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The Kv family of voltage-gated potassium channels regulate neuronal excitability. The biophysical characteristic of Kv channels can be matched to the needs of different neurons by forming homotetrameric or heterotetrameric channels within one of four subfamilies. The cytoplasmic tetramerization (T1) domain plays a major role in dictating the compatibility of different Kv subunits. The only Kv subfamily missing a representative structure of the T1 domain is the Kv2 family. We used X-ray crystallography to solve the structure of the human Kv2.1 T1 domain. The structure is similar to other T1 domains but surprisingly formed a pentamer instead of a tetramer. In solution the Kv2.1 T1 domain also formed a pentamer as determined with in-line SEC-MALS-SAXS and negative stain EM. The Kv2.1 T1-T1 interface involves electrostatic interactions including a salt bridge formed by the negative charges in a previously described CDD motif, and inter-subunit coordination of zinc. We show that zinc binding is important for stability. In conclusion, the Kv2.1 T1 domain behaves differently from the other Kv T1 domains which may reflect the versatility of Kv2.1, the only Kv subfamily that can assemble with the regulatory KvS subunits and scaffold ER-plasma membrane contacts.

Based on recent achievements in phylogenetic studies of the Brassicaceae, a novel infrafamilial classification is proposed that includes major improvements at the subfamilial and supertribal levels. Herein, the family is subdivided into two subfamilies, Aethionemoideae (subfam. nov.) and Brassicoideae. The Brassicoideae, with 57 of the 58 tribes of Brassicaceae, are further partitioned into five supertribes, including the previously recognized Brassicodae and the newly established Arabodae, Camelinodae, Heliophilodae, and Hesperodae. Additional tribus-level contributions include descriptions of the newly recognized Arabidopsideae, Asperuginoideae, Hemilophieae, Schrenkielleae, and resurrection of the Chamireae and Subularieae. Further detailed comments on 17 tribes in need of clarifications are provided.

Generic delimitation in Tillandsioideae has been widely discussed over the last past decades, being frequent changes in generic level. Despite the numerous changes at the generic and infrageneric levels in Tillandsioideae there is not yet a study bringing together the history of systematic of the tillandsioid genera. Given the conservation significance and scientific importance of species of the subfamily, it is time revise the generic status of Tillandsioideae. Therefore the purpose of this paper is to provide an updated phylogenetic and systematic history for the genera of Tillandsioideae. We review the fragmentary literature on the phylogeny, classification and taxonomy of Tillandsioideae, and provide an update of these data.

Mariner-like elements (MLEs) are class II transposons belonging to the Tc1-mariner family that have successfully invaded many insect genomes. In the current study, the availability of the Hessian fly, Mayetiola destructor, genome has enabled us to perform in silico analysis of MLEs using a previously described mariner element (Desmar1) belonging to the mauritiana subfamily. Eighteen mauritiana-like elements were detected and clustered into three main groups: Desmar1-like, MauCons1, and MauCons2. Subsequently, in vitro analysis was carried out to investigate mauritiana-like elements in M. destructor as well as in Mayetiola hordei using primers designed from TIRs of previously identified MLEs. PCR amplifications were successful, and a total of 12 and 17 mauritiana-like elements were detected in M. destructor and M. hordei, respectively. Sequence analyses of mauritiana-like elements obtained in silico and in vitro have shown that MauCons1 and MauCons2 elements share low similarity with Desmar1 ranging from 50% to 55%, suggesting that different groups under the mauritiana subfamily have invaded the genomes of M. destructor and M. hordei. These groups were likely inherited by vertical transmission, which subsequently underwent different evolutionary histories. This work describes new mauritiana-like elements in M. destructor that are distinct from the previously discovered Desmar1 and provides the first evidence of MLEs belonging to the mauritiana subfamily in M. hordei.

