Analysis of environmental selection pressure of superoxide dismutase in deep-sea sea cucumber

Manganese superoxide dismutase(MnSOD)is an antioxidant that exists in mitochondria and can effectively remove superoxide anions in mitochondria.In a dark,high-pressure,and low-temperature deep-sea environment,MnSOD is essential for the survival of sea cucumbers.Six MnSODs were identified from the transcriptomes of deep and shallow-sea sea cucumbers.To explore their environmental adaptation mechanism,we conducted environmental selection pressure analysis through the branching site model of PAML software.We obtained night positive selection sites,and two of them were significant(97F→H,134K→V):97F→H located in a highly conservative characteristic sequence,and its polarity c hange might have a great impact on the function of MnSOD;134K→V had a change in piezophilic a bility,which might help MnSOD adapt to the environment of high hydrostatic pressure in the deepsea.To further study the effect of these two positive selection sites on MnSOD,we predicted the point mutations of F97H and K134V on shallow-sea sea cucumber by using MAESTROweb and PyMOL.Results show that 97F→H,134K→V might improve MnSOD’s efficiency of scavenging superoxide a nion and its ability to resist high hydrostatic pressure by moderately reducing its stability.The above results indicated that MnSODs of deep-sea sea cucumber adapted to deep-sea environments through their amino acid changes in polarity,piezophilic behavior,and local stability.This study revealed the correlation between MnSOD and extreme environment,and will help improve our understanding of the organism’s adaptation mechanisms in deep sea.

Genetic mechanism of body size variation in groupers:Insights from phylotranscriptomics

Animal body size variation is of particular interest in evolutionary biology,but the genetic basis remains largely unknown.Previous studies have shown the presence of two parallel evolutionary genetic clusters within the fish genus Epinephelus with evident divergence in body size,providing an excellent opportunity to investigate the genetic basis of body size variation in vertebrates.Herein,we performed phylotranscriptomic analysis and reconstructed the phylogeny of 13 epinephelids originating from the South China Sea.Two genetic clades with an estimated divergence time of approximately 15.4 million years ago were correlated with large and small body size,respectively.A total of 180 rapidly evolving genes and two positively selected genes were identified between the two groups.Functional enrichment analyses of these candidate genes revealed distinct enrichment categories between the two groups.These pathways and genes may play important roles in body size variation in groupers through complex regulatory networks.Based on our results,we speculate that the ancestors of the two divergent groups of groupers may have adapted to different environments through habitat selection,leading to genetic variations in metabolic patterns,organ development,and lifespan,resulting in body size divergence between the two locally adapted populations.These findings provide important insights into the genetic mechanisms underlying body size variation in groupers and species differentiation.

Comparative transcriptomes reveal the disjunction adaptive strategy of Thuja species in East Asia and North America

The genus Thuja is ideal for investigating the genetic basis of the East Asia-North America disjunction.The biogeographical background of the genus is debatable and an adaptive strategy is lacking.Through the analysis and mining of comparative transcriptomes,species differentiation and positively selected genes(PSGs)were identified to provide information for understanding the environmental adaptation strategies of the genus Thuja.De novo assembly yielded 44,397-74,252 unigenes of the five Thuja species with contig N50length ranging from 1,559 to 1,724 bp.Annotations revealed a similar distribution of functional categories among them.Based on the phylogenetic trees constructed using the transcriptome data,T.sutchuenensis was divided first,followed by T.plicata and T.occidentalis.The final differentiation of T.koraiensis and T.standishii formed a clade.Enrichment analysis indicated that the PSGs of the North American Thuja species were involved in plant hormone signal transduction and carbon fixation of photosynthetic organisms pathways.The PSGs of East Asian Thuja were related to phenolic,alkaloid,and terpenoid synthesis,important stress-resistant genes and could increase plant resistance to external environmental stresses.This study discovered numerous aroma synthetic-related PSGs including terpene synthase(TPS)genes and lipid phosphate phosphatase 2(LPP2),associated with the synthetic aroma of T.sutchuenensis.Physiological indicators,such as the contents of soluble sugars,total chlorophyll,total phenolics,and total flavonoids were determined,which are consistent with the PSGs enrichment pathways associated with adaptive strategies in the five Thuja species.The results of this study provide an important basis for future studies on conservation genetics.

