First Report of Paramyrothecium breviseta Causing Target Spot on Amorphophallus muelleri in Yunnan, China.

Amorphophallus muelleri is an Araceae plant with perennial tuber, widely used in food, pharmaceutical and chemical industry due to its richness in glucomannan. In April 2022, an outbreak of a target spot on A. muelleri plantlets was observed in a nursery in Ruili, Yunnan, China. The leafstalks of the diseased plantlets in the nursery turned brown and decayed (Fig.1 A-B), then gradually some water-soaked spots on the true leaves developed along the veins (Fig.1 A). Subquencely, the spots on the true leaves turned dark green to white-grayish in the center, which formed light to dark brown concentric rings with a target-like appearance surrounded by a yellow halo (Fig.1 C). When the temperature was 20-34℃ and the relatively humidity was 25-80%, dark-green to black sporodochia with white hypha appeared on the lower and upper leaf surfaces. Finally, 5-8% of the plants surveyed on 800 m2 of one-year-old plantlets in the nursery showed the symptoms and some plants with infected leafstalks would be death. Similar symptoms were also observed on about 10% of the transplanted plants surveyed on 12000 m2 (1.2 ha) of two-year-old plantlets in the field. Five diseased leaves from five distinct plantlets in the nursery were collected for pathogen isolation. Leaf pieces(5 x 5 mm) were cut from the edge of necrotic lesions, and surface-sterilized with 2.5% sodium hypochlorite for 1 min, 75% ethanol for 30 s, then rinsed 5 times by sterilized distilled water, finally put the leaf pieces on sterilized filter paper for 3-5 minutes to dry them and transferred onto potato dextrose agar (PDA) in petri dishes at 25℃ for three days. Five pure cultures identical to colony and conidial characteristics were isolated from five individual plants. The representative pure culture (M1) was grayish-white and circular colonies were 7.50 cm in diamter after 15 days at 25℃, with dark green concentric rings of sporodochia, the dorsal view of the colonies were yellowish. Conidia were aseptate, smooth, cylindrical, 5.00-6.25 (5.71) x 1.25-1.67 (1.63) µm (n = 20) rounded at both ends. A spore suspension (1 x 106 spores/ml) was prepared by harvesting spores from 15-day-old cultures grown in the dark at 25℃, then a thirty-ml of spore suspension was sprayed on the healthy leaves of 10 two-year-old plantlets. Thirty-ml of sterile water was sprayed on the healthy leaves of another 10 seedlings and used as the control. All seedlings were placed in a nursery at 20 to 34℃ and a relative humidity of 25 to 80%. Similar symptoms (Fig.1 D-F) to those observed in the nursery and field developed on all the 10 seedlings inoculated with M1 after two days, but not on the control leaves. The pathogenicity tests were repeated for three times. Fungal cultures reisolated from the infected leaves were identical to the original colonies and conidia, completing Koch's postulates. The internal transcribed spacer (ITS, primers ITS1 and ITS4) region of ribosomal DNA (OQ553785), calmodulin (cmdA, primers CAL-228F and CAL2Rd)(OQ559103), RNA polymerase II second largest subunit (rpb2, primers RPB2-5F2 and RPB2-7cR) (OQ559104) and ß-tubulin (tub2, primers Bt2a and Bt2b) (OQ559105) of M1 had 100%, 98.52%, 98.98% and 98.98% identity with the sequences of Paramyrothecium breviseta CBS544.75 (KU846289 for ITS, KU846262 for cmdA, KU846351 for rpb2, and KU846406 for tub2), respectively. In the phylogenic tree based on ITS, cmdA, rpb2 and tub2 gene sequences, the pure culture M1 clustered with P. breviseta CBS544.75, SDBR-CMU387, DRL4 and DRL3, which has been reported as the pathogen of leaf spot of Coffea arabica in China, C. canephora in China and Thailand (Wu et al. 2021; Withee et al. 2022). Molecular and morphological observations showed the pure culture M1 were P. breviseta (Withee et al. 2022), in addition the disease was named as target spot dueing to the typical target symptom on the leaves. To our knowledge, this is the first report of P. breviseta on A. muelleri from Yunnan, China, as well as worldwide. This disease can caused serious economic losses of A. muelleri dueing to that it can result 5-8% death of the plants in the nursery.

Molecular and Morphological Identification of Monilia yunnanensis Causing Brown Rot on Chinese Quince and Peach in Yunnan, China.