The B3 transcription factors (TFs) in plants play vital roles in numerous biological processes. Although B3 genes have been broadly identified in many plants, little is known about their potential functions in mediating seed development and material accumulation. Castor bean (Ricinus communis L.) is a non-edible oilseed crop considered an ideal model system for seed biology research. Here, we identified a total of 61 B3 genes in the castor bean genome, which can be classified into five subfamilies, including ABI3/VP1, HSI, ARF, RAV and REM. The expression profiles revealed that RcABI3/VP1 subfamily genes are significantly up-regulated in the middle and later stages of seed development, indicating that these genes may be associated with the accumulation of storage oils. Furthermore, through yeast one-hybrid and tobacco transient expression assays, we detected that ABI3/VP1 subfamily member RcLEC2 directly regulates the transcription of RcOleosin2, which encodes an oil-body structural protein. This finding suggests that RcLEC2, as a seed-specific TF, may be involved in the regulation of storage materials accumulation. This study provides novel insights into the potential roles and molecular basis of B3 family proteins in seed development and material accumulation.

Fish papillomaviruses form a newly discovered group broadly recognized as the Secondpapillomavirinae subfamily. This study expands the documented genomes of the fish papillomaviruses from six to 16, including one from the Antarctic emerald notothen, seven from commercial market fishes, one from data mining of sea bream sequence data, and one from a western gull cloacal swab that is likely diet derived. The genomes of secondpapillomaviruses are ∼6 kilobasepairs (kb), which is substantially smaller than the ∼8 kb of terrestrial vertebrate papillomaviruses. Each genome encodes a clear homolog of the four canonical papillomavirus genes, E1, E2, L1, and L2. In addition, we identified open reading frames (ORFs) with short linear peptide motifs reminiscent of E6/E7 oncoproteins. Fish papillomaviruses are extremely diverse and phylogenetically distant from other papillomaviruses suggesting a model in which terrestrial vertebrate-infecting papillomaviruses arose after an evolutionary bottleneck event, possibly during the water-to-land transition.

Despite worldwide distribution and rich diversity, the knowledge of mitochondrial genome (mtgenome) characteristics within the family Tenebrionidae (Coleoptera) is still very limited, and phylogenetics remain unresolved for the family. In this study, the mtgenomes for 12 species are newly sequenced and annotated. Three of the species represent the first report of complete mtgenomes from the subfamily Alleculinae. Comparative analysis of 36 tenebrionid mtgenomes finds that gene composition and order are the same as a theoretical ancestral insect mtgenome, and AT bias, length variation, and codon usage are consistent with other reported beetle mtgenomes. Some intergenic overlap and gap sequences may contain phylogenetically informative information, whereas neither the conserved, nor repeat sequences in the control region do. The subfamilies Lagriinae, Pimeliinae, Stenochiinae, and Alleculinae are found to be monophyletic, and the subfamilies Diaperinae and Tenebrioninae polyphyletic in our study. Furthermore, Lagriinae is sister to the rest of the subfamilies. At the tribal level, the tribes Lagriini in the subfamily Lagrrinae; Strongyliini in Stenochiinae; Cteniopodini in Alleculinae; and Triboliini, Opatrini, and Amarygmini in Tenebrioninae are monophyletic; while Diaperini in Diaperinae, and Tenebrionini in Tenebrioninae are polyphyletic. This article is protected by copyright. All rights reserved.

The rapid invasion of the non-native Phragmites australis (Poaceae, subfamily Arundinoideae) is a major threat to native wetland ecosystems in North America and elsewhere. We describe the first reference genome for P. australis and compare invasive (ssp. australis) and native (ssp. americanus) genotypes collected from replicated populations across the Laurentian Great Lakes to deduce genomic bases driving its invasive success. Here, we report novel genomic features including a Phragmites lineage-specific whole genome duplication, followed by gene loss and preferential retention of genes associated with transcription factors and regulatory functions in the remaining duplicates. Comparative transcriptomic analyses revealed that genes associated with biotic stress and defence responses were expressed at a higher basal level in invasive genotypes, but native genotypes showed a stronger induction of defence responses when challenged by a fungal endophyte. The reference genome and transcriptomes, combined with previous ecological and environmental data, add to our understanding of mechanisms leading to invasiveness and support the development of novel, genomics-assisted management approaches for invasive Phragmites.

