Neisseria adhesin A (NadA), which is currently used as a subunit in a licensed vaccine against Neisseria meningitidis. Other TAAs that seem promising vaccine candidates are the Acinetobacter trimeric autotransporter (Ata), the Haemophilus influenzae adhesin (Hia), and TAAs of the genus Bartonella. Here, we review the suitability of various TAAs as vaccine candidates.The current problem of increasing antibiotic resistance and the resurgence of numerous infections indicate the need for novel vaccination strategies more than ever. In vaccine development, the search for and the selection of adequate vaccine antigens is the first important step. In recent years, bacterial outer membrane proteins have become of major interest, as they are the main proteins interacting with the extracellular environment. Trimeric autotransporter adhesins (TAAs) are important virulence factors in many Gram-negative bacteria, are localised on the bacterial surface, and mediate the first adherence to host cells in the course of infection. One example is the  Vaccination against human pathogens was first introduced in medicine in 1796 by Edward Jenner  the in vivo expression of surface epitopes, (ii) a high strain coverage, and (iii) immunogenicity and induction of a protective immune response in the host , 19. In TAAs are a family of obligate homotrimeric, non-fimbrial, non-pilus bacterial adhesins that have numerous biological functions such as bacterial autoagglutination, binding to extracellular matrix (ECM) proteins and host cells, and the induction of distinct host cell responses. They are widespread in α-, β-, and γ-proteobacteria and primarily ensure the initial adhesion to specific molecular components of both abiotic and biotic surfaces Fig. b 16, 2323, 26. FIn general, all TAAs share a common lollipop-like surface structure Fig. a. The C-Type V secretion systems are autotransporters containing a ß-barrel transmembrane domain . Five diEven though the exact secretion mechanism of TAAs is still unclear, the Vc secretion system is a potentially valuable feature in the development of multivalent recombinant bacterial vector vaccines –44. For Escherichia coli autotransporter Hbp (type Va secretion system) both in E. coli and in an attenuated Salmonella enterica serovar Typhimurium vaccine strain  was approved by the European Union (EU) [In general, three types of meningococcal vaccines are available: polysaccharide vaccines, polysaccharide–protein conjugated vaccines and vaccines based on OMPs (developed via reverse vaccinology) , 103. Inunogenic , 107. A unogenic , 98. Nevunogenic . In 2013ion (EU) . Later, ion (EU) , 109.N. meningitidis to epithelial cells via ß1-integrins and in its subsequent invasion during the infection process [NadA is a phase-variable ca. 43 kDa OMP of which the expression is mainly regulated by the transcriptional regulator NadR , 111. Na process , 113. Na process –116. Fur process , 118. Gr process . Variant process . For exa process , 120. Th process , 98, 121nadA gene is present in approximately 30% of N. meningitidis isolated strains and in 75% of hypervirulent N. meningitidis serogroup B lineages [nadA gene [nadA is present in 16.2% of 154 isolates [The lineages , 122. CoadA gene . In caseisolates .N. meningitidis, and the further development as a vaccine component serve as a role model to expedite the development of TAA-based vaccines.Currently, NadA is the only TAA that is used as a component in a licensed vaccine, as NadA3 is a major antigen in the multicomponent vaccine 4CMenB , 96, 98.Haemophilus surface fibril (Hsf) of Haemophilus influenzae, is immunogenic in humans in conjugation with other antigens , and facilitates bacterial attachment to host epithelial cells during infection by binding heparan sulphate and laminin [nhha gene is highly conserved in all meningococcal strains [NhhA was the first vaccine candidate against MenB and was described using whole genome sequencing to identify possible vaccination targets . NhhA sh laminin , 126. Fu laminin , 127. Mo strains , 97.N. meningitidis strains [Moraxella IgD-binding protein providing a more effective vaccine [All these features suggest that NhhA is a promising vaccine candidate , 111. Pe strains . A later vaccine .However, it was found that a subset of clinical isolated MenB strains only  express the monomeric form of NhhA, caused by a single natural mutation (glycine to aspartic acid) in the C-terminal passenger domain. Accordingly, loss in trimerization, surface exposure and adhesive features was observed. These findings question the vaccine candidacy of NhhA because of the need for broad strain coverage .H. influenzae, H. influenzae adhesin (Hia), and H. surface fibril (Hsf) [Two different TAAs are expressed on the outer membrane of il (Hsf) , 130.Haemophilus influenzae is a human specific, Gram-negative pathogen categorised into two different groups, the polysaccharide encapsulated (serotypes a–f), and the unencapsulated group often referred to as non-typeable H. influenzae (NTHi) [H. influenzae serotype b (Hib) encapsulated strains are considered most virulent and are a major agent of respiratory tract systemic infections. Infections can lead to acute epiglottitis, sepsis, acute meningitis, and pneumonia. NTHi mainly causes local diseases such as bronchitis, otitis media, and sinusitis [e (NTHi) , 132. H.inusitis , 134.H. influenzae nor against NTHi, and so research is thus needed [Moraxella catarrhalis surface proteins [Current vaccines are mainly against the most virulent Hib. The earlier polysaccharide-based vaccines showed only short-term protection for children under 18 months after various trials were undertaken in 1977 . The firs needed –140. Forproteins .Two relevant candidate vaccine antigens are the surface proteins Hia and Hsf. Both TAAs contain several repetitive domains, are homologous in their N- and C-termini, and show an overall sequence identity of 81% and 72%, respectively .H. influenzae, and is presented in all encapsulated serotypes and a subset of NTHi [Hsf has a monomeric molecular weight of 243 kDa , represe of NTHi . The bin of NTHi . Further of NTHi , 142.In contrary with Hsf, Hia is only present in 25% of NTHi clinical isolates and has a monomeric molecular weight of 114 kDa , 133. HiHaemophilus ducreyi called the ‘ducreyi serum resistance A’ (DsrA) is a proven virulence factor and thus a potential target as vaccine antigen [H. ducreyi is a pathogen that causes the genital ulcer disease chancroid, for which no vaccines are available [I), administered bi-weekly in Freund’s adjuvant against infection with experimental H. ducreyi in swine [I with alum is highly persistent and of superior quality and quantity compared to subcutaneous administration [H. ducreyi is divided into two clonal populations with varieties in the passenger domain of DsrA, meaning that antibodies recognising class I DsrA do not recognise class II DsrA [The TAA of  antigen . H. ducrvailable . Fusco ein swine . It was stration . Furtherstration . Nonethe II DsrA , 150.A. baumannii, one of the majorcausative agents of hospital-acquired infections worldwide [A. baumannii strains possess the ability to acquire resistance genes rapidly against all commonly used antimicrobial compounds. The dissemination of carbapenem-resistant and in general multidrug-resistant Acinetobacter spp. strains is one of the most urgent health risks of our time and threatens to undo a century of medical progress [A. baumannii is the number one pathogen on the ‘WHO priority pathogens list for R&D of new antibiotics’ [Ata is a TAA present on the bacterial surface of the Gram-negative orldwide , 152. Chprogress . Consequbiotics’ . Effectibiotics’ .A. baumannii are licensed. However, promising vaccine candidates with immunogenic and protective properties have been described, including outer membrane complexes, OmpA and Ata itself [Vaccination can become a valuable alternative for shortcoming antibiotic treatments against multi-resistant pathogenic strains. Currently, no vaccines against a itself –157.A. baumannii [ata gene was detected in 44 out of 75 collected A. baumannii isolates of which 43 showed additional Ata expression on its outer membrane [A. nosocomialis, A. seifertii, and A. baumannii showed presence of the ata gene [Ata was first described in 2012 while searching for novel virulence factors of aumannii . The atamembrane . More reata gene . Ata medata gene , plays aata gene , 159inter alia two heterologous unrelated multidrug resistant A. baumannii strains [A. baumannii were recently re-emphasized [Bentancor et al. demonstrated in a pneumonia infection model in immunocompetent and immunocompromised mice the promising bactericidal, opsonophagocytic, and protective features of Ata-induced antibodies against  strains . Neverthphasized , 156.Moraxella catarrhalis expresses two different TAAs on its outer membrane, the ubiquitous surface protein A1 (UspA1) and the ubiquitous surface protein A2 (UspA2) [ (UspA2) , 161.Moraxella catarrhalis is a Gram-negative and a human-specific bacterium of the respiratory tract [Micrococcus catarrhalis, Neisseria catarrhalis, and Branhamella catarrhalis [M. catarrhalis is a commensal coloniser of the nasopharynx and represents a causative agent of otitis media in (young) children. The role of M. catarrhalis as causative agent of COPD has long been underestimated, however, is a frequent pathogen in the acute exacerbation phase of the disease [M. catarrhalis are a serious burden for health systems worldwide [M. catarrhalis produces ß-lactamases and is thus resistant against various important antibiotics [M. catarrhalis vaccines, are, therefore, highly desirable [ry tract , 163. Itarrhalis . M. cata disease , 165. Ot disease , 162. Diorldwide , 167. Moibiotics . Alternaesirable .M. catarrhalis-associated diseases, but several candidate vaccines are being developed [M. catarrhalis vaccine antigens are adhesive proteins , proteins involved in nutrient acquisition , lipooligosaccharides, or other