 4.92  were assigned to terminal isopropenyl group. Six tertiary methyl singlets were observed at δ  were assumed for methyl protons of . One doublet appeared at δ 3.40 and 3.71  was due to methylene protons of C-28. Compound 3 can be identified as Betulin21.HR-EI-MS spectrum showed [M]+ at 414.3838, correlated with molecular formula C29H50O . Characetristic fragment ions m/z 414 and 396 in EI-MS spectrum also confirmed β-sitosterol pattern. Similarly fragment ion m/z 255 was recorded due to loss of [M+-side chain-H2O] and fragments m/z: 329, 303 confirmed the diagnostic pattern for sterols with ∆5-unsaturation. The 1H-NMR spectrum also confirmed the characteristic steroid pattern, one triplet at δ 5.41 (J = 3.2 Hz) was assigned for olefinic proton of C-6 and a carbinylic signal was recorded at δ 3.45 . Some doublet signals appeared at δ 0.902 (J = 6.4), 0.86 (J = 6.6), 0.82 (J = 6.6), 0.89 (J = 6.8) and were recommended for C-21, 26, 27 and C-29 methyl protons. The 13C-NMR spectrum of compound 4 showed 29 carbons with 6 methyl, 11 methylene, 9 methine and 3 quaternary carbon atoms. The compound 4 was identified as β-sitosterol, also confirmed by reported data in literature22.HR-EI-MS spectrum showed [M]+ at m/z 426.3534, established molecular formula C30H50O, calculated by 426.3860. Eight tertiary methyl protons were identified in the 1H NMR spectrum  δ 0.80, 0.84, 0.88, 0.95, 1.12, 1.34, 1.01 and 1.11, each . C-12 olefinic proton was also indicated at δ 5.22  and a double doublet at δ 3.61  was due to C-3 proton. The 13C NMR (BB and DEPT) spectrum showed presence of 8 –CH3, 10 –CH2 , 5 –CH and 7-quaternary carbons. The spectroscopic data was identical with the data of well-known compound reported in the literature, commonly known as β-amyrin23.HR-EI-MS showed M]+ at m/z 456. 3671 recommended for C30H48O3 calculated for 456.3603. EI-MS spectrum in addition to recording molecular ion peak m/z 456 showed characteristic fragment ions m/z 248, 203 and 133 for ∆12 amyrin skeleton. The 1H NMR spectrum displayed the signal for olefininc proton at δ 5.18  and oxymethine proton germinal to hydroxyl group at δ 3.14 . Seven singlets were also showed for seven methyl group protons were recorded at δ . 13C NMR spectrum showed 30 carbon signals comprising of 7 –CH3, 10 –CH2, 5 –CH and 8 –C carbon atoms. Carbonyl carbon showed prominent signal at δ 180.8, olefinic carbons δ 124.4 and oxymethine carbon δ 78.2. The above spectral data for compound 6 was in complete agreement Oleanolic acid24.HR-EI-MS spectrum showed the [M]+ at 168.1124  for C8H8O4. In EI-MS spectra base peak [M]+ is represented at 168 and other peaks at 151 and 137 represent the loss of –OH and –OCH3 groups. 1H-NMR spectrum  of compound 7 presented a doublets at δ 6.80 (J = 8.2) and a dd at 7.55 with (J = 8.2 and 1.6 Hz) were assigned C-5 and C-6 protons. One proton doublet at δ 7.60 with (J = 1.6 Hz), was assigned as C-2 proton. Singlet, three proton signal at δ 3.88 was attributed due to three protons of methoxy group. Another singlet at δ 4.59 was assigned to hydrogen of hydroxyl group. Compound 7 was thus identified as  commonly known as vanillic acid. The 1H and 13C NMR spectroscopic data was also verified with the previous reported data for vanillic acid25.HREI-MS spectrum showed [M]1H-NMR spectrum of compound 8 showed presence of three aromatic hydrogens, which gives information that three positions of benzene ring might be substituted. Compound 8 gave one double doublet at δ 7.45 with  was assigned to C-6 proton due to possibility of ortho coupling with C-5 proton and meta coupling with C-2 proton. Two doublets appeared for single protons in aromatic region can be assigned to C-5 proton with (J = 8.2 Hz) due to ortho coupling with C-6 proton and to C-2 proton with (J = 1.8 Hz) due to meta coupling with C-6 proton respectively. H-decoupled 13C-NMR spectrum, seven peaks appeared for six non-equivalent carbons and one carboxylic carbon. The calculated mass for compound 18 with molecular formula C7H6O4 was 154.2338 and 154.2331 was observed again confirms the compound as protocatechuic acid26.+ was recorded at 286.0475 which is compatible for molecular formula C15H10O6. EI-MS showed characteristic fragments at 257, 229, 213, 121,104, 93, 77 and 69. 1H NMR spectrum displayed two doublets at δ 8.03 and δ 6.92 representing AA'BB' pattern for para-substituted benzene. Two meta coupled doublets also appeared at δ 6.42 and 6.18 with J = 1.8 Hz. Characteristic flavonoid signals 137.2, 146.2 were shown clearly in 13C NMR (DEPT and BB) spectra. 