The diversity of Asian horned toads is considered highly underestimated and to contain a large number of undescribed cryptic species. In this work, we describe three new species of Boulenophrys from south China, namely, Boulenophrys yaoshanensis sp. nov. from central Guangxi, Boulenophrys yingdeensis sp. nov. from northern Guangdong, and Boulenophrys yunkaiensis sp. nov. from western Guangdong. These three new species can be distinguished from all recognized congeners by a combination of morphological characteristics and significant genetic divergences. These descriptions increase the number of recognized species of Boulenophrys to 61. In addition, an updated checklist of the Asian horned toads of the subfamily Megophryinae is provided in this study.

The genus Dialithoptera Hampson, 1900 of the subfamily Nolinae Hampson, 1894 is newly reported for the first time from Laos, with two species, Dialithoptera gemmata (Hampson, 1896), and D. margarita Laszlo, Ronkay & Witt, 2007. Illustrations of adults and genitalia for the species are provided.

Vietanna Lee & Pham, gen. nov. and Vietanna hanoiensis Pham & Lee, sp. nov. are described based on a cicada specimen from Hanoi, Vietnam and placed in the subtribe Leptopsaltriina Moulton, 1923 of the tribe Leptopsaltriini Moulton, 1923 in the subfamily Cicadinae Latreille, 1802 (Hemiptera: Cicadidae). This new genus is morphologically similar to Tanna Distant, 1905 but distinguished from Tanna by the timbal cover exposing the inner part of timbal, the male abdominal epipleurite 3 with a distinct oblique linear fold, the male abdominal sternites without tubercle-like projections, and the aedeagus bifurcate at the apex. A key to the genera of Leptopsaltriina is provided. www.zoobank.org/urn:lsid:zoobank.org:pub:3B9CBC95-B54A-4BDE-A488-0DEAB69114BD .

Endoglucanases are carbohydrate-degrading enzymes widely used for bioethanol production as part of the enzymatic cocktail. However, family 5 subfamily 5 (GH5_5) endoglucanases are still poorly explored in depth. The Trichoderma reesei representative is the most studied enzyme, presenting catalytic activity in acidic media and mild temperature conditions. Though biochemically similar, its modular structure and synergy with other components vary greatly compared to other GH5_5 members and there is still a lack of specific studies regarding their interaction with other cellulases and application on novel and better mixtures. In this regard, the threedimensional structure elucidation is a highly valuable tool to both uncover basic catalytic mechanisms and implement engineering techniques, proved by the high success rate GH5_5 endoglucanases show. GH5_5 enzymes must be carefully evaluated to fully uncover their potential in biomass-degrading cocktails: the optimal industrial conditions, synergy with other cellulases, structural studies, and enzyme engineering approaches. We aimed to provide the current understanding of these main topics, collecting all available information about characterized GH5_5 endoglucanases function, structure, and bench experiments, in order to suggest future directions to a better application of these enzymes in the industry.

The mitochondrial genetic markers are considered useful tools for discrimination between more closely related lepidopteran taxa. Therefore, the present study aimed to investigate the role of mitochondrial (mt) 16 s rRNA gene in the determination of the taxonomic position for two moth species within Ditrysia clade. Maximum likelihood analysis has indicated a well-supported dendrogram based on the Tamura-Nei model for the recovered lepidopterans. The mt 16 s rRNA query sequences from 24 species within seven families were analyzed. This analysis and bootstrap confidence revealed two major clades representing Glossata suborder within Lepidoptera, with a close relationship of Noctuoidea + (Pyraloidea (Hesperioidea + Papilionoidea)). The subfamily Heliothinae forming a sister group with Risobinae (Noctinae + Hadeninae). In addition, there is a clear observation about the close relation between Phycitinae + Galleriinae within Pyraloidea and Cyrestinae + Limenitidinae within Papilionoidea. The present study supported that the Helicoverpa and Meroptera species are the first accounts of these genera inhabiting Saudi Arabia.

