Isolation and Identification of Pigment Producing Actinomycete Saccharomonospora azurea SJCJABS01

Given the rising demand for biological pigments, especially of microbial origin – the present study was conducted so as to report a potential source for the extraction of microbial pigment. The main objective was to isolate and identify a pigment–producing actinomycete because pigment production is prevelant in this group. A powdery, greenish–blue colony with a chalky azure aerial mass was isolated from one of the many rhizosphere soil samples. Upon preliminary investigation, viz. colony characterization and grams staining, the suspected colony was observed to have a filamentous margin with a slightly raised elevation and gram–positive filamentous hyphae.Biochemical analyses of the organism revealed positive results for carbohydrate fermentation and Triple Sugar Iron (TSI) test with no signs of gas production during the former but gas & H­2­S production during the latter. The identity of the isolate was established via 16S rDNA and phylogeny analysis, which strongly suggested it was Saccharomonospora azurea. Limited research pertaining to morphology, physiology, genomics and secondary metabolite production with no reports on the physicochemical properties of the pigment produced by S. azureaadequately suggests that it is relatively novel. Hence, further studies related to the same could be beneficial to the scientific community.

Unique SARS-CoV-2 variant found in public sequence data of Antarctic soil samples collected in 2018-2019

The COVID-19 pandemic has been going on for two years now and although many hypotheses have been put forward, its origin remain obscure. We investigated whether the huge public sequencing data archives’ samples collected earlier than the earliest known cases of the pandemic might contain traces of SARS-CoV-2. Here we report the bioinformatic analysis of a metagenome sample set collected from soil on King George Island, Antarctica between 2018-12-24 and 2019-01-13. It contains sequence fragments matching the SARS-CoV-2 reference genome with altogether more than half million nucleotides, covering the complete genome on average 17×. Preliminary phylogeny analysis places the sample close to the known earliest cases. The high sequence coverage rules out chance alignments from other species but possible laboratory contamination cannot be excluded. The sequence harbours a unique combination of mutations, unseen in other samples, so whatever its origin, it can add important piece of information to the puzzle of the ongoing pandemic.

Evidences of Colletotrichum fructicola Causing Anthracnose on Passiflora edulis Sims in China

Passion fruit (Passiflora edulis) is a tropical and subtropical plant that is widely cultivated in China due to its high nutritional value, unique flavor and medicinal properties. In August 2020, typical anthracnose symptoms with light brown and water-soaked lesions on Passiflora edulis Sims were observed, which result in severe economic losses. The incidence of this disease was approximately 30%. The pathogens from the infected fruit were isolated and purified by the method of tissue isolation. Morphological observations showed that the colony of isolate BXG-2 was gray to celadon and grew in concentric circles. The orange conidia appeared in the center after 14 days of incubation. The pathogenicity was verified by Koch’s postulates. The internal transcribed spacer (ITS), chitin synthase (CHS-1), actin (ACT), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were amplified by relevant PCR programs. The multi-gene (ITS, GAPDH, ACT, CHS-1) phylogeny analysis confirmed that isolate BXG-2 belongs to Colletotrichum fructicola. The inhibitory effect of six synthetic fungicides on the mycelial growth of the pathogen was investigated, among which difenoconazole 10% WG showed the best inhibitory effect against C. fructicola with an EC50 value of 0.5579 mg·L−1. This is the first report of anthracnose on Passiflora edulis Sims caused by Colletotrichum fructicola in China.

Phylogenetic and full-length genome mutation analysis of SARS-CoV-2 in Indonesia prior to COVID-19 vaccination program in 2021

Background Indonesia has started the big project of COVID-19 vaccination program since 13 January 2021 by employing the first shot of vaccine to the President of Indonesia as the outbreak and rapid transmission of COVID-19 have endangered not only Indonesian but the global health and economy. This study aimed to investigate the full-length genome mutation analysis of 166 Indonesian SARS-CoV-2 isolates as of 12 January 2021. Results All data of the isolates were extracted from the Global Initiative on Sharing All Influenza Data (GISAID) EpiCoV database. CoVsurver platform was employed to investigate the full-length genome mutation analysis of all isolates. This study also focused on the phylogeny analysis in unlocking the mutation of S protein in Indonesian SARS-CoV-2 isolates. WIV04 isolate that was originated from Wuhan, China was used as the virus reference according to the CoVsurver default. The result showed that a full-length genome mutation analysis of 166 Indonesian SARS-CoV-2 isolates was successfully generated. Every single mutation in S protein was described and then visualized by utilizing BioRender platform. Furthermore, it also found that D614G mutation appeared in 103 Indonesian SARS-CoV-2 isolates. Conclusions To sum up, this study helped to observe the spread of COVID-19 transmission. However, it also proposed that the epidemiological surveillance and genomics studies might be improved on COVID-19 pandemic in Indonesia.