More than 30% of fruits of Chinese Quince (Chaenomeles speciosa) and peach (Prunus persica) showed circular, water-soaked and brown spots in July 2022 in Kunming, Yunnan, China. The center of these spots was covered by a large number of earthy brown and oblate sporogeneous mycelium containing conidiophore and conidia, which were one-celled, limoniform, hyaline (13.73 to 22.77 x 8.17 to 12.84 µm, n=50). By September 2022, almost 100% of fruits showed symptoms. Later, most of them fell or a few stiff, black and mummified fruits were left on the trees. Fungal isolates were isolated by single-spore technique on Potato Dextrose agar (PDA) from the diseased fruits, and incubated at room temperature (20-28 °C) in darkness for 14 days. The colony was gray, smooth at margins, 7.6-8.0 cm in diameter. To fullfill Koch's postulates, mycelial plugs of one representative isolate YHD611 from Chinese Quince and another YHD610 from peach were used to inoculate three wounded and three non-wounded surface-disinfected fruits of both hosts at room temperature (19-27 °C), respectively. Three wounded and three non-wounded fruits inoculated with sterile PDA plugs served as the control. The wounded peaches appeared water-soaked and had brown lesions after three days of inoculation, then completely decayed after nine days, while non-wounded fruits showed symptoms after five days. The wounded fruits of Chinese Quince developed similar symptoms after eight days of inoculation, and completely decayed after 13 days, while non-wounded fruits showed obvious symptoms after 15 days. In a subsequent study, isolate YHD611 was inoculated to peach while isolate YHD610 was inoculated to Chinese Quince to understand host specificity of the isolates. The results showed that when peaches were infected with YHD611, symptoms were observed on wounded fruits after three days while on non-wounded fruits after five days. When Chinese Quince was infected with YHD610, symptoms were observed on wounded fruits after 14 days while on non-wounded fruits after 21 days. Fungal isolates from symptomatic fruits were identical to the original isolates. There were no symptoms on the control fruits of both hosts. Molecular identification was confirmed based on the sequences of internal transcribed spacer (ITS, primers ITS1 and ITS4) and ß-tubulin (TUB2, primers Bt2a and Bt2b) genes (Niu et al. 2016). BLASTn analysis of the ITS (OQ15519and OQ155196) and TUB2 (OQ185202 and OQ185201) of YHD611 and YHD610 revealed a 100% sequence identity, respectively, to Monilia yunnanensis AH7-2 (KT735924.1 for ITS, KT736008.1 for TUB2). In the phylogenetic analyses based on ITS and TUB2 sequence data, the isolates YHD611 and YHD610 belonged to the M. yunnanensis clade. Based on morphological and molecular identification, both isolates were identified as M. yunnanensis, which was reported as the pathogen causing brown rot of plum, peach, apple and pear in Yunnan, China (Hu et al. 2011; Yin et al. 2015). To our knowledge, this is the first report of M. yunnanensis causing brown rot on the fruits of Chinese Quince in Yunnan, China. This study also reports that M. yunnanensis from Chinese Quince can infect peach, and the pathogen from peach can infect Chinese Quince. These findings suggest that M. yunnanensis can transfer from one host to another and causing serious economic losses in multiple fruit crops in Yunnan, China. References: Hu, M. J., et al. 2011. PLoS One. 6:e24990. Niu, C. W., et al. 2016. Mycosystema, 35(10):1. Yin, L. F., et al. 2015. Plant Dis. 99:1775.

Bacillus tamaricis sp. nov., an alkaliphilic bacterium isolated from a Tamarix cone soil.

A Gram-stain-positive, alkaliphilic bacterium, designated EGI 80668T, was isolated from a Tamarix cone soil in Xinjiang, north-west China. Cells were facultatively anaerobic, terminal endospore-forming and motile by means of peritrichous flagella. Colonies were yellowish and the cells showed oxidase-negative and catalase-positive reactions. Strain EGI 80668T grew at pH 8.0-10.0 and with 0-10 % (w/v) NaCl (optimally at pH 9.0 and with 1-2 % NaCl) on marine agar 2216. The predominant menaquinone was MK-7. The major fatty acids were anteiso-C17 : 0 and anteiso-C15 : 0. The cellular polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, four unknown phospholipids and one unknown aminophospholipid. The G+C content of the genomic DNA was 38.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EGI 80668T was affiliated to the genus Bacillus. The highest 16S rRNA gene sequence similarity between strain EGI 80668T and a member of the genus Bacillus was 96.83 % with Bacillus cellulosilyticus JCM 9156T. A polyphasic taxonomic study based on morphological, physiological, biochemical and phylogenetic data indicated that strain EGI 80668T represents a novel species of the genus Bacillus, for which the name Bacillus tamaricis sp. nov. (type strain EGI 80668T=KCTC 33703T=CGMCC 1.15917T) is proposed.

