Microcerotermes eugnathus Silvestri  showed structural damage in Bir al-Shaghala cemeteries located in the oasis of Dakhla, Egypt. The mud tubes of this termite spread inside and over the mural painted floors of the tombs. Extracts from Lavandula latifolia, Origanum vulgare, and Syzygium aromaticum were tested for their anti-termitic activity and compared with the bio-insecticide, Bacillus thuringiensis var. kurstaki (Protecto 9.4% WP) and Dursban (Chlorpyrifos 48%). The bioassay experimental showed that the extracts have low activity against M. eugnathus compared to Protecto and Dursban, but the extract from O. vulgare showed promising natural termiticides.The termite Lavandula latifolia (Spike lavender), Origanum vulgare (Marjorum), and Syzygium aromaticum (Clove) against the termite Microcerotermes eugnathus. Plant extract results were compared to two commercially used termite pesticides, the bio-insecticide, Bacillus thuringiensis var. kurstaki (Protecto 9.4% WP) and Dursban (Chlorpyrifos 48%). Gas chromatography–mass spectrometry (GC-MS) analysis was used to identify the main compounds in the three plant extracts. The main compounds in Lavandula Latifolia were linalool (21.49%), lavandulol (12.77%), β-terpinyl acetate (10.49%), and camphor (9.30%). Origanum vulgare extract contained thymol (14.64%), m-cymene (10.63%), linalool (6.75%), and terpinen-4-ol (6.92%) as main compounds. Syzygium aromaticum contained eugenol (99.16%) as the most abundant identified compound. The extract of O. vulgare caused the highest termite death rate, with an LC50 of 770.67 mg/L. Exposure to lavender extract showed a high death rate with an LC50 of 1086.39 mg/L. Clove extract did not show significant insecticidal activity with an LC50 > 2000 mg/L. Significant termiticide effects were found, with LC50 values of 84.09 and 269.98 mg/L for soldiers and workers under the application of Dursban and Protecto, respectively. The LC50 values reported for nymphs were <120, <164.5, and 627.87 mg/L after exposure to Dursban, Protecto, and O. vulgare extract, respectively. The results of the study show that some of the extracts have low toxicity compared to the bioagent and Dursban, and may show promise as natural termiticides, particularly as extracts from O. vulgare.A trend towards environmentally friendly chemicals for use in termite management has been occurring globally. This study examined three naturally occurring plant extracts from  Dakhla Oasis, Egypt, are the site of heavy infestation by the termite Microcerotermes eugnathus Silvestri  [Termites, although important in ecosystems, are economically important structural pests to wood and cellulosic material in service ,3,4,5,6.mitidae) ,8,9,10. M. eugnathus [In Egypt, this termite was reported along the northwestern coast  and in tugnathus .Treatments used to control this termite include bioagents, chemical pesticides, and natural products. Natural products investigated for termite management include plant extracts. In many parts of the world, chemical pesticides are considered poor for the environment and new research is examining the use of naturally occurring, less environmentally toxic, compounds ,12,13.Bacillus thuringiensis var. kurstaki and Stenotrophomonas maltophilia Palleroni & Bradbury have potential application as biological control agents of termites [B. thuringiensis is an example of this [S. maltophilia has shown potential application as a biocontrol agent and could degrade chitin in the termites, Coptotermes heimi (Wasmann) (Blattodea: Rhinotermitidae) and Heterotermes indicola (Wasmann) (Blattodea: Rhinotermitidae) [Bioagents such as termites ,15,16. T of this ,18. ChitDursban (Chlorpyrifos 48%) is an organophosphate pesticide that acts on the insect nervous system by inhibiting acetylcholinesterase (AChE) ,20,21. DC. intermedius Silvestri (Blattodea: Rhinotermitidae) died within 30, 40, and 110 min when exposed to 70% ethanol, aqueous, and acetone extracts from seeds of Parkia biglobosa (Jacq) Benth, respectively, at the concentration of 4 gm/L [Khaya senegalensis (Desr.) A. Juss., K. ivorensis A. Chev. and Swietenia mahagoni (L.) Jacq. showed potential insecticidal efficacy towards the dry wood termite Kalotermes flavicollis   [Larix leptolepis (Lamb.) Carr. Wood, with its large quantities of flavonoids, showed strong feeding deterrent activity against the subterranean termite Coptotermes formosanus Shiraki (Insecta: Blattodea: Rhinotermitidae) [Jatropha curcas L. oil showed maximum wood protection against Microcerotermes beesoni Snyder (Termitidae: Amitermitinae) at a concentration of 20% [Natural products are being studied as potential termiticides. The termite f 4 gm/L . Seed oimitidae) . Water emitidae) . Jatrophn of 20% .Tectona grandis L.f., Dalbergia sissoo Roxb., Cedrus deodara (Roxb.) G.Don and Pinus roxburghii Sarg. rapidly lowered numbers of protozoans in the hindgut of Reticulitermes flavipes (Kollar) (Insecta: Blattodea: Rhinotermitidae) and H. indicola workers, which were closely correlated with the mortality of worker [Morus alba L. heartwood resulted in 100% mortality in R. flavipes after feeding [Microcerotermes beesoni Snyder (Termitidae: Amitermitinae) [D. sissoo showed termite mortality at 10 mg/mL against H. indicola and R. flavipes with LC50s at 5.54 and 3.89 mg/mL, respectively [H. indicola mortality reached more than 75% after feeding on Populus deltoides W.Bartram ex Marshall wood treated with extract from M. alba [Some heartwoods show resistance to termite attack, which is attributed to the presence of toxic compounds within the heartwood ,30. For  feeding . Methanomitinae) . Heartwoectively . H. indi M. alba . Cymbopogon citratus (DC.) Stapf, Eucalyptus globulus Labill., Syzygium aromaticum (L.) Merr. and L. M. Perry, Origanum vulgare L., Rosmarinus officinalis L., Cinnamomum verum J.Presl, and Thymus vulgaris L. showed strong termiticidal activity against Odontotermes assamensis Holmgren (Blattodea: Termitidae) [Cananga odorata (Lam.) Hook.f. and Thomson flower extract of 2 mg/filter paper showed cumulative mortalities of 18% and 94%, after 2 and 7 days of exposure, respectively, against the termite Reticulitermes speratus (Kolbe) (Rhinotermitidae: Blattodea) [K. flavicollis pseudergates fed Casuarina sp. wood wafers, previously treated separately with Taxodium distichum (L.) Rich., E. citriodora Hook. and Cupressus sempervirens L. extracts, showed reduced numbers and vigour in spirochaete and flagellate populations [Tagetes erecta L. leaf extract, caused 100% mortality in Odontotermes obesus (Rambur) (Blattodea: Termitidae) after 24 h of exposure [Microcerotermes eugnathus.Many plants contain bioactive compounds such as phenols, terpenoids , aldehydes, ketones, and other compounds that have a strong effect against microorganisms and termites ,39,40,41mitidae) . Insectiattodea) . K. flavulations . At a doexposure . TherefoLavandula latifolia Medik.) belongs to the family Lamiaceae and is native to the Mediterranean. It is also cultivated in different regions of the world. The bioactivity of L. angustifolia Mill. and L. latifolia extracts from Spain were found to be associated with the presence of linalool, camphor, p-cymene, and limonene [α-pinene, thujene, β-pinene, sabinene, myrcene, p-cymene, limonene, camphene, camphor, 1,8-cineole, (Z)- and (E) ocimene, terpinene, terpinene-4-ol, lavandulol, lavandulylacetate, β-caryophyllene, and (Z)- and (E) farnesene [Lavender  species are characterized by the presence of two main chemotypes, thymol and carvacrol, while others have γ-terpinene, p-cymene, cis-sabinene hydrate, terpinen-4-ol, α-terpineol, α-terpinene, γ-terpinene, sabinene, and linalool as their main compounds [O. vulgare L. ssp. vulgare extract showed the presence of caryophyllene, spathulenol, germacrene-D, and α-terpineol and had antimicrobial properties [γ-terpinene, α-terpineol, thymol, carvacrol, cis-sabinene-hydrate, linalool, p-cimene, 1,8-cineole were identified as the main compounds in the extract of O. vulgare L. [O. vulgare extract and its major compounds carvacrol, p-cymene, and γ-terpinene showed potent larvicidal activity against the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae: Heliothinae) [Several ompounds ,50,51,52operties . Terpinelgare L. . O. vulgothinae) . Syzygium aromaticum (L.) Merr. and L.M.Perry, Myrtaceae family) bud extract has shown the presence of eugenol, eugenol acetate and β-caryophyllene as main compounds with percentages of 88.61, 8.89 and 1.89%, respectively. It exhibited strong contact toxicity against Cacopsylla chinensis (Yang and Li) (Hemiptera: Psyllidae) with LD50 values of 0.730 µg/adult and 1.795 µg/nymph, respectively [S. aromaticum extract gave LC50 values of 13.871 and 15.551 μL against larvae and adult Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) 48 h after fumigation [50 values of 9.45 and 10.15 μL/g, against Acanthoscelides obtectus (Say) (Coleoptera: Chrysomelidae: Bruchinae) and Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), respectively [Clove  and the chemical compound Dursban (chlorpyrifos 48%). In addition, the chemical composition of the three extracts were analyzed with using GC-MS apparatus.This research aimed to (1) determine the chemical composition of three plant extracts  with potential anti-termitic activities; (2) evaluate the anti-termitic activity of the three plant extracts against Microcerotermes eugnathus foraging tubes. Termite tunnels could be observed on the floors and tomb substructure, causing cracks and separation of mortar. This termite species produces winged alates during the months of April and January, the remnants of which (wings) could often be observed around the tombs [Termite infestation in the cemeteries of Bir al-Shaghala located at the oases of Dakhla, Egypt was noted by the presence of he tombs . Figure Lavandula latifolia Medik., Lamiaceae) dry flower, Marjoram  dry leaves and Clove (Syzygium aromaticum (L.) Merrill and Perry, Mytaceae) dry buds were selected for their potential effect on termites. The extraction process was carried out at the National Research Center in Cairo, Egypt. Plant material was shade air-dried in the laboratory at room temperature for one week, then ground into a fine powder in an electrical blender. About 100 g of the fine powdered material was put into a 2-L flask containing 1000 mL of distilled water (DW) and extracted by the hydrodistillation method using a Clevenger-type apparatus for 3 h [Three plant extracts from Spike Lavender  with a TG–5MS direct capillary column (30 m × 0.25 mm × 0.25 µm film thickness). Extracts were diluted in n-hexane solvent in the ratio of 3:1 (3 n-heaxane: 1 extract sample) before being injected to the GC-MS. The column oven temperature was initially held at 50 °C and then increased by 5 °C/min to 250 °C and held for 2 min, then increased to a final temperature of 310 °C by 25 °C/min and held for 2 min. The injector and MS transfer line temperatures were kept at 270 and 260 °C, respectively; helium was used as a carrier gas at a constant flow rate of 1 mL/min. The solvent delay was 3 min, and diluted samples of 2 µL were injected automatically using an Autosampler AS1310 coupled with GC in the splitless mode. EI mass spectra were collected at 70 eV ionization voltages over the m/z range of 50–650 in full scan mode. The ion source temperature was set at 250 °C. The components were identified by comparison of their retention times and mass spectra with those in the WILEY 09 and NIST 11 mass spectral databases [The chemical composition of extracts from atabases . The matatabases ,40,62,63Bacillus thuringiensis var. kurstaki” (Protecto 9.4% WP) was diluted in distilled water to obtain the following