13C NMR signals at δ 161.4, 160.2 and 114.2 represent the para-substituted benzene ring moiety. Similarly carbon atoms directly linked with oxygen displayed signals at 161.4, 103.4 and 160.2. The spectroscopic data was also matched with the reported data in literature and compound (9) was identified as 3,4′,5,7-tetrahydroxyflavone commonly known as kaempferol27.Molecular ion peak [M]10) HR-EI-MS spectra depicted molecular ion peak at m/z 302.0285 with relevant molecular formula C15H10O6. The 1H-NMR spectrum of (10) showed five signals each for five different aromatic protons. A doublet signal at 7.72 represents single proton in meta coupling with H-6′, 2.1 Hz might be attributed due to H-2′. A double doublet at 7.62 represents H-6′, in meta coupling with H-2′, J = 2.1 Hz and ortho coupling with H-5′, J = 8.4 Hz. Another doublet at 6.87 might be due to H-5′, J = 8.4. It suggested an ABC type-system having flavonol moiety with 3′, 4′-disubstituted B-ring. Similarly a pair of meta coupling protons signals at 6.17  and 6.38  were recorded for ring A. 13C-NMR spectra showed 15 signals having carbonyl signal at δ 170.5 for C-4. Five carbon nuclei attached to hydroxyl group gave five signals recorded at δC 137.1 (C-3), 160.9 (C-5), 165.1 (C-7), 146.4 (C-3′), 150.8 (C-4′) also supported the presence of 3,5,7,3′,4′-oxygenated flavone nucleus. The detailed studies on compound 10 were in consistent with the reported spectroscopic data, commonly known as quercetin28.Compound  showed the highest antifungal against M.phaseolina, causing 79–81% followed by betulin (3) 77–79% reduction in biomas of the target fungus, respectively. Other compounds showing pronounced antifungal activities were β-amyrin (5), oleanolic acid (6), vanillic acid (7) and protocatechuic acid (8) that reduced fungal biomass by 68–71%, 68–71%, 74–79% and 67–72%, respectively. Compounds namely lupeol (1), β-sitosterol (4), kaempferol (9) and quercetin (10) were comparatively less antifungal and reduced fungal biomass by 40–43%, 57–64%, 46–53% and 47–55%, respectively over corresponding control.Antifungal activity of mencozeb and the ten isolated compounds against M.buxifolia methanolic extract ten compounds were isolated from various sub-fractions. Various physical analysis and spectroscopic data co-ordinated from the already reported data and compound 1 was identified as lupeol, compound 2 as lupeol acetate, compound 3 as betulin, compound 4 as β-sitosterol, compound 5 as β-amyrin, compound 6 as oleanolic acid, compound 7 as vanillic acid, compound 8 as protocatechuic acid, compound 9 as kaempferol and compound 10 as quercetin28 Structures of these compounds are presented in Fig. From 2) 79–81% and betulin (3) 77–79% were found very close to that of fungicide mancozeb 83–84%. Some earlier studies also reported antifungal activity of lupeol acetate (2) against Aspergillusflavus. However, most of the previous studies showed antibacterial activity of this compound against Bacillussubtilis, Staphylococcusaureus and Escherichiacoli30. In contrast to the present study, Manzano et al.31 reported no antifungal activity of 2 against Penicilliumchrysogenum and Fusariumoxysporum, showing specificity of this compound against M.phaseolina. Betulin (3) has been reported fungicidal against a number of fungi including Fusariumsolani and Aspergillusniger and its effect was equal to standard antifungal drug miconazole32.All the concentrations of methanolic extracts and its various fractions significantly reduced fungal biomass. Antifungal activity of both compounds lupeol acetate (5) is known to inhibit growth of various clinical fungal species namely Candidastellatoidea,Candidakrusei,Microsporum sp. and Trichophytonrubrum and Ascochytarabiei, the cause of blight disease of chickpea34 Oleanolic acid (6) is a pentacyclic triterpenoid that is widely spread in plant kingdom, with members of Oleaceae family as its main source. Apart from its various pharmaceutical properties, it also possesses antifungal activity against yeast and dermatophyte species36. Protocatechuic acid (8) is a type of phenolic acid that is found in many food plants and possesses a number of pharmaceutical properties and antifungal activity against Microsporumaudouinii38.