Genome Wide Analysis of GH Gene Family Reveals Vvgh9 Positively Regulates Sugar Accumulation under Low Sugar Content in Grape

Sugar content directly affects grape (Vitis vinifera L.) berry quality and the resulting wine. Therefore, it is of great importance to study and explore novel genes that affect sugar accumulation in grapes. Glycosyl hydrolases (GHs) are key enzymes hydrolyzing polysaccharides into monosaccharides and play important roles in the regulation of carbohydrate metabolism. Nevertheless, the impact of GHs on the regulation of sugar accumulation in plants has rarely been investigated. In this study, we identified 11 putative GH genes in grapevines by phylogeny analysis. RNA-seq and quantitative real-time PCR results demonstrated that the expression level of VvGH9 was higher during the fruit set stage, which had lower sugar content than the véraison and ripe stages. Treatment of grape berries with exogenous sugar two weeks before véraison revealed that VvGH9 was rapidly induced by sucrose, fructose, and glucose. When ‘41B’ calli was treated with different concentrations of glucose, VvGH9 expression increased at first and then decreased with the increase of glucose concentration. Overexpression of VvGH9 in grape calli and tomatoes also confirmed that this gene could contribute to sugar accumulation. All the above results demonstrated that VvGH9 promotes sugar accumulation under low sugar content in plants.

Contributions to Molecular Phylogeny of Lichen-Forming Fungi 2. Review of Current Monophyletic Branches of the Family Physciaceae