Moraxella surface proteins [M. catarrhalis and play an important role in the first adherence to the epithelial host cells, during the infection process, and the subsequent disease development [Currently, no licensed vaccines are available to prevent eveloped , 169. Poproteins , 171. Nuelopment .uspA1 and uspA2 genes was demonstrated [uspA genes can swap between other uspA genes from the same strains [UspA1 and UspA2 are TAAs with a predicted molecular weight of ca. 83.5 and ca. 59.5 kDa, respectively . They arnstrated , 178 resnstrated . In addi strains . Consequ strains . A possi strains .Escherichia coli, in particular the E. coli immunoglobulin binding (Eib) proteins [E. coli auto-agglutinating adhesin (Saa) [E. coli adhesin G (UpaG) [E. coli adhesin G (EhaG) [Four different TAAs have been characterised from pathogenic proteins , the Shiin (Saa) , the uroG (UpaG) , and, moG (EhaG) .E. coli are available[E. coli is done for the enterotoxigenic E. coli (ETEC) expressing enterotoxins and colonisation factors  upon infection [Currently, no broadly protective vaccines against pathogenic vailable, 186, buvailable–189. Mosnfection , as thisnfection . ETEC vanfection , 192. Twnfection . Promisinfection , 194–196E. coli (EHEC) strains [E. coli (STEC) causes severe diseases in humans such as haemorrhagic colitis or haemolytic–uremic syndrome (HUS) [Eib genes occur in various pathogenic and multidrug-resistant E. coli strains, for example enteroaggregative E. coli (EAEC), extraintestinal pathogenic E. coli (ExPEC), and verotoxigenic E. coli (VTEC) [Eib proteins are mostly found in intimin-negative, shiga toxin-producing enterohaemorrhagic  strains , 198. Shme (HUS) . Eib geni (VTEC) –201. No i (VTEC) .Currently, six homologous Eib proteins are described  , 198. Thsaa-positive STEC strains was inhibited upon application of a polyclonal antiserum that was raised against purified Saa, emphasizing its potential as a vaccine antigen [In 2001, the gene for Saa was isolated from a large, virulence-related plasmid in a STEC strain negative for the locus for enterocyte effacement. Saa mediates autoaggregation and adherence to human epithelial type 2 cells, shows variation in size for different STEC strains, and has just ca. 25% identity with the Eib proteins. Furthermore, Saa was not proven to contribute to serum resistance. Nevertheless, in vitro adherence of  antigen .Escherichia coli UpaG, was identified by Durant et al. via reverse vaccinology [E. coli (UPEC) strain CFT073, mediates binding to ECM proteins and bladder epithelial cells, and promotes bacterial cell aggregation and biofilm formation [upaG gene in UPEC shows extensive sequence variation with the upaG gene in ExPEC strains [cinology . UpaG, cormation . The upa strains .E. coli [Furthermore, UpaG was proven to induce protective antibodies in a mouse model against lethal sepsis due to virulent extraintestinal isolates of E. coli . UpaG shE. coli , suggestE. coli .E. coli strains [The most recently identified TAA is EhaG which occurs in EHEC strains. EhaG is a positional orthologue of UpaG , 204, bu strains . Some ofSalmonella adhesin A (SadA) is a TAA expressed in vivo on the bacterial surface of the pathogenic Salmonella enterica (serovar Typhimurium) during infections [fections , 205.Salmonella enterica causes significant morbidity and mortality worldwide in humans and cattle [Salmonella is the most frequent bacterial cause of foodborne disease in the US and is responsible for the majority of foodborne outbreaks in the European Union [S. enterica can result in enteric salmonellosis and sometimes manifests as septicaemia. When not self-limiting, Salmonella-infected patients are treated via antimicrobial therapy. Consequently, multidrug-resistant S. enterica are on the rise [d cattle . Moreovean Union . Infectithe rise , 208.Salmonella vaccines exist: (i) a whole-cell live-attenuated vaccine (Vivotif®); (ii) a polysaccharide unconjugated vaccine; and (iii) a polysaccharide-conjugated vaccine , all against one S. enterica serovar Typhi [Salmonella spp. does exist for livestock breeding; for instance, an attenuated S. enterica serovar Typhimurium was designed providing higher cross protection against Salmonella serovars in swine [Salmonella vaccines.Currently, three types of licensed ar Typhi . Furtherin swine . DespiteS. enterica and SadA-deficient S. enterica. SadA, nonetheless, plays an important role in adherence to and invasion of intestinal epithelial cells. Large surface structures such as LPS or fimbria inhibit the function of SadA, suggesting a specific role during certain conditions in colonisation and infection of epithelial cells. Moreover, SadA is highly conserved within S. enterica strains and is considered as a positional orthologue of UpaG and EhaG in E. coli, but with some different functions [S. enterica might involve multiple antigens in parallel [The TAA SadA has an approximate trimeric size of 426 kDa and promotes biofilm formation and autoaggregation, but does not mediate serum resistance and does not bind ECM proteins. In addition, no distinction in virulence was observed between wild-type unctions , 204. Anparallel .Bartonella quintana, B. bacilliformis, and B. henselae are clinically the three most important Bartonella species each expressing one or more TAAs [B. quintana expresses four variably expressed outer membrane proteins (VompA–D) [B. bacilliformis the B. bacilliformis adhesins A, B, and C  and B. henselae the B. henselae adhesin A (BadA). Antimicrobial treatment of Bartonella spp.-associated diseases depends solely on the clinical situation and immunological status of the patient and less on the infective species. Consequently, no general treatment recommendation does exist for all Bartonella spp.-associated diseases [ore TAAs , 212. B.VompA–D) , B. bacidiseases .Bartonella quintana is transmitted via the human body louse and is the causative agent of trench fever. Infections with B. quintana can lead to endocarditis, bacillary angiomatosis and peliosis hepatis in immunocompromised patients [B. quintana infections [patients , 215–217fections .Bartonella quintana expresses four TAAs called VompA–D, which are encoded by four genes  and are tandemly arranged in a 12.8 kb gene locus [B. quintana [ne locus . The domne locus . This requintana . Vomps aquintana , but do quintana . Vomps aquintana , 218.B. quintana antigens, 24 immunoreactive membrane proteins were identified of which, among others, VompA and VompB were most frequently recognised by sera from B. quintana-infected patients [The immunogenicity of Vomps and their suitability as a vaccine antigen have been described. While analysing protective and diagnostically relevant patients . FurtherBartonella bacilliformis is the causative agent of Carrion’s disease, a biphasic illness restricted to the South American Andes [B. bacilliformis infects endothelial cells and stimulates cell proliferation which results in the formation of blood-filled nodular haemangioma-like lesions in the skin known as ‘verruga peruana’ [an Andes . The patperuana’ .B. bacilliformis. However, vaccines against B. bacilliformis infections should be effective, as indigenous people in B. bacilliformis endemic regions seem less susceptible to infections and hemolytic diseases compared to non-indigenous people [Currently, no vaccine is available for s people . In addis people .B. bacilliformis strains. Twenty-two Peruvian guards working in a region notorious for frequent incidents of Carrion’s disease were subcutaneously vaccinated. The vaccine did not prevent infection, but alleviated the severity of the Carrion’s disease [One of the few attempts to prepare a vaccine against the Carrion’s disease was performed in 1943 by Howe and Hertig. The vaccine contained a formalin suspension of four  disease , 224. No disease , 226.B. bacilliformis, three genes encoding for putative TAAs were identified [B. bacilliformis adhesins A–C (BbadA–C), originally called Bartonella repeat proteins (Brps), share common domain features with other TAAs of the genus Bartonella. The 130 kDa monomeric BbadA shows a highly similar head structure compared to BadA of B. henselae. In contrast, BbadB and the much shorter BbadC have more in common with the VompA–C of B. quintana. The role of B. bacilliformis adhesins during the infection process remains unclear.In entified . The B. Among other candidates , 227, thBartonella henselae is the etiologic agent of cat scratch disease (CSD) and vasculoproliferative disorders. CSD is a self-limiting disease, but can be life-threatening for immunocompromised patients [B. henselae, and the role of ticks in the transmission of B. henselae to humans remains unclear [patients , 211. Ca unclear , 229.B. henselae [B. henselae infections. One possible obstacle in this research is the variable gene pool of B. henselae strains promoting antigenic variation and defining the specific immune response [Unlike research towards clinical serodiagnostic tools including immunogenic proteins of henselae –233, andhenselae –237, theresponse .B. henselae is the TAA BadA. BadA is a large (ca. 240 nm and ca. 328 kDa monomeric) outer membrane protein primarily responsible for the first interaction of the pathogen with endothelial host cells and ECM proteins  infected with henselae , 240. A henselae , 238.Brucella suis trimeric autotransporter F (BtaF) and E (BtaE) are described as a promising immunogenic target for vaccination against mucosal B. suis infections [Burkholderia pseudomallei and Burkholderia mallei, respectively, is ongoing and describes various expressed TAAs with immunogenic properties such as BPSL2063 [The fections . Other rBPSL2063  and BimABPSL2063 , 245.ahsA) of Mannheima haemolytica A1, the principal cause of bovine pneumonic pasteurellosis, promotes colonisation and subsequent infection via its ability to bind collagen and, more importantly, is suggested to be immunogenic in calves [Avibacterium paragallinarum, which can cause infectious coryza in poultry. A variable