β-Amyrin (1), a pentacyclic triterpenoid, has also been isolated from a number of plant species including green pepper, mangoes, strawberry, grapes, white cabbage and olive, and is known to have useful effect as a preventive and therapeutic agent against a number of ailments39. Although in the present study, it found comparatively less effective against M.phaseolina, however, it was highly effective against Penicilliumnotatum causing 90% growth inhibition when used in 200 µg mL−1 concentration31. It was also inhibitory to the growth of Fusariumsolani,Aspergillusniger,Rhizoctoiaphaseoli,Candidaalbicans,Penicilliumchrysogenum,Cantharellusflavus and Microsporumcanis, and its antifungal activity was comparable with miconazole40. β-Sitosterol (4) is known to inhibit growth of Aspergillusniger and Cladosporiumcladosporioides at a concentration of 0.01 mg mL−1. It also inhibited growth of germ tube of Fusariumverticillioides by 82% at 50 mg mL−141. Quercetin (10) is a polyphenolic flavonoid and showed a low activity against different Candida species42. This compound is also known to increase efficacy of a fungicidal compound amphotericin B against a clinical fungal species Cryptococcusneoformans43. Kaempferol (9) is a phenolic compounds generally showing antibacterial activity44. Information regarding its antifungal activity are very rare. The present study concludes that aerial parts of M.buxifolia contain potentent antifungal constituents especially lupeol acetate (2) causing 79–81% reduction followed by betulin (3) 77–79% and vanillic acid (7) 74–79% for the management of M.phaseolina, a highly problematic phytopathogen for which there is not any registered fungicides so far.Lupeol . Silica and Flash column silica (70–230 mesh) and (230–400 mesh) were assimilated from E. Merck, Darmstadt, Germany.max in nm. Infrared spectra (IR) were recorded on Shimadzu IR-460 in nujol or KBr pellets and reported in cm−1. NMR spectra were recorded on Bruker AMX-500 and AM-400 for1H (300–500 MHz) and13C (125–150 MHz) spectrometers. "spectra were recorded on Bruker Avance spectrometers ranging from 7.05 up to 14.09 T". Mass spectrometer Jeol-JMS H X-110 was employed to record high resolution electron ionization mass spectra (HR-EIMS).Melting point apparatus Buchi 535 was used to record melting point. Digital Polarimeter; Jasco DIP-360 was used to record optical rotation. Shimadzu UV-240 spectrophotometer was used to record λM.buxifolia is wild and commonly grown plant species in mountains of Swat, Pakistan. Aerial parts viz., stem and leaves (11 kg) of M.buxifolia were collected from Swat, Pakistan according to prescribed rules in The Pakistan Trade Control of Wild Fauna and Flora Act, 2012. Species was identified by Dr. Arshad Javaid , assigned voucher no. GC. Bot. Herb. 815 (Monothecabuxifolia) and was deposited in the Dr. Sultan Ahmed Herbarium, Department of Botany, GC University, Lahore, Pakistan.n-hexane, chloroform and ethyl acetate (10 × 3 L) each to afford 151, 102 and 152 g of the fractions respectively. Aqueous fraction was collected 161 g and all the portions were refrigerated17.Aerial parts were dried under shade, pulverized and weighed (4.5 kg). Powdered plant material was soaked in MeOH 95% (16 L × 4). Extracts were assembled and methanol was removed under reduced pressure at 50 °C temperature. Gummy mass (586 g) having dark brown colour was collected. This MeOH extract was added in water (410 mL) and further fractionation was done using M.phaseolina (FCBP-0751) on the basis of characteristic black-colored oblong microsclerotia18.Diseased root pieces of mungbean were collected, washed and cut into small pieces of 0.5 cm. Root pieces were surface sterilized using 0.1% sodium hypochlorite solution, transferred aseptically on autoclaved malt extract agar plates and incubated for 7 days at 27 °C. Grey fungal colonies appeared on root pieces which became darken with time. The colonies were further purified by growing on fresh malt extract agar plates. The fungus was isolated and identified as M.phaseolina in vitro. An amount of 1.2 g of methanolic extract and its various fractions was dissolved in 1 mL of dimethyl sulphoxide (DMSO) and added to 5 mL autoclaved malt extract broth to make stock solution of 200 mg mL−1 concentration. Six lower concentrations viz. 100, 50, 25, 12.5, 6.25 and 3.125 mg mL−1 were prepared from stock solution in series of double dilution. Similarly, control with respect to each concentration was prepared by dissolving 1 mL DMSO in 5 mL malt extract broth