Seven genera new to science, i.e.: Helmutiopsis, Huriopsis, Johnsheardia, Klauskalbia, Kudratovia, Kurokawia and Poeltonia of the Physciaceae are proposed for the ‘Rinodina’ atrocinerea, the ‘Rinodina’ xanthophaea, the ‘Rinodina’ cinnamomea, the ‘Heterodermia’ obscurata, the ‘Rinodina’ straussii, the ‘Anaptychia’ isidiata and the ‘Physconia’ grisea groups consequently that all form strongly supported monophyletic branches in a phylogeny analysis based on a combined matrix of nrITS and mtSSU sequences. Phylogenetic positions of species belonging to the genera Kashiwadia s. l., Leucodermia, Mischoblastia,Oxnerella, Phaeorrhiza s. l., Polyblastidium and Rinodinella s. l. are discussed. Oxnerella afghanica which for the first time recorded as parasitic lichen species from both epiphytic and saxicolous crustose lichens is designated as type species for the genus Oxnerella. Sequences of the recently described Physcia orientostellaris as well as Huriopsis xanthophaea and additional sequences of Kashiwadia aff. orientalis and Mischoblastia aff. oxydata are submitted to the GenBank. The positions of Polyblastidium casaterrinum from Costa Rica, ‘Rinodina’ efflorescens from Białowieża, Poland, and ‘Mischoblastia’ confragosula from Cambodia in the Physciaceae are confirmed in a phylogeny analysis based on the nrITS sequences. The presence of ‘extraneous mycobiont DNA’ in lichen associations is exemplified with earlier incorrect identifications of Heterodermia, Kashiwadia, Kurokawia,Oxnerella and Poeltonia specimens. Fifty-six new combinations are presented: Helmutiopsis alba (for Rinodina alba Metzler ex Arn.), Helmutiopsis aspersa (for Lecanora aspersa Borrer), Helmutiopsis atrocinerea (for Parmelia atrocinerea Fr.), Huriopsis chrysidiata (for Rinodina chrysidiata Sheard), Huriopsis chrysomelaena (for Rinodina chrysomelaena Tuck.), Huriopsis lepida (for Lecanora lepida Nyl.), Huriopsis luteonigra (for Rinodina luteonigra Zahlbr.), Huriopsis plana (for Rinodina plana H. Magn.), Huriopsis thiomela (for Lecanora thiomela Nyl.), Huriopsis xanthomelana (for Rinodina xanthomelana Müll. Arg.), Huriopsis xanthophaea (for Lecanora xanthophaea Nyl.), Johnsheardia cinnamomea (for Rinodina mniaroea var. cinnamomea Th. Fr.), Johnsheardia herteliana (for Rinodina herteliana Kaschik), Johnsheardia jamesii (for Rinodina jamesii H. Mayrhofer), Johnsheardia reagens (for Rinodina reagens Matzer et H. Mayrhofer), Johnsheardia zwackhiana (for Lecanora zwackhiana Kremp.), Kashiwadia austrostellaris (for Physcia austrostellaris Elix), Kashiwadia jackii (for Physcia jackii Moberg), Kashiwadia littoralis for Physcia littoralis Elix), Kashiwadia nubila (for Physcia nubila Moberg), and Kashiwadia tropica (for Physcia tropica Elix), Klauskalbia crocea (for Heterodermia crocea R. C. Harris), Klauskalbia flabellata (for Parmelia flabellata Fée), Klauskalbia obscurata (for Physcia speciosa (Wulfen) Nyl. *obscurata Nyl.), Klauskalbia paradoxa (for Heterodermia paradoxa Schumm et Schäfer-Verwimp), Kudratovia bohlinii (for Rinodina bohlinii H. Magn.), Kudratovia candidogrisea (for Rinodina candidogrisea Hafellner, Muggia et Obermayer), Kudratovia luridata (for Buellia luridata Körb.), Kudratovia metaboliza (for Rinodina metaboliza Vain.), Kudratovia pycnocarpa (for Rinodina pycnocarpa H. Magn.), Kudratovia roscida (for Lecanora roscida Sommerf.), Kudratovia straussii (for Rinodina straussii J. Steiner), Kudratovia terrestris (for Rinodina terrestris Tomin), Kurokawia bryorum (for Anaptychia bryorum Poelt), Kurokawia isidiata (for Anaptychia isidiata Tomin), Kurokawia mereschkowskii (for Physcia mereschkowskii Tomin), Kurokawia palmulata (for Psoroma palmulatum Michx.), Kurokawia runcinata (for Lichen runcinatus With.), Kurokawia stippea (for Parmelia aquila var. stippea Ach.), Lecania safavidiorum (for Oxnerella safavidiorum S. Y. Kondr., Zarei-Darki, Lőkös et Hur), Leucodermia erinacea (for Lichen erinaceus Ach.), Mischoblastia confragosula (for Lecanora confragosula Nyl.), Mischoblastia destituta (for Lecidea destituta Nyl.), Mischoblastia moziana (for Lecanora moziana Nyl.), Mischoblastia moziana subsp. parasitica (comb. et stat. nova for Rinodina moziana var. parasitica Kaschik et H. Mayrhofer), Mischoblastia ramboldii (for Rinodina ramboldii Kaschik), Mischoblastia vezdae (for Rinodina vezdae H. Mayrhofer), Oxnerella afghanica (for Rinodina afghanica M. Steiner et Poelt), Oxnerella castanomelodes (for Rinodina castanomelodes H. Mayrhofer et Poelt), Physciella nigricans (for Lecanora nigricans Flörke), Poeltonia elegantula (for Physconia elegantula Essl.), Poeltonia grisea (for Lichen griseus Lam.), Poeltonia isidiomuscigena (for Physconia isidiomuscigena Essl.), Poeltonia perisidiosa (for Physcia perisidiosa Erichsen), Poeltonia venusta (for Parmelia venusta Ach.), and Polyblastidium albicans (for Parmelia albicans Pers.) are proposed.

Complete Genome Analysis of a Novel Alcaligenes Faecalis Phage vB_Af_QDWS595

A novel lytic phage named vB_Af_QDWS595 against Alcaligenes faecalis was isolated and characterized in this study. The genome of phage vB_Af_QDWS595 was sequenced and analyzed, and the result revealed that the phage contained a 88,795 bp of circular double-stranded DNA with 41.12% of GC content. There were 74 putative open reading frames (ORFs) and 11 tRNAs predicted in genome of phage vB_Af_QDWS595. Phenotype and phylogeny analysis indicated that this phage might be a new member within the family Schitoviridae. Phage vB_Af_QDWS595 is the first sequenced phage against Alcaligenes faecalis to the best of our knowledge.