region within HMTp210 is proposed as a candidate for recombinant vaccine production [Actinobacillus pleuropneumoniae adhesin 1 (Apa1) and 2 (Apa2) are two TAAs expressed on the bacterial surface of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. The main functional head domain, Apa2H1, activates dendritic cells and provides effective protection in mice against lethal infections with A. pleuropneumoniae by both reducing bacterial colonisation and dissemination [Finally, vaccine development against animal pathogenic bacteria expressing TAAs is of high veterinary importance. For example, AhsA units may be that research on TAAs itself is still fairly new, especially research towards their applicability as potential vaccines. The current trend to use OMVs in vaccines (as in 4CMenB) and to apply reverse vaccinology to identify new vaccine antigens might give a boost for the usage of TAAs as vaccine antigens.H. influenzae, DsrA of H. ducreyi, Ata of A. baumannii, UpaG of uropathogenic E. coli and EhaG of enterohaemorrhagic E. coli. TAAs that are no longer of major interest as vaccine targets are NhhA of N. meningitidis and UspA1 and UspA2 of M. catarrhalis due to their irregular expression patterns and high degree of diversity, respectively. Other TAAs, including YadA of Y. enterocolitica, Saa of Shiga toxin-producing E. coli, BbadA–C of B. bacilliformis, and VompA and VompB of B. quintana show promising results as potential future vaccine candidates. In conclusion, more extensive research bringing more insights in the functionality and effectiveness of TAAs as vaccines is necessary.TAAs showing the highest potential as vaccine targets are Hia of


Characteristics of wild peas and their habitats at the periphery of the range are interesting with respect to their potential importance for pre-breeding programs aimed at selection for different environmental conditions. However, wild pea diversity in peripheral regions is insufficiently represented in the existing germplasm collections. In such regions, wild pea populations are rare, small in size and suffer from climatic change and land exploitation, hence their focused search is strongly desirable. A two-week-long expedition to Iran in May 2017 revealed two small populations of the wild pea (Pisum sativum subsp. elatius) in the Zagros Mts, in Aligudarz and Khorramabad Districts of Lorestan Province, Iran, at elevations of 1841 and 1971 m a.s.l., respectively. Their habitats are briefly described. Two pea accessions derived from them, CE9 and CE10, were characterised for some visible and molecular characters. These peas appeared to belong to the evolutionary lineage B, recognised by us earlier in P. sativum as opposed to the so-called lineage AC. They contain a unique non-conservative substitution in subtype 5 of histone H1 and turned to be most related to some wild pea accessions originating from southern and south-eastern Turkey and Golan Heights. Scarce information available on wild pea occurrence in Iran suggests their existence in the south-western principal slope of Zagros Mts and southern principal slopes of Elborz and Kopet Dagh Mts. It was found that wild peas representing the evolutionary lineage B produce poorly open and poorly coloured flowers (as reported by us earlier) only in the greenhouse conditions but normally pigmented and open flowers in the wild and mesh houses at open air in Israel. Some issues of pea taxonomy are discussed. The pea (Pisum sativum L.) is an important crop of higherlatitudes useful as vegetable, grain, fodder and natural soilfertiliser in crop rotation and was among the founder crops firstdomesticated in the Near East in the course of the so-called‘Neolithic revolution’ . The cultivated pea was the first genetic object andhas accumulated enormous genetic and phenotypic variation. At the sametime, representatives of the same species, P. sativum, still existin the wild, enjoying a broad range in the Mediterranean in thebroad sense, stretching from Portugal in the west (ca 9° W) toTurkmenistan in the east (ca 60°30′ E) and from Normandy inthe north (48°44′ N) to Sinai in the south (ca 34° N).Taxonomical attribution of wild representatives of P. sativumwas equivocal until some temporary stabilisation undera compromise system by Maxted and Ambrose (2001), wholumped all them under subspecies P. sativum L. subsp. elatius(Bieb.) Aschers. & Graebn. in a broad sense, defined solely bythe fact of being wild and thus inevitably paraphyletic sincethe cultivated pea, P. sativum L. subsp. sativum, was derivedfrom a wild representative of the same species. This treatment,however, disregards the fact that according to the rulesof the botanical nomenclature ,the correct name of wild peas in a subspecies rank should beP. sativum L. subsp. biflorum (Rafin.) Soldano rather thanP. sativum subsp. elatius . Moreover, the comprehensivemolecular phylogenetic analysis of the tribe Fabeaeby Schaefer et al. (2012) suggested that the genera Pisum L.and Vavilovia A. Fed. form a branch inside the genus LathyrusL. so making the latter paraphyletic. As a consequence,Coulot and Rabaute (2016) made an attempt to revise theFabeae taxonomy to make it phylogenetically consistent and,in particular, downgraded Pisum to the section Lophotropis(Jaubert et Spach) H. Schaefer, Coulot et Rabaute of the genusLathyrus L. Kosterin (2017) pointed out that the section nameLophotropis was incorrect and corrected its name to Lathyrussectio Pisum (L.) Kosterin, which was accepted by Coulot andRabaute (2017). Although phylogenetically consistent, thisrevised system ,where the pea gets the name Lathyrus oleraceus Lamarckand its wild representatives the name L. oleraceus Lamarcksubsp. biflorus (Rafinesque) H. Schaefer, Coulot et Rabaute,is practically inconvenient as downgrading the small generaPisum L. and Lens L. which contained such important cropsas pea and lentil, respectively. Hence scholars whose interestto these two groups is motivated, at least to some extent, bypractical agricultural aspects are either reluctant to adopt soradically revised a system or, more frequently, have no idea ofit. In this paper we will keep to the habitual although somewhatoutdated system by Maxted & Ambrose (2001) and denotewild representatives as P. sativum subsp. elatius.As crop wild relatives, wild peas are practically importantas a source of genetic diversity potentially valuable for peabreeding, first of all genes for resistance to various pests,diseases and draught . No doubt, any informationon the wild pea natural populations and theirhabitats is of importance, both theoretical and practical, i. e.for preliminary conjectures about selectively useful genes ina given population.In spite of the great natural range of wild peas, in mostparts of it they are rare plants with small populations , strongly affected by sheep and goat grazing and,supposedly, by global warming . Informationon their habitat and ecology is scattered over local floraswhere it is provided in few general words at most. At the sametime, it may be useful for at least a preliminary evaluationof usefulness of particular wild pea stocks for pre-breedingfocused on certain traits. Two works from Israel, nearly thecore of diversity of wild peas and the country where they areperhaps most common, contain more detailed information.Ben-Ze’ev and Zohary (1973) provided information on thehabitat from where each wild pea accession involved intotheir study originated. Abbo et al. (2008) provided most detailedinformation on Israeli habitats of wild peas includingthe rock and soil types. Zlatcović et al. (2010) characterisedthe habitat and population of P. sativum subsp. elatius at thePčinja River in SE Serbia.We found populations of the wild pea subspecies P. sativumsubsp. elatius in a number of regions at the periphery of itsrange. For the time being these are, from west to east, Portugal,Crimea, the Caucasus within Krasnodarskiy Kray, and Iran.The information on the wild pea habitat and population foundin NE Portugal is published in ‘Materials and methods’ inZaytseva et al. (2015) and a photo of a withered pea plant ofthat population is published in Kosterin . Thispaper concerns the wild pea findings in the Zagros Mts in Iran.Populations of crop wild relatives from Zagros are of specialinterest since these mountains are considered the eastern partof the so-called Fertile Crescent, the area of origin of the‘Neolithic Revolution’ in the Near East .On May 18–31, 2017, the first author had an opportunity tojoin a dipterological expedition to Iran focused at long-leggedflies (Dolichopodidae) by Igor Y. Grichanov from All-RussianInstitute of Plant Protection, Saint-Petersburg, Russia, andAzam Ahmadi from Baran Plant Protection Institute, Arak,Ostan-e Markazi, Iran. The entomological results were publishedin Grichanov et al. (2017) and Kosterin, Ahmadi (2018);the latter source contains detailed descriptions of the localitiesexamined.Search for wild pea populations. The expedition was basedat Arak City, the capital of Markazi  Ostan (province) and visited Markazi, Lorestan and Esfahan Ostans. In total,33 localities were examined, mostly associated with runningwater . Four times, onMay 23, 25, 26 and 31, the first author had an opportunity tovisit Lorestan and to examine the first outposts of the Mediterraneanvegetation in the valleys of the Higher Zagros (thepeaks of which were still covered by snow). The Silakhor Plainlying north-west of the Higher Zagros is in its rain shadow andboth the valley itself (except for the floodplains of few rivers)and the bordering mountain slopes lack natural arboreal vegetation(only that associated with human activity is present).At the same time, in the upper parts of the valleys dividingthe south-western slope of the Higher Zagros at ca 2000 mabove sea level (a.s.l.), an open stand of the Persian Oak(Quercus brantii Lindl.) appears, with the participation of theMontpellier Maple (Acer monspessulanum L.), Christ’s Thorn and Prunus sp. At lower levelsof those valleys this