Current status of genetic transformation technology developed in cucumber(Cucumis sativus L.)

Genetic transformation is an important technique for functional genomics study and genetic improvement of plants. Until now, Agrobacterium-mediated transformation methods using cotyledon as explants has been the major approach for cucumber, and its frequency has been up to 23%. For example, significantly enhancement of the transformation efficiency of this plant species was achieved from the cotyledon explants of the cultivar Poinsett 76 infected by Agrobacterium strains EHA105 with efficient positive selection system in lots of experiments. This review is to summarize some key factors influencing cucumber regeneration and genetic transformation, including target genes, selection systems and the ways of transgene introduction, and then to put forward some strategies for the increasing of cucumber transformation efficiency. In the future, it is high possible for cucumber to be potential bioreactor to produce vaccine and biomaterials for human beings.

GCH1 plays a role in the high-altitude adaptation of Tibetans

Tibetans are welt adapted to high-altitude hypoxia. Previous genome-wide scans have reported many candidate genes for this adaptation, but only a few have been studied. Here we report on a hypoxia gene （GCH1, GTP-cyclohydrolase I）, involved in maintaining nitric oxide synthetase （NOS） function and normal blood pressure, that harbors many potentially adaptive variants in Tibetans. We resequenced an 80.8 kb fragment covering the entire gene region of GCH1 in 50 unrelated Tibetans Combined with previously published data, we demonstrated many GCHI variants showing deep divergence between highlander Tibetans and lowlander Han Chinese. Neutrality tests confirmed a signal of positive Darwinian selection on GCH1 in Tibetans. Moreover, association analysis indicated that the Tibetan version of GCH1 was significantly associated with multiple physiological traits in Tibetans, including blood nitric oxide concentration, blood oxygen saturation and hemoglobin concentration. Taken together, we propose that GCH1 plays a role in the genetic adaptation of Tibetans to high altitude hypoxia.

Genome-wide evolution of MAPKs family and their expression in response to bacterial infection in seahorse Hippocampus erectus

Seahorses have evolved many unique biological traits,including a male brood pouch,the absence of caudal and pelvic fins,and the lack of spleen and gut-associated lymphatic tissue.The mitogenactivated protein kinases(MAPKs)are known to be involved in various important biological processes including growth,differentiation,immunity,and stress responses.Therefore,we hypothesized that the adaptive evolution and expression of the MAPK gene family in seahorse may differ from those of other teleost species.We identified positive selection sites in the erk2,erk5,jnk1,and p38αMAPK genes of the lined seahorse Hippocampus erectus and tiger-tailed seahorse Hippocampus comes.A novel expression profile of MAPK cascade genes was found in seahorse larvae during the first day after birth based on the RNA-seq data of H.erectus,which refl ected vital signs of immune response to its parental immune system.The expression patterns of the four positively selected MAPK genes were analyzed following the bacterial challenge of Vibrio fortis,revealing their upregulation pattern in brood pouch and other immune tissues.This study enriched our knowledge of the evolution of the H.erectus MAPK subfamilies,and could help better understanding the functional role of MAPKs in teleosts.

Genes for the high life： New genetic variants point to positive selection for high altitude hypoxia in Tibetans

People living on the high plateaus of the world have long fascinated biological anthropologists and geneticists because they live in ＂thin air＂ and epitomize an extreme of human biological adaptation.