Transglutaminases (TGases) are widely known to play critical roles in innate immunity, in particular, TGase2, which involves in autophagy process to help degrade protein aggregates under stressful conditions in mammals. Nevertheless, the function of the TGase2 counterpart whether involves in invertebrate autophagy is largely unknown. In this study, a novel TGase2-like homologous gene from the sea cucumber Apostichopus japonicus (named as AjTGase2-like) was cloned using RACE technology and its biological functions were also investigated. The AjTGase2-like gene encoded a peptide of 750 amino acids with the representative domains of Transglut_N domain, TGc domain, and two Transglut_C domains, which exhibited highly conservative with vertebrate TGase2. Multiple sequence alignments and phylogenetic analysis both supported that AjTGase2-like belonged to a new member of TGase2 subfamily. AjTGase2-like was pervasively expressed in all examined tissues, with the largest transcription in muscle, followed by respiratory trees, and intestine. After immersion infection with Vibrio splendidus, the mRNA and protein levels of AjTGase2-like were both significantly induced and reached the highest levels at 24 h, indicating AjTGase2-like plays a key role in immune response. Further functional analysis showed that the ubiquitinated protein level was significantly increased by 1.65-fold (p 0.01) after silencing of AjTGase2-like, and the protein levels of AjLC3-II/I and AjBeclin1 were both obviously decreased by 0.49-fold (p 0.01) and 0.64-fold (p 0.01) at the same time, while the authophagy receptor of Ajp62 was signally up-regulated by 1.40-fold (p 0.01) under same condition. Moreover, the immunofluorescence signals of AjLC3 and Ajp62 were consistent with their protein levels, suggesting knockdown of AjTGase2-like causes a blockage in autophagy. More importantly, the AjLC3 positive signal was not increased after adding with chloroquine in the case of AjTGase2-like interference, indicating AjTGase2-like might play pivotal role in the early step of autophagosome formation. Besides, our results showed that the fluorescence signal of AjTGase2-like was largely co-localized with Ajp62 around the cytoplasm in vivo, and rAjp62 could directly combine with rAjTGase2-like in vitro, indicating AjTGase2-like interacts with Ajp62 during autophagy. Overall, our findings supported that AjTGase2-like served as a positive regulator in sea cucumber authophay.

Pollen micromorphology is not only used to check the functional and structural evolution in plants but also to solve the taxonomic problem related to the classification of plants. Therefore, keeping in view the significance of pollen traits, selected taxa of the subfamily Caesalpiniaceae was collected from different geographical regions of Pakistan. The species were then analyzed under both light microscopy and scanning electron microscopy techniques to investigate the importance of micromorphological characters of pollen in the identification and classification of species. Great variation was recorded in equatorial shape, surface ornamentation, tectum, polar diameter, equatorial diameter, and exine thickness. However, little variation was observed in pollen type, polar shape, and fertility of pollen. The equatorial shape of five types was observed: prolate, prolate-spheroidal, spheroidal-subprolate, subspheroidal-prolate, and subspheroidal. Four types of surface ornamentation, psilate, granulate, clavate, and perforate, were recorded. Tectum of five types, intactate, reticulate regulate, medium reticulate, tactate, and striate, was observed. Sexine was thicker than nexine in all studied species. The largest polar diameter was observed in Caesalpinia pulcherrima 64.1 μm while the smallest in Parkinsonia aculeata 26.1 μm. The largest equatorial diameter was found in C. pulcherrima of 70.25 μm whereas the smallest in P. aculeata 27.57 μm. All the pollens analyzed were tricolporate. All studied species have a fertility ratio of more than 90%. A taxonomic key was developed to show the variation in pollen features and delimit species for the correct identification. In conclusion, the pollen traits were found useful to define species boundaries at various taxonomic ranks and will strengthen the taxonomy of this subfamily. Besides, this study also explored the palynological traits and their implication in the taxonomy of the subfamily Caesalpiniaceae.