oak parkland covers the slopes entirely.The first author examined four valleys and found wild peapopulations in two of them.Plant growing and derivation of wild pea accessions.Seeds collected in nature in two populations found in Iran  weresowed in the greenhouse in autumn generation (October–December)2017. One plant from each of these two populationswas chosen, their progenies were propagated in the samegreenhouse in spring generation (February–May) 2018, andgave rise to accessions CE9 and CE10, respectively.The prefix ‘CE’ was at first introduced by us for wild peaaccessions derived from wild populations found in 1991 inCrimea , abbreviated from ‘Crimeanelatius’. It appeared convenient to adopt it for theentire collection of confirmed wild peas of our Laboratoryof Geneticsand Evolution of Legumes at ICG SB RAS withits meaning reconsidered as certa exempla, that means ‘true,reliable specimens’ in Latin. , with a synonymJI3557 in John Innes Centre collection, gets in thissystem the accession number CE11.) This collection is a partof GENAGRO collection at this institute.Molecular procedures. DNA isolation, PCR and CAPSanalysis of the plastid gene rbcL and mitochondrial gene cox1were carried out as described in Kosterin, Bogdanova (2008).The nuclear gene His5 of histone H1 subtype 5 was sequencedaccording to Zaytseva et al. (2012); the plastid spacer psbAtrnH,according to Zaytseva et al. (2017).DNA sequences obtained in these works are stored in publicdatabases with the following accession numbers: MK933283,MK933284 (psbA-trnH spacer), MK952766Natural populations of wild pea(Pisum sativum subsp. elatius s. l.) in ZagrosA wild pea population was found on May 23, 2017 in Iran,Ostan-e [Province of] Lorestan, Shakhrestan-e [County of]Aligudarz, Bakhsh-e [District of] Besharat, 700 m N of thecentre of Kagelestan-e Bar Aftab village, at 33°02′13″ N,49º39′23″ E, 1841 m a.s.l. The wild peas were found in thelower part of the steep NW slope of the left (opposite to thevillage) board of the valley of the Rudbar-e Aligudarz River(a Dez River tributary). The slope had large rock (supposedlydolomite) outcrops and was covered with annual Gramineavegetation and sparse bushes of a wild almond Prunus scopariaSchneider . About 40–50 plants were foundon an area ca 10 × 10 m at the bases of spiny almond bushesseemingly protecting them from being grazed by cattle, thepaths of which were numerous on that slope. The plants wereat the final vegetation stage, with the vegetative parts withered,pods ripen, about half of them dried out and about quarter ofthem dehisced . In total 193 seeds were collected, 48 ofwhich later appeared to be infested by the pea weevil (Bruchuspisorum L.). One of those seeds gave rise to accession CE9(= W6 56889 in the USDA GRIN).In the same habitat and also at the almond bush basesanothercrop wild (distant) relative, Cicer anatolicum Alef. occurred,at the stage of flowering and young pods. The second wild pea population was found on May 31,2017 in Iran, Ostan-e Lorestan, Shakhrestan-e Khorramabad, Bakhsh-e Papi, 6.7 km NW of Istgah-e Bisheh village(broadly known as simply Bisheh), 33°22′12″ N, 48°49′34″ E,1971 m a.s.l. , in arocky dell with a stony/detritous bottom with rock outcrops,its upper part becomes a small gorge between large cliffs. The vegetation was dry Persian Oak stand; annualPoaceae, already withered, predominated in the grass layer.Wild pea plants occurred on the detritous bottom and at thebase of the rocky right slope of the dell, in a stripe ca 110 mlong but not more than 10 m wide; they alternate with plantsof some perennial vetch (Vicia sp.). Not less than a hundredplants were found. They were completely withered ,with all normally developed pods already open, with onaverage one seed per plant found captured in the rolled podwalls . Only some pods which dried underdeveloped,had the walls not opened. Also about a dozenof ripen but not yet dried pods were found, bearing traces ofpea weevil eggs, traces of burrowed young larvae (up to 20 perpod), and solitary neoplastic pustules caused by the Np gene and being the plant’s defense reaction to weevil oviposition . In total 87 seeds were collected, ofthem 24 appeared infested by the pea weevil. One of those seeds gave rise to accessionCE10 (= W6 56890 in the USDA GRIN).It should be noted that on May 25, 2017 in the environs of Hayan village (Shakhrestan-e Borujerd), also in Lorestanbut at the NE foothills of the Inner Zagros Range facing the Silakhor Plain, in aroadside herbaceous vegetation under a stripe of poplars, the first author foundseveral plants of obviously feral peas escaped from cultivation. They had two small non-dehiscing pods per inflorescence, seeds with anthocyanincoloration (phenotype Fs), marblepattern (phenotype M), and non-grittytesta (phenotype gty). Locals told thatin that place peas had been grown forfodder 7–8 years ago.Characters of wild peas from ZagrosWe scored accessions CE9 and CE10for some molecular characters involvedin our previous studies. Both lackedthe recognition sites for HspAI endonucleasein the plastidic rbcL and thesite for PsiI restriction endonucleasein the mitochondrial cox1 gene. Thesequence of psbA-trnH plastidic spacerof CE9 was identical to its consensusin peas  whilethat of CE10 had a substitution A→Tin position 128. The sequences of theHis5 gene coding for histone H1 subtype5 obtained from CE9 and CE10were compared to those obtained in thecourse of our earlier works . The His5 sequences in CE9 andCE10 appeared identical to each otherand most close to those of JI1794 (GolanHeights) and P012  ,differing from them only by an A→Csubstitution in position 452 resulting ina non-conservative substitution of lysineto threonine in the globular domain ofthe molecule (protein position 111). Thissubstitution was not found in any otherpea accession. One more His5 sequencevery close to the above mentioned, thatfrom accession JI3233 (Syria), differedfrom them by the T→C substitution inposition 722 leading to the valine→alanine amino acid substitution in theC-terminal domain.The seeds of both accessions CE9and CE10  have gritty testa, blackhilum (Pl), a brownish marble pattern(M), violet specks (Fs), no furcapattern (rf). Those of CE9 in additionhave conspicuous violet stripes (Ust). The young seed groundcolour is rather pale greenish-grey inCE9  and brownish-greyin CE10 ; the shape isslightly irregular, not perfectly sphaeric.If scarified and sowed after 1–2 monthafter formation, they readily germinated,earlier than most other peas.The plants found in the Kagelestan-e Bar Aftab population(CE9) in nature were up to 70 cm in height, those in theBisheh population (CE10) only up to 40 cm. In the intraspeciestaxonomy of wild representatives of P. sativum used in thepast and based on the plant height, they would be classifiedas ‘Pisum humile’  or P. sativumsubsp. syriacum Berger . In the conditionsof the greenhouse spring vegetation the difference in plantheight retained but had a lesser magnitude of ca 30 %: theCE9 plants were 69–122 cm tall ,flowered from 14th–19th node (mean 16.7 ± 1.0) and totallyhad 18–23 nodes on the main stem (mean 21.0 ± 1.2); the sameparameters of CE10 had the following values: 40–94 , 14–18 (mean 15.7 ± 1.0) and 17–21 (mean19.2 ± 1.1), respectively. The plants were elegant, with longinternodes, narrow stipulae only moderately dentate at base,rather narrow rhomboid-oval very slightly dentate leaflets(larger in CE10), had numerous aerial cameras on both stipulaeand leaflets and outer anthocyanin rings (but no inner rings)at the base of stipulae . Besides, CE9 (but not CE10) had tinybut conspicuous violet specks on leaflets . The plantsmoderately branched at the base and the main stem belowflowering nodes, had medium-long peduncles, with one or two(at the middle of the range of flowering nodes) flowers. Thepods dehisced explosively upon ripening ; their walls bore sparse and small neoplasticpustules caused by the Np gene in the greenhouse conditions(while in the wild as a response to the weevil oviposition).The flowers of both Zagros accessions grown in the greenhousenever opened fully and were greenish .At the same time in the natural-like conditions of an outdoormesh house at Agronomy Faculty of the Hebrew Universityof Jerusalem in Rehovot, Israel, these accessions providedlarge, fully open and well coloured flowers .Wild peas in IranMost part of the huge territory of Iran is in the rain shadow ofZagros Mts, which captures precipitation from the Mediterranean,and hence is too arid for wild peas. The Iranian Plateauand the north-eastern (inner) principal slope of Zagros havemostly Irano-Turanian rather than Mediterranean vegetation,mostly its desert versions . The Mediterraneanvegetation and flora, to which wild peas belong, is presentonly in the sea-facing outer principal slopes of the Iranianmountain systems: the south-western slope of Zagros Mtsand the northern slope of Elborz Mts. These are the regionswhere wild pea populations should be sought for.  parkland extendingto ca 2000 m a.s.l., while in the north-eastern part any naturalarboreal vegetation is missing even as low as at 1700 m a.s.l.)The presence of wild peas in the Iranian southern slope ofKopet Dagh is also possible but is probably marginal in thoserather hostile arid mountains.Reports of wild peas from Iran are found in two multivolumedFloras: “Flora Iranica” published in Latin/Englishand “Flora of Iran” published in Farsi; curiously, data in thesereports do not overlap. The volume devoted to the tribe Vicieae(currently Fabeae)  contains seven locationsfor P. sativum subsp. elatius: three in Lorestan in westernIran  andfour in northern Iran, one in Gorgan Province (now GolestanProvince) (Ziarat env.) and three in Gilan Province ; Lake Mordab westerly ofBandar-e Pahlavi, 26 m a.s.l.; Astara). The 33th volume of“Flora of Iran”  contains differentlocalities for the same taxon: one in Ostan-e Kurdistan , three in Ostan-e Azerbaijan , one inOstan-e Golestan  and two in Ostan-eFars . Occurrence ofwild peas as southerly as Fars, if true, was unexpected.Both sources do not report for Iran P. sativum subsp. elatiusvar. pumilio Meikle or its synonyms. At the same time the mapin Maxted, Kell  shows in western Iran fivelocalities of P. sativum subsp. elatius var. pumilio but none ofP. sativum subsp. elatius var. elatius. As the source of informationfor that map an unpublished thesis by A.S. Mumtazis indicated, which was defended in 2005 in BirminghamUniversity. Most probably here we face an equivocal treatmentof intra-species taxonomy of P. sativum by different authors.World germplasm collections hitherto contained only fouraccessions claimed to represent wild peas from the huge territoryof Iran. Accession IG65050  is from ICARDA collection and originated formZagros Mts. The coordinates adduced refer to a south-westernslope at 1800 m a.s.l. in the northern environs of Beyravand-e Jonubi village in Khorramabad Shahrestan. AccessionPI143673 was derived from plants collected in 1940 in Dorud,Lorestan. For accession JI1030 (=PI140295), coordinates34° N and 56° E and provenance “Khorassan” were indicated,but the coordinates are most probably erroneous as referringto the low and deserted Tebess Mts between Dash-e Kavir andDasht-e Lut Deserts in South Khorasan Province.  