Comparative analysis of the complete chloroplast genomes of six white oaks with high ecological amplitude in China

Quercus L.has significant societal,ecological and economic benefits in the Northern Hemisphere.However,species identification among oaks is notoriously difficult.China harbours highly diverse oaks,of which the diversity of white oaks is the most extensive;however,to date,the evolution of chloroplast(cp)genomes in white oaks in China has not been comprehensively studied.Thus,we sequenced the complete cp genomes(161,254 bp,161,229 bp and 161,254 bp in size)of three white oak species(Quercus serrata Thunb.var.brevipetiolata A.DC.Nakai,Quercus wutaishansea Mary and Quercus mongolica Fischer ex Ledebour,respectively).Six white oak species(Quercus aliena Blume,Quercus dentata Thunb.,Quercus aliena Blume var.acutiserrata Maximowicz ex Wenzig,Q.serrata var.brevipetiolata,Q.wutaishanica and Q.mongolica)and five other Fagaceae species(Quercus rubra L.,Quercus variabilis Bl.,Quercus aquifolioid.es Rehd.et Wils.,Fagus engleriana Seem.and Castanea henryi Skan Rehd.et Wils.)were retrieved for comparative analyses.We detected11 highly divergent regions(psbA,matK/rps16,rps16,trnSGCU/trnG-GCC,trnR-UCU/atpA,trnT-GGU/psbD,ndhJ,ndhJ/ndhK,accD,ndhF and ycfl)through comparative analyses and these regions might be used as molecular markers.Theωratio of the rps12,rpoC2 and ycf1 genes was greater than 1 in several comparison groups between white oaks and the petA gene was subjected to significant positive selection between the comparison of six white oaks and Q.variabilis.Phylogenetic analyses revealed that six white oaks were grouped with Q.rubra,forming a single clade.

New insights into the evolutionary history of Megacodon:Evidence from a newly discovered species

Megacodon is an ideal genus to study speciation and ecological adaptation in the Sino-Himalayan region.The genus contains two species distributed at different elevations and in two separate areas.However,studies of this genus have long been impeded by a lack of fieldwork on one of its species,Megacodon venosus.In this study,we collected specimens of two Megacodon species and found an extraordinary new species of Megacodon in Lushui county of north-west Yunnan province,which we have since named Megacodon lushuiensis.We propose new species based on both morphological and molecular evidence.The finding of this new species emphasized the importance of ecological divergence in the divergence of Megacodon stylophorus and its parapatric low-elevation Megacodon species.To identify genetic determinants that underlie adaptations to different elevations,we characterized transcriptomes of the new species M.lushuiensis,which is distributed at low elevations,and M.stylophorus,which is distributed at high elevations.Comparative transcriptome analysis identified 8926 orthogroups containing single-copy genes,and 370 orthogroups containing significantly positively selected genes.The set of positively selected genes was enriched into 25 Gene Ontology terms,including "response to water deprivation","response to osmotic stress",and "cellular response to external stimulus".Our results provide new insights into how ecological adaptation and speciation occurred in Megacodon and highlight the role of heterogeneous habitats in the speciation of plants in the Sino-Himalayan region.

Transcriptome assembly of Modiolus modiolus and comparative analysis with Bathymodiolus platifrons

The genetic basis for bivalves＇ adaptation and evolution is not well understood. Even few studies have focused on the mechanism of molluscan molecular evolution between the coastal intertidal zone and deep-sea environment.In our studies, we first conducted the transcritpome assembly of Modiolus modiolus mussels living in coastal intertidal zones. Also, we conducted transcriptome comparison analyses between M. modiolus and Bathymodiolus platifrons living in hydrothermal vents and cold methane/sulfide-hydrocarbon seeps. De novo assemblies of the clean reads yielded a total of 182 476 and 156 261 transcripts with N50 values of 1 769 and 1 545 in M. modiolus and B. platifrons. A total of 27 868 and 23 588 unigenes were identified, which also displayed the similar GO representation patterns. Among the 10 245 pairs of putative orthologs, we identified 26 protein-coding genes under strong positive selection（Ka/Ks〉1） and 12 genes showing moderate positive selection（0.5

Identification of genes under positive selection reveals evolutionary adaptation of Ulva mutabilis

Ulvophytes are attractive model systems for understanding the evolution of growth,development,and environmental stress responses.They are untapped resources for food,fuel,and high-value compounds.The rapid and abundant growth of Ulva species makes them key contributors to coastal biogeochemical cycles,which can cause significant environmental problems in the form of green tides and biofouling.Until now,the Ulva mutabilis genome is the only Ulva genome to have been sequenced.To obtain further insights into the evolutionary forces driving divergence in Ulva species,we analyzed 3905 single copy ortholog family from U.mutabilis,Chlamydomonas reinhardtii and Volvox carteri to identify genes under positive selection(GUPS)in U.mutabilis.We detected 63 orthologs in U.mutabilis that were considered to be under positive selection.Functional analyses revealed that several adaptive modifications in photosynthesis,amino acid and protein synthesis,signal transduction and stress-related processes might explain why this alga has evolved the ability to grow very rapidly and cope with the variable coastal ecosystem environments.