Diacylglycerol acyltransferase (DGAT) is a crucial enzyme in the triacylglycerol (TAG) biosynthesis pathway. The oleaginous fungus Mortierella alpina can accumulate large amounts of arachidonic acid (ARA, C20:4) in the form of TAG. Therefore, it is important to study the functional characteristics of its DGAT. Two putative genes MaDGAT1A/1B encoding DGAT1 were identified in M. alpina ATCC 32222 genome by sequence alignment. Sequence alignment with identified DGAT1 homologs showed that MaDGAT1A/1B contain seven conserved motifs that are characteristic of the DGAT1 subfamily. Conserved domain analysis showed that both MaDGAT1A and MaDGAT1B belong to the Membrane-bound O-acyltransferases superfamily. The transforming with MaDGAT1A/1B genes could increase the accumulation of TAG in Saccharomyces cerevisiae to 4·47 and 7·48% of dry cell weight, which was 7·3-fold and 12·3-fold of the control group, respectively, but has no effect on the proportion of fatty acids in TAG. This study showed that MaDGAT1A/1B could effectively promote the accumulation of TAG and therefore may be used in metabolic engineering aimed to increase TAG production of oleaginous fungi.

ZupT fromEscherichia coliis a member of the Zrt-/Irt-like Protein (ZIP) transporter family, which is responsible for zinc uptake during zinc-sufficient conditions. ZIP transporters have been shown to transport different divalent metal ions including zinc, iron, manganese, and cadmium. In this study, we show that ZupT has an asymmetric binuclear metal center in the transmembrane domain; one metal-binding site, M1, binds zinc, cadmium, and iron, while the other, M2, binds iron only and with higher affinity than M1. Using site-specific mutagenesis and transport activity measurements in whole cells and proteoliposomes, we show that zinc is transported from M1, while iron is transported from M2. The two sites share a common bridging ligand, a conserved glutamate residue. M1 and M2 have ligands from highly conserved motifs in transmembrane domains 4 and 5. Additionally, M2 has a ligand from transmembrane domain 6, a glutamate residue, which is conserved in the gufA subfamily of ZIP transporters, including ZupT and the human ZIP11. Unlike cadmium, iron transport from M2 does not inhibit the zinc transport activity but slightly stimulates it. This stimulation of activity is mediated through the bridging carboxylate ligand. The binuclear zinc-iron binding center in ZupT has likely evolved to enable the transport of essential metals from two different sites without competition; a similar mechanism of metal transport is likely to be found in the gufA subfamily of ZIP transporter proteins.

The present paper adds to the knowledge of the family Mantispidae from India. Herein we describe two new Euclimacia species: E. similis sp. nov. from Madhya Pradesh and E. nicobarica sp. nov. from Andaman & Nicobar Islands. A new combination is also proposed: Mantispilla coorgensis (Ohl, 2004a) comb. nov. With these additions, the family is now represented in India by 22 species in eight genera.

The grasshopper Subfamily Gomphomastacinae (Orthoptera: Acridoidea: Eumastacidae) is widely distributed in the Qinghai-Tibetan Plateau and Central Asia. This paper describes three new species: Ptygomastax nihilsulcus Ge, sp. nov. Phytomastax pentaspinula Ge, sp. nov. and Pentaspinula unispinula Ge, sp. nov. of the subfamily from the eastern Qinghai-Tibetan Plateau, along with an updated identification key of species of the related genera. Detail figures of the body and genitalia of the new species are also provided.