For accession JI2105 only coordinates 32.659° N,51.671° E are provided by the online database of John InnesCentre collection; they refer to the Esfahan City environs.However, we grew out the latter accession and found it torepresent a cultivated pea with non-dehiscing pods . The two untested accessions from Lorestanperhaps represent true wild peas. At least their provenance is close to our findings: CE9 was collected 120 km SE and CE1043 km SE of Beyravand-e Jonubi, the presumed provenanceof IG65050.Thus, wild peas are reliably known in Iran from the southwesternprincipal slope of Zagros and the western and easternparts of Elborz, although in general Iran plus Turkmenistanoccupy about one third of the Pisum natural range by longitude.It is broadly accepted that in the western Eurasia,productive farming arose and plant domestication occurred inthe so-called Tauro-Zagros Arch or Fertile Crescent, a mountainbelt including Golan Heights, Taurus and Anti-TaurusRanges and Zagros Mts . Formally theZagros Mts comprise about one third of the Arch and mostof these mountains are in the territory of Iran, from whichthey only protrude to the Iraqi Kurdistan. However, only thesouth-western principal slope of Zagros has the Mediterraneanvegetation and can be attributed to the Fertile Crescent. Basedon literature, the southern border of the wild pea range in Irancan be extrapolated to cross Ostan-e Fars.Wild pea habitatsThe CE10 locality is 75 km NW and the CE9 locality47 km SE of the Oshtorankuh Mountain (4050 m a.s.l.), thehighest summit of Lorestan. At the same time, the mountainssituated to the south and south-east of these localities do notexceed 2000–2500 m a.s.l. Thus the wild pea habitats foundare not situated in the main rain shadow of the Higher Zagros,that is the reason of the appearance of arboreal vegetationof the Mediterranean type, which is absent to the north andnorth-east of them at any elevations. One can note that all wildpea findings in Lorestan were made at close elevations a. s. l.:2100 m , 1971 m, 1841 m (our findings)and 1800 m (reconstructed from coordinates of accessionIG65050). The former value refers to the upper limit of arborealvegetation in this area.Occurrence of wild peas in oak parkland was expectablesince this is one of the primary habitats of at least some wildpea ecotypes elsewhere, e. g. in Israel . It remains unclear if a slope withspiny almond bushes is a regular habitat of wild peas (andC. anatolicum) in Iran since the first author failed to examinemore examples of such habitats.The analysis of regional literature suggests that wild peasare associated with calcareous habitats on limestone or dolomitesthroughout their vast range but also with igneous rocksand volcanic slag in Israel  and East Turkey . The HigherZagros is composed mostly by the Mesozoic dolomites thatconforms the notion of predominant calciphily of P. sativumsubsp. sativum.Relationships of Zagros wild peasSuch characters as dehiscing pods (Dpo) and gritty seedtesta (Gty) evidence that both small populations found amongMediterranean vegetation represent genuine wild peas. Absenceof the target restriction sites in the plastidic rbcL andmitochondrial cox1 genes  inCE9 and CE10 suggests their belonging to the so-calledevolutionary lineage B of wild P. sativum. This lineage wasrevealed by us earlier, as a monophyletic clade opposed to the so-called lineage AC, in the phylogenetic reconstruction basedon the histone H1 genes and plastid genomes  and is identifiableby convenient molecular markers from different cellulargenomes .The lineage B includes a great number of wild peas as well asthe cultivated pea subspecies (P. sativum subsp. sativum). Atthe same time, as expected for wild peas, CE9 and CE10 hadno 7-bp deletion in the plastid psbA-trnH spacer which, withonly two known exceptions, is specific to (a synapomorphyof) the cultivated subspecies . The A→Tsubstitution in position 128 of this spacer in CE10 is sharedby wild pea accessions JI1794 (Golan Heights) and P017 ). The His5 gene ofboth CE9 and CE10 contains the same nucleotide substitutionresulting in the lysine→threonine replacement in the globulardomain not found in any other pea accession; otherwise theirHis5 sequence is identical to those of accessions JI1794 andP012 . One more very closeHis5 sequence, differing from the above mentioned ones inone substitution, belongs to accession JI3233 (Syria).We may conclude that the wild peas found by us in Zagrosrepresent some subtle evolutionary branch of the lineage Balso occurring at least in southern and southeastern Turkey,Golan Heights and Syria. It is noteworthy that JI1794 is a lowplant, P017, CE9 and CE10 are moderately high and JI3233 isa high plant, which once again stresses the inapplicability ofplant height to evaluate relatedness of wild peas . Also some genetic difference already foundbetween CE9 and CE10 originating from the populations85 km apart is noteworthy: the substitution in position 128 ofpsbA-trnH in CE10 and the violet stripes (Ust) on the seed testa,violet specks on leaflets and a greater plant height in CE9.Flowers of peas of evolutionary lineage BZaytseva et al. (2017) claimed that most of wild peas of theevolutionary lineage B have flowers poorly pigmented andopened, in contrast to well open and coloured flowers of cultivatedpeas, belonging to the same lineage, and wild peas ofthe ‘evolutionary lineage AC’. It turned out that this statementis true only of our greenhouse conditions. Accessions CE9and CE10 grown in our greenhouse and in the mesh house in Rehovot, where the conditions were close to natural, have,respectively, poorly open greenish  versuswell open, coloured and large  flowers.Analogously, the photos taken by S.A. Litvinskaya (pers.comm.) of wild peas of the lineage B in two populations at theBlack Sea Coast of Krasnodarskiy Kray, Russia, show large,well open and coloured flowers, while the flowers of the plantsgrown in our greenhouse from seeds from these populationsare poorly opened and coloured, as in the case of Iranianwild peas. Finally, accession JI1794 originating from the TellAbu Nida Hill, Golan Heights, northern Israel, also producedpoorly open greenish flowers in the greenhouse ,while plants in the natural habitat at Tell Abu Nida have wellopened and strikingly coloured flowers, deep purple includingthe standard, most saturated among peas . , Pe 013 (Turkey) and sometimesalso JI1096 (Greece) and JI3557 .)The factor that prevents the normal development of theflower corolla of wild representatives of the linage B (butnot affecting the pods) in our greenhouse is still unclear.Our preliminary experiment ruled out the involvement of thetemperature of germination. An edaphic factor can also beexcluded, since the Black Sea natural habitats of wild peasare on limestone while the Tel Abu Nida habitat is on thebasalt and volcanic ash. For the time being the temperatureand illumination regime at the onset of flowering is the mostprobable candidate.In this paper we report findings of two wild pea populationin the Zagros Mts comprising the eastern part of the FertileCrescent which is considered to be an area where the foundercrops were domesticated and the productive farming appearedin the Near East. Indeed, these populations represented thesame evolutionary lineage of the species P. sativum to whichthe cultivated pea also belongs. However, another result of thisstudy is the observed paucity of wild peas in these mountainsand their occurrence only in their south-western principalslopes which offers suitable habitats. This is in contrast totheir frequent occurrence and diversity in the western (e. g.Israel) and northern (SE Turkey) parts of the Fertile Crescent.The authors declare no conflict of interest.Abbo S., Lev-Yadun S., Heun M. Gopher A. On the ‘lost crops’ of theneolithic Near East. J. Exp. Bot. 2013;64:815-822. DOI 10.1093/jxb/ers373.Abbo S., Zesak I., Schwartz E., Lev-Yadun S., Gopher A. Experimentalharvesting of wild peas in Israel: implications for the origins of NearEast farming. J. Archaeol. Sci. 2008;35:922-929. DOI 10.1016/j.jas.2007.06.016.Ben-Ze’ev N., Zohary D. Species relationship in the genus Pisum L.Israel J. Botany. 1973;2:73-91.Berdnikov V.A., Trusov Y.A., Bogdanova V.S., Kosterin O.E., RozovS.M., Nedel’kina S.V., Nikulina Y.N. The neoplastic podgene (Np) may be a factor for resistance in pea to the pest Bruchuspisorum L. Pisum Genetics. 1992;24:37-39.Blixt S. Mutation genetics in Pisum. Agri Hortique Genetica. 1972;30:1-294.Bogdanova V.S., Mglinets A.V., Shatskaya N.V., Kosterin O.E., SolovyevV.I., Vasiliev G.V. Cryptic divergences in the genus Pisum L.(peas), as revealed by phylogenetic analysis of plastid genomes.Mol. Phylogenet. Evol. 2018;129:280-290.Coulot P., Rabaute P. Monographie de Leguminosae de France. 