Shao-Lan Li

Ten-Membered Lactones from Phomopsis sp., an Endophytic Fungus of Azadirachta indica

Four new 10-membered lactones ( 1- 4) and one known one ( 5) were isolated from the broth extract of an endophytic fungus, Phomopsis sp., obtained from the stem of Azadirachta indica. Their structures were assigned by analysis of spectroscopic data, and the structures of 1 and 4 were also confirmed by X-ray analysis. Compounds 1- 5 were tested for antifungal activity against several plant pathogens. Compound 4 demonstrated antifungal activity in the MIC value range 31.25-500 microg/mL.

Two New Solanapyrone Analogues from the Endophytic Fungus Nigrospora sp. YB-141 of Azadirachta indica

Two new solanapyrone analogues, solanapyrones N and O (1 and 2, resp.), and three known compounds, solanapyrone C (3), nigrosporalactone (4), and phomalactone (5), were isolated from the fermentation culture of Nigrospora sp. YB-141, an endophytic fungus isolated from Azadirachta indica A. Juss. The structures of the new compounds were elucidated on the basis of spectroscopic analysis. The antifungal activities of 1-5 towards seven phytopathogenic fungi were tested. Most of the compounds exhibited no or only weak antifungal activities.

Chemical constituents from the stem bark of Trewia nudiflora

Trewia nudiflora L. is the only member of the genus Trewia (Euphorbiaceae), which is mainly distributed in India, Malaysia, and southwest of China. Previous studies have shown that the seed of T. nudiflora is a rich source of maytansinoid tumor inhibitors [1, 2]. Phytochemical studies are mainly focused on the seed and pericarp of the plant; however, there are only few reports on its stem bark [3, 4]. The air-dried powdered stem bark of T. nudiflora (8.8 kg) was extracted with 95% EtOH three times at room temperature. The EtOH extract was concentrated in vacuum to give a residue. The residue was suspended in water and successively treated with EtOAc. The EtOAc extract (32 g) was subjected to chromatography on silica gel eluting with CHCl3–MeOH gradient (1:0–0:1) to give nine fractions (I–IX). Fraction III was repeatedly subjected to column chromatography on RP-18 silica gel with MeOH-H2O (2:3) and Sephadex LH-20 with MeOH to give compound 1 (9 mg). Repeated chromatography of fraction IV on silica gel with petroleum ether–Me2CO gradient (4:1, 7:3, 6:4) and RP-18 silica gel with MeOH–H2O gradient (2:3, 1:1) afforded compounds 2 (6 mg) and 3 (10 mg). Fraction V was submitted to repeated column chromatography on silica gel with CHCl3–MeOH gradient (15:1, 10:1) and RP-18 silica gel with MeOH–H2O gradient (3:7, 2:3) to afford compounds 4 (13 mg), 5 (18 mg), and 6 (10 mg). Fraction VI was chromatographed on silica gel column with CHCl3–MeOH (8:1) to yield compounds 7 (16 mg) and 8 (13 mg). Trewiasine was isolated from the stem bark of T. nudiflora for the first time. Compounds 2–8 were isolated from this plant for the first time. The structures of these compounds were confirmed using a combination of spectral analyses, including NMR and mass spectrometry, and by comparison with reported spectroscopic data in the literature. Trewiasine (1). C37H52ClN3O11, colorless crystals, mp 180−182°C. ESI-MS m/z: 772 [M+Na]+. 1H NMR (500 MHz, CDCl3, δ, ppm, J/Hz): 2.19 (1H, dd, J = 14.5, 3.1, H-2a), 2.56 (1H, dd, J = 14.5, 12.0, H-2b), 4.77 (1H, dd, J = 12.0, 3.1, H-3), 0.78 (3H, s, 4-CH3), 3.02 (1H, d, J = 9.7, H-5), 1.28 (3H, d, J = 6.3, 6-CH3), 4.29 (1H, m, H-7), 3.53 (1H, d, J = 9.0, H-10), 5.74 (1H, dd, J = 15.1, 9.0, H-11), 6.46 (1H, dd, J = 15.1, 11.2, H-12), 6.99 (1H, d, J = 11.2, H-13), 1.54 (3H, s, 14-CH3), 4.87 (1H, s, H-15), 6.55 (1H, d, J = 1.5, H-17), 7.24 (1H, d, J = 1.4, H-21), 3.35 (3H, s, 10-OCH3), 3.37 (3H, s, 15-OCH3), 4.01 (3H, s, 20-OCH3), 3.18 (3H, s, 18-NCH3), 5.39 (1H, m, H-2′), 1.29 (3H, d, J = 6.8, 2′-CH3), 2.89 (3H, s, 2′-NCH3), 2.79 (1H, m, H-4′), 1.13 (3H, d, J = 6.8, 4′-CH3), 1.08 (3H, d, J = 6.5, 4′-CH3), 6.28 (1H, s, 9-NH). 13C NMR (125 MHz, CDCl3, δ, ppm): 32.3 (C-2), 78.1 (C-3), 59.9 (C-4), 67.4 (C-5), 38.8 (C-6), 74.1 (C-7), 36.0 (C-8), 80.7 (C-9), 85.3 (C-10), 129.6 (C-11), 132.5 (C-12), 127.8 (C-13), 142.0 (C-14), 86.6 (C-15), 141.3 (C-16), 120.2 (C-17), 139.2 (C-18), 119.2 (C-19), 156.2 (C-20), 108.7 (C-21), 176.7, 170.8, 168.7, 152.2 (4 × C=O), 56.3, 56.5, 56.7 (3 × OCH3), 14.5, 13.0, 11.9, 9.9 (4 × CH3), 35.2 (18-NCH3), 30.6 (2′-NCH3), 52.4 (C-2′), 30.4 (C-4′), 19.4, 18.8 (2 × 4′-CH3) [1, 2]. Balanophonin (2). C20H20O6, pale yellow oil. EI-MS (70 eV) m/z (%): 356 [M] + (82), 338 (100), 326 (55), 306 (18), 295 (7), 152 (24), 137 (22), 115 (12), 77 (14). 1H NMR (500 MHz, CD3COCD3, δ, ppm, J/Hz): 7.04 (1H, d, J = 1.9, H-2), 6.81 (1H, d, J = 8.1, H-5), 6.88 (1H, dd, J = 8.1, 1.9, H-6), 5.65 (1H, d, J = 6.7, H-7), 3.64 (1H, m, H-8), 3.85 (2H, d, J = 4.9, H-9), 7.29 (1H, d, J = 1.7, H-2′), 7.31 (1H, d, J = 1.7, H-6′), 7.58 (1H, d, J = 15.8, H-7′), 6.66 (1H, dd, J = 15.8, 7.8, H-8′), 9.63 (1H, d, J = 7.8, H-9′), 3.82 (3H, s, 3-OMe), 3.89 (3H, s, 5′-OMe). 13C NMR (125 MHz, CD3COCD3, δ, ppm): 133.7 (C-1), 110.5 (C-2), 148.4 (C-3), 147.5 (C-4), 115.7 (C-5), 119.7 (C-6), 89.4 (C-7), 54.2 (C-8), 64.1 (C-9), 128.9 (C-1′), 119.6 (C-2′), 131.2 (C-3′), 152.4 (C-4′), 145.6 (C-5′), 113.5 (C-6′), 154.1 (C-7′), 127.1 (C-8′), 193.8 (C-9′), 56.2 (3-OMe), 56.4 (5′-OMe) [5].