concentrations: 164.5, 258.5, 376, 756, and 1410 mg/L. Dursban (Chlorpyrifos 48%) was diluted in distilled water and prepared in concentrations of 120, 240, 360, 480, and 960 mg/L.Extract concentrations for laboratory testing were prepared by mixing with 0.01% of Tween 80 and diluting with distilled water to obtain the following concentrations: 125, 250, 500, 1000, and 2000 mg/L. The bioagent “Microcerotermes eugnathus castes were collected from soil around cemeteries and kept in a container with a portion of termite nesting material. Briefly, from the experimental location, termites were placed inside a special box (4-L in capacity) for preservation and the box was covered with wool textile to maintain the temperature, because the temperatures in the interior oases are as high as 50 °C. During the transfer to the laboratory in Cairo, the death of the termites occurs due to the different environmental conditions, so it was necessary to warm the termites. Termites’ death was also observed during transportation to the laboratory or even in the laboratory during the preparation for the laboratory experiment. Therefore, 10% glucose sugar solution [solution ,65 was pA no-choice bioassay method ,67 was e50) expressed as mg/L was calculated from log-concentration mortality regression lines [Untreated control replicates (five replicates) contained 10 workers, five soldiers and five nymphs, who were supplied with pieces of untreated cardboard paper only. Each concentration tested was replicated five times. All the treatments were kept in darkness at 48 ± 3 °C. After seven days, the test was halted, the number of surviving termites was counted, and percent mortality was calculated. The lethal concentration , lavandulol (12.77%), β-terpinyl acetate (10.49%), camphor (9.30%), linalyl acetate (7.87%), (−)-β-fenchol (5.12%), isobornyl acetate (5.00%), α-pinene (3.81%), terpineol (3.64%), and L-α-terpineol (2.62%).O. vulgare dry leaves. The main components were thymol (14.64%), m-cymene (10.63%), terpinen-4-ol (6.92%), linalool (6.75%), estragole (5.42%), γ-terpinene (5.39%), anethole (5.00%), eucalyptol (3.61%), borneol (3.29%), β-caryophyllene (3.29%), α-terpinene (2.89%), carvacrol (2.58%), camphor (2.36%) and carvacryl methyl ether (2.34%). S. aromaticum extract. The main component was eugenol (99.16%).Microcerotermes eugnathus. After a seven-day exposure to this extract, the LC50 was 770.67 mg/L against soldiers and workers and 627.87 mg/L against nymphs. Lavender extract had less of toxic effect against M. eugnathus than marjoram, with an LC50 of 1086.39 mg/L for the group of soldiers and workers and 1140.74 mg/L for nymphs. Clove extract showed the lowest insecticidal activity against M. eugnathus for both nymphs and soldier/workers, with an LC50 > 2000 mg/L  extract from Macedonia [p-cymen-8-ol and bornyl acetate with values of 32.3%, 12.4%, 11.7%, 8.7%, 7.7%, and 4.2%, respectively, were the main compounds in L. latifolia extract from the Northern Tunisia [L. latifolia extracts from Spain, characterized by 1,8-cineole with high amounts (36.3–33.65%) [L. latifolia from Iran showed the presence of linalool (31.9–30.6%) as the main compound [Lavandula, 1,8-cineole, 2-β-pinene, α-thujone, camphor and cis-α-bisabolene were the main compounds isolated from L. angustifolia extract [Linalool, borneol and terpinene-4-ol, 1,8-cineole, camphor, and linalyl acetate were reported in lavandin  ,82, whilcompound . In othe extract .α-terpinene, γ-terpinene, cis-sabinene-hydrate, linalool, terpinen-4-ol, terpineol, thymol, and carvacrol as the main compounds in O. vulgare extract with percentages of 5.58, 12.32, 3.35, 3.47, 21.43, 3.17, 9.45, and 11.67%, respectively [O. vulgare extract contained p-cymene, linalool, thymol, caryophyllene, and carvacrol. These authors showed insecticidal activity against Schistocerca gregaria (Forskål) (Orthoptera: Acrididae) [O. vulgare extract. Other studies reported this same compound at 9.45% [Coleus amboinicus Lour. extract caused 100% mortality to the termite Odontotermes obesus at a dose of 2.5 × 10−2 mg/cm3. This study found that thymol was the main compound, with 94.3% [Pittosporum undulatum Vent., Lippia sidoides Cham. and Lippia gracilis Schauer showed 100% mortality to the termite Heterotermes sulcatus Mathews (Blattodea: Rhinotermitidae) after 14 days of exposure, where the activity was related to the major compounds, limonene (80.8%), carvacrol (45.6%) and thymol (78.55%), that were identified in each extract, respectively [Previous work identified ectively . Other wrididae) . In the at 9.45% , 44.55% at 9.45% , 26.75% at 9.45% , and 58.at 9.45% . After 5th 94.3% . Extractectively .O. vulgare extract [O. vulgare extract with its main compounds of thymol (40.35%), p-cymene (17.32%), γ-terpinene (15.66%), and carvacrol (12.15%), observed 72% and 88% mortality at 48 and 72 h, respectively, against Sitophilus oryzae (L.) (Coleoptera: Curculionidae) [O. vulgare extract showed toxicity in higher doses against Plutella xylustella L.  [O. vulgare extract at 40% exhibited 100% repellency against Cimex lectularius Linnaeus (Hemiptera: Cimicidae) [Terpineol (22.85%), α–terpinene (20.60%), caryophyllene (6.75%) and thymol (4.53%) were the main compounds isolated from  extract . O. vulgionidae) . O. vulgralidae) . After amicidae) .Origanum were shown to have insecticidal activity against larvae of Culex pipiens L. (Diptera: Culicidae) [Lippia sidoides contained 44.55% thymol and showed high insecticidal activity against the termite Nasutitermes corniger (Blattodea: Termitidae) with 48 h exposure at a dose of 0.27 μg/mg [Oxygenated monoterpenes such as thymol, eucalyptol, linalool and carvacrol found in the extracts from the present study are reported to be more toxic against termite workers . For exalicidae) . The ext27 μg/mg .Microcerotermes eugnathus with an LC50 value of >2000 mg/L. Eugenol has been shown to be an effective fumigant and feeding deterrent against the termite C. formosanus [γ-aminobutyric acid, causing inhibitory symptoms in the nervous system [Odontotermes assamensis Holmgren (Blattodea: Termitidae) at 2.5 mg/g after 8 days [Reticulitermes messperatus Kolbe (Blattodea: Rhinotermitidae) nymphs [S. aromaticum extract at 0.5 mL/L after 10 min of exposure showed 100% mortality against Coptotermes formosanus Shiraki (Insecta: Blattodea: Rhinotermitidae) [R. speratus [In this study, clove extract showed the lowest activity against rmosanus . Carvacrrmosanus . It causs system . Carvacrr 8 days . At 1.5 ) nymphs ,97. S. amitidae) . Clove bsperatus .M. eugnathus against termiticides than those of other termites. In addition, our results suggest that O. vulgare extract caused moderate activity against the termite M. eugnathus compared to commercial termiticides tested. Further research may show that O. vulgare extract or its derivatives could potentially be used to control and manage some termite infestations and lessen or limit the amount of more toxic pesticides currently in use. Our present results show a higher resistance of M. eugnathus. However, due to the highly toxic nature of Dursban and the fact that it has been withdrawn for use in some countries, the search for less toxic and natural termiticides from plants is of interest. Extract from the plant Origanum vulgare showed some promise as a plant-based toxicant for this termite. Chemical pesticides had the greatest impact on the mortality of the termite


Bacillus sp. strains EKM417B and EKM420B (from Citrullus lanata [watermelon]) and EKM501B (from Cucurbita moschata [butternut squash]) and Paenibacillus sp. strain EKM301P (from Cucurbita pepo L. var. pepo L. [pumpkin]). These strains previously demonstrated biostimulant and biocontrol activities.Here, we announce the draft genome sequences of four endophytic bacilli isolated from surface-sterilized seeds of three cucurbit species,  Bacillus sp. strains EKM417B and EKM420B (from Citrullus lanata [watermelon]) and EKM501B (from Cucurbita moschata [butternut squash]) and Paenibacillus sp. strain EKM301P (from Cucurbita pepo L. var. pepo L. [pumpkin]). These strains previously demonstrated biostimulant and biocontrol activities.Here, we announce the draft genome sequences of four endophytic bacilli isolated from surface-sterilized seeds of three cucurbit species,  Bacillus/Paenibacillus genera) (Bacillus sp. strains EKM417B and EKM420B (from Citrullus lanata [watermelon]), Bacillus sp. strain EKM501B (from Cucurbita moschata [butternut squash]), and Paenibacillus sp. strain EKM301P (Cucurbita pepo L. var. pepo L. [pumpkin]) using 16S rRNA gene primer pair 799F and 1492R and then submitted to GenBank   in planta; EKM501B grew on N2-free medium, produced indole-3-acetic acid (IAA/auxin) and ribonucleases, and suppressed Rhizoctonia solani; and EKM301P secreted cellulase and antagonized Fusarium graminearum and Rhizoctonia solani (Plant microbiomes have evolved to perform defensive/growth-promoting functions –3. 16S I– genera) , consist genera) . We isolde novo assembled using the EvoCAT pipeline (Evogene Clustering and Assembly Toolbox) and identified using KmerFinder 3.2 (Bacillus velezensis strain KD1 (GenBank accession number NZ_CP014990.2) (EKM417B and EKM420B), Bacillus cereus strain FORC087 (NZ_CP029454.1) (EKM501B), and Paenibacillus polymyxa strain SQR-21 (NZ_CP006872.1) . Genomic DNA was extracted using DNeasy UltraClean microbial kits  and adjusted to 50 ng/μl. Libraries were constructed using TruSeq DNA Nano library prep kits  and then sequenced using the Illumina NovaSeq 6000 system to produce 1,461,384 (EKM417B), 1,659,509 (EKM420B), 1,823,824 (EKM501B), and 2,124,086 (EKM301P) raw reads in 150-bp paired-end format. Quality-trimmed reads  were 06872.1) . The pro06872.1) . Peptide06872.1) . DefaultGenome mining identified candidate genes involved in biofertilizer/biocontrol metabolic pathways, including those discussed above. These genes encode proteins involved in nitrogen fixation, phytase, alkaline phosphatase, carbon-nitrogen hydrolase, trehalose-6-phophate hydrolase, and tryptophan synthase (IAA/auxin production) –14. Bioc–14–This whole-genome shotgun project and the Illumina raw reads have been deposited in DDBJ/EMBL/GenBank and the SRA, respectively, under the accession numbers provided in


Y.Tsiang 7712) when he described Tsugalongibracteata W.C.Cheng. Later, researchers suggested that the type is either in NAS or in PE. However, we found more than one duplicate of the type collection in both NAS and PE. Following the Shenzhen Code, we lectotypify the name T.longibracteata with Y.Tsiang 7712 (PE00003223) that bears a handwritten identification of W.C.Cheng.W.C.Cheng did not clearly indicate the herbarium repository of the type specimen ( Tsuga (Endl.) Carrière: Tsugalongibracteata W.C.Cheng. This species differs from all known species of Tsuga in both vegetative and reproductive characters , two in NAS (NAS00070064 and NAS00070063), two in HUH (A00052508 and A00052510), one each in E (E00215871), IBSC (IBSC0012857), K (K000288277), NY (NY00001279), and S (S-C-4796) respectively. Shenzhen Code .1A55C046-BD56-5079-A68C-4A134EA9F396Nothotsugalongibracteata (W.C.Cheng) Hu ex C.N.Page, Notes Roy. Bot. Gard. Edinburgh 45(2): 390 .  ≡Y.Tsiang (蒋英) 7712 .China. Guizhou (贵州): Yinjiang Tujiazu Miaozu Zizhixian , Fanjing Shan , in densely shaded ravine, alt. 400–500 m, 19 December 1930,


Correction to: Arthritis Res Ther 22, 98 (2020)https://doi.org/10.1186/s13075-020-02185-0Following publication of the original article , the aut1. In Fig. 2. Supplementary Figure The original article has been corrected.Additional file 1:Figure S1. Distribution of FOI PVM enhancement in joints with increasing degree of osteoarthritis.