4. Tribusdes Fabeae, des Cicereae et des Genisteae. Bulletin de la SociétéBotanique du Centre-Ouest. 2016;46:1-902.Coulot P., Rabaute P. Deuxièmes compléments à la Monographie desLeguminosae de France. Le Monde des Plantes. 2017;516:11-35.Coyne C.J., McGee R.J., Redden R.J., Ambrose M.J., Furman B.J., MilesC.A. Genetic adjustment to changing climate: pea. In: Yadav S.S.,Redden J.H., Lotze-Campen H., Hall A.J. (Eds.). Crop Adaptation toClimate Change. New York: John Wiley & Sons, 2011;238-250.Grichanov I.Y., Ahmadi A., Kosterin O.E. New records of long-leggedflies  from Central and North-EasternIran. Acta Biologica Sibirica. 2017;3(4):99-112.International Code of Nomenclature for algae, fungi, and plants (MelbourneCode). Oberreifenberg: Koeltz Scientific Books, 2012.Kosterin O.E. Under the reign of the Pea King :1-14.DOI 10.1134/S2079059716010081.Kosterin O.E. Prospects of the use of wild relatives for pea breeding.Russ. J. Genet.: Appl. Res. 2016b;6(3):233-243. DOI 10.1134/S2079059716030047.Kosterin O.E. Abyssinian pea (Lathyrus schaeferi Kosterin nom. nov.pro Pisum abyssinicum A. Br.) is a problematic taxon. VavilovskiiZhurnal Genetiki i Selektsii = Vavilov Journal of Genetics andBreeding. 2017;21(2):158-169. DOI 10.18699/VJ17.234. (in Russian).Kosterin O.E., Ahmadi A. Odonata observed in Central Zagros, Iran,in late May 2017. International Dragonfly Fund Report. 2018;117:1-65.Kosterin O.E., Bogdanova V.S. Relationship of wild and cultivatedforms of Pisum L. as inferred from an analysis of three markers, ofthe plastid, mitochondrial and nuclear genomes. Genet. Res. CropEvol. 2008;55:735-755. DOI 10.1007/s10722-007-9281-y.Kosterin O.E., Zaytseva O.O., Bogdanova V.S., Ambrose M. New dataon three molecular markers from different cellular genomes in Mediterraneanaccessions reveal new insights into phylogeography ofPisum sativum L. subsp. elatuis (Beib.) Schmahl. Genet. Res. CropEvol. 2010;57:733-739. 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Hyriposis species, H. schlegelii and H. cumingii  from Lake Kasumigaura, Japan, and inferred the Unioninae phylogeny. Complete mitochondrial genomes  contain 13 protein-coding genes (PCGs), 2 rRNA genes, and 22 tRNA genes. Molecular phylogenetic analyses using the 13 PCGs including the two species were performed. This study should be basic data to investigate the evolution of Gonideinae and genetic diversity of Hyriposis species in local populations.We have sequenced the female type (F-type) complete mitochondrial genomes of two  Hyriopsis schlegelii is an endangered species, endemic to Lake Biwa, Japan. Since the 1930s, it was transplanted to Lake Kasumigaura, Japan. Hyriopsis cumingii inhabits in China, and produces freshwater pearls  were collected at Lake Kasumigaura, Japan . DNA extraction and sequence analyses were performed as described by Fukata and Iigo (H. schlegelii (AP018551) and H. cumingii (HM347668) identified the F-type complete mitochondrial genomes of the two species . The complete mitogenomes contain 13 protein-coding genes (13PCGs), 2 rRNA genes, and 22 tRNAs. The sequences have been submitted to DDBJ/EMBL/Genbank with accession numbers LC498622 (H. schlegelii) and LC498621 (H. cumingii).and Iigo . Local BH. cumingii) might be H. schlegelii; NC_015110 and HQ641407 (both registered as H. schlegelii) might be the F-type mitogenomes of a new Hyriposis species .Male and female mitogenomes of Gonideinae are known to have different gene orders (Breton et al. Hyriposis species.This research would provide valuable information for investigating the evolution of Gonideinae and the genetic diversity of local populations of


Sargentodoxa cuneata is used as traditional Chinese medicine. In this study, we report its complete chloroplast genome by Illumina pair-end sequencing. The total chloroplast (cp) genome size was 158,094 bp in length, containing a pair of inverted repeats of 26,132 bp, separated by large single-copy and small single-copy regions of 86,508 bp and 19,322 bp, respectively. The chloroplast genome of S. cuneata encodes 113 different genes, including 79 protein-coding genes, 30 transfer RNAs, and 4 ribosomal RNAs. A total of 84 perfect chloroplast microsatellites were analyzed in the S. cuneata. The majority of the SSRs in this chloroplast genome are mononucleotides (66.67%). The reconstructed phylogeny revealed that S. cuneata was sister to the remaining Lardizabalaceae. Sargentodoxa, a monotypic genus of the Lardizabalaceae has often been placed in its own family, Sargentodoxaceae  Rehder and E. H. Wilson, mostly confined to subtropical China . Wild resources of this species have been seriously deteriorated due to years of over-harvesting, indicating an urgent need for reasonable conservation strategies. It is necessary to develop genomic resources for S. cuneata to provide intragenic information for its utilization and conservation. Because of the small and relatively constant size, conserved genome structure, and uniparental inheritance, chloroplast genomes provide a valuable genetic resource for phylogenetic analysis and species conservation . The voucher specimen (Specimen accession number: HNRC010701) was deposited at the herbarium of the National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences. Total genomic DNA was extracted using mCTAB protocol , and separated large single-copy  and small single-copy  regions. The GC content of the chloroplast DNA is 38.2%. The chloroplast genome of S. cuneata encodes 113 different genes, including 79 protein-coding genes, 30 transfer RNAs (tRNA), and 4 ribosomal RNAs (rRNA). Fifteen distinct genes contain a single intron while ycf3 and clpP each contain two. Simple sequence repeats in the S. cuneata chloroplast genomes were detected using GMAT (Wang and Wang S. cuneata . The majority of the SSRs in this chloroplast genome were mononucleotides (66.67%) and almost all of the mononucleotides (88.09%) are composed of A/T.The size of the chloroplast genome of S. cuneata with other Lardizabalaceae species. Phylogenetic analysis was performed using the whole chloroplast genome sequence. The chloroplast genome sequences were aligned using MAFFT v7 (Katoh and Standley S. cuneata was sister to the remaining Lardizabalaceae (To estimate phylogenetic relationships of balaceae . The who


Phascolarctobacterium faecium is a strict anaerobe belonging to the Firmicutes phylum that is found abundantly in the human gastrointestinal tract. Here, we report the complete genome sequence of P. faecium G 104, a strain isolated from a fresh stool sample from a healthy lean donor. Phascolarctobacterium faecium is a strict anaerobe belonging to the Firmicutes phylum that is found abundantly in the human gastrointestinal tract. Here, we report the complete genome sequence of P. faecium G 104, a strain isolated from a fresh stool sample from a healthy lean donor. Phascolarctobacterium faecium is a Gram-negative, nonsporulating, and strictly anaerobic bacterium that was first isolated from a healthy human donor  . The single-molecule real-time (SMRT) Link software v.6.0.0 was used to filter and assemble the subreads. The HGAP4 algorithm (https://github.com/sanger-pathogens/circlator) was used for circularization and removal of the overlaps, as described in the PacBio assembly sequencing pipeline  fermentation into butyrate and three genes encoding a bile acid sodium symporter and bile acid-inducible protein F were detected but not functionally determined.an donor  and was an donor . This baan donor  and posian donor  and to lan donor . P. faecum (IBM)  supplemelgorithm  was usedlgorithm  (https:/pipeline . ContigsCM 30894 . The genP. faecium G 104 was deposited in GenBank under accession number CP061002.1, BioProject accession number PRJNA659610, BioSample accession number SAMN15921107, and SRA accession number SRR13005647.The complete genome sequence of


Helicobacter pylori and reflux esophagitis (RE) remains controversial. This study aimed to prospectively evaluate the effect of H. pylori eradication on RE and gastroesophageal reflux (GERD) symptoms in H. pylori-positive patients who underwent endoscopic resection of gastric neoplasm.The association between H. pylori-positive patients underwent follow-up at least once. We evaluated the prevalence of RE and GERD symptoms in these patients following H. pylori eradication.Of the 244 patients enrolled in this study, 173 H. pylori did not increase the incidence of RE . H. pylori status was also not associated with the development of GERD symptoms . In the univariate analysis for RE, present smoking history , present alcohol consumption history , and diabetes mellitus  were found to be associated with RE. Multivariate analysis showed that present smoking history  was a significant risk factor for RE.There were 75.7% (131/173), 78.6% (125/159), and 78.9% (105/133) subjects who were successfully eradicated after 6, 12, and 18–24 months, respectively. During the 2-year follow-up period, the eradication of H. pylori eradication did not increase the incidence of RE or GERD symptoms in patients who underwent endoscopic resection of gastric neoplasm. Helicobacter pylori plays a key role in the pathogenesis of various diseases such as peptic ulcers, gastric mucosa-associated tissue lymphoma, and intestinal-type gastric cancer. Since Schutze et al. first mentioned in 1995 that reflux esophagitis (RE) was developed after H. pylori eradication [H. pylori with RE or gastroesophageal reflux (GERD) symptoms have been reported. Reports have shown that H. pylori infection is negatively associated with erosive esophagitis and Barrett’s esophagus but not with GERD symptoms [symptoms –5.H. pylori eradication significantly lowers the incidence of metachronous lesion in patients who underwent endoscopic resection of early gastric cancer, we evaluated the effect of H. pylori eradication on RE and GERD symptoms in a large cohort of patients who underwent endoscopic resection of gastric epithelial