Starmerella jinningensis sp. nov., a yeast species isolated from flowers of Erianthus rufipilus.

Five yeast strains (Ym24403, Ym24404, Ym24408, Ym24409 and Ym24410(T)) were isolated from different flowers of Erianthus rufipilus (Gramineae), a wild plant growing in the phosphorus-rich region in Yunnan Province, south-western China, and were found to be phenotypically and genetically divergent from currently recognized yeast species. Sequence analysis of the D1/D2 domain of the large subunit rRNA gene revealed that the five strains represented a novel species described as Starmerella jinningensis sp. nov. The type strain is Ym24410(T) (= CBS 11864(T) =CCTCC AY 2011002(T)). Phylogenetic analysis based on the D1/D2 region of the large subunit rRNA gene suggested that S. jinningensis sp. nov. is placed within the Starmerella clade.

Phytoactinopolyspora endophytica gen. nov., sp. nov., a halotolerant filamentous actinomycete isolated from the roots of Glycyrrhiza uralensis F.

A novel endophytic actinomycete, designated strain EGI 60009T, was isolated from the roots of Glycyrrhiza uralensis F. collected from Xinjiang Province, north-west China. The isolate was able to grow in the presence of 0-9% (w/v) NaCl. Strain EGI 60009T had particular morphological properties: the substrate mycelia fragmented into rod-like elements and aerial mycelia differentiated into short spore chains. ll-2, 6-Diaminopimelic acid was the cell-wall diamino acid and rhamnose, galactose and glucose were the cell-wall sugars. MK-9(H4) was the predominant menaquinone. The major fatty acids of strain EGI 60009T were iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0, iso-C17 : 0, iso-C17 : 1 and I/anteiso-C17 : 0 B. Mycolic acids were absent. The DNA G+C content of strain EGI 60009T was 70.4 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain EGI 60009T belongs to the family Jiangellaceae and formed a distinct clade in the phylogenetic tree. 16S rRNA gene sequence similarities between strain EGI 60009T and other members of the genera Jiangella and Haloactinopolyspora were 96.1-96.4 and 95.7-96.0%, respectively. Based on these results and supported by morphological, physiological and chemotaxonomic data and numerous phenotypic differences, a novel species of a new genus, Phytoactinopolyspora endophytica gen. nov., sp. nov., is proposed. The type strain of Phytoactinopolyspora endophytica is EGI 60009T ( = KCTC 29657T = CPCC204078T).