Bacillus capparidis sp. nov., an endophytic bacterium isolated from roots of Capparis spinosa L.

A novel endophytic bacterium, designated strain EGI 6500252T, was isolated from the surface-sterilized roots of a medicinal plant (Capparis spinosa L.) collected from Urumqi city, Xinjiang, north-west China. Cells were Gram-stain-positive, non-motile, aerobic, catalase- and oxidase-positive, rod-shaped and did not display spore formation. Strain EGI 6500252T grew at 10-40 °C (optimum 25-30 °C), at pH 6.0-8.0 (optimum pH 7.0) and in the presence of 0-10 % (w/v) NaCl (optimum 0-3 %). The major cellular fatty acids (>10 %) were identified as iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and summed feature 4. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, four unknown phospholipids, one unknown glycolipid and one unknown lipid. The dominant isoprenoid quinone was menaquinone 7 (MK-7). The DNA G+C content was 39.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain EGI 6500252T belonged to the genus Bacillus, and exhibited a highest 16S rRNA gene sequence similarity (96.2 %) that was lower than the suggested threshold (97.0 %) for separating bacterial species. On the basis of the phylogenetic analysis, chemotaxonomic data and physiological characteristics, strain EGI 6500252T represents a novel species of the genus Bacillus, for which the name Bacillus capparidis sp. nov. is proposed. The type strain is EGI 6500252T (=CGMCC 1.12820T=KCTC 33514T).

Nocardiopsis rhizosphaerae sp. nov., isolated from rhizosphere soil of Halocnermum strobilaceum (Pall.) Bieb.

An alkalitolerant actinomycete strain, designated EGI 80674T, was isolated from a rhizosphere soil of Halocnermumstrobilaceum (Pall.) Bieb in Xinjiang, north-west China and subjected to a taxonomic characterization using a polyphasic approach. Strain EGI 80674T formed white aerial hyphae with long spore chains. Whole-cell hydrolysates of the isolate contained meso-diaminopimelic acid as the diagnostic diamino acid with no diagnostic sugars. The major fatty acids identified were iso-C16 : 0, anteiso-C17 : 0 and 10-methyl-C18 : 0TBSA. The predominant menaquinones detected were MK-10(H8) and MK-10(H6). The G+C content of the genomic DNA of strain EGI 80674T was 70.9 mol%. Strain EGI 80674T showed the highest 16S rRNA gene sequence similarity (97.24 %) to Nocardiopsis nikkonensis NBRC 102170T. The DNA-DNA relatedness value of strain EGI 80674T and N. nikkonensis NBRC 102170T was 18.4±1.3 %. Phenotypical, chemotaxonomic and phylogenetic characteristics and DNA-DNA hybridization data suggest that strain EGI 80674T represents a novel species of the genus Nocardiopsis, for which the name Nocardiopsis rhizosphaerae sp. nov. is proposed. The type strain is EGI 80674T (=CGMCC 4.7228T=KCTC 39673T).

Pseudoclavibacter endophyticus sp. nov., isolated from roots of Glycyrrhiza uralensis.

A Gram-stain-positive, aerobic, rod-shaped, non-motile actinomycete strain, designated EGI 60007T, was isolated from healthy roots of Glycyrrhiza uralensis F. collected from Yili County, Xinjiang Province, north-west China. A polyphasic approach was applied to study the taxonomic position of the new isolate. The 16S rRNA gene sequence of strain EGI 60007T had highest similarities with members of the genus Pseudoclavibacter, including Pseudoclavibacter chungangensis CAU 59T (96.98 % 16S rRNA gene sequence similarity), Pseudoclavibacter helvolus DSM 20419T (96.43 %) and Pseudoclavibacter terrae THG-MD12T (96.14 %). The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain EGI 60007T clustered with members of the genus Pseudoclavibacter, and formed a distinct clade with P. chungangensis CAU 59T. The polar lipids detected for strain EGI 60007T were phosphatidylglycerol, diphosphatidylglycerol, one unidentified glycolipid and one unidentified lipid. The DNA G+C content was determined to be 63.3 mol%. The chemotaxonomic features of strain EGI 60007T showed typical characteristics of the genus Pseudoclavibacter, with MK-9 as the respiratory quinone, 2,4-diaminobutyric acid as the diamino acid in the peptidoglycan, and anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0 as the major fatty acids. The sugars of whole-cell hydrolysates were mainly mannose, rhamnose, ribose and glucose, and a minor amount of xylose. Based on the results of the phylogentic analysis supported by morphological, physiological, chemotaxonomic and other differentiating phenotypic characteristics, strain EGI 60007T is considered to represent a novel species of the genus Pseudoclavibacter, for which the name Pseudoclavibacter endophyticus sp. nov. is proposed. The type strain is EGI 60007T ( = CGMCC 1.15081T = KCTC 39112T = DSM 29943T).