and double diluted in series. Experiment was performed in glass test tubes (10 mL), each containing 1 mL of growth medium. Tubes were inoculated aseptically with 20 µL of M.phaseolina suspension and each treatment was replicated three times. After 7 days of incubation at room temperature, fungal biomass was filtered, dried and weighed.The methanolic extract and its four fractions were tested against the pathogenic fungus M.buxifolia lipophilic hexane fraction was subjected to vacuum liquid chromatography over silica gel and eluted with increasing order of solvent polarity as hexane–EtOAc (0 → 10). Hexane first sub-fraction, collected using n-hexane: EtOAc (7:3) was again chromatographed to isolate compound 1 (10 mg) using same polarity system. Hexane second sub-fraction collected using n-hexane–EtOAc (6:4) was again chromatographed to isolate compound 2 (8 mg) eluted with same solvent system. CHCl3 fraction was subjected to silica gel column and eluted with n-hexane-chloroform (10:0 → 0:0) to chloroform–methanol (0:0 → 0:10). Chloroform fraction lead to isolation of three main sub-fractions. CHCl3 sub-fraction 1 was further purified and compound 3 and 4  were obtained with n-hexane-CHCl3 (7:3) n-hexane-dichloromethane (5:5) and (6:4) as eluent, respectively. Sub-fraction 2 eluted with n-hexane-chloroform (6:4) was purified with n-hexane:dichloromethane (5:5) to obtain compound 5 (15 mg). CHCl3 sub-fraction 3 eluted with chloroform–methanol (9.5:0.5) was rechromatographed using dichloromethane-methanol (9:1) to afford compound 6 (30 mg). The EtOAc fraction was chromatographed using silica gel and eluted with solvent system of increasing polarity n-hexane, n-hexane: DCM and DCM:MeOH and three sub-fractions were collected. EtOAc sub-fraction 1, eluted by n-hexane: DCM (3:7) was refilled on silica gel and eluted by n-hexane:DCM (1:9) to attain compound 4 (4 mg) and 7 (6 mg). EtOAc sub-fraction 2 eluted by DCM 100%, was reloaded and eluted by same solvent system to separate compound 8 (8 mg). EtOAc sub-fraction 3 was isolated using DCM:MeOH (9:1) was rechromatographed and eluted with identical solvent system to collect compound 9 and 10 .1H-NMR , δ ppm: 4.20 and 4.21 , 3.30 , 0.74, 0.81, 0.85, 0.89, 0.91, 0.97, 1.06 . 13C-NMR  δ ppm: 150.7 (C-20), 109.7 (C-29), 79.2 (C-3), 55.2 (C-5), 50.3 (C-9), 48.3 (C-18), 47.8 (C-19), 43.2 (C-17), 42.7 (C-14), 40.7 (C-8), 39.9 (C-22), 38.6 (C-4), 38.2 (C-1), 38.0 (C-13), 37.3 (C-10), 35.9 (C-16), 35.0 (C-7), 29.7 (C-21), 28.4 (C-22), 28.2 (C-23)27.9 (C-2), 27.2 (C-15), 25.3 (C-12), 21.1 (C-11), 19.3 (C-30), 18.4 (C-6), 18.2(C-28), 15.8 (C-25), 15.5 (C-26), 15.3 (C-27), 15.1 (C-24). EI-MS m/z [M]+ at : 426.Melting point, 215 °C was recorded. −1. 1H-NMR , δ ppm: 4.47 and 4.44 , 4.03 , 0.76, 0.81, 0.83, 0.86, 0.92, 1.01, 1.31  and 2.32 . 13C- NMR  δ ppm: 172.2 (C-1′), 150.2 (C-20), 110.5 (C-29), 81.1 (C-3), 50.7 (C-9), 48.7 (C-18), 48.4 (C-19), 38.5 (C-1), 22.7 (C-2), 37.2 (C-4), 55.3 (C-5), 18.3 (C-6), 34.8 (C-7), 41.2 (C-8), 40.1 (C-22), 37.2 (C-10), 21.2 (C-11), 24.6 (C-12), 37.2 (C-13), 43.0 (C-14), 26.4 (C-15), 35.8 (C-16), 43.1 (C-17), 29.9 (C-21), 28.2 (C-23), 20.4 (C-2′), 19.3 (C-30),16.1 (C-24), 16.3 (C-25), 15.4 (C-26), 14.2 (C-27), 14.8 (C-28). EI-MS m/z [M]+ at : 468.IR spectrum showed absorption at 1730 cm3 showed IR spectrum at 3457 cm−1. 1H-NMR , δ ppm: 4.46 and 4.92 , 3.40 and 3.71 , 3.25 , 0.83 (H3-24), 0.84 (H3-25), 0.86 (H3-23), 0.87 (H3-26), 1.02 (H3-27), 1.04 (H3-30) . 13C- NMR  δ ppm: 150.8 (C-20), 109.8 (C-29), 79.1 (C-3), 60.4 (C-28), 38.4 (C-1), 55.3 (C-5), 50.6 (C-9), 48.6 (C-19), 46.6 (C-17), 42.5 (C-18), 42.8 (C-14), 27.5 (C-2), 39.2 (C-4), 37.5 (C-13), 36.2 (C-22), 30.2 (C-21), 18.4 (C-6), 34.3 (C-7), 39.5 (C-8), 37.2 (C-10), 21.0 (C-11), 24.2 (C- 12), 27.2 (C-15), 30.3 (C-16), 28.1 (C-23), 19.1 (C-30), 16.1 (C-25), 16.2 (C-26), 15.2 (C-24), 14.8 (C-27). EI-MS m/z [M]+ at : 442.Compound 1H-NMR , δ ppm: 5.41 , 3.45 , 1.034 , 0.902 , 0.89 , 0.86 , 0.82 , 0.68 . 13C-NMR  δ ppm: 141.2 (C-5), 122.1 (C-6), 72.2 (C-3), 56.8 (C-14), 56.2 (C-17), 51.9 (C-9), 46.4 (C-24), 42.4 (C-4), 41.8 (C-13), 40.6 (C-12), 36.8 (C-1), 35.8 (C-10), 34.4 (C-20), 32.2 (C-22), 31.8 (C-7), 31.6 (C-8), 31.2 (C-2), 30.4 (C-25), 28.6 (C-16), 25.4 (C-23); 24.8 (C-15), 23.8 (C-28), 22.8 (C-11), 21.2 (C-26), 20.6 (C-19), 19.8 (C-27), 18.9 (C-21), 12.8 (C-29), 12.6 (C-18). EI-MS m/z [M]+ at : 414.1H-NMR , δ ppm: 5.22 , 3.61 , 0.80 , 0.84 , 0.88 , 0.95 , 1.12 , 1.34  , 1.01  and 1.11 . 