The present corrigendum corrects errors that occurred in Gómez S., Corgosinho P.H.C. & Rivera-Sánchez K.I. 2021. Proposal of new genera and species of the subfamily Diosaccinae (Copepoda : Harpacticoida : Miraciidae). European Journal of Taxonomy 759 : 1–62. https://doi.org/10.5852/ejt.2021.759.1433

The current knowledge on the diversity of the genus Plexaurella is based on a series of dated revisions, often with no examination of types. Although being common octocorals in western Atlantic reefs, there is no consensus on an exact number of valid species. Furthermore, phylogenetic reconstructions do not support the current classification of Plexaurella within the family Plexauridae. Thus, this study reviews the genus based on examination of available types and assesses monophyly using mitochondrial (COI+igr, mtMutS) and nuclear (28S) markers, mostly from available molecular data. Until now, up to six species were considered valid. Our results show that the group is composed of at least seven previously described species: P. dichotoma, P. nutans, P. grisea, P. teres, P. grandiflora, P. regia and P. obesa; and one new species: Plexaurella rastrera sp. nov. An illustrated key to the valid species and a list of all available names are provided and the current classification of the genus is discussed. Based on congruent phylogenetic reconstructions and genetic distances, we propose the elevation of the former plexaurid subfamily Plexaurellinae to family level. Finally, based on examination of types, we propose the synonymy between Pseudoplexaura crucis and Plexaurella tenuis under Pseudoplexaura tenuis new comb.

Although the Mesozoic Era played an important role in the evolution and diversification of Elateridae, the Cretaceous click-beetle fauna remains very poorly known. Here we describe Cretopachyderes burmitinus gen. et sp. nov. based on a single specimen from the mid-Cretaceous Burmese amber. This species is remarkable for its extremely long posterior angles of pronotum, which is a unique character among fossil Elateridae. We discuss the diagnostic characters of Cretopachyderes gen. nov. and tentatively place it to subfamily Agrypninae close to extant genus Pachyderes Guérin-Méneville, 1829.

Made available in DSpace on 2022-04-28T19:46:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-03-01 Agencia Nacional de Investigación e Innovación American University University of Arizona University of Vermont Junta de Andalucía Comisión Sectorial de Investigación Científica Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Ministerio de Educación y Cultura The subfamily Triatominae (Hemiptera-Reduviidae) includes more than 150 blood-sucking species, potential vectors of the protozoan Trypanosoma cruzi, causative agent of Chagas disease. A distinctive cytogenetic characteristic of this group is the presence of extremely stable chromosome numbers. Unexpectedly, the analyses of the chromosomal location of ribosomal gene clusters and other repetitive sequences place Triatominae as a significantly diverse hemipteran subfamily. Here, we advance the understanding of Triatominae chromosomal evolution through the analysis of the 45S rDNA cluster chromosomal location in 92 Triatominae species. We found the 45S rDNA clusters in one to four loci per haploid genome with different chromosomal patterns: On one or two autosomes, on one, two or three sex chromosomes, on the X chromosome plus one to three autosomes. The movement of 45S rDNA clusters is discussed in an evolutionary context. Our results illustrate that rDNA mobility has been relatively common in the past and in recent evolutionary history of the group. The high frequency of rDNA patterns involving autosomes and sex chromosomes among closely related species could affect genetic recombination and the viability of hybrid populations, which suggests that the mobility of rDNA clusters could be a driver of species diversification. Sección Genética Evolutiva Facultad de Ciencias Universidad de la República Department of Experimental Biology Genetics University of Jaén Department of Biomedicine and Biotechnology University of Alcalá Laboratório de Entomologia em Saúde Pública Departamento de Epidemiologia Faculdade de Saúde Pública Universidade de São Paulo Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista “Júlio de Mesquita Filho” (Unesp) Fundação Oswaldo Cruz Laboratorio de Investigación en Genética Evolutiva – LIGE Universidad Pedagógica y Tecnológica de Colombia Programa de Biología Celular y Molecular ICBM Facultad de Medicina Universidad de Chile Laboratorio de Entomología Aplicada y Parasitología Escuela de Biología Facultad de Farmacia Universidad de San Carlos de Guatemala Department of Biological Sciences Loyola University New Orleans Departamento de Microbiología y Parasitología Facultad de Medicina Universidad Nacional Autónoma de México Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista “Júlio de Mesquita Filho” (Unesp) Comisión Sectorial de Investigación Científica: 160 FAPESP: 17/05015-7 FAPESP: 19/02145-2 FAPESP: 307 398/2018-8 Ministerio de Educación y Cultura: II/FVF/2019/054