neoplasm and had H. pylori infection. These patients, who are less likely to lose follow-up, are relatively homogenous. The aim of our study was to evaluate the effect of H. pylori eradication on RE and GERD symptoms in a large cohort of patients who underwent endoscopic resection of gastric epithelial neoplasm and had H. pylori infection.Based on the study showing that H. pylori-negative, (2) presence of upper gastrointestinal bleeding, (3) presence of esophageal or gastric varices, (4) presence of esophageal or pyloric stenosis, (5) previous history of gastrectomy, (6) history of H. pylori eradication, (7) presence of tumor at the gastroesophageal junction or pylorus resulting in outlet obstruction, (8) past or present history of taking H2 receptor blocker or proton pump inhibitor (PPI) within at least 1 month, (9) alcohol or drug abuse, and (10) being pregnant. Written informed consent was obtained from all participants. The study was conducted according to the guidelines of the Declaration of Helsinki and was approved by the Institutional Review of Board of Asan Medical Center (IRB number: 2014-2-10).We conducted a single-center prospective study. Enrolled patients were aged 20–70 years who underwent endoscopic resection including endoscopic mucosal resection and endoscopic submucosal dissection of gastric neoplasm at Asan Medical Center, Seoul, Korea, from September 2012 to March 2014. Patients were excluded on the basis of the following criteria: (1) Patients who agreed to participate in the study answered questionnaires regarding GERD symptoms initially, at 6 months, and at 12 months after endoscopic resection. The questionnaire  which weThe endoscopist was blinded to the result of the questionnaires. The presence or absence of RE was evaluated initially when the patient underwent endoscopic resection. The severity of RE on EGD was graded in accordance with the Los Angeles classification system . Minor cThe serum levels of pepsinogens I (PGI) and II (PGII) were evaluated using Pepsinogen I and Pepsinogen II EIA TEST kits . When PGI levels were < 25 μg/L and PGI to PGII ratio was < 3, serological atrophy was defined to be present , 10.H. pylori infection was determined using a rapid urease test, a 13Curea breath test, and histology. For histological confirmation, two biopsy specimens were obtained from the greater curvature of the body, and the antrum and the specimens were examined after Giemsa staining. At least one positive result of the above tests was assumed as present H. pylori infection. If the patients showed positive results for H. pylori, eradication therapy was provided. As a first-line therapy, full-dose PPI, 500-mg clarithromycin, and 1000-mg amoxicillin, twice for 7–14 days, were prescribed. 13Curea breath test or rapid urease test was performed for confirming the success of eradication therapy after at least 4 weeks from the completion of eradication therapy. In the event that the patient failed triple therapy, bismuth-based quadruple therapy was initiated that consisted of full-dose PPI b.i.d., 500-mg metronidazole t.i.d., 500-mg tetracycline q.i.d., and 120-mg bismuth t.i.d. for 7–14 days.H. pylori status using the rapid urease test.Full-dose PPI was usually prescribed for 4 weeks after endoscopic resection. Follow-up EGD was performed after 6 months for 2 years, and we evaluated the presence of RE and P-values were two-sided, and P-values of < 0.05 were considered statistically significant. For analyzing the probability of RE and GERD symptoms, generalized estimating equation with logistic regression was used. Risk factors for RE and GERD symptoms were assessed using univariate and multivariate analyses with logistic regression analysis. All statistical analyses were performed using the statistical package for the social sciences software .Continuous variables were reported as means ± standard deviations. Depending on the distribution, the two-sample t-test or the Mann–Whitney U test was used for comparing continuous variables. All H. pylori results were excluded. Therefore, we analyzed the data of 173 patients who initially showed H. pylori-positive results. Baseline characteristics of this study population are presented in Table Of the 244 patients who were enrolled during the study period, 25 patients underwent surgery, 18 patients were lost to follow-up, and 28 patients who initially showed negative At 6, 12, and 18–24 months after endoscopic resection, 173, 159, and 133 patients, respectively, were followed up. During the study period, 32 patients failed the first line eradication therapy, two patients refused for further eradication, and 30 patients underwent second line regimens. Among four patients who showed sustained positive result after the second line therapy, three underwent third line regimen .H. pylori-positive and H. pylori-negative patients is not significantly different at 6, 12, and 18–24 months after endoscopic resection. The probabilities of RE that failed to show a difference in both groups were as follows: 0.18 in H. pylori-positive vs. 0.14 in H. pylori-negative at 6 months, 0.22 vs. 0.18 at 12 months, and 0.17 vs. 0.17 at 18–24 months after endoscopic resection, respectively , 78.6% 125/159), and 78.9% (105/133) subjects who were successfully eradicated after 6, 12, and 18–24 months, respectively. The prevalence of RE and GERD symptoms during follow-up period is presented in Table 5/159, an31/173, 7P < 0.012), present smoking status, alcohol consumption, and diabetes mellitus were associated with RE. In multivariate analysis, present smoking was noted to be the only significant risk factor for RE . Otherwise, risk factors such as BMI and the presence of hiatal hernia were not associated with reflux symptoms.In univariate analysis for factors associated with reflux symptoms, H. pylori who underwent endoscopic resection of gastric neoplasm. During the follow-up period, no significant difference was noted in the development for RE and reflux symptoms between H. pylori-positive and H. pylori-negative patients. We assumed that eradication therapy did not result in the development of RE or GERD symptoms.In the present study, we evaluated the probability of RE and reflux symptoms among patients with positive H. pylori infection protects against the development of erosive esophagitis [H. pylori reduced the requirement of PPI [H. pylori eradication than that in H. pylori–positive patients [H. pylori eradication does not have a clinical impact on RE. In a meta-analysis involving 11 randomized controlled trials, no difference in erosive esophagitis and reflux symptoms (reflux and heartburn) was noted between H. pylori eradication and persistent groups. In another meta-analysis including 10 trials, no statistically significant difference was found for endoscopic evidence of RE  or symptomatic GERD  between H. pylori treatment and the no-treatment groups [H. pylori eradication was even lower than those who were never infected [It has been reported that phagitis –14. The t of PPI . In a stpatients . Howevert groups . A recenH. pylori eradication does not lead to the development of GERD symptoms [H. pylori eradication did not result in the development of new symptomatic GERD or worsening of symptoms in patients with preexisting GERD [GERD symptoms are relatively subjective and are not usually correlated with an endoscopic finding or a result from 24-h pH monitoring. Reports have demonstrated that symptoms , 18. In ing GERD .H. pylori infection is associated with gastric acid output. In the early stage of the study, we postulated that our study population who primarily had severe atrophic gastritis and acid hyposecretion state would develop RE or GERD symptom after H. pylori eradication. Actually, many patients (76.3%) showed open-type gastric atrophy and all patients except one showed atrophic changes. However, RE or GERD symptoms were not different between the two groups. In a prospective double-blind study measuring 24-h esophageal pH for established GERD patients, esophageal acid exposures were not different between H. pylori-positive and H. pylori-negative patients [H. pylori infection did not affect esophageal acid exposure [H. pylori infection, pathophysiological studies show that transient lower esophageal sphincter relaxations are the predominant mechanism of reflux [Multiple pathophysiological factors influence the development and course of RE, and gastric acid secretion is known to be a key factor in the pathophysiology of the disease. The possible underlying mechanism of patients . Anotherexposure . In patif reflux .H. pylori; male sex, BMI ≥ 25 kg/m2, the use of calcium channel blocker, and hiatal hernia were associated with the development of RE [H. pylori eradication on the prevalence of RE, male sex, higher BMI, habitual alcohol consumption, habitual smoking, larger diaphragmatic hiatal size, and milder gastric mucosal atrophy were noted to be risk factors for RE [In our study, smoking was the only risk factor for the development of RE. A recent study evaluated the risk factors for RE after the eradication of nt of RE . In anots for RE .H. pylori eradication among patients who underwent endoscopic resection of gastric neoplasm.Our study has several limitations. First, the prevalence of gastric atrophy was relatively higher in our study population and patients who have gastric neoplasm are not representative of the screened population. Second, the study was performed in a single center in Korea and ethnic variation could not be considered. Third, we did not perform functional study of gastroesophageal junction, which could have provided additional physiological evidence. Fourth, the questionnaire for GERD symptom evaluation was not validated. Despite these limitations, this is a prospective study that analyzes RE and GERD symptoms after H. pylori eradication did not increase the incidence of endoscopic RE or typical GERD symptoms in patients who underwent endoscopic resection of gastric neoplasm.Additional file 1.