Food and Drug Administration in 2019 for the treatment of community-acquired bacterial pneumonia (CABP). In this study we evaluated the The test strains were isolated from patients across China during the period from 2017 to 2019, including 634 strains of respiratory pathogens. The minimum inhibitory concentrations (MICs) of lefamulin and comparators were determined by broth microdilution method.Streptococcus pneumoniae and Staphylococcus evidenced by 100% inhibition at 0.25 mg/L, and favorable MIC50/90 (0.125/0.125 mg/L) against S. pneumoniae (penicillin MIC ≥ 2 mg/L), MIC50/90 (≤0.015/0.125 mg/L) against methicillin-resistant S. aureus, and MIC50/90 (≤0.015/0.06 mg/L) against methicillin-resistant S. epidermidis. Lefamulin also had good activity against Streptococcus pyogenes and Streptococcus agalactia (MIC50/90: ≤0.015/≤0.015 mg/L), β-lactamase-producing Haemophilus influenzae (MIC50/90: 0.5/1 mg/L), β-lactamase-negative H. influenzae (MIC50/90: 1/1 mg/L), Moraxella catarrhalis (MIC50/90: 0.25/0.25 mg/L), and Mycoplasma pneumoniae (MIC50/90: 0.03/0.03 mg/L) regardless of resistance to azithromycin. Lefamulin was generally more active than the comparators against the test strains.Lefamulin showed potent activity against Staphylococcus, S. pneumoniae, β-hemolytic Streptococcus, H. influenzae, M. catarrhalis and M. pneumoniae). In vitro activity supports the use of lefamulin in the treatment of CABP in China.In summary, lefamulin has good and broad-spectrum coverage of respiratory pathogens (methicillin-sensitive and -resistant  Clitopilus scyphoides, Clitopilus passeckerianus, or other Clitopilus species in basidiomycota. Lefamulin is the first-in-class semi-synthetic pleuromutilin antibiotic for systemic use. Its molecular formula is C28H45NO5S (molecular weight 567.79 g). Lefamulin inhibits bacterial protein synthesis by binding to “A” and “P” sites of the peptidyl transferase center (PTC) of the 23s rRNA of the 50S ribosomal subunit of bacterial cell. The binding is through the mutilin core and C-14 side chain in the forms of hydrogen bonds, hydrophobic interactions, and conformational change to prevent correct orientation of tRNA’s 3’-CCA ends for peptide transfer  coded by the transposon Tn5406 and vga(A) carried by plasmids (encoding ABC transporter)  . So far,Streptococcus pneumoniae (PRSP), macrolide-resistant Mycoplasma pneumoniae, and methicillin-resistant S. aureus (MRSA)  . In Auguin vitro activity of lefamulin against a broad range of respiratory pathogens.The antibacterial spectrum and activity of lefamulin have been studied in the United States and Europe , 2019, bMycoplasma pneumoniae. These strains were isolated from 29 hospitals across China, representing 23 provinces and municipalities, during the period from October 2017 to July 2019. Specifically, the test strains included S. aureus (n = 121), S. epidermidis (n = 30), β-lactamase-producing Haemophilus influenzae (n = 48), β-lactamase-negative H. influenzae (n = 48), Haemophilus parainfluenzae (n = 10), Moraxella catarrhalis (n = 54), S. pneumoniae (n = 172), Streptococcus pyogenes (n = 30), and Streptococcus agalactiae (n = 13). All the strains were re-identified before susceptibility testing. Species identification was confirmed by MALDI-TOF/MS system , and antimicrobial susceptibility testing were controlled with reference strains S. aureus ATCC29213, S. pneumoniae ATCC49619, H. influenzae ATCC49247, and M. pneumoniae ATCC 29342.A total of 634 non-duplicate strains of respiratory pathogens were tested, including 580 strains of bacteria and 54 strains of M. pneumoniae were measured according to the methods for antimicrobial susceptibility testing for human mycoplasmas described in CLSI document M43-A (2011) (The minimum inhibitory concentrations (MICs) of lefamulin and the comparators were determined by broth microdilution method according to the Clinical and Laboratory Standards Institute  2018) M M2018) MA (2011) . The antth Edition .Staphylococcus strains , 0.06 mg/L against methicillin-sensitive S. aureus (MSSA), and 0.03 mg/L against methicillin-sensitive S. epidermidis (MSSE). Lefamulin displayed MIC values ranging from ≤0.015 mg/L to 0.25 mg/L (MIC90: ≤0.25 mg/L) against 172 strains of S. pneumoniae, including penicillin-susceptible (PSSP) strains (penicillin MIC ≤0.06 mg/L), penicillin-intermediate (PISP) strains (penicillin MIC: 0.125 mg/L–1 mg/L), and penicillin-resistant (PRSP) strains (penicillin MIC ≥ 2 mg/L). Lefamulin inhibited the growth of all PSSP strains at ≤0.015 mg/L and all PISP and PRSP strains at 0.25 mg/L. The MIC50/90 values of lefamulin were ≤0.015/≤0.015 mg/L against S. pyogenes and ≤0.015/0.06 mg/L against S. agalactiae. Lefamulin inhibited the growth of all the S. pyogenes and S. agalactiae strains at 0.06 mg/L . Its activity was comparable to moxifloxacin and significantly superior to erythromycin and azithromycin  in this respect.In the present study, lefamulin displayed excellent antimicrobial activity against all the respiratory pathogens, including MRSA, MSSA, MRSE, MSSE, 015–2016 , 2019. THaemophilus and M. catarrhalis. Lefamulin was comparable to ceftriaxone in activity against S. pneumoniae strains  and β-hemolytic Streptococcus, but better than ceftriaxone against PRSP, better than penicillin against PISP and PRSP, and similar to penicillin against β-hemolytic Streptococcus. Lefamulin had similar activity as moxifloxacin, vancomycin, and linezolid against Streptococcus. It inhibited the growth of all Streptococcus species at 0.125 mg/L, which was lower than the above mentioned three agents. Lefamulin was significantly better than erythromycin and azithromycin in the activity against S. pneumoniae and β-hemolytic Streptococcus.Lefamulin also displayed high antimicrobial activity against H. influenzae and M. catarrhalis. As for the β-lactamase-negative strains, lefamulin provided significantly better activity than azithromycin. Lefamulin was comparable to tigecycline, ceftriaxone, and levofloxacin, and significantly superior to azithromycin and trimethoprim-sulfamethoxazole in the activity against M. catarrhalis. These results are consistent with those reports from other countries  (It has been reported that the 02 mg/L) .90 value of lefamulin was 0.5 μg/mL against the 50 strains of S. pneumoniae isolated from the patients in phase III clinical trial LEAP 1  was 0.12–0.25 μg/mL. The post-treatment bacterial clearance rate was up to 100%. Research results at home and abroad have shown that lefamulin had similar antimicrobial activity against S. epidermidis and S. aureus  over the MIC (24 h AUC/MIC). Lefamulin achieves rapid and predictable penetration into human tissues, with a mean 5.7-fold higher concentration in the pulmonary epithelial lining fluid compared with plasma. Percent probabilities of attaining the median AUCELF/MIC ratio targets associated with a 1-log10 CFU reduction from baseline by MIC were 97.0% at a MIC of 0.5 μg/mL for S. pneumoniae and 99.4% at a MIC of 0.25 μg/mL for S. aureus .DZ and FH designed the study. SW, YZ, YG, and DY performed the experimental work. SW and YZ collected the data. FH analyzed the data. All authors read and approved the final manuscript, contributed to the article, and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


T(temperature), Moisture content (MC), sediment total phosphorus (STP), ion-exchangeable form of ammonia (IEF-NH4+-N), weak-acid extractable form of ammonia (WAEF-NH4+-N), weak-acid extractable form of nitrate (WAEF-NO3−-N), and strong-alkali extractable form of ammonia (SAEF-NH4+-N) were the dominant environmental factors and explained 11.1%, 8.2%, 10.7%, 6.9%, 9.3%, 8.1%, 10.5%, 7.5%, and 7% variation, respectively, of the total variation in the microbial community. Furthermore, the network analysis showed that symbiotic relationships accounted for a major percentage of the microbial networks. The keystone aerobic denitrifying bacteria belonged to Comamonas, Rhodobacter, Achromobacter, Aeromonas, Azoarcus, Leptothrix_Burkholderiales, Pseudomonas, Thauera, unclassified_Burkholderiales, and unclassified_bacteria. The composition of the keystone aerobic denitrifying microbial community also exhibited significant differences  on the basis of the adonis analysis. T, STP, IEF-NH4+-N, ion-exchangeable form of nitrate (IEF-NO3−-N), WAEF-NO3−-N, SAEF-NH4+-N, and TN were the dominant environmental factors that explained 8.4%, 6.2%, 4.6%, 5.9%, 5.9%, 4.5%, and 9.4% variation, respectively, of the total variation in the keystone aerobic denitrifying microbial community. The systematic investigation could provide a theoretical foundation for the evolution mechanism of the aerobic denitrifying microbial community in Baiyangdian Lake.Here, the ion-exchangeable form of nitrogen (IEF-N), weak-acid extractable form of nitrogen (WAEF-N), strong-alkali extractable form of nitrogen (SAEF-N), strong-oxidant extractable form of nitrogen (SOEF-N), residue nitrogen (Res-N), and total nitrogen (TN) showed spatial differences, and most of the sediment nitrogen fractions exhibited positive correlations in Baiyangdian Lake. High-throughput sequencing analysis revealed that the aerobic denitrification microbial community was composed of proteobacteria (42.04%–99.08%) and unclassified_bacteria (0.92%–57.92%). Moreover, the microbial community exhibited significant differences (R Thiosphaera pantotropha) [Excessive nitrogen concentrations present problems for water quality and cause eutrophication . An incrotropha)  has beenotropha) , Wadden otropha) , wetlandotropha) , coastalotropha)  and reseotropha) ,10. Therotropha) ,12.napA is the biomarker of aerobic denitrifying bacteria [napA) in Baiyangdian Lake.Currently, some full-scale experiments on bioaugmentation with aerobic denitrification bacteria have been conducted successfully for the remediation of wastewater, urban river, and river sediment. Such as, Duan et al. 2015) explored the nitrogen removal performance of halophilic heterotrophic nitrification-aerobic denitrification SF-16 in saline wastewater ; Du et a5 explorebacteria ,21. TherIn this study, our objectives were to (1) investigate the characteristics of environmental factors; (2) examine the composition of the aerobic denitrifying microbial community and keystone operational taxonomic unit (OTU); (3) evaluate the differences in abundance, diversity, and community structure; (4) analyze the relationship between the microbial community structure and environmental driving factors; and (5) estimate the relative contributions of the environmental driving factors.Baiyangdian Lake, located in Xiong’an New Area, is the largest freshwater lake in the North China Plain a. Baiyan3−-N, NO2−-N, and NH4+-N were measured using DR6000  [T, DO, pH, ORP, and EC of the water samples were determined using Hydrolab DS5 . According to the standard methods, TN, NOny, USA) . The difny, USA) ,24.napA [https://submit.ncbi.nlm.nih.gov/subs/sra/) database with the accession number PRJNA623955.The whole DNA of the sediment was extracted using the Soil DNA Kit . After DNA purification, the extracted DNA was stored at −80 °C for PCR amplification. The extracted DNA was amplified using primer V66F, 5′-TAYTTYYTNHSNAARATHATGTAYGG-3′, and V67R, 5′- DATNGGRTGCATYTCNGCCATRTT-3′, for aerobic denitrifying bacterial napA ,25. PCR napA . High-thOn the basis of the OTU data, the richness index, Shannon index, Simpson index, Pielou index, Chao1 index, ACE index, and Good’s Coverage were calculated to evaluate the alpha diversity. The differences in the microbial communities were evaluated using principal co-ordinate analysis (PCoA) and permutational MANOVA (adonis) in the vegan package (R.3.5.3). Redundancy analysis (RDA) was used to determine the correlation between the microbial community and predominant environmental factors with variance inflation factor (VIF) < 10 in the vegan package (R.3.5.3). Indicator species analysis was conducted using labdsv package (R.3.5.3). Hierarchical partitioning (HP) analysis was performed to quantitatively evaluate the relative influences of the environmental factors on the aerobic denitrifying bacterial community by using the rdacca.hp package (R.3.5.3) .Network analysis was performed to investigate the biotic interactions between the microbial populations, and the results were visualized with Gephi software (0.9.2). In this study, a positive correlation implied a mutualistic interaction, whereas a negative correlation may indicate competition . The “keNatural area. The IEF-N, WAEF-N, SAEF-N, SOEF-N, and Res-N values were 100.61, 2831.38, 1487.38, 995.42, and 1093.82 mg/kg, respectively , whereas