This article focuses on the sciurid (Rodentia, Sciuridae) remains from the early Miocene (MN4) Mokra-Quarry sites, Moravia, Czech Republic. The sciurid assemblage in the different fissures ranges from one genus in MWQ4/2018 to three different genera in MCQ3/2005. Regarding subfamily Sciurinae, the dentognathic remains are referred to Palaeosciurus aff. fissurae, which shows an intermediate stage of evolution between Palaeosciurus fissurae and Palaeosciurus sutteri according to the development of conules in both upper and lower molars. Although the sciurid postcranial remains are left unassigned, several calcanei show strong morphological features belonging to ground squirrels, and, are therefore attributed to cf. Palaeosciurus. As for Pteromyinae subfamily, three genera were recovered (Miopetaurista, Blackia and Aliveria), including a new species: Aliveria mojmiri sp. nov. The new species shows a more specialised dental pattern and could further represent the ancestral species from which several middle Miocene genera evolved. Furthermore, some of the recovered calcanei have been here identified as cf. Aliveria, representing the first postcranial remains ever belonging to the genus, and confirm its attribution as a flying squirrel. The presence of both ground and flying squirrels is in concordance with an open landscape with patches of woodland proposed for Mokra-Quarry sites.

Yuan-Jie Liu,1– 3,* Sheng-Yan Yin,2,3,* Shu-Hong Zeng,2,3 Yi-Dou Hu,1 Meng-Qi Wang,2,3 Pan Huang,1 Jie-Pin Li1– 3 1Department of Oncology, Zhangjiagang TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Zhangjiagang, Jiangsu, 215600, People’s Republic of China; 2Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Chinese Medicine, Nanjing, Jiangsu, 210029, People’s Republic of China; 3No. 1 Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, 210023, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jie-Pin Li; Pan HuangDepartment of Oncology, Zhangjiagang TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Chang ‘an South Road No. 77, Zhangjiagang, Jiangsu, 215600, People’s Republic of ChinaTel +8615150224526; +8613921976647Email zjgzy027@njucm.edu.cn; yezipan2020@126.comPurpose: The identification of biomarkers and effective therapeutic targets for gastric cancer (GC), the most common cause of cancer-related deaths around the world, is currently a major focus in research. Here, we examined the utility of LHFPL6 as a prognostic biomarker and therapeutic target for GC.Methods: We explored the clinical relevance, function, and molecular role of LHFPL6 in GC using the MethSurv, cBioPortal, TIMER, Gene Expression Profiling Interactive Analysis, ONCOMINE, MEXPRESS, and EWAS Atlas databases. The GSE118919, GSE29272, and GSE13861 datasets were used for differential expression analysis. Using The Cancer Genome Atlas, we developed a Cox regression model and assessed the clinical significance of LHFPLs. In addition, we used the “CIBERSORT” algorithm to make reliable immune infiltration estimations. Western blot and immunohistochemistry were used to examine protein expression. Cell migration and invasion were assessed using transwell experiments. THP-1-derived macrophages and GC cells were co-cultured in order to model tumor–macrophage interactions in vitro. The levels of CD206 and CD163 were measured using immunofluorescence assays. The results were visualized with the “ggplot2” and “circlize” packages.Results: Our results showed that in GC, LHFPL6 overexpression was significantly associated with a poor prognosis. Our findings also suggested that LHFPL6 may be involved in the activation of the epithelial–mesenchymal transition. Furthermore, LHFPL6 expression showed a positive correlation with the abundance of M2 macrophages, which are potent immunosuppressors.Conclusion: LHFPL6 could be a prognostic biomarker and therapeutic target for GC.Keywords: gastric cancer, LHFPL6, biomarker, EMT, M2 macrophages