Ornithinicoccus halotolerans sp. nov., and emended description of the genus Ornithinicoccus.

A halotolerant actinobacterial strain, designated EGI 80423T, was isolated from a desert soil of Xinjiang, north-west China, and subjected to a polyphasic taxonomic characterization. Strain EGI 80423T grew at pH 7.0-10.0 and with 0-14.0% (w/v) NaCl, optimally at pH 8.0-9.0 and with 2.0-4.0% (w/v) NaCl. Cells of strain EGI 80423T were Gram-stain-positive, non-motile cocci with diameters of 0.6-0.8 µm. The diagnostic diamino acid of the peptidoglycan was ornithine, and the interpeptide bridge was Orn ← Glu. The major fatty acids identified were iso-C17:1ω9c, iso-C15:0 and iso-C17:0. The predominant menaquinone was MK-8(H4), while the polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown phospholipids, two unknown glycolipids, six unknown phosphoglycolipids and five unknown polar lipids. The G+C content of the genomic DNA was 72.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EGI 80423T clustered with the single member of the genus Ornithinicoccus. Sequence similarity between strain EGI 80423T and Ornithinicoccus hortensis NBRC 16434T. Because the type strain has been provided by NBRC, Japan was 97.7%. The DNA-DNA relatedness value between strain EGI 80423T and O. hortensis NBRC 16434T was 36.84%. Based on morphological, chemotaxonomic and phylogenetic characteristics, and DNA-DNA hybridization data, strain EGI 80423T represents a novel species of the genus Ornithinicoccus, for which the name Ornithinicoccus halotolerans sp. nov. is proposed. The type strain is EGI 80423T (=CGMCC 1.14989T=KCTC 39700T). The description of the genus Ornithinicoccus has also been emended.

Phytoactinopolyspora alkaliphila sp. nov., an alkaliphilic actinomycete isolated from a saline-alkaline soil.

An alkaliphilic, filamentous actinomycete, designated EGI 80629T, was isolated from a soil sample of Xinjiang, north-west China. Strain EGI 80629T grew at pH 6.0-11.0 (optimum pH 9.0-10.0) and in the presence of 0-13.0 % NaCl (optimum 3.0-5.0 %). The isolate formed fragmented substrate mycelia, and aerial hyphae with short spore chains with rod-like spores. Whole-cell hydrolysates of the isolate contained ll-diaminopimelic acid as the diagnostic diamino acid, and mannose and rhamnose as diagnostic sugars. The major fatty acids identified were iso-C15 : 0, iso-C16 : 0, anteiso-C15 : 0 and iso-C17 : 0. The predominant menaquinone was MK-9(H4), while the polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two phosphatidylinositol mannosides, five unknown phospholipids, three unknown phosphoglycolipids, one unknown glycolipid, four unknown polar lipids and one unknown aminophospholipid. The G+C content of the genomic DNA was 67.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EGI 80629T clustered with the genus Phytoactinopolyspora. The 16S rRNA gene sequence similarity between strain EGI 80629T and Phytoactinopolyspora endophytica EGI 60009T was 96.8 %. Based on morphological, chemotaxonomic and phylogenetic characteristics, strain EGI 80629T represents a novel species of the genus Phytoactinopolyspora, for which the name Phytoactinopolyspora alkaliphila sp. nov. is proposed. The type strain is EGI 80629T ( = CGMCC 4.7225T = KCTC 39701T).

Nocardiopsis ansamitocini sp. nov., a new producer of ansamitocin P-3 of the genus Nocardiopsis.