it was negatively correlated with IEF-NO3−-N , SAEF-NH4+-N , IEF- N , and IEF-NH4+-N  , IEF-N , SAEF-N , IEF-NH4+-N , SAEF-NO3−-N , and TAN   and IEF-NO3−-N . IEF-NO3−-N was positively correlated with IEF-N  , SAEF-NO3−-N , and TAN  (p < 0.05), IEF-NH4+-N , SAEF-NO3−-N , TAN , SOEF-NO3−-N , SOEF-N , and SOEF-NH4+-N  , SAEF-NO3−-N , and TAN  , TAN , SOEF-NO3−-N , SOEF-N , and SOEF-NH4+-N  , SOEF-NO3−-N , SOEF-N , and SOEF-NH4+-N  , SOEF-N , and SOEF-NH4+-N   and SOEF-NH4+-N  .WAEF-NO < 0.05) . Res-N w< 0.001) . WAEF-NH < 0.01) . SAEF-NH < 0.01) . IEF-N w < 0.05) b. SAEF-N< 0.001) . IEF-NH4 < 0.05) . SAEF-NO < 0.05) . TAN was < 0.01) b. SOEF-N< 0.001) . SOEF-N After quality trimming, a total of 223754 sequences with an average length of 373 bp were obtained for the 14 sediment samples. MiSeq revealed a total of 1886 OTUs with 97% similarity . The Shaα-diversity was influenced by PCoA1 , MC (R = 0.70), STP (R = 1.00), IEF-NH4+-N (R = 0.97), and WAEF-NH4+-N (R = 0.80) were the dominant environmental parameters  , unclassified_bacteria (0.92%–57.92%), 0–0.25%) . ProteobAt the class level , BetaproComamonas (0–83.37%), unclassified_bacteria (1.44–58.39%), Rhodobacter (0.01–50.84%), unclassified_Burkholderiales (0.46–28.84%), Aeromonas (0.08–23.42%), Azoarcus (1.19–22.78%), Thauera (0.31–18.17%), Achromobacter (0.02–11.86%), Pseudomonas (0.11–10.78%), Sulfuritalea (0.09–9.75%), unclassified_betaproteobacteria (0.28–9.28%), Cedecea (0–8.16%), Dinoroseobacter (0–7.61%), Sulfurospirillum (0–7.54%), Leptothrix_Burkholderiales (0.29–5.61%), Magnetospirillum (0.19–4.59%), Azospirillum (0.03–3.52%), Azospira (0–3.30%), Rhodopseudomonas (0.16–3.09%), and Rhizobium (0–2.10%) (Comamonas (BH), unclassified_Betaproteobacteria (BGYH), Achromobacter (BGYH), Sulfurospirillum (PH), Azospira (BGYH), and Azospirillum (PH); for the breeding area, unclassified_bacteria (FYD) and Thauera (SHD); for the tourist area, Sulfuritalea (YYD) and Dinoroseobacter (YYD); for the living area, unclassified_Burkholderiales (BTZXD), Aeromonas (ZLZ), Magnetospirillum (ZLZ), Cedecea (PYD), Leptothrix_Burkholderiales (BTZXD), and Rhizobium (BTZXD); and for the natural area, Azoarcus (ZZD), Rhodobacter (ZZD), Pseudomonas (ZZD), and Rhodopseudomonas (ZZD). Previous studies have shown that these dominant genera are aerobic denitrifying bacteria. For example, Comamonas is capable of heterotrophic nitrification-aerobic denitrification nitrogen removal, and it has been successfully used for bioaugmentation of municipal wastewater [Rhodobacter is a typical denitrifying bacterium in alkaline copper mine drainage [Aeromonas sp. HN-02 [Aeromonas, Pseudomonas, and Acinetobacter) could be successfully enriched and conducted denitrification to remove 90% of TN in a wetland test system [Azoarcus was an important genus in a denitrifying quinoline-removal bioreactor [Thauera was identified as the key potential heterotrophic nitrification and aerobic denitrification genus for principal nitrogen removal in granular reactors [Achromobacter xylosoxidans CF-S36, a typical heterotrophic nitrification-aerobic denitrification bacterium, were investigated through molecular analysis [Pseudomonas indoloxydans YY-1, through physiological and transcriptomic analyses [Pseudomonas putida strain NP5 [Azospira was the main dentrifier in a tidal flow constructed wetland [Halomonas was the dominant heterotrophic nitrifying/aerobic denitrifying genus under hypersaline conditions [Rhodopseudomonas is a potential autotrophic denitrifying genus that can use electrons and reducing power from cathodes [At the genus level, the top 20 abundant genera are as follows: 0–2.10%) . Moreovestewater  and enristewater . Liu et nas 0–83.%, unclasdrainage . Chen et 1.44–58.%, Rhodobt system . Azoarcuoreactor , and it  0.08–23.%, Azoarcreactors . The dynanalyses  and Yangriales 0.–28.84%,  wetland , reservo1.44–58.3%, Rhodob wetland . Halomonnditions . Rhodopster 0.01–.84%, uncDifferences in the composition of the aerobic denitrifying microbial community in the five areas were investigated with PCoA and NMDS analysis. RDA was used to demonstrate the links between the environmental parameters and microbial community. HP analysis was performed to investigate the relative influence of the environmental driving factors on microbial community composition.For the whole aerobic denitrifying microbial community. The PCoA results showed that PCoA1 and PCoA2 accounted for 42.91% and 17.42%, respectively, of the variability in the microbial community composition  on the basis of the adonis analysis. The stress result (stress = 0.06 < 0.1) of NMDS showed that the NMDS analysis exhibited good representation  on the basis of the adonis analysis. The stress result (stress = 0.04 < 0.05) of NMDS showed that NMDS analysis exhibited excellent representation , module 4 , module 6 , module 7 , and module 8 ; STP and WAEF-NO3−-N exhibited a significantly negative correlation with modules 1–8, and the correlation reached −0.56 to −0.58 (p < 0.05) and −0.64 (p < 0.05); WAEF-NH4+-N, Res-N, and TN were positively correlated with modules 1–8, and the correlation reached 0.54–0.64 (p < 0.05), 0.65–0.70 (p < 0.05), and 0.67–0.71 (p < 0.01). WAEF-N was positively correlated with module 1 , module 2 , module 5 , and other modules . Especially, the keystone aerobic denitrifying microbial community exhibited a significantly positive correlation with T , WAEF-NH4+-N , Res-N , and TN  and a significantly negative correlation with STP . On the basis of all the results, T, STP, WAEF-NH4+-N, WAEF-NO3−-N, and TN were important environment factors, which is consistent with the RDA and HP analysis results.The correlations between the modules and environmental factors were investigated in Baiyangdian Lake c. The ennapA). In this study, the environmental parameters  exhibited significantly spatial differences among the different functional areas, and most of the sediment nitrogen fractions exhibited positive correlations in Baiyangdian Lake. MiSeq revealed a total of 1886 OTUs identified as proteobacteria (42.04–99.08%), unclassified_bacteria (0.92–57.92%), and Deinococcus-Thermus (0.00–0.25%). The unclassified genus accounted for an important part in Baiyangdian Lake. The RDA and VIF results showed that T, MC, STP, IEF-NH4+-N, and WAEF- NH4+- N were the dominant environment parameters and explained 11%, 13.6%, 27.6%, 16.3%, and 2.8% variation in α-diversity. Moreover, the microbial community exhibited significantly spatial differences . T, MC, STP, IEF-NO3−-N, WAEF-NH4+-N, WAEF-NO3−-N, and SAEF-NH4+-N were the dominant environmental factors and explained the 11.1%, 8.2%, 10.7%, 9.3%, 8.1%, 10.5%, and 7.5% variation of the total variation in the whole aerobic denitrifying microbial community, respectively. The network analysis showed that symbiotic relationships accounted for a major percentage of the microbial networks, and 40.18% nodes belonged to betaproteobacteria. The keystone OTUs belonged to Comamonas, Rhodobacter, Achromobacter, Aeromonas, Azoarcus, Leptothrix_Burkholderiales, Magnetospirillum, Pseudomonas, Sulfuritalea, and Thauera. Furthermore, the composition of the keystone aerobic denitrifying microbial community also exhibited significantly spatial differences , and T, STP, IEF-NH4+-N, IEF-NO3−-N, WAEF-NO3−-N, SAEF-NH4+-N, and TN were the dominant environmental factors. From all the results, this study supplied a new view on investigating the distribution characteristics and driving factors of the aerobic denitrifying microbial community in Baiyangdian Lake.This is the first report of the characteristics and driving factors of the aerobic denitrifying microbial community, especially, the “keystone species” and the dominant environment factor in natural water ecosystem through the aerobic denitrification functional gene (


Correction to: Isr J Health Policy Res 10, 12 (2021)https://doi.org/10.1186/s13584-021-00449-xFollowing publication of the original article , the autBelow the incorrect and correct table description:Incorrect: Table 1 Cumulative COVID-19 vaccination doses administered per 100 people by Feb. 1, 2021, or latest date availableCorrect: Table 1: Cumulative COVID-19 vaccination doses administered per 100 people by Feb. 11, 2021, or latest date availableThe original article has been updated.


Stem rust in recent years has acquired an epiphytotic character, causing significant economic damagefor wheat production in some parts of Western Siberia. On the basis of a race composition study of the stem rustpopulations collected in 2016–2017 in Omsk region and Altai Krai, 13 pathotypes in Omsk population and 10 inAltai population were identified. The race differentiation of stem rust using a tester set of 20 North AmericanSr genes differentiator lines was carried out. The genes of stem rust pathotypes of the Omsk population are avirulentonly to the resistance gene Sr31, Altai isolates are avirulent not only to Sr31, but also to Sr24, and Sr30. A lowfrequency of virulence (10–25 %) of the Omsk population pathotypes was found for Sr11, Sr24, Sr30, and for Altaipopulation – Sr7b, Sr9b, Sr11, SrTmp, which are ineffective in Omsk region. Field evaluations of resistance to stemrust were made in 2016–2018 in Omsk region in the varieties and spring wheat lines from three different sources.The first set included 58 lines and spring bread wheat varieties with identified Sr genes – the so-called trap nursery. The second set included spring wheat lines from the Arsenal collection,that were previously selected according to a complex of economically valuable traits, with genes for resistanceto stem rust, including genes introgressed into the common wheat genome from wild cereal species. The thirdset included spring bread wheat varieties created in the Omsk State Agrarian University within the framework ofa shuttle breeding program, with a synthetic wheat with the Ae. tauschii genome in their pedigrees. It was establishedthat the resistance genes Sr31, Sr40, Sr2 complex are effective against stem rust in the conditions of WesternSiberia. The following sources with effective Sr genes were selected: (Benno)/6*LMPG-6 DK42, Seri 82, Cham 10,Bacanora (Sr31), RL 6087 Dyck (Sr40), Amigo , Siouxland , Roughrider , Sisson, and Fleming , Pavon 76 (Sr2 complex) from the ISRTN nursery; No. 1 BC1F2(96 × 113) × 145 × 113 , No. 14а F3 (96 × 113) × 145 , No. 19 BC2F3 (96 × 113) × 113 , and No. 20 F3 (96 × 113) × 145  from the Arsenal collection; and the Omsk State AgrarianUniversity varieties Element 22 , Lutescens 27-12, Lutescens 87-12 , Lutescens 70-13, andLutescens87-13 . These sources are recommended for inclusion in the breeding process for developingstem rust resistant varieties in the region. Stem rust of wheat caused by Puccinia graminis f. sp. triticiErikss. for a long time had a weak manifestation in the territoryof Western Siberia and only in the recent years acquiredan epiphytotic nature, causing significant economic damagefor wheat production in the region. First of all, this is due tothe deterioration of the phytosanitary situation in the region,the general trend of climate warming and cultivation of susceptiblewheat varieties on large area . The threat of stem rust race Ug99 appearance and theemergence of new pathotypes of this race, affecting varietieswith genes Sr24 and Sr36 present a serious threat for wheatproduction in West Siberian region. Genetic diversity of cultivatedwheat varieties for resistance to Ug99 and stem rustin general is very limited .Enhancement of genetic resistance to pathogens can besolved germplasm exchange, and also cultivation of varietieswith different level of resistance to diseases and to differentraces. Crop protection is necessary to restrain the evolutionof pathogens and the emergence of new virulent races.Such programs are widely used in Europe and America. Theduration of the variety cultivation in advanced countries is3–4 years, while in Russia – 7–10 years . Inthis regard, the breeding of spring wheat varieties, whichhave a diverse genetic basis of resistance to stem rust, is veryrelevant.Amigo .Since the 1950s, many resistance genes introduced intobread wheat have lost their effectiveness .The most significant genes for breeding practice are Sr2,Sr23, Sr24, Sr25, Sr31, Sr33, Sr36, Sr38, Sr45, Sr50, SrTmp,Sr1RSIntrogression of resistance genes of wild and cultivatedwheat relatives allows to expand the genetic diversity ofvarieties and contributes to their long-term protection . To date, about 86 Sr genes have beenidentified,of which 26 stem rust resistance genes havebeen transferred into bread wheat from other cereal species. For example, T. turgidum