Background Anthocyanins, which belong to flavonoids, are widely colored among red-purple pigments in the Asiatic hybrid lilies (Lilium spp.). Transcription factor (TF) LhMYBSPLATTER (formerly known as LhMYB12-Lat), identified as the major kernel protein, regulating the anthocyanin biosynthesis pathway in ‘Tiny Padhye’ of Tango Series cultivars, which the pigmentation density is high in the lower half of tepals and this patterning is of exceptional ornamental value. However, the research on mechanism of regulating the spatial and temporal expression differences of LhMYBSPLATTER, which belongs to the R2R3-MYB subfamily, is still not well established. To explore the molecular mechanism of directly related regulatory proteins of LhMYBSPLATTER in the anthocyanin pigmentation, the yeast one-hybrid (Y1H) cDNA library was constructed and characterized. Results In this study, we describe a yeast one-hybrid library to screen transcription factors that regulate LhMYBSPLATTER gene expression in Lilium, with the library recombinant efficiency of over 98%. The lengths of inserted fragments ranged from 400 to 2000 bp, and the library capacity reached 1.6 × 106 CFU of cDNA insert, which is suitable to fulfill subsequent screening. Finally, seven prey proteins, including BTF3, MYB4, IAA6-like, ERF4, ARR1, ERF WIN1-like, and ERF061 were screened by the recombinant bait plasmid and verified by interaction with the LhMYBSPLATTER promoter. Among them, ERFs, AUX/IAA, and BTF3 may participate in the negative regulation of the anthocyanin biosynthesis pathway in Lilium. Conclusion A yeast one-hybrid library of lily was successfully constructed in the tepals for the first time. Seven candidate TFs of LhMYBSPLATTER were screened, which may provide a theoretical basis for the study of floral pigmentation.

BackgroundGRAS transcription factors perform indispensable functions in various biological processes, such as plant growth, fruit development, and biotic and abiotic stress responses. The development of whole-genome sequencing has allowed theGRASgene family to be identified and characterized in many species. However, thorough in-depth identification or systematic analysis ofGRASfamily genes in foxtail millet has not been conducted.ResultsIn this study, 57GRASgenes of foxtail millet (SiGRASs) were identified and renamed according to the chromosomal distribution of theSiGRASgenes. Based on the number of conserved domains and gene structure, theSiGRASgenes were divided into 13 subfamilies via phylogenetic tree analysis. TheGRASgenes were unevenly distributed on nine chromosomes, and members of the same subfamily had similar gene structures and motif compositions. Genetic structure analysis showed that mostSiGRASgenes lacked introns. SomeSiGRASgenes were derived from gene duplication events, and segmental duplications may have contributed more toGRASgene family expansion than tandem duplications. Quantitative polymerase chain reaction showed significant differences in the expression ofSiGRASgenes in different tissues and stages of fruits development, which indicated the complexity of the physiological functions ofSiGRAS. In addition, exogenous paclobutrazol treatment significantly altered the transcription levels of DELLA subfamily members, downregulated the gibberellin content, and decreased the plant height of foxtail millet, while it increased the fruit weight. In addition,SiGRAS13andSiGRAS25may have the potential for genetic improvement and functional gene research in foxtail millet.ConclusionsCollectively, this study will be helpful for further analysing the biological function ofSiGRAS. Our results may contribute to improving the genetic breeding of foxtail millet.

The subfamily Nothopuginae is revised. Titanopuga salinarum gen. et sp. nov., Cuyanopuga bilobata gen. et sp. nov. and Nothopuga telteca sp. nov. are described. We performed a cladistic analysis based on 144 morphological characters and 24 terminals, including representatives of all ammotrechid subfamilies. Nothopuginae is recovered monophyletic with moderate support, but its sister group relation with Oltacolinae is well supported. The genus Mortola is considered a junior synonym of Pseudocleobis, and Pseudocleobis mortola comb. nov. is considered nomen dubium. The subfamily Mortolinae is represented by a monophyletic clade including Chinchippus, Dasycleobis and Pseudocleobis. A dichotomous key for the genera and species of Nothopuginae is provided.