13C-NMR  δ ppm: 145.6 (C-13), 121.4 (C-12), 78.8 (C-3), 47.2 (C-18), 46.6 (C-19), 37.8 (C-1), 28.1 (C-2), 38.4 (C-4), 55.6 (C-5), 18.8 (C-6), 33.1 (C-7), 38.2 (C-8), 47.8 (C-9), 37.2 (C-10), 23.4 (C-11), 41.2 (C-14), 26.8 (C-15), 27.3 (C-16), 32.2 (C-17), 31.1 (C-20); 34.2 (C-21), 37.3 (C-22), 28.9 (C-23), 15.5 (C-24), 15.6 (C-25), 16.7 (C-26), 26.4 (C-27), 28.8 (C-28), 33.4 (C-29), 23.8 (C-30). EI-MS m/z [M]+ at : 426.Absence of absorption in UV region was indicated. - respectively.1H-NMR , δ ppm: 5.18 , 3.14 , 1.19 , 1.02 , 0.92 , 0.88 , 0.80 , 0.74  and 0.71 . 13C-NMR  δ ppm: 180.8 (C-28), 145.8 (C-13), 124.4 (C-12), 78.2 (C-3), 55.9 (C-5), 48.5 (C-9), 47.2 (C-17), 46.6 (C-19), 42.5 (C-18), 42.4 (C-14), 39.4 (C-4), 38.8 (C-1), 33.8 (C-29), 33.3 (C-7), 40.1 (C-8), 37.1 (C-10), 31.4 (C-20); 34.8 (C-21), 33.1 (C-22), 29.8 (C-23), 24.1 (C-11), 28.8 (C-15), 28.2 (C-2), 26.8 (C-27), 24.2 (C-16), 23.6 (C-30), 19.1 (C-6), 17.2 (C-26), 16.4 (C-24), 16.1 (C-25). EI-MS m/z [M]+ at : 456.IR spectrum showed absorption band 3430–2650, 1680 and 1650–850 cmmax showed absorption at 282 nm. The infrared spectrum displayed absorption bands at 3400–2700 and 1670 cm−1 indicated the presence of conjugated carboxylic acid. 1H-NMR , δ ppm: 7.60 , 7.55 , 6.80 , 4.59 , 3.88 . 13C- NMR  δ ppm: 55.9 (CH3O-3), 112.3 (C-2), 114.3 (C-5), 121.9 (C-6), 124.0 (C-1), 146.6 (C-3), 150.5 (C-4). EI-MS m/z [M]+ at : 168.The UV λ−1. 1H-NMR , δ ppm: 7.45 , 7.42 , 6.82 , 13C- NMR  δ ppm: 123.4 (C-1), 120.9 (C-2), 147.2 (C-3), 148.9 (C-4), 117.4 (C-5), 118.5 (C-6), 170.5 (C-7). EI-MS m/z [M]+ at : 154.UV absorption was recorded at 274 nm, IR spectrum displayed absorption bands at 3210–2610 cmmax at 265, 212 and 336 (4.0) nm while IR spectrum showed absorption at 3412, 2926, 1660, 1610, 1600–1525 and 1380 cm−1. 1H-NMR , δ ppm: 8.03 , 6.92 , 6.42 , 6.18 .13C- NMR  δ ppm: 178.1 (C-4), 161.4 (C-5), 160.2 (C-7), 159.4 (C-4′), 155.9 (C-9), 146.2 (C-2), 137.2 (C-3), 128.3 , 121.3 (C-1′), 114.2 , 103.4 (C-10), 99.8 (C-6), 93.4 (C-8). EI-MS m/z [M]+ at : 286.Melting point at 276–278 °C was recorded. UV spectrum recorded λm/z 302.0285 with relevant molecular formula C15H10O6. IR spectrum showed two characteristic bands at 3402 and 1610 cm−1. 1H-NMR , δ ppm: 7.72 , 7.62 , 6.87 , 6.38 , 6.17 . 13C-NMR  δ ppm: 170.5 (C-4), 165.1 (C-7), 160.9 (C-5), 160.0 (C-9), 150.8 (C-4′), 146.4 (C-3′),146.2 (C-2), 137.1 (C-3), 124.2 (C-1′), 121.4 (C-6′)117.0 , 104.2 (C-10), 99.2 (C-6), 94.1 (C-8). EI-MS m/z [M]+ at : 302.Melting point was recorded at 314–316 °C. Two characteristic absorption bands in UV spectrum at 250 and 360 nm were showed. HR-EI-MS spectra depicted molecular ion peak at  were isolated from n-hexane fraction, four compounds (3–6) from chloroform fraction and four compounds (7–10) from ethyl acetate fraction of the methanolic extract of M.buxifolia were tested for MIC values by microdilution assay. MIC values of mancozeb as reference synthetic fungicide  and isolated compounds were tested by serial dilution in culture tubes. Six milligrams of each of the ten isolated compounds and mancozeb (active ingredient) were dissolved in 20 µL DMSO and added to autoclaved malt extract to raise the volume up to 3 mL, to prepare a growth medium of 2 mg mL−1 concentration. Further serial double dilutions viz. 1, 0.5, 0.25…0.0312 mg mL−1 were made using malt extract broth in culture tubes. Spore suspension was prepared by adding 10 day old fungal culture in double distilled water. DMSO (20 µL mL) was added in malt extract broth to prepare 3 mL of control that serially double diluted to make corresponding control treatments for each concentration. Each treatment was replicated thrice. Fugal suspension (20 µL) was added to each concentration (0.5 mL) of the growth medium and incubated at 27 °C. Fungal biomass was collected on filter papers after 3 days growth, dried and weighed.Two compounds All the data were analyzed by analysis of variance followed by Tukey’s HSD Test using computer software Statistix 8.1.This article does not contain any studies with human participants or animal experiments.