Functional analysis of multiple nifB genes of Paenibacillus strains in synthesis of Mo-, Fe- and V-nitrogenases

Background Biological nitrogen fixation is catalyzed by Mo-, V- and Fe-nitrogenases that are encoded by nif, vnf and anf genes, respectively. NifB is the key protein in synthesis of the cofactors of all nitrogenases. Most diazotrophic Paenibacillus strains have only one nifB gene located in a compact nif gene cluster (nifBHDKENX(orf1)hesAnifV). But some Paenibacillus strains have multiple nifB genes and their functions are not known. Results A total of 138 nifB genes are found in the 116 diazotrophic Paenibacillus strains. Phylogeny analysis shows that these nifB genes fall into 4 classes: nifBI class including the genes (named as nifB1 genes) that are the first gene within the compact nif gene cluster, nifBII class including the genes (named as nifB2 genes) that are adjacent to anf or vnf genes, nifBIII class whose members are designated as nifB3 genes and nifBIV class whose members are named as nifB4 genes are scattered on genomes. Functional analysis by complementation of the ∆nifB mutant of P. polymyxa which has only one nifB gene has shown that both nifB1 and nifB2 are active in synthesis of Mo-nitrogenase, while nifB3 and nifB4 genes are not. Deletion analysis also has revealed that nifB1 of Paenibacillus sabinae T27 is involved in synthesis of Mo-nitrogenase, while nifB3 and nifB4 genes are not. Complementation of the P. polymyxa ∆nifBHDK mutant with the four reconstituted operons: nifB1anfHDGK, nifB2anfHDGK, nifB1vnfHDGK and nifB2vnfHDGK, has shown both that nifB1 and nifB2 were able to support synthesis of Fe- or V-nitrogenases. Transcriptional results obtained in the original Paenibacillus strains are consistent with the complementation results. Conclusions The multiple nifB genes of the diazotrophic Paenibacillus strains are divided into 4 classes. The nifB1 located in a compact nif gene cluster (nifBHDKENX(orf1)hesAnifV) and the nifB2 genes being adjacent to nif or anf or vnf genes are active in synthesis of Mo-, Fe and V-nitrogenases, but nifB3 and nifB4 are not. The reconstituted anf system comprising 8 genes (nifBanfHDGK and nifXhesAnifV) and vnf system comprising 10 genes (nifBvnfHDGKEN and nifXhesAnifV) support synthesis of Fe-nitrogenase and V-nitrogenase in Paenibacillus background, respectively.

Genome-Wide Analysis of PEBP Genes in Dendrobium huoshanense: Unveiling the Antagonistic Functions of FT/TFL1 in Flowering Time

Dendrobium is a semi-shade epiphytic Orchidaceae herb with important ornamental and medicinal value. Parts of the cultivation of Dendrobium germplasm resources, as well as the identification of medicinal components, are more studied, but the functional characterization of the flowering regulation in Dendrobium plants is less reported. Here, six PEBP family genes (DhFT3, DhFT1, DhMFT, DhTFL1b, DhFT2, and DhTFL1a) were identified from the Dendrobium huoshanense genome. The chromosome-level mapping showed that these genes were sequentially distributed on chromosomes 6, 9, 15, and 17. The paralogous gene DhTFL1b corresponded to DhTFL1a, which was determined through tandem duplication. The gene structure and conserved motif of DhPEBP indicated five PEBP genes apart from DhMFT contained four exons and three introns entirely. The phylogeny analysis showed that the PEBP gene family in A. thaliana, O. sativa, Z. mays, S. lycopersicum, and P. equestris were classified into three subclades, FT, TFL, and MFT, which maintained a high homology with D. huoshanense. The conserved domain of the amino acid demonstrated that two highly conserved short motifs (DPDXP and GXHR) embed in DhPEBPs, which may contribute to the conformation of the ligand binding bag. The 86th position of DhFTs was tyrosine (Y), while the 83th and 87th of DhTFL1s belonged to histidine (H), suggesting they should have distinct functions in flowering regulation. The promoter of six DhPEBPs contained several cis-elements related to hormone induction, light response, and abiotic stress, which indicated they could be regulated by the environmental stress and endogenous signaling pathways. The qRT-PCR analysis of DhPEBPs in short-term days induced by GA indicated the gene expressions of all DhFTs were gradually increased, whereas the expression of DhTFL1 was decreased. The results implied that DhPEBPs have various regulatory functions in modulating flowering, which will provide a scientific reference for the flowering regulation of Dendrobium plants.