An alkalitolerant actinomycete strain, designated EGI 80425T, capable of producing ansamitocin P-3, was isolated from a saline-alkali soil sample of Xinjiang province, north-west China, and subjected to a polyphasic taxonomic characterization. Strain EGI 80425T formed non-fragmented substrate mycelia and white aerial hyphae with long spore chains. Whole-cell hydrolysates of the isolate contained meso-diaminopimelic acid as the diagnostic diamino acid and rhamnose as the major sugar. The major fatty acids were anteiso-C17 : 0, iso-C16 : 0 and C18 : 1ω9c. The predominant menaquinones were MK-10(H4), MK-10(H6), MK-10(H8) and MK-9(H4). The G+C content of the genomic DNA of strain EGI 80425T was 70.2 mol%. Strain EGI 80425T showed highest 16S rRNA gene sequence similarity to Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111T (96.44 %). Phylogenetic analysis showed that strain EGI 80425T clustered with the members of the genus Nocardiopsis. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain EGI 80425T represents a novel species of the genus Nocardiopsis, for which the name Nocardiopsis ansamitocini sp. nov. is proposed. The type strain is EGI 80425T ( = CGMCC 9969T = KCTC 39605T).

Egicoccus halophilus gen. nov., sp. nov., a halophilic, alkalitolerant actinobacterium and proposal of Egicoccaceae fam. nov. and Egicoccales ord. nov.

A novel Gram-stain-positive, non-motile, moderately halophilic and alkalitolerant actinobacterium, designated EGI 80432T, was isolated from a saline-alkaline soil of Xinjiang province, north-west China. Cells were non-endospore-forming cocci with a diameter of 0.5-0.8 µm. Strain EGI 80432T grew in the presence of 0-9 % (w/v) NaCl (optimum at 3-5 %), and also grew within the pH range 6.0-10.0 (optimum at pH 8.0-9.0) on marine 2216E medium. The peptidoglycan type was A1γ. The whole-cell hydrolysates contained glucose, galactose, mannose and three unknown sugars as major sugars. The predominant menaquinone was MK-9(H4). The major fatty acids were C17 : 1ω8c, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), C18 : 1ω9c and iso-C15 : 0 The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, one unknown phosphoglycolipid, three unknown phospholipids and four unknown polar lipids. The genomic DNA G+C content was 75.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain EGI 80432T clustered within the radius of the class Nitriliruptoria. Levels of sequence similarity between strain EGI 80432T and its phylogenetic neighbours Nitriliruptor alkaliphilus ANL-iso2T and Euzebya tangerina F10T were 94.1 and 88.1 %, respectively. Based on morphological, physiological and chemotaxonomic characteristics and phylogenetic analysis, a novel species of a new genus, Egicoccus halophilus gen. nov., sp. nov., is proposed, within the new family and new order Egicoccaceae fam. nov. and Egicoccales ord. nov. in the class Nitriliruptoria. The type strain of Egicoccus halophilus is EGI 80432T ( = CGMCC 1.14988T = KCTC 33612T).

Lipingzhangella halophila gen. nov., sp. nov., a new member of the family Nocardiopsaceae.

An alkaliphilic and halophilic actinomycete strain, designated EGI 80537T, was isolated from a saline-alkali soil sample of Xinjiang, north-west China and subjected to a taxonomic characterization using a polyphasic approach. Strain EGI 80537T formed reticulate long aerial hyphae. Whole-cell hydrolysates of the isolate contained meso-diaminopimelic acid as the cell-wall diamino acid and mannose as the diagnostic sugar. The major fatty acids identified were iso-C16 : 0, anteiso-C17 : 0 and 10-methyl-C18 : 0 (TBSA). The predominant menaquinones detected were MK-10(H8) and MK-10(H6). The G+C content of the genomic DNA of strain EGI 80537T was 67.6 mol%. Strain EGI 80537T showed the highest 16S rRNA gene sequence similarity to Allosalinactinospora lopnorensis CA15-2T (96.7 %). Phylogenetic analysis showed that strain EGI 80537T clustered with the members of the family Nocardiopsaceae. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain EGI 80537T represents a novel species of a new genus in the family Nocardiopsaceae, for which the name Lipingzhangella halophila gen. nov., sp. nov. is proposed. The type strain of the type species is EGI 80537T(=CGMCC 4.7224T= DSM 102030T).

Novosphingobium endophyticum sp. nov. isolated from roots of Glycyrrhiza uralensis.