According to French Paradox, red wine was famous for the potential effects on coronary heart disease (CHD), but the specific compounds against CHD were unclear. Therefore, screening and characterization of bioactive compounds from red wine was extremely necessary. In this paper, the multi-activity integrated strategy was developed and validated to screen, identify and quantify active compounds from red wine by using ultra high performance liquid chromatography-fraction collector (UHPLC-FC), ultra fast liquid chromatography-quadrupole-time-of-flight/mass spectrometry (UFLC-Q-TOF/MS) and bioactive analysis. UHPLC-FC was employed to separate and collect the components from red wine, which was further identified by UFLC-Q-TOF/MS to acquire their structural information. Furthermore, the active fractions were tested for antioxidant activity, inhibitory activity against thrombin and lipase activities in vitro by the activity screening kit. As the results, there were 37 fractions had antioxidant activity, 22 fractions had thrombin inhibitory activity and 28 fractions had lipase inhibitory activity. Finally, 77 active components from red wine were screened and 12 ingredients out of them were selected for quantification based on the integration of multi-activity. Collectively, the multi-activity integrated strategy was helpful for the rapid and effective discovery of bioactive components, which provided reference for exploring the health care function of food. As a nutritious drink recommended by World Health Organization (WHO), red wine was widely recognized for its role in health care. According to Ben Cao Gang Mu (Compendium of Materia Medica) record, the famous classic book of traditional Chinese medicine, red wine could treat cold coagulation and blood stasis. Modern pharmacological studies showed that red wine had the function of antioxidant, anticoagulant and lipid-lowering ,2, whichIt was widely believed that the mechanisms of CHD were the result of the interaction of many complex factors, among them the “response to injury hypothesis” had been recognized by researchers ,6, whichThe famous French paradox proved that red wine can prevent CHD . AccumulIn this study, UHPLC-FC was used to separate, collect and enrich the fractions from red wine. Then antioxidant activity, inhibitory activity against thrombin and lipase activities of all fractions were performed for evaluating activities. Furthermore, UFLC-Q-TOF/MS was used to identify the components in fractions from red wine . As the In the study, an ultra high performance liquid chromatography-photo-diode array-fraction collector (UHPLC-PDA-FC) system was used to establish multi-activity integrated strategy for screening and evaluating the potentially bioactive components from red wine. The sample was divided into sixty fractions (60 s per fraction) by UHPLC-PDA-FC. Sixty fractions were used to screen the antioxidant activity, thrombin inhibitory activity and lipase inhibitory activity. The multi-activity integrated chromatogram consisted of chromatogram and multi-targets integrated column diagram of antioxidant activity, inhibitory activity of thrombin and lipase were established  and the 5H9NO2) with MS/MS fragment of 70.0647 (C4H7N) [M+H-HCOOH]+, which was detected to be D-proline [4H9N) [M+H-HCOOH]+ and 14 (C5H3N) [M+H-HCOOH]+ were based on the same regular. Therefore, peaks 11 and 14 were identified as L-valine (C5H11NO2) [6H5NO2) [6H14N4O2) with MS/MS fragment of 70.0652 [M+H-HCOOH-2NH2-CN]+, which was detected to be L-arginine [6H12O6) with fragments of 99.0087 (C4H4O3) [M-H-3H2O-C2HO]−, 71.0137 (C3H4O2) [M-H-3H2O-C2HO-CO]− and 59.0139 (C2H4O2) [M-H-3H2O-C2HO-CO-CH2]−. Therefore, it was identified as D-glucose. Peak 4 had mass spectrum ion at 181.072 (C6H14O6) with MS/MS experiment gave fragmentation information for 83.0143 [M-H-C3H2O3]− and 71.0140 [M-H-C3H6O3-H2O]−, which could be identified as D-mannitol [6H8O7) and the MS/MS fragments were 85.0296 (C4H6O2) [M-H-2CO2-H2O]− and 59.0134 (C3H8O) [M-H-3CO2]−. Therefore, it was identified as citric acid [3H6O4) [M-H-CO2]− and 87.0085 (C3H4O3) [M-H-CO2-H2O]− were observed [9H8O3) had the MS/MS fragment of m/z 119.0502 (C8H8O) [M-H-CO2]−. Therefore, Peak 40 was identified as 2-hydroxycinnamic acid [7H7NO2) and the fragment ions observed included m/z 120.0433 (C6H7N) [M+H-H2O]+, 78.0339 (C6H6) [M+H-HCOOH-NH2]+, which was identified to be 4-aminobenzoic acid [12H22O11) with MS/MS fragments of 161.0459 (C6H9O5) [M-H-C6H11O5-H2O]− and 71.0137 (C3H4O2) [M-H-C9H14O8-H2O]−, which was detected to be trehalose − and 59.0133 (C2H2O2) [M-H-C3H8O2]−. By comparing the characteristic fragmentation pattern with the reported in references, it can be identified as D-ribose. Peaks 10 (C4H6O5) was identified as L-malic