Positive selection analysis reveals the deep-sea adaptation of a hadal sea cucumber ( Paelopatides sp.) to the Mariana Trench

The Mariana Trench,the deepest trench on the earth,is ideal for deep-sea adaptation research due to its unique characters,such as the highest hydrostatic pressure on the Earth,constant ice-cold temperature,and eternal darkness.In this study,tissues of a the hadal holothurian(Paelopatides sp.)were fi xed with RNA later in situ at~6501-m depth in the Mariana Trench,which,to our knowledge,is the deepest in-situ fi xed animal sample.A high-quality transcript was obtained by de-novo transcriptome assembly.A maximum likelihood tree was constructed based on the single copy orthologs across nine species with their available omics data.To investigate deep-sea adaptation,113 positively selected genes(PSGs)were identifi ed in Paelopatides sp.Some PSGs such as microphthalmia-associated transcription factor(MITF)may contribute to the distinct phenotype of Paelopatides sp.,including its translucent white body and degenerated ossicles.At least eight PSGs(transcription factor 7-like 2[TCF7L2],ETS-related transcription factor Elf-2-like[ELF2],PERQ amino acid-rich with GYF domain-containing protein[GIGYF],cytochrome c oxidase subunit 7a,[COX7A],type I thyroxine 5′-deiodinase[DIO1],translation factor GUF1[GUF1],SWI/SNF related-matrix-associated actin-dependent regulator of chromatin subfamily C and subfamily E,member 1[SMARCC]and[SMARCE1])might be related to cold adaptation.In addition,at least nine PSGs(cell cycle checkpoint control protein[RAD9A],replication factor A3[RPA3],DNA-directed RNA polymerases I/II/III subunit RPABC1[POLR2E],putative TAR DNA-binding protein 43 isoform X2[TARDBP],ribonucleoside-diphosphate reductase subunit M1[RRM1],putative serine/threonine-protein kinase[SMG1],transcriptional regulator[ATRX],alkylated DNA repair protein alkB homolog 6[ALKBH6],and PLAC8 motif-containing protein[PLAC8])may facilitate the repair of DNA damage induced by the high hydrostatic pressure,coldness,and high concentration of cadmium in the upper Mariana Trench.

Transcriptomic comparison to identify rapidly evolving genes in Braya humilis

The Brassicaceae species Braya humilis shows broad adaptation to different climatic zones and latitudes. However, the molecular adaptation mechanism of B. humilis is poorly understood. In China, B. humilis is mainly distributed on the QinghaiTibetan Plateau(QTP) and in the adjacent arid region. Previous transcriptome analysis of B. humilis has revealed that 39 salt and osmotic stress response genes are subjected to purifying selection during its speciation. To further explore the adaptation mechanism of B. humilis to an arid environment, OrthoMCL program was employed in this study and 6,268 pairs of orthologous gene pairs with high confidence were obtained between B. humilis and Arabidopsis thaliana. A comparative evolutionary analysis based on nonsynonymous to synonymous substitution ratio(Ka/Ks) was then conducted. There were 64 pairs exhibiting a Ka/Ks ratio more than 0.5 and among which, three instrumental candidate genes, T20487,T22576, and T23757, were identified with strong selection signatures(Ka/Ks >1). The corresponding A. thaliana orthologs are double-stranded RNA-binding domain protein, MADS-box family protein, and NADH-dehydrogenase subunit6, which is encoded by mitochondria genome. This report not only demonstrates the adaptation contribution of fast evolving nuclear genes, but also highlights the potential adaptive value of mitochondria gene to the speciation and adaptation of B. humilis toward the extreme environment in an arid region.