was thesource of the stem rust resistance genes Sr2, Sr9d, Sr9e, Sr9g,Sr11, Sr12, Sr13, Sr14, and Sr17, of which the Sr2, Sr13, andSr14 genes are effective against Ug99 race; T. monococcumwas the source of Sr21, Sr22, and Sr35 genes .Genes that caused the resistance to stem rust have beenintroduced into wheat gene pool from the genome of variousAegilops L. species: Ae. speltoides – Sr32, Sr39, Sr47;Ae. comosa – Sr34; Ae. ventricosa – Sr38 . Ae. tauschii contributed genes Sr33, Sr45, Sr46 . Direct hybridization of T. aestivum withAe. tauschii and following backcrosses allowed introductionof new resistance genes SrTA1662, SrTA1017, and SrTA10187effective against Ug99 race . The searchof new resistance genes in wild wheat relatives continues, forexample, G. Yu et al. (2017) identified two new Sr genes inAe. sharonesis.One of the objectives of Kazakh-Siberian Spring WheatImprovement Network (KASIB) is expanding of the geneticpolymorphism of new varieties, including resistance to harmfuldiseases . This is based onshuttle breeding with CIMMYT (Mexico). Varieties andbreeding lines developed through shuttle breeding with participationof Ae. tauschii and T. dicoccum, as well as linesof the “Arsenal” collection, which have wild species in theirpedigree are of interest for breeding for resistance to stemrust in the region.The aim of the research was analysis of the racial compositionof the Western-Siberian stem rust population, resistanceassessment of spring bread wheat lines and varieties with identified resistance genes and identification of the sourceswith effective Sr genes for breeding under Western Siberianconditions.http://agro.au.dk/forskning/internationaleplatforme/wheatrust).The racial composition of Puccinia graminis f. sp. tritici populationscollected in 2016–2017 in Omsk region ) and Altai region  were analyzed in the Global Rust Reference Center was carriedout. Monopustule isolates were reproducted to identifyrace Ug99 with usage of the test PCR-Stage 1. A total of19 single pustule isolates were selected from Omsk populationand 20 – from Altai population (Table 1).Selection of single pustule isolates according to requirementsof GRRC protocols (Differentiation of stem rust races was performed with useof the set of 20 North American differentiator lines containingSr genes: Sr5 (ISr5-Ra), Sr21 (CnS_Triticum monoc.Deriv.), Sr9e (Vernstein), Sr7b (ISr7b-Ra), Sr11 (ISr11-Ra), Sr6 (ISr6a-Ra), Sr8a (ISr8a-Ra), Sr9g (CnSr9g), Sr36(W2691SrTt-1), Sr9b (W2691Sr9b), Sr30 (BtSr30Wst),Sr17+13 (Combination VII), Sr9a (ISr9a-Ra), Sr9d (ISr9d-Ra), Sr10 (W2691Sr10), SrTmp (CnsSrTmp), Sr24 (LcSr24Ag),Sr31 (Benno Sr31/6*LMPG), Sr38 (VPM-1), SrMcN(McNair 701). Infected plants were evaluated in 14–16 daysafter inoculation according to modified E.C. Stakman scale. Virulence phenotypes were classifiedaccording to North American system .The varieties and lines of bread wheat from three germplasmsets were evaluated in Omsk at least 4–5 times for reaction tostem rust on scales recommended by Koyshibaev et al. (2014).The type of reaction on E.B. Mains and H.S. Jacksonscale(1926) and severity – on modified Peterson scale  were considered: 0 – immunity, uredopustulesnot formed; R (Resistance – high resistance), 1 score, severity 5–10 %; MR (Moderately resistant – average resistance),2 score, severity 10–25 %; M (heterogeneous type), pustulesof different sizes, surrounded by chlorotic and necrotic spotsor without them; MS (Moderately susceptible – averagesusceptibility), 3 score, severity 40–50 %; S (Susceptible –susceptibility), 4 score, severity more than 60 %.In 2016–2018, International Stem Rust Trap Nursery with58 genotypes with identified Sr genes was evaluated to Omskstem rust population (Table 2). Varieties and lines of nurserytrapwere sown manually in 100 cm-long rows with stem rustresistant (Element 22) and susceptible checks (Chernyava 13)alternating every entries.1F2 (96 × 113) × 145 × 113]; No. 13, 14а [F3 (96 × 113) × 145];No. 16, 17, 17а [BC1F4 (96 × 113) × 113]; No. 19 [BC2F3(96 × 113) × 113]; No. 20, 22а [F3 (96 × 113) × 145]. The lineswere studied in 2016–2018 in un-replicated trial with the plotsize of 2 m2.In 2015, 9 spring wheat lines originating from wide crosses“Arsenal” collection were kindly provided by I.F. Lapochkinafor evaluation in Omsk. These lines carry a pyramid ofstem rust resistance genes  – No. 1[BCNine spring wheat varieties and breeding lines from advancedyield trial at Omsk SAU developed through utilizationof synthetic wheat with the Ae. tauschii genome (Lutescens24-12 (Kasibovskaya), Lutescens 27-12, Lutescens87-12, Lutescens 70-13, Lutescens 87-13, Lutescens 88-13(Silantiy), Lutescens 124-13, Lutescens 53-15, Lutescens 128-15) were evaluated for stem rust resistance and other traits in2016–2018. The plot size was 25 m2 with four replications.The checks were Pamyati Azieva (early maturing), Duet (mediummaturing), and Element 22 (late maturing).http://maswheat.ucdavis.edu/protocols/StemRust/index.htm). The resistance genes of spring bread wheat lines and varietiesfrom nursery-trap and from collection “Arsenal” were identifiedearlier .Sr genes of Omsk SAU varieties were identified usingmolecular markers: Xsts638 – Sr15, Xcfa2123 – Sr22,Xgwm210 – Sr23, Xscs73 – Sr24, Xwmc221 – Sr25,BE518379 – Sr26, Xscm09 – Sr31, SCS421 – Sr34,Xcfa2170 – Sr35, Xstm773-2 – Sr36, Ventriup-LN2 – Sr38,Lr34plus – Sr57, according to established protocol . In all studiedWestern-Siberian populations of P. graminis Ug99 and Sicilianraces were not identified. Genes of stem rust pathotypesof Omsk population were avirulent only to Sr31 gene, whileAltai pathotypes were avirulent to Sr31, Sr24, and Sr30.The race composition analysis of stem rust populationsidentified a significant number of pathotypes: in the Omskpopulation – 13 and in Altai population – 10 (see Table 1).Unlike many regions of the world where stem rust is a harmfuldisease for decades, for example in Krasnodar region of Russia, for Western Siberia this is surprisingresult considering a short period of time since its appearance.Most of the identified pathotypes of stem rust population inOmsk and Altai regions were not identical in virulence to thepathotypes, which were found in recent years in Asia andAfrica (Low frequency of virulence (10–25 %) of Omsk populationpathotypes was established for Sr11, Sr24, Sr30 genes, forAltai population – for Sr7b, Sr9b, Sr11, SrTmp genes, whichwere ineffective in Omsk region. The results of laboratoryevaluation of virulence of P. graminis pathotypes collectedin Omsk region were confirmed by field of trap nursery withidentified Sr genes (see Table 2).Genotypes with Sr31: Sr31(Benno)/6*LMPG-6 DK42,Seri 82, PBW343=Attila with Sr31, Cham 10=Kauz//Kauz/star, Bacanora=Kauz’s’ showed high level of resistance toOmsk stem rust population in all years of study (2016–2018).Line 28 LcSr24Ag + BTSr24Ag with Sr24 gene was characterizedby moderate resistance. For some Sr genes, resistanttype of reaction under epiphytotic conditions was observedon the stage of adult plants, and susceptible type – on theseedling stage in the laboratory conditions.For example, variety Trident (entries 46 and 47) withSr38 gene had high resistance (R–5MR) in the field; varietyEinkorn (entry 25) with Sr21 gene, and line W2691SrTt-1CI 17385 (entry 44) with Sr36 gene had moderate resistance(10M) in the field conditions. In the laboratory conditionsthe seedlings plants with above mentioned genes were classifiedas susceptible. Genotypes of ISRTN nursery with agene pyramid had high resistance to stem rust in all years ofresearch: entry 50 Amigo (Sr24 + 1RS-Am), entry 51 Siouxland(Sr24 + Sr31), entry 52 Roughrider (Sr6 + Sr36), entry 53Sisson (Sr6 + Sr31 + Sr36), entry 55 Fleming (Sr6 + Sr24 +Sr36 + 1RS-Am). The results of stem rust resistance evaluationof “Arsenal” collection and Omsk SAU germplasm arepresented in Table 3.Lines from “Arsenal” collection are of great interest assources of resistance to pathogen since they possess thegene pyramid: Sr2 (T. turgidum), Sr36, Sr40 (T. timopheevii),Sr44 (Th. intermediate). The pedigree of selected linescontains spring wheat line 13/00/i-4 with 7 resistance genes: Sr2, Sr36, Sr39, Sr40, Sr44, Sr47, Sr15, and winter lineGT 96/90 with genes Sr15, Sr24, Sr31, Sr36, Sr40, Sr47.In Omsk SAU varieties 3 resistance genes were identified:Sr23, Sr31, Sr36. Variety Element 22, which has winter wheatAurora in its pedigree also possesses wheat-rye translocation1BL.1RS with Sr31 gene . Thecombination of effective resistance genes Sr31 and Sr35 inthis variety results a high level of resistance to stem rust. Element22 is one of the few varieties with combined resistanceto stem and leaf rust. It was included into State register ofbreeding achievements in Western Siberian region. This varietyis the check of the late maturity group at the State VarietyTrials in Omsk region.Stem rust resistant breeding lines Lutescens 27-12, Lutescens70-13, Lutescens 87-13, Lutescens 88-13 were selectedfrom a cross Lutescens 30-94*2/3/T. dicoccon PI 94625/Ae. squarrosa (372)//3*Pastor involving Kazakhstan springwheat line Lutescens 30-94 and CIMMYT line developedby hybridization of synthetic wheat with variety Pastor. Theline Lutescens 87-12 originated from a cross Kazakhstanskaya25/2*Attila/3/T. dicoccon PI 94625/Ae. squarrosa alsoinvolving synthetic wheat. Omsk SAU germplasm possesseddifferent combinations of genes Sr23, Sr31, and Sr36.In modern conditions, stem rust is the most dangerous diseasefor grain production in Western Siberia. In the epiphytoticyears the grain losses of wheat in the region were about 2 milliontons. Unfortunately, stem rust resistant varieties includedinto the State register occupy about 10–15 % of the totalwheat sowing area in the region. In 2015–2016, evaluationof spring wheat varieties at Moskalenskiy State Variety Trialof Omsk region (southern forest-steppe zone) demonstratedthat out of 57 varieties tested only Element 22 (Sr31 + Sr35),Omskaya 37, Sigma, Uralosibirskaya (Sr31), and Sigma 2(Sr31 + Sr25) were resistant to stem rust (5–15MR). The othervarieties were affected by pathogen in medium and high degreerequiring the use of chemical protection . Previously, Shamanin et al. (2016b) identified thestem rust resistance genes in the germplasm developed bybreeding institutions of Western Siberia. High frequency ofgenes Sr25, Sr31, and their combination was observed. Highvariability of the race composition of the pathogen population,as shown in our studies, and the uniformity of resistance genesto stem rust in cultivated varieties, threaten grain productionstability in Western Siberia.The breeding strategy should focus on limiting diseasedevelopment in the region. The study of the populations ofP. graminis, formed on wheat in the different regions, is veryessential to guide the breeding efforts. There were no clonesavirulent to Sr24 gene in Omsk population of P. graminiswhile in Altai region there were no clones virulent to Sr24,which remains its effectiveness in Novosibirsk region . The results of the population compositioncomparison suggest that Omsk and Altai subpopulations haverelatively independent sources of genetic diversity and the contact zone. Western Siberian population of P. graminis hasquite complex structure. Two subpopulations are assumed toexist: Omsk and Altai – with independent sources of geneticdiversity, and zone of genotypic exchange on wheat crop inNovosibirsk region .Omsk stem rust population analysis showed that the spectrumof effective resistance genes has narrowed due to lossesof some genes to the local population of P. graminis.Highly resistant varieties and lines of ISRTN nurserywere identified: Sr31 (Benno)/6*LMPG-6 DK42, Seri 82,Cham 10, Bacanora (Sr31), RL 6087 Dyck (Sr40), Amigo, Siouxland , Roughrider , Sisson , Fleming , Pavon 76 (Sr2 complex). Selected varieties andlines are recommended for using as sources of resistance inbreeding programs to create resistant wheat varieties to stemrust. Effective resistance genes Sr31, Sr40, Sr2 complex,and their combinations with ineffective genes are recommendedfor use in breeding, taking into account the constantrotation, combination of genes of nonspecific resistance, aswell as the possibility of infection threat from neighboringterritory.The resistance gene Sr2, widely used in breeding for resistanceto virulent stem rust races, is common in commercial varietiesin a number of countries around the world, particularlyin the United States, Australia, India, and Mexico. This geneis practically absent in the commercial varieties of RussianFederation, however, for effective protection against stem rust,its pyramiding