acid − and 71.0137 (C3H4O2) [M-H-H2O-CO2]−. Peaks 12 (C7H10O5) was identified as shikimic acid. MS/MS fragment was 137.0225 (C7H6O3) [M-H-2H2O]−. The molecular weight of peak 13 was 150.0586 (C5H11NO2S) and ion m/z 74.0243 (C3H8O2) [M+H-NH3-C2S]+ was observed in MS/MS spectra, which could be identified as L-methionine − and 73.0291 (C3H4O) [M-H-CO2-H2O]−. Hence, it can be identified as succinic acid [3H5O2]−) with peak 15. The other fragment was 55.0191 [M-H-C3H5O2-H2O]−, which can be identified as dimethyl succinate − and 91.0298 [M-H-CO2-H2O]−), 23 (135.0560 [M-H-CO2]− and 89.0194 [M-H-CO2-CH3CO]−), 24 (135.0454 [M-H-CO2]− and 117.0351 [M-H-CO2-H2O]−), 25 (109.0298 [M-H-CO2]− and 91.0298 [M-H-CO2-H2O]−), 29 (93.0340 [M-H-CO2]−), 31 (163.0400 [M-H-2CO2]− and 119.0503 [M-H-3CO2]−), 32 (119.0498 [M-H-CO2]− and 93.0347 [M-H-CO2-H2O]−), 33 (163.0400 [M-H-2CO2]− and 119.0498 [M-H-3CO2]−), 39 (108.0212 [M-H-CO2-CH3]−), 42 (193.0490 [M-H-2CO2-C2H5O]− and 117.0341 [M-H-3CO2-C2H5O-CH3-H2O]−), 43 (135.0449 [M-H-CO2]−), 49 (119.0527 [M-H-CO2]− and 103.0565 [M-H-CO2-H2O]−), 50 (93.0470 [M-H-CO2]− and 65.0473 [M-H-CO2-CO]−), 52 (123.0436 [M-H-CO2]− and 108.0212 [M-H-CO2-H2O]−), 55 (119.0503 [M-H-CO2]− and 93.0347 [M-H-CO2-H2O]−), 56 (123.0450 [M-H-CO2]−), 60 (257.0110 [M-H-CO2]− and 229.0126 [M-H-CO2-CO]−) and 61 (109.0286 [M-H-CO2-C2H2]−) were the same regular, which were identified as gallic acid [11H12O4). The MS/MS fragments were the same as peak 43 of 179.0433 [M-H-CH3]− and 135.0446 [M-H-CH3-CO2]−. Therefore, it was identified as ethyl caffeate [9H8O3) and the product ion was 119.0479 [M+H-HCOOH]+, which could be identified as trans-4-hydroxycinnamic acid [8H11NO) with the MS/MS fragments at 121.0616 [M+H-NH3]+ and 95.0561 [M+H-NH3-C2H4]+, which was detected to be tyramine − ion at m/z 153.0558. In MS/MS mode, the produced ions were m/z 123.0085 [C8H10O3-CH2O]− and 108.0221 [C8H10O3-CH2OH-CH3]−, which can be identified as hydroxytyrosol [− in the first-order mass spectrum. They had the same prominent ion at 125.0246 [C15H14O7-C9H8O4]−. They were speculated to be (−)-epigallocatechin [− ion at m/z 137.0255. The product ion was 108.0213 [M-H-CHO]−. Therefore, peak 26 was identified as protocatechualdehyde. Peak 27 had the mass spectrum ion at 181.0506 (C9H10O4) with MS/MS fragment of 135.0453 [M-H-CH3-H2O]−, which was identified as homovanillic acid [8H8O5). The product fragment ions were 124.0234 [M-H-CO2-CH3]− and 95.0136 [M-H-CO2-CH3-CHO]−. Hence, it was identified as methyl gallate [− ion at m/z 353.0877 and MS/MS produced ions were at m/z 191.0616 [M-H-C6H5O5]− and 135.0439 [M-H-C6H5O5-CO]−, it can be identified as chlorogenic acid [− and 108.0212 [M-H-CH]−. Peaks 36, 38, 46 and 57 had same molecular weight of C30H26O12, which illustrated that they were isomers. Base on the MS/MS spectra, peak 36 , 38 , 46  and 57  were detected, which could be identified as procyanidin B1 [15H14O6) with fragments of 245.0843 [M-H-CO2]−, 203.0713 [M-H-H2O-C3O2]− and 151.0401 [M-H-C8H10O2]−, therefore it was identified as catechin [7H2O4]−, 151.0386 [M-H-C7H4O4-C8H10O2]−), 51 , 58 , 64 (285.0413 [M-H-C6H11O5]− and 151.0041 [M-H-C14H8O7]−), 66 , 67 , 68 (151.0020 [M-H-C14H16O7]−), 69 (285.0413 [M-H-C6H11O5]− and 151.0041 [M-H-C14H8O7]−), 70 (301.0379 [M-H-C6H10O5]− and 151.0039 [M-H-C6H10O5-C7H6O2-CO]−), 72 , 73 (245.0504 [M-H-H2O-2CO]− and 151.0035 [M-H-C8H6O4]−), 75 , 83 (151.0034 [M-H-C8H6O3]−), 85 (316.0231 [M-H-CH3]− and 151.0016 [M-H-C9H8O4]−), 86 (151.0034 [M-H-C8H8O]−), 88 (151.0034 [M-H-C8H6O2]−) and 91 (300.0270 [M-H-CH3]− and 151.0016 [M-H-C9H8O3]−) were identified as the type of ion of 151.0037 (C7H4O4), which were the characteristic fragment of flavonoid components. Indeed, they were epicatechin gallate [9H10O5) with MS/MS ions at 182.0256 [M-H-CH3]−, 167.0014 [M-H-CH3-CO2]−, 138.0317 [M-H-CH3-CO2-CH3]− and 123.0090 [M-H-CH3-2CO2-CH3]−, which can be identified as syringic acid [− at m/z 325.0926 with the molecular formula C15H18O8. A loss of C6H10O5 in MS/MS was confirmed to be a hexose neutral loss. The other ions were 145.0294 [M-H-C6H10O5-H2O]− and 117.0343 [M-H-C6H10O5-CO]−. Peak 47 was tentatively identified as p-coumaric acid glucoside [8H14O5) and the MS/MS fragment was 71.0503 [M-H-C2H5-CO2-C2H5O]−. Therefore, it was identified as diethyl malate [9H10O5-C2H4]− and 125.0239 [C9H10O5-C2H4-CO2]− were observed [6H10O5]− and 316.0553 [M-H-C6H10O5-CH3]−. Hence, peak 54 was tentatively