Hannaella dianchiensis sp. nov., a basidiomycetous yeast species isolated from lake water

Three strains (YIM-HL1107T, YIM-HL1045, YIM-HL1112) representing a novel yeast species were isolated from surface water samples collected from the Caohai region of Dianchi Lake in Yunnan, south-western China. On the basis of morphological, physiological and biochemical characteristics and sequence analysis of the D1/D2 region of the LSU rRNA gene and the internal transcribed spacer (ITS) region, they were assigned to a novel species of the genus Hannaella. The closest relative to the novel species was Hannaella pagnoccae, but it showed 6.3 % nucleotide differences (34 nt substitutions out of 541 nt) in the D1/D2 region of the LSU rRNA gene and 9.3-9.6 % nucleotide differences (40-41 substitutions and 7-8 gaps out of 430 nt) in the ITS region. The name Hannaella dianchiensis sp. nov. is proposed. The type strain is YIM-HL1107T (=CBS 14191T=CCTCC AY 2015009T), and the MycoBank number is MB 816297.

Chemical constituents from the root bark of Paeonia delavayi

The genus Paeonia is the only member of the family Paeoniaceae. The plants in this genus are rich in monoterpene glycosides, which have been established as the main biologically active constituents [1–3]. The root bark of Paeonia delavayi Franch., one of the main sources of Chinese traditional medicine “mudanpi”, is an important herb known for its analgesic, sedative, and antiinflammatory properties. It is also used as a remedy for female diseases in traditional oriental medicine [4–6]. Previous studies on this plant led to the isolation of monoterpene glycosides [7, 8]. In continuation of our investigation into the chemical constituents of this plant, we isolated nine compounds 1−9. The root bark of P. delavayi was collected from Lijiang County, Yunnan Province, the People′s Republic of China and was identified by Prof. Zheng-Wei Lu of Kunming Institute of Botany, Chinese Academy of Science. A voucher specimen has been deposited in the Herbarium of Kunming Institute of Botany. The air-dried powdered root bark of P. delavayi (5 kg) was extracted with 95% EtOH three times at room temperature. The EtOH extract was concentrated in vacuum to give a residue. The residue was suspended in water and successively treated with EtOAc. The EtOAc extract (53 g) was subjected to chromatography on eluting with CHCl3–MeOH gradient (1:0−0:1) to give eight fractions (I−VIII). Fraction I was repeatedly subjected to silica gel column chromatography with petroleum ether−EtOAc (9:1) to give compounds 7 (11 mg) and 9 (325 mg). Fraction II was subjected to silica gel column chromatography with petroleum ether−acetone (9:1) to give compounds 1 (27 mg) and 3 (18 mg). Fraction III was subjected to silica gel column chromatography with chloroform−acetone gradient (9:1, 4:1) to give compounds 2 (25 mg) and 6 (21 mg). Repeated chromatography of fraction V on silica gel with CHCl3−MeOH gradient (95:5, 9:1, 85:15) and RP-18 silica gel with MeOH−H2O gradient (3:7, 4:6, 1:1) afforded compounds 4 (12 mg) and 5 (35 mg). Fraction VI was submitted to silica gel column chromatography with CHCl3–MeOH (4:1) to afford compound 8 (68 mg). The compounds identified as oleanolic acid (1) [9, 10], 3β,23-dihydroxy-30-norolean-12, 20(29)-dien-28-oic acid (2) [9, 10], akebonic acid (3) [11], arjunglucoside II (4) [12], 3-O-β-D-glucopyranoside β-sytosterine (5) [13, 14], syringic acid (6) [15], p-hydroxybenzoic acid (7) [16], gallic acid (8) [17], and benzoic acid (9) [18]. Oleanolic Acid (1). C30H48O3, mp 202–204°C, colorless crystals (acetone). IR (KBr, ν, cm–1): 3440, 2938, 2870, 1695, 1461, 1385, 1363, 1271, 1183, 1030, 996). 3β,23-Dihydroxy-30-norolean-12,20(29)-dien-28-oic Acid (2). C29H44O4, mp 241–243°C, colorless crystals (acetone). IR (KBr, ν, cm–1): 3438, 2940, 1688, 1465, 1384, 1298, 1215, 1105, 885. Akebonic Acid (3). C29H44O3, mp 152–154°C, colorless crystals (acetone). IR (KBr, ν, cm–1): 3424, 2935, 1691, 1653, 1463, 1384, 1297, 1212, 1102, 996, 886. Arjunglucoside II (4). C36H58O10, white powder. UV (MeOH, λmax, nm): 207.0 (log ε 3.71). IR (KBr, ν, cm –1): 3414, 2941, 1731, 1556, 1391, 1259, 1177, 1072, 893, 472. The structures of these compounds were confirmed using a combination of spectral analyses, including NMR and mass spectrometry and by comparison with reported spectroscopic data in the literature. Compounds 1−8 were isolated from P. delavayi Franch. for the first time.