with other resistance genes is recommended.For the development of varieties with long-term resistance,the strategy of combining genes responsible for differenttypes of resistance in one genotype is used. Pyramiding ofspecific resistance genes  withAPR gene Sr2, which causes the slow development of thedisease (slow rusting), will provide longer protection ofwheat crops from stem rust in Western Siberia in the presentphytosanitary situation.1F2 (96 × 113) × 145 × 113 ; No. 14а F3(96 × 113) × 145 ; No. 19 BC2F3 (96 × 113) × 113; No. 20 F3 (96 × 113) × 145  represent a promising starting material for breedingand creation of varieties with long-term resistance.In this regard, the lines from “Arsenal” collection – No. 1BCIt is justified to include resistance sources to stem rust withminimum number of negative traits that reduce their breedingvalue. In this regard, stem rust resistant germplasm fromOmsk SAU with identified effective genes Element 22 , Lutescence 27-12, Lutescence 87-12 ,Lutescence 70-13, Lutescence 87-13 ,Lutescence 88-13 (Sr23) are valuable starting material forbreeding in the region.1F2 (96 × 113) × 145 × 113 , No. 14а F3(96 × 113) × 145 , No. 19 BC2F3 (96 × 113) × 113, No. 20 F3 (96 × 113) × 145 , varieties of Omsk Agrarian University – Element 22, Lutescens 27-12, Lutescens 87-12 ,Lutescens 70-13, Lutescens 87-13  arerecommendedfor inclusion into breeding process of thecreation of resistant to stem rust varieties in the region. Furthermonitoring of the virulence of stem rust pathogen andcoordination strategy of breeding programs in Western Siberia,and neighboring regions of the Kazakhstan Republic isrecommended. Incorporation of effective resistance genes, inparticular Sr2 and Sr40, will improve the phytosanitary situationand expand the segment of resistant varieties in the region.Thus, the genetic similarity of spring wheat varieties on stemrust resistance genes cultivated over large areas in WesternSiberia, and the predominance of varieties with race specific resistance genes contribute to spreading and high variabilityof the pathogen. The lines from collection “Arsenal” – No. 1BCThe authors declare no conflict of interest.Ablova I.B., Bespalova L.A., Kolesnikov F.A., Nabokov G.D.,Kovtunenko V.Ya., Filobok V.A., Davoyan R.O., KhudokormovaZh.N., Mokhova L.M., Levchenko Yu.G., Tarkhov A.S. Principlesand methods of wheat breeding on tolerance to diseasesin KRIA named after P.P. Lukiyanenko. Zernovoe KhozjaistvoRossii = Grain Economy of Russia. 2016;5:32-36. (in Russian)Baranova O.A., Lapochkina I.F., Anisimova A.V., Gajnullin N.R.,Iordanskaya I.V., Makarova I.Yu. Identification of Sr genes innew common wheat sources of resistance to stem rust race Ug99using molecular markers. Russ. J. Genet.: Appl. Res. 2016;6(3):344-350.Gomez-Becerra H., Morgounov A., Abugalieva A. Evaluation ofyield grain stability, reliability and cultivar recommendation inspring wheat (Triticum aestivum) from Kazakhstan and Siberia.Central Eur. J. Agriculture. 2006;6:649-660.Jin Y., Szabo L.J., Pretorius Z.A., Singh R.P., Ward R., Fetch T.,Jr. Detection of virulence to resistance gene Sr24 within raceTTKS of Puccinia graminis f. sp. tritici. Plant Dis. 2008;92:923-926.Kerber E.R., Dyck P.L. Resistance to stem and leaf rust in Aegilopssquarrosa and transfer of a gene for stem rust resistanceto hexaploid wheat. In: Ramanujam S. (Ed.). Proceeding of the5th International Wheat Genetics Symposium. New Delhi: Ind.Soc. of Genetics and Plant Breeding, Ind. Agric. Res. Institute,1979;358-364.Koyshybaev M., Shamanin V.P., Morgunov A.I. Screening of Wheatfor Resistance to Major Diseases. Ankara: FAO-SEK, 2014. (inRussian)Lapochkina I.F., Baranova O.A., Shamanin V.P., Volkova G.V.,Gainullin N.R., Anisimova A.V., Galinger D.N., Lazareva E.N.,Gladkova E.V., Vaganova O.F. The development of the initialmaterial of spring common wheat for breeding for resistance tostem rust (Puccinia graminis Pers. f. sp. tritici), including theUg99 race, in Russia. Russ. J. Genet.: Appl. Res. 2017;7(3):308-317. DOI 10.1134/S207905971703008X.Leonova I.N., Orlovskaya O.A., Röder M.S., Nesterov M.A., BudashkinaE.B. Molecular diversity of common wheat introgressionlines (T. aestivum/T. timopheevii). Russ. J. Genet.: Appl.Res. 2015; 5(3):191-197. DOI 10.1134/S2079059715030090.Mains E.B., Jackson H.S. Physiologic specialization in the leaf rustof wheat, Puccinia triticina Erikss. Phytopathology. 1926;16(2):89-120.McIntosh R.A., Dubcovsky J., Rogers W.J., Morris C., Xia X.C.Catalogue of gene symbols for wheat: 2017 Supplement. Availableat: http://shigen.nig.ac.jp/wheat/komugi/genes/macgene/supplement 2017.pdfMcIntosh R.A., Yamayaki Y., Dubcovsky J., Rogers W.J., MorrisC., Appels R., Xia X. Catalogue of Gene Symbols for Wheat:2013–2014 Supplement. Available at: http://www.shigen.nig.ac.jp/wheat/komugi/genes/macgene/supplement2013.pdfOlson E.L., Rous M.N., Pumphrey M.O., Bowden R.L., Gill B.S.,Poland J.A. Introgression of stem rust resistance genesSrTA10187 and SrTA10171 from Aegilops tauschii to wheat.Theor. Appl. Genet. 2013;126:2477-2484. DOI 10.1007/s00122-013-2148-z.Peterson R.F., Campbell A.B., Hannah A.E. A diagrammatic scalefor estimating rust intensity on leaves and stems of cereals. Can.J. Res. (Sect. C). 1948;26:496-500.Roelfs A.P., Martens J.W. An international system of nomenclaturefor Puccinia graminis f. sp. tritici. Phytopathology. 1988;78:526-533.Sanin S.S. Problems of phytosanitary of grain production. In: Protectionof Cereal Crops Against Diseases, Pests, and Weeds:Progress and Problems: Proc. of the Int. sci. and pract. conf.Bolshye Vyazemy, 5–9 Dec. 2016. Moscow, 2016;4-15. (in Russian)Schneider A., Molnár I., Molnár-Láng M. Utilisation of Aegilops(goatgrass) species to widen the genetic diversity of cultivatedwheat. Euphytica. 2008;163:1-19. DOI 10.1007/s10681-007-9624-y.Shamanin V.P., Morgunov A.I., Petukhovskiy S.L., Likhenko I.E.,Levshunov M.A., Salina E.A., Pototskaya I.V., Trushchenko A.Ju. Breeding of spring bread wheat for resistance to stem rustin West Siberia. Omsk: FGOU VPO OmGAU Publ., 2015. (inRussian)Shamanin V.P., Pototskaya I.V., Klevakina M.V. Assessment of theSiberian collection of spring wheat on resistance to stem rustin the southern forest-steppe of Western Siberia. Vestnik KazanskogoGAU = The Herald of Kazan State Agrarian University.2016a;2(40):55-59. (in Russian)Shamanin V., Salina Е., Wanyera R., Zelenskiy Yu., Morgounov A.Genetic diversity of spring wheat from Kazakhstan and Russiafor resistance to stem rust Ug99. Euphytica. 2016b;12:287-296.DOI 10.1007/s10681-016-1769-0.Singh R.P., Hodson D.P., Huerta-Espino J., Jin Y., Njau P., WanyeraR., Herrera-Foessel S.A., Ward R.W. Will stem rust destroythe world’s wheat crop? Adv. Agron. 2008;98:271-309. DOI10.1016/S0065-2113(08)00205-8.Singh R.P., Hodson D.P., Jin Y., Lagudah E.S., Ayliffe M.A., BhavaniS., Rouse M.N., Pretorius Z.A., Szabo L.J., Huerta-EspinoJ., Basnet B.R., Lan C., Hovmøller M.S. Emergence andspread of new races of wheat stem rust fungus: Continued threatto food security and prospects of genetic control. Phytopathology.2015;105:872-884.Singh R.P., Huerta-Espino J., Bhavani S., Herrera-Foessel S.A.,Singh D., Singh P.K., Velu G., Mason R.E., Jin Y., Njau P., CrossaJ. Race non-specific resistance to rust diseases in CIMMYTspring wheats: breeding and advances. Euphytica. 2011;179:175-186. DOI 10.1007/s10681-010-0322-9.Skolotneva E.S., Bukatich E.Ju., Bojko N.I., Piskarev V.V., SalinaE.A. Screening of an international stem rust nursery trapfor Ug99 in the Priobie forest-steppe in 2017. In: Gene Pooland Plant Breeding: Proc. IV Int. sci. and pract. conf., 4–6 April2018. Novosibirsk, 2018;313-318. (in Russian)Skolotneva E.S., Kel’bin V.N., Morgunov A.I., Bojko N.I., ShamaninV.P., Salina E.A. Races composition of the Novosibirskpopulation of Puccinia graminis f. sp. tritici. Mikologiya iFitopatologiya= Mycology and Phytopathology. 2020;54(1):49-58. (in Russian)Yu G., Champouret N., Steuernagel B., Olivera P.D., Simmons J.,Williams C., Johnson R., Moscou M.J., Hernández-Pinzón I.,Green P., Sela H., Millet E., Jones J.D.G., Ward E.R., SteffensonB.J., Wulff B.B.H. Discovery and characterization of twonew stem rust resistance genes in Aegilops sharonensis. Theor.Appl. Genet. 2017;130:1207-1222.


The F2 populations derived from these crosses were genotyped using InDel marker specific to crtRB1. Severe marker segregation distortion was observed. Seventeen crtRB1 inbreds developed through marker-assisted pedigree breeding and seven inbreds generated using marker-assisted backcross breeding were characterized using 77 SSRs. Wide variation in gene diversity (0.08 to 0.79) and dissimilarity coefficient (0.28 to 0.84) was observed. The inbreds were grouped into three major clusters depicting the existing genetic diversity. The crtRB1-based inbreds possessed high β-carotene (BC: 8.72μg/g), β-cryptoxanthin (BCX: 4.58μg/g) and proA (11.01μg/g), while it was 2.35μg/g, 1.24μg/g and 2.97μg/g in checks, respectively. Based on their genetic relationships, 15 newly developed crtRB1-based inbreds were crossed with five testers (having crtRB1 gene) using line × tester mating design. 75 experimental hybrids with crtRB1 gene were evaluated over three locations. These experimental hybrids possessed higher BC (8.02μg/g), BCX (4.69μg/g), proA (10.37μg/g) compared to traditional hybrids used as check . Environment and genotypes × environment interaction had minor effects on proA content. Both additive and dominance gene action were significant for proA. The mean proportion of proA to total carotenoids (TC) was 44% among crtRB1-based hybrids, while 11% in traditional hybrids. BC was found to be positively correlated with BCX (r = 0.68) and proA (r = 0.98). However, no correlation was observed between proA and grain yield. Several hybrids with >10.0 t/ha grain yield with proA content >10.0 μg/g were identified. This is the first comprehensive study on development of diverse proA rich maize hybrids through marker-assisted pedigree breeding approach. The findings provides sustainable and cost-effective solution to alleviate vitamin-A deficiency.Malnutrition has emerged as one of the major health problems worldwide. Traditional yellow maize has low provitamin-A (proA) content and its genetic base in proA biofortification breeding program of subtropics is extremely narrow. To diversify the proA rich germplasm, 10 elite low proA inbreds were crossed with a proA rich donor (HP702-22) having mutant  DeficieVarious avenues namely, food-fortification, medical-supplementation and food-diversification are implemented to alleviate VAD ,7. HowevZea mays L.) is an important cereal crop grown in almost all parts of the world and cultivated across diverse climatic spectrum [CrtRB1 gene that codes for β-carotene hydroxylase is associated with higher accumulation of proA especially BC and BCX in maize. Rare natural variation in crtRB1 gene limits the hydroxylation of BC and BCX [crtRB1, while the TE is absent in mutant version [Maize  characterize the newly developed crtRB1-based inbreds using microsatellite markers, (iii) study combining ability of the crtRB1-based inbreds for different carotenoid fractions, and (iv) identify promising hybrids with high proA and grain yield.Diverse proA rich inbreds have been developed in the tropics –19. Howeermplasm ,20. Therviz., UMI-1200, UMI-1230, BML-6Q, BML-7Q, LM-11Q, LM-12Q, LM-13Q, LM-14Q, PDM-4341 and PDM-4251, possessing good general combining ability for greater yield but low in proA were selected as recipient parents. To incorporate favourable allele of crtRB1 into these inbreds, a CIMMYT-HarvestPlus bred inbred i.e., HP704-22 was used as donor. All these recipient inbreds represent a great extent of adaptation range as parental lines of released/promising hybrids while the donor HP704-22 has poor adaptation in Indian conditions (1s were raised and selfed during winter season (December-April) of 2015–16 at the Winter Nursery Centre (WNC), ICAR-Indian Institute of Maize Research (ICAR-IIMR), Hyderabad (17°21´50.39ʺN and 78°29´42.31ʺE). Ten F2 populations consisting of 99–111 plants of each cross were then grown at ICAR-IARI, New Delhi during rainy season (2016)  .1s were tested for hybridity using crtRB1-based InDel marker present in 3’UTR. The true F1s were selfed to derive F2 populations. Genomic DNA was extracted from three weeks old seedlings of F2 progenies using standard procedure of CTAB with minor modification [3’TE-InDel-based marker for crtRB1 [viz., Forward (F): 5’ACACCACATGGACAAGTTCG3’, Reverse1 (R1):5’ACACTCTGGCCCATGAACAC3’ and Reverse2 (R2): 5’ACAGCAATACAGGGGACCAG3’. The primers were custom