identified as malvidin-3-O-glucoside [6H10O5]− and 316.0553 [M-H-C6H10O5-2H2O-CO]−), 65 (303.0510 [M-H-2C6H10O5]−), 74 (317.0648 [M-H-C6H10O5]− and 302.0429 [M-H-C6H10O5-CH3]−), 78 (303.0767 [M-H-C6H10O5-CO2]−) and 80 (477.1196 [M-H-C9H10O3]− and 331.0874 [M-H-C9H10O3-C6H10O5]−) were displayed in MS/MS spectra, which could be identified as delphinidin-3-O-glucoside [− ion at the m/z 389.1242 with MS/MS spectra ions m/z 227.0994 [M-H-C6H10O5]− and 143.0505 [M-H-C6H10O5-C2H2O-C2H2O]−. Peak 59 was tentatively identified as trans-piceid [6H10O5]− and 185.0606 [M-H-C6H10O5-C2H2O]−), 76 (185.0587 [M-H-C6H10O5-C2H2O]− and 143.0501 [M-H-C6H10O5-C2H2O-C2H2O]−) and 84 (143.0504 [M-H-C6H10O5-C2H2O-C2H2O]−) were displayed in MS/MS spectra, which can be identified as cis-piceid [21H20O12). The product fragment ions were 301.0379 [M-H-C6H10O5]−, 300.0277 [M-H-C6H11O5]−, 271.0245 [M-H-C6H10O5-CHO]−, 243.0291[M-H-C6H10O5-CHO2]− and 151.0031 [M-H-C6H10O5-C7H6O2-CO]−. Therefore, it was identified as hyperoside [21H24O10) with MS/MS fragments of 273.0769 [M-H-C6H12O6]−, 179.0349 [M-H-C15H14O5]− and 167.0352 [M-H-C6H12O6-C7H6O]−, which was detected to be phloridzin [15H10O7) with fragments of 285.0398 [M+H-H2O]+, 257.0456 [M+H-H2O-CO]+, 229.0492 [M+H-H2O-2CO]+ and 153.0188 [M+H-C8H5O3]+, therefore it was identified as morin [2O]− and 153.0188 [M-H-C8H6O3]−,which can be identified as delphinidin [2O-CO] and 137.0233 [M-H-C8H6O3]) and 90 (229.0498 [M-H-2H2O-CO-CH3] and 153.0188 [M-H-CH3-C8H6O3]) were showed in MS/MS spectra, which can be identified as cyanidin [3]− and 315.0153 [M-H-C2H6]− [13H20O) and the fragment ion was observed at m/z 56.9419 [M+H-C10H15]+, which was identified as ionone [10H20O2) with fragment of 76.0221 [M+H-HCOOH]+, therefore it was identified as ethyl caprylate (C10H20O2). The molecular weight of peak 94 was 195.1748. MS/MS ions of m/z 177.1634 [M+H-H2O]+ and 163.0394 [M+H-H2O-CH2]+ were observed, which revealed that it was theaspirane. Moreover, the MS spectrums of all compounds were shown in The components from red wine with antioxidant activity or inhibitory potential against thrombin and lipase activities were analyzed by UFLC-Q-TOF/MS to obtain structural information. The molecular compositions of ingredients were identified according to the literature and exact molecular weight. The peak number, retention time, MS/MS fragmentation ions information and identification of 94 components from red wine were listed in -proline ,15,16. P × 100%, where Asample was the fluorescence of the reactions with added fraction, Acontrol was the control, and Acontrol sample was the fluorescence of the solvent of the fraction [In this study, the inhibition of each fraction was assayed by measuring the fluorescence intensity of 4-methylumbelliferyl oleate (4-MU) (the hydrolytic product of 4-MUO). The assay was performed according to the method described with slight modifications. Initially, the concentrations of trypsin  and time were investigated to establish linear range with the concentration of 4-MUO at 0.1 mM. Then, 25 μL fractions and 25 μL lipase solutions were mixed together and 50 μL 4-MUO was added to the mixture. Finally, the amount of 4-MU was measured by microplate reader at an excitation wavelength of 355 nm and an emission wavelength of 460 nm after incubating at 25 °C for 30 min. The inhibition of pancreatic lipase activity was calculated as follows: inhibition (%) = [(Afraction ,54.The results of precision and recovery have been measured in triplicates and the stability have been measured in sextuplicate, which were expressed as mean and RSD. The content of 12 compounds have been measured in triplicates and expressed as mean.In this study, the multi-activity integrated strategy was proposed to screen, identify and quantify components from red wine for prevention of CHD via UHPLC-FC and UFLC-Q-TOF/MS. The multi-activity integrated strategy of red wine was established and verified the availability of screening the preventing CHD compounds from the complex sample. Red wine exhibited potential antioxidant activity and inhibitory activity against thrombin and lipase, which could be used to prevent CHD. In addition, the system had advantages over traditional methods in screening potential active compounds. The multi-activity integrated strategy may help to discover bioactive components rapidly and efficiently, which provided reference for exploring the health care function in food.