Molecular evolution of Toll-like receptors in rodents

Toll-like receptors(TLRs),the key sensor molecules in vertebrates,trigger the innate immunity and prime the adaptive immune system.The TLR family of rodents,the largest order of mammals,typically contains 13 TLR genes.However,a clear picture of the evolution of the rodent TLR family has not yet emerged and the TLR evolutionary patterns are unclear in rodent clades.Here,we analyzed the natural variation and the evolutionary processes acting on the TLR family in rodents at both the interspecific and population levels.Our results showed that rodent TLRs were dominated by purifying selection,but a series of positively selected sites(PSSs)primarily located in the ligand-binding domain was also identified.The numbers of PSSs differed among TLRs,and nonviralsensing TLRs had more PSSs than those in viral-sensing TLRs.Gene-conversion events were found between TLR1 and TLR6 in most rodent species.Population genetic analyses showed that TLR2,TLR8,and TLR12 were under positive selection in Rattus norvegicus and R.tanezumi,whereas positive selection also acted on TLR5 and TLR9 in the former species,as well as TLR1 and TLR7 in the latter species.Moreover,we found that the proportion of polymorphisms with potentially functional change was much lower in viral-sensing TLRs than in nonviral-sensing TLRs in both of these rat species.Our findings revealed the first thorough insight into the evolution of the rodent TLR genetic variability and provided important novel insights into the evolutionary history of TLRs over long and short timescales.

KaKs_Calculator 2.0:A Toolkit Incorporating Gamma-Series Methods and Sliding Window Strategies

We present an integrated stand-alone software package named KaKs_Calculator 2.0 as an updated version. It incorporates 17 methods for the calculation of nonsynonymous and synonymous substitution rates; among them, we added our modified versions of several widely used methods as the gamma series including y-NG, y-LWL, ）,-MLWL, y-LPB, y-MLPB, y-YN and y-MYN, which have been demonstrated to perform better under certain conditions than their original forms and are not implemented in the previous version. The package is readily used for the identification of positively selected sites based on a sliding window across the sequences of interests in 5＇ to 3＇ direction of protein-coding sequences, and have improved the overall performance on sequence analysis for evolution studies. A toolbox, including C＋＋ and Java source code and executable files on both Windows and Linux platforms together with a user instruction, is downloadable from the website for academic purpose at https：//sourceforge.net/projects/kakscalculator2/.

Molecular evolution and functional divergence of HAK potassium transporter gene family in rice(Oryza sativa L.)

The high-affinity K＋ （HAK） transporter gene family is the largest family in plant that functions as potassium transporter and is important for various aspects of plant life. In the present study, we identified 27 members of this family in rice genome. The phylogenetic tree divided the land plant HAK transporter proteins into 6 distinct groups. Although the main characteristic of this family was established before the origin of seed plants, they also showed some differences between the members of non-seed and seed plants. The HAK genes in rice were found to have expanded in lineage-specific manner after the split of monocots and dicots, and both segmental duplication events and tandem duplication events contributed to the expansion of this family. Functional divergence analysis for this family provided statistical evidence for shifted evolutionary rate after gene duplication. Further analysis indicated that both point mutant with positive selection and gene conversion events contributed to the evolution of this family in rice.

Natural selection maintains the transcribed LTR retrotransposons in Nosema bombycis

Eight intact LTR retrotransposons （Nbr1-Nbr8） have been previously characterized from the genome of Nosema bombycis, a eu- karyotic parasite with a compact and reduced genome. Here we describe six novel transcribed Nbr elements （Nbr9-Nbr14） identified through either cDNA library or RT-PCR. Like previously determined ones, all of them belong to the Ty3/Gypsy superfamily. Retrotransposon diversity and incomplete domains with insertions （Nbr12）, deletions （Nbrll） and in-frame stop codons in coding regions （Nbr9） were detected, suggesting that both defective and loss events of LTR retrotransposon have happened in N. bornbycis genome. Analysis of selection showed that strong purifying selection acts on all elements except Nbr11. This implies that selective pressure keeps both these Nbrs and their functions in genome. Interestingly, Nbrll is under positive selection and some positively selected codons were identified, indicating that new functionality might have evolved in the Nbrll retrotransposon. Unlike other transposable elements, Nbrll has integrated into a conserved syntenic block and probably resulted in the inversion of both flanking regions. This demonstrates that transposable element is an important factor for the reshuffling and evolution of their host genomes, and may be maintained under natural selection.