The prognostic value of immune-related genes and lncRNAs in neuroblastoma has not been elucidated, especially in subgroups with different outcomes. This study aimed to explore immune-related prognostic signatures.Immune-related prognostic genes and lncRNAs were identified by univariate Cox regression analysis in the training set. The top 20 C-index genes and 17 immune-related lncRNAs were included in prognostic model construction, and random forest and the Least Absolute Shrinkage and Selection Operator (LASSO) regression algorithms were employed to select features. The risk score model was constructed and assessed using the Kaplan-Meier plot and the receiver operating characteristic curve. Functional enrichment analysis of the immune-related lncRNAs was conducted using the STRING database.In GSE49710, five immune genes  were included in the risk score five genes (RS5_G) signature, and eleven immune-related lncRNAs  were brought into risk score LncRNAs (RS_Lnc) signature. Patients were divided into high/low-risk score groups by the median. Overall survival and event/progression-free survival time were shortened in patients with high scores, both in training and validation cohorts. The same results were found in subgroups. In grouping ability assessment, the area under the curves (AUCs) in distinguishing different groups ranged from 0.737 to 0.94, better in discriminating MYCN status and high risk in training cohort (higher than 0.9). Multivariate Cox analysis demonstrated that RS5_G and RS_Lnc were the independent risk factors for overall and event/progression-free survival . Correlation analysis showed that RS5_G and RS_Lnc were negatively associated with aDC, CD8+ T cells, but positively correlated with Th2 cells. Functional enrichment analyzes demonstrated that immune-related lncRNAs are mainly enriched in cancer-related pathways and immune-related pathways.We identified the immune-related prognostic signature RS5_G and RS_Lnc. The predicting and grouping ability is close to being even better than those reported in other studies, especially in subgroups. This study provided prognostic signatures that may help clinicians to choose optimal treatment strategies and showed a new insight for NB treatment. These results need further biological experiments and clinical validation. Neuroblastoma (NB), one of the most common malignant solid tumors in children, accounts for nearly 15% of childhood cancer-related deaths . At presThanks to the development of microarrays and high-throughput technologies, an increasing number of abnormalities have been discovered in both driver and non-driver genes in NB . Unlike The tumor microenvironment (TME) has been confirmed to play an important role in various cancer types, including NB . The TMELiu Z depicted the immune gene expression profiles in high risk NB and found an ultra-high risk NB (UHR-NB) group that showed the worst prognosis. Besides, the authors identified four up-regulated immune genes  among the UHR-NB patients . HoweverThe present study aimed to explore the prognostic immune-related genes and lncRNAs, and to discuss their feasibility as prognostic and follow-up markers in NB to help clinicians optimize individual treatment protocols. This study focused on the significance of immune-related prognostic signatures in predicting outcomes for NB patients in each group.https://www.ncbi.nlm.nih.gov/gds/?term=), validation cohorts were downloaded from R2 database (https://hgserver1.amc.nl/cgi-bin/r2/main.cgi), including Neuroblastoma public - Versteeg - 88 , TARGET- Asgharzadeh - 249, and Neuroblastoma Primary - NRC - 283 . We excluded 5 patients with MYCN amplification status NA, and included 493 patients as a training cohort (n=493), the validation sets were GSE16476, TARGET – 249, and GSE85047.Neuroblastoma training set GSE49710  was downloaded from the GEO database (https://www.ncbi.nlm.nih.gov/gene/).If one gene corresponded to multiple probes, the highest expression value was taken as the representative and log2 transformed. For lncRNAs, we confirmed them as “ncRNAs” from the NCBI database (https://immport.niaid.nih.gov/), including 1534 genes, after excluding duplicates and mismatches, a total of 993 genes remained. Univariate Cox proportional model survival analysis  regression analysis  was perfhttps://string-db.org/) was performed. FDR <0.05 was considered significant.To explore the biological functions of prognostic immune genes and immune-related lncRNAs, KEGG  Kyoto E databasehttp://xcell.ucsf.edu/). The xCell scores of cell types represented a fraction of cells in tumor tissues.To describe the cell composition of the tumor microenvironment (TME) in neuroblastoma, we employed the xCell algorithm   was calculated to assess the performance of the model. The ggforest plots were generated by R package “survminer”. Kaplan-Meier algorithm was used to evaluate the survival time of high/low-risk groups, and a Log-rank test was employed. To test the predictive ability of the prognostic signature, the area under the curves (AUC) of receiver operating characteristic (ROC) was calculated and plot by R package “pROC”. The random forest algorithm was carried out by R package “randomForest”. The Lasso regression analysis was implemented by R package “glmnet”. The Spearman Correlation Coefficient was calculated by R method “cor.test ”, and the correlation heatmaps were plotted by GraphPad Prism 8.0. The Mann-Whitney U test was used to compare the xCell score of cells in different groups. All statistical tests were two-sided and a p-value < 0.05 was considered statistically significant.Univariate Cox regression analysis was performed in the GSE49710 dataset. Altogether 681 immune-related prognostic genes associated with OS time were identified, and a p-value <0.01 indicated statistical significance. Among these identified genes, 597 were positively correlated whereas 84 were negatively correlated with survival. As suggested by functional enrichment analysis, these genes were mainly enriched in tumor-related pathways  and immune-related pathways , as shown in The immune-related genes in Patients in the GSE49710 dataset (training cohort) were divided into high- or low-risk score groups according to the median RS5_G prognostic score. As shown in Moreover, the grouping ability of the prognostic signature was also evaluated by the receiver operating characteristic (ROC) curve analysis. As a result, the area under the curve (AUC) values of age group, high risk, MYCN status, vital status, INSS_h1and progression were 0.794 (0.752-0.836), 0.939 (0.918-0.96), 0.929 (0.907-0.952), 0.879 (0.848-0.91), 0.829(0.792-0.865) and 0.804 (0.764-0.845), respectively . The results showed that the above-mentioned features were all risk factors. The _G high, age ≥18 months, high risk, MYCN amplified, and INSS stage 3/4 were correlated with unfavorable outcomes. However, the RS5_G signature had the highest C-index Table 2.The SCCs were calculated between lncRNAs and genes in the RS5_G signature. The lncRNAs with SCC >0.5 and univariate Cox regression p-value <0.01 were selected, as the immune-related lncRNAs. Finally, 17 lncRNAs were identified, as presented in Survival analysis was subsequently implemented for RS_Lnc, and the results showed that patients with high scores had a significantly shorter OS and EFS time than those with low scores. The results of subgroup analysis showed that the OS time of patients in high risk groups for all subgroups was significantly shortened, except for MYCN amplified groups Figure 4Thereafter, the ROC curve analysis was adopted to assess performance in discriminating different prognostic groups by RS_Lnc. As a result, the AUC values of age group, high risk, MYCN status, vital status, INSS_h1 and progression were 0.775 (0.732-0.818), 0.907 (0.879-0.934), 0.94 (0.918-0.962), 0.874 (0.84-0.909), 0.804 (0.764-0.844) and 0.766 (0.722-0.811), respectively . Multivariate Cox analysis identified that RS_Lnc and high risk were the independent prognostic factors for OS, whereas RS_Lnc, high risk, and INSS_h1 were the independent prognostic factors for EFS. However, RS_Lnc had the most significant p-value  network. The KEGG pathways are shown in In this study, the xCell algorithm was employed to calculate the xCell scores of different cell types in the tumor tissues of NB patients from the GSE49710 dataset, which could represent the proportions of various cells in the tumor microenvironment (TME). Typically, the activated dendritic cells (aDC), T helper cells (Th2 cells), and CD8+ T cells, and their correlations with the prognostic signatures RS5_G and RS_Lnc were analyzed, respectively. These three cell types were chosen because of their higher xCell scores and their associations with tumor prognosis. As shown in To further confirm the robustness of the prognostic signature RS5_G, its performance was validated in another three independent datasets, GSE16476, TARGET- Asgharzadeh – 249, and GSE85047. Furthermore, the performance of RS_Lnc was also validated in GSE16476.The OS and progression-free survival (PFS) time were significantly shorter in high-risk score groups than in low-risk groups of the GSE16476 dataset, both for RS5_G and RS_Lnc prognostic signatures, as shown in Univariate Cox regression analysis identified that RS5_G and RS_Lnc were the risk factors for OS time in GSE16476, which had higher C-indexes than the other risk factors   is a malignant solid tumor in children, which is linked with highly heterogeneous outcomes ranging from spontaneous regression without treatment to rapid progression after intensive multidisciplinary therapy , 29. NB At present, NB treatment is mainly based on the INRG risk stratification system . ResearcFive genes, including CDK4, PIK3R1, THRA, MAP2K2, and ULBP2, were included in the immune-related gene signature. The use of CDK4 inhibitor alone or in combination with other drugs in the treatment of NB has entered a phase I clinical trial –43. MAP2THRA is a gene encoding the specific high-affinity nuclear receptor that mediates thyroid hormone. Studies have shown that THRA and THRB (another gene that encodes this receptor), may be involved in human cancer . For insThe RS5_G signature constructed in this study performed well in predicting prognosis, both in training and validation cohorts, especially in subgroups. Taking the performance of RS5_G in distinguishing the survival differences in INRG high risk group as an example, our signature achieved comparable results with fewer genes compared with previous studies. Liu identified an ultra-high risk NB (UHR-NB) group by employing the spectral co-clustering algorithm based on 283 immune-related genes and the two-year survival rates were 48.8% and 43.2% for TARGET and GSE49710, respectively; whereas the two-year survival rates were 79% and 75% in HR-NB group, respectively . In our The results of multivariate Cox regression analysis indicated that RS5_G, high risk, and INSS_h1 can be independent prognostic factors for OS and EFS in GSE49710, and RS5_G had the most significant p-value. It demonstrated that the RS5_G signature might help clinicians to choose the appropriate therapeutic regimen for different patients. For example, we can choose a stronger plan at the time of initial treatment for patients with high-risk scores, rather than the intensive treatment at the time of relapse. By comparing RS5_G with the characteristics previously included in NB risk stratification through multivariate Cox analysis, we believe that the immune-related prognostic signature screened by gene expression profiles can help clinicians distinguish patients with different outcomes in INRG high risk groups, to give different treatment regimens. Of course, further experimental verification is needed for accurate application in clinical practice, in particular, the cut-off value is needed to confirm by moderate methods.Long non-coding RNAs (lncRNAs) participate in the pathological process of tumors including NB in a variety of ways , 49, andvia different pathways  and immune-related pathways . This indicates that in-depth research from the aspects of signaling pathways and immune response can be considered in the treatment of NB.Immunocytes, including CD8+ T cells, NK cells, Th2 cells, etc., may assist patients in killing cancer cells or help tumor cells to evade immune killing , 20. In Moreover, the grouping ability of our signature was verified in three datasets, GSE49710, GSE16476, and GSE85047. The AUC values in distinguishing different groups ranged from 0.737 to 0.94. Typically, the signature performed well in discriminating MYCN status and vital status compared with other groups. In the future, further research is warranted to examine the relationship between genes/lncRNAs and MYCN amplified and to explore the possibility of these genes as therapeutic targets.Even though our prognostic signatures performed well in both the training set and validation cohorts, certain limitations should be noted in this study. Firstly, the datasets came from different platforms, and there were differences in testing, which might cause some bias in the results. Secondly, some lncRNAs were not tested in all datasets, and the performance of the RS_Lnc signature was not verified in all validation cohorts. Thirdly, our study was based on data mining, and these findings should be further validated through biological experiments and clinical validations.Despite these shortcomings, the RS5_G and RS_Lnc signatures constructed in this study performed well at predicting prognosis and grouping different prognostic groups. In conclusion, an immune-related prognostic signature incorporating fewer genes was constructed in this study, which has comparable and even better at predicting and grouping abilities than those reported in other studies. Some immune-related lncRNAs are found to be correlated with the survival of NB patients, which are used to build the prognostic signature RS_Lnc, and it achieved good robustness. Collectively, this study not only provides prognostic signatures to help clinicians to choose the optimal therapeutic strategies but also sheds new light on research on NB treatment.https://www.ncbi.nlm.nih.gov/gds/?term=), survival information and validation datasets were downloaded from R2 database (https://hgserver1.amc.nl/cgi-bin/r2/main.cgi), including Neuroblastoma public—Versteeg—88 , TARGET—Asgharzadeh—249, and Neuroblastoma Primary—NRC—283 .Publicly available datasets were analyzed in this study. This data can be found here: The expression data of training cohort GSE49710  was downloaded from GEO database  and the national key research and development program (Grant No. [2020]151).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.