A novel Gram-staining-negative, non-motile, rod-shaped and aerobic bacterial strain, designated EGI 60015(T), was isolated from healthy roots of Glycyrrhiza uralensis F. collected from Yili County, Xinjiang Province, Northwest China. The 16S rRNA gene sequence of strain EGI 60015(T) was found to show 97.6% sequence similarity with Novosphingobium pentaromativorans US6-1(T). The phylogenetic analysis based on 16S rRNA gene sequences showed that the strain formed a clade with N. pentaromativorans US6-1(T) in the neighbor-joining tree. Q-10 was identified as the respiratory quinone of strain EGI 60015(T). The major fatty acids were summed feature 8 (C18:1 ω6c and/or C18:1 ω7c; 55.04%), summed feature 4 (C17:1 anteiso B and/or iso I; 18.34%) and summed feature 3 (C16:1 ω6c and/or C16:1 ω7c; 8.53%). The polar lipids detected were phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and sphingoglycolipids. The DNA G+C content of strain EGI 60015(T) was determined to be 66.6 mol%. The genomic DNA relatedness value between EGI 60015(T) and N. pentaromativorans US6-1(T) (54%) was below the 70% limit for species identification. Based on the result of the molecular studies supported by its morphological, physiological, chemotaxonomic and other differentiating phenotypic characteristics, strain EGI 60015(T) was considered to represent a novel species within the genus Novosphingobium, for which the name Novosphingobium endophyticum sp. nov. is proposed. The type strain is EGI 60015(T) (=CGMCC 1.15095(T) = KCTC 42486(T) = DSM 29948(T)).

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).

Frigoribacterium endophyticum sp. nov., an endophytic actinobacterium isolated from the root of Anabasis elatior (C. A. Mey.) Schischk.

A novel endophytic actinobacterium, designated EGI 6500707(T), was isolated from the surface-sterilized root of a halophyte Anabasis elatior (C. A. Mey.) Schischk collected from Urumqi, Xinjiang province, north-west China, and characterized using a polyphasic approach. Cells were Gram-stain-positive, non-motile, short rods and produced white colonies. Growth occurred at 10-45 °C (optimum 25-30 °C), at pH 5-10 (optimum pH 8) and in presence of 0-4% (w/v) NaCl (optimum 0-3%). The predominant menaquinone was MK-9. The diagnostic phospholipids were diphosphatidylglycerol and phosphatidylglycerol. The major fatty acids were anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0). The DNA G+C content of strain EGI 6500707(T) was 69.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain EGI 6500707(T) should be placed in the genus Frigoribacterium (family Microbacteriaceae , phylum Actinobacteria ), and that the novel strain exhibited the highest 16S rRNA gene sequence similarity to Frigoribacterium faeni JCM 11265(T) (99.1%) and Frigoribacterium mesophilum MSL-08(T) (96.5%). DNA-DNA relatedness between strain EGI 6500707(T) and F. faeni JCM 11265(T) was 47.2%. On the basis of phenotypic and chemotaxonomic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain EGI 6500707(T) represents a novel species of the genus Frigoribacterium , for which the name Frigoribacterium endophyticum sp. nov. is proposed. The type strain is EGI 6500707(T) ( = JCM 30093(T) = KCTC 29493(T)).

Okibacterium endophyticum sp. nov., a novel endophytic actinobacterium isolated from roots of Salsola affinis C. A. Mey.

A white bacterial strain, designated EGI 650022(T), was isolated from the roots of Salsola affinis C. A. Mey, collected from Urumqi City, Xinjiang, north-western China. The strain was found to be aerobic, Gram-stain positive, oxidase-positive and catalase-positive. Cells were non-motile and irregular rods. Growth occurred at NaCl concentrations between 0 and 7 % (w/v), temperatures between 5 and 45 °C, and pH 6.0-9.0. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain EGI 650022(T) belongs to a clade with the genera Okibacterium and Plantibacter in the family Microbacteriaceae. The novel strain EGI 650022(T) showed highest levels of 16S rRNA gene sequence similarity with members of the genera Okibacterium and Plantibacter (97.2-98.0 %). The cell-wall peptidoglycan contained glutamate, homoserine, glycine, alanine and lysine. The predominant menaquinones (MKs) were MK-11, MK-12 and MK-12 (H4). The polar lipid pattern comprised phosphatidylglycerol, diphosphatidylglycerol, two unknown glycolipids and two unknown phospholipids. The major fatty acids were anteiso-C15:0 and anteiso-C17:0. The DNA G+C content was 66.0 mol%. The DNA-DNA relatedness values of strain EGI 650022(T) with Okibacterium fritillariae DSM 12584(T), Plantibacter flavus DSM 14012(T) and Plantibacter auratus DSM 19586(T) were 39.7, 19.7 and 22.0 %. Based on phylogenetic, phenotypic, chemotaxonomic and DNA-DNA hybridization data, strain EGI 650022(T) is considered to represent a novel species of the genus Okibacterium, for which the name Okibacterium endophyticum sp. nov. is proposed; the type strain is EGI 650022(T) (=JCM 30086(T) = KCTC 29492(T)).