synthesized from Macrogen Inc., Seoul, South Korea. Polymerase chain reaction (PCR) protocol for crtRB1 [In-vitro amplification using ready-to-use master mix OnePCRTM  including Taq buffer, MgCl2, dNTPs and Taq Polymerase was used to perform PCR reaction in 96 well microtiter plate (M/s Genaxy) using GenePro thermal cycler (M/s Hangzhou Bioer Technology Co. Ltd.). The amplified product was resolved on 1.5% Seakem LE agarose gel .The Ffication . The quar crtRB1 , viz., Fr crtRB1  standard2 segregants with homozygous crtRB1 were selfed and derived F3 progenies were raised at ICAR-IARI, New Delhi during rainy season (2017). The F4s were raised during winter season (2017–18) at WNC, ICAR-IIMR, Hyderabad, and F5 progenies during rainy season (2018) at ICAR-IARI, New Delhi. To avoid chances of out-crossing, the presence of favourable allele of crtRB1 in each generation was validated using crtRB1-based InDel marker. Desirable segregants were advanced to further generation based on plant-, ear- and grain- characteristics.The FcrtRB1-based inbreds (MGU-PVMAS-1 to MGU-PVMAS-15) from 10 F2 populations were selected for molecular characterization  earlier developed by marker-assisted backcross breeding (MABB) at ICAR-IARI were also included. PMI-PV-1 and PMI-PV-2 are parental inbreds of India’s first proA rich maize hybrid (Pusa Vivek QPM9 Improved). PMI-PV-5, PMI-PV-6, PMI-PV-7, PMI-PV-8 and PMI-PV-9 are the parents of MABB-derived proA rich hybrids . Further, HP465-41, a CIMMYT-derived proA rich inbred derived through marker-assisted pedigree breeding was also included for characterization. Two low proA elite inbreds  were also included as control for quality analysis. These 26 inbreds were planted in randomized complete block design (RCBD) at ICAR-IARI, New Delhi in rainy season of 2018. Each inbred was planted with two replications, in rows of 3 m with a plant-to-plant distance of 20 cm. The rows were spaced 75 cm apart. Recommended cultural practices were followed to raise a good experimental crop. To avoid contamination by foreign pollens, 2–3 plants in each row were selfed for estimation of carotenoids.Fifteen rization . The doncrtRB1 were used for molecular analysis using simple sequence repeats (SSRs) markers. DNA was isolated from seeds using standard sodium dodecyl sulphate (SDS) extraction protocol [http://www.maizegdb.org). PCR was carried out as per Choudhary et al. (2016) [Selfed seeds of 24 inbreds with favourable allele of protocol . A totalviz., gene diversity, major allele frequency, total number of alleles detected, heterozygosity and polymorphism information content (PIC) were estimated using PowerMarker v3.0 [For each allele, presence of a band in a genotype was indicated by 1 and absence of the band as 0. Five parameters, ker v3.0 . An alleker v3.0 . Principker v3.0 .crtRB1 and two inbreds with unfavourable allele of crtRB1) were extracted from maize endosperm through protocol of Kurilich and Juvik (1999) [30 column  and detected with a diode array detector-3000 (RS). The mobile phase comprised of methanol: tert-butyl methyl ether  with flow rate at 1 ml/min and peaks were detected at 450 nm. For each carotenoid component, viz., BC, BCX, LUT and ZEA, six dilutions of standards were used to construct the regression curve. To estimate proA concentration, amount of BC was added to one-half of BCX amount, while sum of LUT and ZEA gave the non-proA fractions [Carotenoids from the selfed seeds of 26 inbreds   with modractions . Total cractions .crtRB1-based inbreds developed under the current study were crossed with five crtRB1-based tester inbreds  as per line × tester mating design [crtRB1-based promising inbred. These five testers belonged to two different groups viz., -A (PMI-PV-5 and HP465-41), -B . The 75 hybrid combinations and five commercial check hybrids  were evaluated using RCBD at three locations, viz. ICAR-IARI, New Delhi; CCS-HAU Regional Station, Uchani ; and ICAR-IARI Regional Research Centre, Dharwad  in rainy season of 2018. Each entry was evaluated in two replications, and was grown in a single row of 3 m length, with a row-to-row distance of 75 cm and plant-to-plant distance of 20 cm. Combining ability of the inbreds for carotenoids was calculated as per Singh and Choudhary (1985) [A set of 15 g design  at WNC, y (1985) .In each of the 75 experimental hybrid combinations generated from crosses, 2–3 plants were selfed to avoid contamination by the foreign pollen. The selfed seeds were used for estimation of carotenoids using UHPLC as per the procedure depicted for the inbreds.Grain yield (YLD) per plot was converted to t/ha as per the standard procedure. Magnitude of heterosis in hybrids over five commercial checks was estimated following Singh and Choudhary (1985) .The statistical analyses on ANOVA, correlation coefficients and combining ability were computed using Windostat 8.0.1s had both 296 and 543 bp amplicons. The 10 F2 populations were genotyped using crtRB1-specific 3’TE-InDel-based marker  were 407 and 435, respectively. Thus, crtRB1 showed severe marker segregation distortion both cumulatively as well as in individual populations. Out of 201 favourable homozygotes, 75 segregants were selected based on ear- and grain- characteristics, and advanced to generate F3 progenies. Finally, 15 locally adapted F4 progenies  to 270 bp (bnlg1537). The average major allele frequency was 0.62 with a range from 0.29 (phi126) to 0.96 (umc2167 and umc1068). A set of 24 SSRs showed major allele frequency of ≤0.5. The average gene diversity was 0.49, ranging from 0.08 (umc2167 and umc1068) to 0.79 (bnlg1740) (umc2167 and umc1068) to 0.75 (bnlg1740) with an average of 0.43. Of the 77 SSRs, 29 loci had PIC ≥0.5. The current study also detected 14 unique alleles and 23 rare alleles. The heterozygosity existing among the inbreds varied from 0.00 to 0.25 with a mean of 0.04.Some loci such as umc1446 (0.25) and bnlg1346 (0.25) showed high heterozygosity  to 0.84 (MGU-PVMAS-13 and MGU-PVMAS-8) with mean of 0.67  . The PICzygosity . Genetic of 0.67 . Cluster of 0.67 . PCoA di of 0.67 .crtRB1-based inbreds and two check inbreds . Among 15 inbreds developed under the study, MGU-PVMAS-11 (14.63 μg/g), MGU-PVMAS-5 (13.09 μg/g), MGU-PVMAS-4 (12.53 μg/g), MGU-PVMAS-12 (12.52 μg/g) and MGU-PVMAS-2 (12.28 μg/g) were the most promising ones (crtRB1-based inbreds had significantly low mean LUT (12.16 μg/g), ZEA (5.86 μg/g) and non-proA (18.02 μg/g) compared to check inbreds . However,TC was nearly comparable among the crtRB1-based (31.31 μg/g) and check inbreds (36.34 μg/g).ANOVA revealed significant variation for BC, BCX, proA, LUT, ZEA, non-proA and total carotenoid (TC) among 24  inbreds . The meaectively , Fig 4. ing ones . Six inbcrtRB1-based inbreds, BC and BCX contributed 28% and 15% of TC, while LUT and ZEA contributed 39% and 18%, respectively. The contribution of BC and BCX to TC was only 6% and 3% in check inbreds, while the same for LUT and ZEA was 58% and 33%, respectively.Among crtRB1-based hybrids had significant variation for BC, BCX, proA, LUT, ZEA, non-proA and TC (× environment (G × E) interaction was also low with 8% (non-proA) to 22% (BCX).Pooled ANOVA revealed that A and TC . The proThe mean BC and BCX among experimental hybrids were 8.02 μg/g and 4.69 μg/g, as compared to 2.36 μg/g (BC) and 1.53 μg/g (BCX) among low-proA checks . High-prThe experimental crosses possessed low LUT  and ZEA  relative to low-proA checks . High-proA checks also possessed low LUT (5.22 μg/g) and ZEA (10.56 μg/g). The non-proA fraction among experimental hybrids varied from 10.78 to 23.50 μg/g, with an average of 16.47 μg/g. The non-proA fraction in low-proA checks was higher (31.88 μg/g) than that of high-proA checks (15.78 μg/g). TC in experimental hybrids varied from 21.92 to 37.86 μg/g, with a mean of 29.18 μg/g. Low-proA checks showed a mean of 35.77 μg/g, while high-proA commercial checks showed a mean of 29.80 μg/g .In case of experimental hybrids, the contribution of BC and BCX towards TC was 27% and 17%, while LUT and ZEA contributed 38% and 18%, respectively . In caseCorrelation analysis revealed that BC was positively correlated with BCX (r = 0.68**). Whereas, BC and BCX were negatively correlated with LUT  and ZEA , respectively. LUT and ZEA, however, were positively correlated (r = 0.70**). ProA and non-proA carotenoids were also negatively correlated (r = -0.57**). BC, BCX and proA were not correlated with TC, while LUT (r = 0.72**), ZEA (r = 0.72**) and non-proA (r = 0.78**) showed strong positive association with TC. The present study revealed that kernel carotenoid components and grain yield exhibited non-significant relationships (r = -0.18 to 0.15) except for ZEA (r = 0.29**).Pooled ANOVA revealed that environment was highly significant for all the carotenoids except for ZEA . Line efThe proportion of additive and dominance variance, and the contribution of lines, testers and line × testers for pooled dataset are presented in GCA effect for proA varied from -1.52 to 1.27. Nine lines and one tester showed significant positive GCA effects for proA . Among tat par with CoMH-08-292. Among high proA checks, ‘Pusa HQPM-5 Improved’ emerged as the best check with 8.6 t/ha. A total of 26 experimental hybrids were significantly better than ‘Pusa HQPM-5 Improved’ for grain yield. These hybrids showed heterosis of 8.04% to 31.63% over the best high proA check.Heterosis of the 75 experimental hybrids was estimated over commercial low-proA and high proA checks. CMH08-292 and DHM-121 were medium maturing hybrids with low-proA. ‘Pusa Vivek QPM-9 Improved’ is an early maturing proA rich hybrid and was released as country’s first proA rich maize hybrid during 2017. ‘Pusa HQPM-5 Improved’ and ‘Pusa HQPM-7 Improved’ are the medium maturing proA rich hybrids released in 2020. All the experimental hybrids matured in 92–100 days, thus had medium maturity. The mean grain yield among experimental hybrids was 8.9 t/ha, with the highest yield of 11.4 t/ha  Tables. crtRB1 significantly enhances proA in maize kernel [crtRB1 in the Indian maize germplasm is quite low (3.38%) [crtRB1 inbreds have been developed in India. Thus, targeted breeding approach for selection of crtRB1 is essential for broadening the genetic base of proA rich maize germplasm [Plant based-food is the major source of nutrition especially in developing world. Traditional yellow maize lacks the required level of proA . The mute kernel . Diversee kernel ,19. Howe (3.38%) . So far,ermplasm .2 populations indicated that crtRB1 did not segregate as per the expected 1:2:1. This observation drives strength from earlier results of Lu et al. (2002) [crtRB1 gene could be precisely selected due to the reliable gene-based marker. In case of linked marker, there is always chance of selection of false positive individuals due to crossing over between the gene and marker [crtRB1-based inbreds using marker-assisted pedigree breeding. Earlier, Muthusamy et al (2014) [crtRB1 into the elite inbreds using MABB approach. In case of MABB, the improved lines are genetically similar to the recurrent parents except for gene under introgression [crtRB1-based inbreds were developed from F2 populations, the genetic makeup remains novel leading to development of new and diverse crtRB1-based inbreds. The MAS-based selection of crtRB1 is quite cost-effective, as selection of genotypes for high proA using UHPLC involves US$30–35 per sample. On the contrary, PCR marker-based selection of crtRB1 employed here costs only US$0.5–1.0 per sample. Molecular breeding is now a preferred choice among the maize breeders to develop proA rich germplasm [Genotyping of F. 2002)  and Babu. (2002) . The seg. (2002) . Since, d marker . The prel (2014) , Liu et l (2014) , Zunjarel (2014)  and Goswl (2014)  have intgression . In the 002  and Knowledge of the genetic relationship among inbreds is essential for efficient exploitation in breeding programme. Molecular markers have been successfully employed to derive the genetic distance in maize . High dicrtRB1-based inbreds and hybrids recorded nearly 3–4 folds more proA (8–14 μg/g) over the traditional checks (2–3 μg/g). Muthusamy et al., (2014) [crtRB1-based maize genotypes, respectively. However, traditional yellow maize possesses low concentration of proA (<2.5 ppm) [crtRB1, while low proA genotypes harbour the wild-type allele [The , 2014)  and Zunj, (2014)  also rep14  and Z2.5 ppm) . The acce allele ,17.phytoene synthase 1 (psy1) or Yellow 1 (Y1) gene condenses two geranyl-geranyl pyrophosphate molecules into one molecule of phytoene [y1 allele is unable to catalyze the reactions and white