Genomic, Transcriptomic, and Phenomic Variation Reveals the Complex Adaptation of Modern Maize Breeding

The temperate-tropical division of early maize germplasms to different agricultural environments was argu- ably the greatest adaptation process associated with the success and near ubiquitous importance of global maize production. Deciphering this history is challenging, but new insight has been gained from examining 558 529 single nucleotide polymorphisms, expression data of 28 769 genes, and 662 traits collected from 368 diverse temperate and tropical maize inbred lines in this study. This is a new attempt to systematically exploit the mechanisms of the adaptation process in maize. Our results indicate that divergence between tropical and temperate lines apparently occurred 3400-6700 years ago. Seven hundred and one genomic selection signals and transcriptomic variants including 2700 differentially expressed individual genes and 389 rewired co-expression network genes were identified. These candidate signals were found to be functionally related to stress responses, and most were associated with directionally selected traits, which may have been an advantage under widely varying environmental conditions faced by maize as it was migrated away from its domestication center. Our study also clearly indicates that such stress adaptation could involve evolution of protein-coding sequences as well as transcriptome-level regulatory changes. The latter process may be a more flexible and dynamic way for maize to adapt to environmental changes along its short evolutionary history.

Role of Positive Selection Pressure on the Evolution of H5N1 Hemagglutinin

The surface glycoprotein hemagglutinin （HA） helps the influenza A virus to evade the host immune system by antigenic variation and is a major driving force for viral evolution. In this study, the selection pressure on HA of H5N1 influenza A virus was analyzed using bioinformatics algorithms. Most of the identified positive selection （PS） sites were found to be within or adjacent to epitope sites. Some of the identified PS sites are consistent with previous experimental studies, providing further support to the biological significance of our findings. The highest frequency of PS sites was observed in recent strains isolated during 2005-2007. Phylogenetic analysis was also conducted on HA sequences from various hosts. Viral drift is almost similar in both avian and human species with a progressive trend over the years. Our study reports new mutations in functional regions of HA that might provide markers for vaccine design or can be used to predict isolates of pandemic potential.

The chromosome-scale genome of Phoebe bournei reveals contrasting fates of terpene synthase (TPS)-a and TPS-b subfamilies

Terpenoids,including aromatic volatile monoterpenoids and sesquiterpenoids,function in defense against pathogens and herbivores.Phoebe trees are remarkable for their scented wood and decay resistance.Unlike other Lauraceae species investigated to date,Phoebe species predominantly accumulate sesquiterpenoids instead of monoterpenoids.Limited genomic data restrict the elucidation of terpenoid variation and functions.Here,we present a chromosome-scale genome assembly of a Lauraceae tree,Phoebe bournei,and identify 72 full-length terpene synthase(TPS)genes.Genome-level comparison shows pervasive lineage-specific duplication and contraction of TPS subfamilies,which have contributed to the extreme terpenoid variation within Lauraceae species.Although the TPS-a and TPS-b subfamilies were both expanded via tandem duplication in P.bournei,more TPS-a copies were retained and constitutively expressed,whereas more TPS-b copies were lost.The TPS-a genes on chromosome 8 functionally diverged to synthesize eight highly accumulated sesquiterpenes in P.bournei.The essential oil of P.bournei and its main component,b-caryophyllene,exhibited antifungal activities against the three most widespread canker pathogens of trees.The TPS-a and TPS-b subfamilies have experienced contrasting fates over the evolution of P.bournei.The abundant sesquiterpenoids produced by TPS-a proteins contribute to the excellent pathogen resistance of P.bournei trees.Overall,this study sheds light on the evolution and adaptation of terpenoids in Lauraceae and provides valuable resources for boosting plant immunity against pathogens in various trees and crops.