Labedella endophytica sp. nov., a novel endophytic actinobacterium isolated from stem of Anabasis elatior (C. A. Mey.) Schischk.

A Gram-stain positive, yellow-coloured, aerobic, non-motile, catalase-positive and oxidase-negative, endophytic actinobacterium, designated strain EGI 6500705(T), was isolated from the surface-sterilized stem of a halophyte Anabasis elatior (C. A. Mey.) Schischk collected from Urumqi, Xinjiang province, north-west China. The organism had ornithine as the diagnostic cell-wall diamino acid. The major fatty acids were anteiso-C15:0, anteiso-C17:0 and iso-C16:0. The predominant menaquinones were MK-9, MK-10 and MK-11. The DNA G + C content of strain EGI 6500705(T) was 69.2 mol%. Phylogenetic analyses based on 16S rRNA gene sequence showed that strain EGI 6500705(T) is clearly affiliated with the genus Labedella and most closely related to Labedella gwakjiensis KCTC 19176(T), with 99.0 % sequence similarity. DNA-DNA relatedness between strain EGI 6500705(T) and L. gwakjiensis KCTC 19176(T) was 27.4 %. On the basis of phenophytic, chemotaxonomic and phylogenetic analysis, the strain EGI 6500705(T) represents a novel species of the genus Labedella, for which the name Labedella endophytica sp. nov. is proposed. The type strain is EGI 6500705(T) (=KCTC 29494(T) = CPCC 203961(T) = JCM 30092(T)).

Treatment of oilfield fracturing wastewater by a sequential combination of flocculation, Fenton oxidation and SBR process.

In this study, a combined process was developed that included flocculation, Fenton oxidation and sequencing batch reactor (SBR) to treat oilfield fracturing wastewater (FW). Flocculation and Fenton oxidation were applied to reduce chemical oxygen demand (COD) organic load and to enhance biodegradability, respectively. For flocculation, the optimum conditions were: polymeric aluminium chloride dosage, 40 mg/L; polyacrylamide dosage, 4 mg/L; dilution ratio, 1:2 and stirring time, 30 min. For Fenton oxidation, a total reaction time of 60 min, a H2O2dosage of 2 m mol/L, with a [H2O2]/[FeSO4] ratio of 2 were selected to achieve optimum oxidation. Under these optimum flocculation and Fenton oxidation conditions, the COD removal efficiency was found to be 76.6%. Following pretreatment with flocculation and Fenton oxidation, the FW was further remediated using a SBR. Results show that COD was reduced to 92 mg/L, and the overall water quality of the final effluent could meet the class I national wastewater discharge standard of petrochemical industry of China.

Establishment of a nomogram for potential prediction of lung metastasis in patients with primary limb bone tumors: a study based on the SEER database.

Background: The prognosis of lung metastasis in primary limb bone tumors represents a pivotal yet challenging aspect of oncological management. Despite advancements in diagnostic modalities, the predictive accuracy for metastatic spread remains suboptimal. This study aims to bridge this gap by leveraging the Surveillance, Epidemiology, and End Results (SEER) database to construct a nomogram that forecasts the risk of lung metastasis, thereby enhancing clinical decision-making processes. Methods: A retrospective cohort, including 1,822 patients with primary limb bony tumors from 2010 to 2015 in the SEER database, was extracted. Using precise inclusion and exclusion criteria, variables essential for predicting lung metastasis were identified through univariate and multivariate analyses, along with least absolute shrinkage and selection operator (LASSO) regression. These variables provided a solid basis for creating the multivariable nomogram, of which the discriminating power and utility were verified using receiver operating characteristic (ROC) curves, calibration plots, and decision curve analysis. Results: The model incorporated seven key predicting variables, including age, histological type, surgery, radiation, chemotherapy, T stage, and N stage. The nomogram emerged as a cohesive whole with good discriminative power. The area under the curve (AUC) was 0.806 in the training cohort and 0.767 in the validation cohort. The calibration curves demonstrated the model's validity by showing a good match between the actual outcomes and the model-predicted probabilities of lung metastasis. Conclusions: This study showed for the first time the reliability of the predictive model in translating the hard-to-interpret demographic, clinical, and pathologic data into a very usable predictive model. Thus, it represents a significant step toward demystifying the risk of lung metastasis in primary limb bone tumors. It is an invitation for a paradigm shift of oncology, to evidence-based, person-based oncology that is taking a new metric for cancer prognosis.