grains are formed due to no synthesis of carotenoids. However, when the Y1 is functional, crtRB1 causes hydroxylation of BC and BCX into ZEA. CrtRB1 located on chromosome 10 codes for β-carotene hydroxylase. The mutant version of crtRB1 drastically slows down the conversion, leading to more accumulation of proA carotenoids [crtRB1 allele, a large variation in proA was observed. This could be due to variation in the activity of other key genes such as psy1, lycopene β-cyclase (lcyB), lycopene ε-cyclase (lcyE), phytoene desaturase (pds) and ζ-carotene desaturase (zds) catalyzing the carotenoid biosynthesis [lcyE present on chromosome 8 has been observed [lcyE along with crtRB1 is beneficial in enhancing proA in maize. Besides, several modifier loci/QTLs alone or in combination with other pathway genes could also influence the accumulation of proA in maize [In the carotenoid biosynthesis pathway, phytoene . The mutotenoids . Though ynthesis . Allelicobserved . Zunjareobserved  reportedin maize ,48.crtRB1-based and check- genotypes, thereby, suggesting greater flux of lycopene towards α-branch than the β-branch of pathway [crtRB1-based genotypes, the conversion of BC to ZEA is partially blocked, leading to higher proportion of BC next to LUT. ProA content among the crtRB1-based genotypes constituted 43–44% of TC as against 9–11% in the check genotypes. The non-proA component was less (56–57%) among crtRB1-based genotypes compared to 89–91% in checks [Among carotenoids, proportion of LUT was higher in both  pathway . LUT ser pathway . Howevern checks ,49. In sn checks .BC, BCX and proA showed strong positive correlations, as BCX is produced from BC, while both contribute to proA . Since, crtRB1 possesses much higher proA compared to heterozygote [crtRB1 in all hybrids. Since, all the hybrids were homozygous for favourable allele of crtRB1, other modifier loci could be the reason for dominance effects for further influencing the proA. Several lines including testers were identified as the best general combiners for proA. MGUH-57 (14.90 μg/g), MGUH-52 (14.60 μg/g), MGUH-27 (14.36 μg/g) and MGUH-32 (14.26 μg/g) possessed high proA, and had both the parents being high in GCA effects as well. Besides, MGUH-1, MGUH-14, MGUH-15, MGUH-18, MGUH-19, MGUH-28, MGUH-31, MGUH-48, MGUH-50 and MGUH-75 had one of the parents (used as line) having high GCA effects for proA. The inbreds with high GCA for proA thus serve as a promising inbreds in the future breeding programme [The current study revealed that environments had minor effect on BC, BCX and proA. Muthusamy et al. (2015b)  also reprozygote ,17,22. Crogramme .at par with CoMH08-292  and significantly better than DHM-121  for grain yield potential.Several promising experimental hybrids with >10.0 t/ha grain yield and >10.0 μg/g proA were identified. MGUH-15 , MGUH-50 , MGUH-72 , MGUH-54  and MGUH-55  were the most promising combinations. Most of these promising hybrids belongs to different clusters. These hybrids are much higher yielding than the ‘Pusa Vivek QPM-9 Improved’ , the first proA rich hybrid released in India. This is primarily due to extra-early maturity of ‘Pusa Vivek QPM-9 Improved’. Besides, two of the proA hybrids, ‘Pusa HQPM5 Improved’  and ‘Pusa HQPM7 Improved’  in the medium maturity groups have been recently released. Thus, these identified hybrids are higher yielding than the proA check hybrids as well. These selected hybrids also possessed higher proA than the normal best check (CoMH08-292 and DHM-121), but were crtRB1-donor was only 1.4t/ha, thereby suggesting its poor adaptability in subtropical conditions. The high grain yield of new crtRB1-based inbreds depicts better adaptability. The high yielding proA rich hybrids identified here would thus provide more productivity and profit to the farmers, and offer higher vitamin-A to the consumers [Further, parents of these proA rich hybrids also produce high yield from seed production perspective. The average grain yield among parental inbreds (used as lines) developed under this programme was 3.1t/ha with a range of 2.6–3.5t/ha. The same in onsumers . The procrtRB1-based inbreds have been developed in the study through marker-assisted pedigree breeding. These new inbreds possessed significantly higher proA than the checks. Molecular characterization of the inbreds depicted their diverse genetic nature. Genetic analysis revealed that both additive and non-additive variances were important. The crtRB1-based inbreds were successfully used in development of proA rich hybrids. Promising high yielding hybrids with very high concentration of proA have been identified. These proA rich hybrids are higher yielding than the exiting proA checks and at par with the normal checks in grain yield but with higher proA. The present study demonstrated the successful application of markers-assisted pedigree breeding in broadening the genetic base, and developing promising hybrids with higher grain yield as well as improved nutritional quality.Diverse S1 Tableopaque2 versions, TNAU: Tamil Nadu Agricultural University; ANGRAU: Acharya N. G. Ranga Agricultural University; PAU: Punjab Agricultural University.Q: Represents the (DOC)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table1. MGU-PVMAS-13, 2. PMI-PV-8, 3. PMI-PV-7, 4. MGU-PVMAS-6, 5. MGU-PVMAS-5, 6. MGU-PVMAS-11, 7. MGU-PVMAS-9, 8. MGU-PVMAS-4, 9. MGU-PVMAS-8, 10. MGU-PVMAS-15, 11. MGU-PVMAS-14, 12. MGU-PVMAS-7, 13. HP465-41, 14. PMI-PV-1, 15. MGU-PVMAS-3, 16. MGU-PVMAS-2, 17. MGU-PVMAS-12, 18. MGU-PVMAS-10, 19. HP704-22, 20. PMI-PV-6, 21. PMI-PV-9, 22. PMI-PV-5, 23. PMI-PV-2, 24. MGU-PVMAS-1, Inb. = Inbred.(DOCX)Click here for additional data file.S4 TableBC: β- Carotene, BCX: β-Cryptoxanthin, ProA: Provitamin A, LUT: Lutein, ZEA: Zeaxanthin, Non-ProA: Non-Provitamin A, TC: Total carotenoids, YLD: Yield.(DOCX)Click here for additional data file.S5 Table*Significant at p = 0.05; **Significant at p = 0.01, BC: β- Carotene, BCX: β-Cryptoxanthin, ProA: Provitamin-A, LUT: Lutein, ZEA: Zeaxanthin, Non-ProA: Non-Provitamin-A, TC: Total carotenoids, SE: Standard Error.(DOCX)Click here for additional data file.S6 Table*Significant at p = 0.05; **Significant at p = 0.01.(DOCX)Click here for additional data file.S1 File(DOCX)Click here for additional data file.


Coronavirus disease-2019 (COVID-19) is now an unprecedented worldwide pandemic. However, there are no specific antiviral drugs available for its treatment. A handful of studies have summarized convalescent plasma (CP) transfusion in severe or critical cases ,4,5,6, w9/L with 78% neutrophils, 9.2% lymphocytes, 2.3% basophils, 0.4% eosinophils, and 10.1% monocytes; hemoglobin of 51 g/L; platelet count of 79x109/L; high-sensitivity C-reactive protein of 57.43 mg/L; and interleukin-6 level of 59.25 pg/mL. The real-time polymerase chain reaction (RT-PCR) assay of the throat swab was positive for severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection. A chest CT obtained on February 21 revealed bilateral ground-glass opacities primarily distributed along the pleura  starting in November 2017, but a drug-related hematologic adverse event occurred quickly. As a result, dasatinib (150 mg/day) was given instead. She had not achieved complete hematological remission at the diagnosis of COVID-19 due to poor responses to these therapies. At admission , the most relevant clinical findings included white blood cell count of 4.93x10e pleura  and 1d. e pleura  that expe pleura . RT-PCR e pleura  accordin2) oscillating between 90% and 93%, after receiving conventional antiviral therapy including arbidol (200 mg three times daily), oseltamivir (75 mg twice daily), ribavirin (500 mg every 12 hours), and interferon-alpha-2b inhalation (5 million units twice daily). A follow-up chest CT scan showed increased consolidation and extended opacities . Three repetitive RT-PCR tests were negative from the 6th to 8th day after CP transfusion. Chest images showed absorption of opacities within 10 days (th day of admission.The patient developed worsening hypoxemia, with oxyhemoglobin saturation (SaOpacities  and 1e.  10 days  and 1f. A recent study showed a 10% case rate of COVID-19 among 128 patients with hematological cancers in Wuhan . The tre


Tepidicella baoligensis strain B18-50T, isolated from a high-temperature oil well in Baolige Oilfield, China. The estimated genome is 2.87 Mb, with 2,653 protein-coding sequences.We report the draft genome sequence of  Tepidicella baoligensis strain B18-50T, isolated from a high-temperature oil well in Baolige Oilfield, China. The estimated genome is 2.87 Mb, with 2,653 protein-coding sequences.We report the draft genome sequence of  Tepidicella species are slightly thermophilic Gram-negative rods that are distributed in geothermal areas. These organisms do not assimilate carbohydrates or polyols but use organic acids as carbon and energy sources and oxidize thiosulfate to sulfate in the presence of an assimilable carbon source  in China. The liquid sample was diluted with sterilized enrichment medium (EN), plated onto EN agar, and incubated at 50°C , and 1.0 g liter−1 tryptone (Oxoid). Genomic DNA was prepared from an overnight culture by using the phenol-chloroform extraction method (Strain B18-50 at 50°C . A pure 2 medium  supplemehttps://github.com/aquaskyline/SOAPdenovo2)  libraries were prepared from the sheared fragments using the NEXTflex rapid DNA-Seq kit  and sequenced in 150-bp paired-end mode using the Illumina HiSeq X Ten platform at Majorbio Bio-Pharm Technology, Inc. . The sequencing generated 3,449,944 pairs of raw reads totaling 1,041,883,088 bp, giving approximately 362× coverage. The reads were quality trimmed with Trimmomatic v0.36 (http://ccb.jhu.edu/software/glimmer/index.shtml) (http://trna.ucsc.edu/software/) (https://github.com/tseemann/barrnap) was used for rRNA prediction. The predicted CDSs were annotated from the nonredundant (NR), Swiss-Prot, Pfam, GO, COG, and KEGG databases using the sequence alignment tools BLAST+ v2.3.0   was usedftware/)  was used+ v2.3.0 , Diamond+ v2.3.0 , and HMM+ v2.3.0 . A totalJACCDB000000000; the raw reads are available under SRA accession number SRR12179937. This announcement represents the first version of the genome.This whole-genome shotgun sequencing project has been deposited in DDBJ/ENA/GenBank under the accession number


Orthobunyavirus, family Peribunyaviridae) is an arthropod-borne virus that causes congenital abnormalities in ruminants. Here, we report the complete genome sequences of two AKAV strains causing nonsuppurative encephalomyelitis in cattle by postnatal infection in Japan.Akabane virus (AKAV)  is an arthropod-borne virus that causes congenital abnormalities in ruminants. Here, we report the complete genome sequences of two AKAV strains causing nonsuppurative encephalomyelitis in cattle by postnatal infection in Japan.Akabane virus (AKAV) (genus  Akabane orthobunyavirus . Since it was first isolated from mosquitos in Gunma, Japan, in 1959, it has been widely detected in Australia, Asia, and Africa (Culicoides spp.) and causes spontaneous abortions, stillbirths, and congenital malformations in pregnant ruminants (Akabane virus (AKAV) is an enveloped, tripartite, negative-sense RNA virus of the species AKAV strains are clustered into four genogroups based on phylogenetic analysis of their medium (M) segments , 2. Genohttps://jorgensen.biology.utah.edu/wayned/ape), with the OBE-1 strain (the genogroup II strain that is the progenitor of the vaccine strain used in Japan) as the reference .Both virus strains were propagated in HmLu-1 cells. Their genomic RNA was extracted using the QIAamp virus RNA minikit . Reverse transcription-PCR was performed using SuperScript IV reverse transcriptase  and PrimeSTAR GXL DNA polymerase . Primer sequences are available on request. The 5ʹ- and 3ʹ-terminal sequences of all of the genomic RNA segments were determined using 5′ and 3′ rapid amplification of cDNA ends (RACE) . The seqThe full-length S, M, and L segments of KM-2/Br/06 are 856, 4,309, and 6,869 nucleotides (nt), respectively, whereas those of FI-1/Br/06 are 867, 4,308, and 6,867 nt, respectively. The GC contents of the segments were also determined . The nucThe sequences determined in this study will be useful in identifying the virulence factors implicated in bovine postnatal encephalomyelitis.LC552047.1 (KM-2/Br/06 S segment), LC552048.1 (KM-2/Br/06 M segment), LC552049.1 (KM-2/Br/06 L segment), LC552050.1 (FI-1/Br/08 S segment), LC552051.1 (FI-1/Br/08 M segment), and LC552052.1 (FI-1/Br/08 L segment).The sequences are available in GenBank under the accession numbers