Barley farmland harbors a highly homogeneous soil bacterial community compared to wild ecosystems in the Qinghai-Xizang Plateau.

Introduction: Understanding patterns and processes of microbial biogeography in soils is important for monitoring ecological responses to human activities, particularly in ecologically vulnerable areas such as the Qinghai-Xizang Plateau. Highland barley is the staple food of local people and has mainly been cultivated along the Yarlung Zangbo River valley in Xizang. Methods: Here we investigated soil bacterial communities from 33 sampling sites of highland barley farmland in this region and compared them to those from wild ecosystems including alpine tundra, meadow, forest, and swamp. Additionally, the effects of environmental factors on bacterial communities, as well as the relative importance of stochastic and deterministic processes in shaping the beta diversity of soil bacterial communities in alpine ecosystems were assessed. Results: In contrast to soils of wild ecosystems, these farmland samples harbored a highly homogeneous bacterial community without significant correlations with geographic, elevation, and edaphic distances. Discriminant bacterial taxa identified for farmland samples belong to Acidobacteria, with Acidobacteria Gp4 as the dominant clade. Although Acidobacteria were the most abundant members in all ecosystems, characterized bacterial taxa of meadow and forest were members of other phyla such as Proteobacteria and Verrucomicrobia. pH and organic matter were major edaphic attributes shaping these observed patterns across ecosystems. Null model analyses revealed that the deterministic assembly was dominant in bacterial communities in highland barley farmland and tundra soils, whereas stochastic assembly also contributed a large fraction to the assembly of bacterial communities in forest, meadow and swamp soils. Discussion: These findings provide an insight into the consequences of human activities and agricultural intensification on taxonomic homogenization of soil bacterial communities in the Qinghai-Xizang Plateau.

Effects of ulinastatin combined with dexmedetomidine on cognitive dysfunction and emergence agitation in elderly patients who underwent total hip arthroplasty.

BACKGROUND: With the continuous growth of the modern elderly population, the risk of fracture increases. Hip fracture is a common type of fracture in older people. Total hip arthroplasty (THA) has significant advantages in relieving chronic pain and promoting the recovery of hip joint function. AIM: To investigate the effect of ulinastatin combined with dexmedetomidine (Dex) on the incidences of postoperative cognitive dysfunction (POCD) and emergence agitation in elderly patients who underwent THA. METHODS: A total of 397 patients who underwent THA from February 2019 to August 2022. We conducted a three-year retrospective cohort study in Shaanxi Provincial People's Hospital. Comprehensive demographic data were obtained from the electronic medical record system. We collected preoperative, intraoperative, and postoperative data. One hundred twenty-nine patients who were administered Dex during the operation were included in the Dex group. One hundred fifty patients who were intravenously injected with ulinastatin 15 min before anesthesia induction were included in the ulinastatin group. One hundred eighteen patients who were administered ulinastatin combined with Dex during the operation were included in the Dex + ulinastatin group. The patients' perioperative conditions, hemodynamic indexes, postoperative Mini-Mental State Examination (MMSE) scores, Ramsay score, incidence of POCD, and serum inflammatory cytokines were evaluated. RESULTS: There was a significant difference in the 24 h visual analogue scale score among the three groups, and the score in the Dex + ulinastatin group was the lowest (P < 0.05). Compared with the Dex and ulinastatin group, the MMSE scores of the Dex + ulinastatin group were significantly increased at 1 and 7 d after the operation (all P < 0.05). Compared with those in the Dex and ulinastatin groups, incidence of POCD, levels of serum inflammatory cytokines in the Dex + ulinastatin group were significantly decreased at 1 and 7 d after the operation (all P < 0.05). The observer's assessment of the alertness/sedation score and Ramsay score of the Dex + ulinastatin group were significantly different from those of the Dex and ulinastatin groups on the first day after the operation (all P < 0.05). CONCLUSION: Ulinastatin combined with Dex can prevent the occurrence of POCD and emergence agitation in elderly patients undergoing THA.