Molecular Evolutionary History Research Articles

Characterization of HSP70 and HSP90 Gene Family in Takifugu fasciatus and Their Expression Profiles on Biotic and Abiotic Stresses Response

Background: Heat shock proteins (HSPs) play crucial roles in response to temperature changes and biotic stresses. However, the HSP gene family in the pufferfish (Takifugu fasciatus) herring has not been comprehensively investigated. Methods and Results: This study presents a systematic analysis of the HSP70 and HSP90 gene families in T. fasciatus, focusing on gene characterization, conserved structural domains, molecular evolutionary history, and expression patterns of the HSP gene family under stress conditions. The findings reveal that 16 HSP genes are evolutionarily conserved, while hspa4 and hsp90aa appear specific to teleost fish. HSP genes exhibit widespread expression across 11 examined tissues, with most demonstrating high expression levels in the heart, brain, and liver. Furthermore, T. fasciatus was subjected to cryogenic and biotic stresses, revealing distinct expression patterns of HSPs under various stress conditions. The response of HSPs to cold stress and Aeromonas hydrophila infection was sustained. In contrast, gene expression of HSPs significantly changed only in the pre-infection period following Ichthyophthirius multifiliis infection, gradually returning to normal levels in the later stages. Conclusions: These experimental results provide a foundation for further in-depth investigations into the characteristics and functions of HSPs in T. fasciatus.

Genome-Wide Identification and Molecular Evolutionary History of the Whirly Family Genes in Brassica napus.

Whirly transcription factors are unique to plants, playing pivotal roles in managing leaf senescence and DNA repair. While present in various species, their identification in Brassica napus L. (B. napus) and their differences during hybridization and polyploidy has been elusive. Addressing this, our study delves into the functional and evolutionary aspects of the Whirly gene family during the emergence of B. napus, applying bioinformatics and comparative genomics. We identified six Whirly genes in B. napus. In Brassica rapa L. (B. rapa), three Whirly genes were identified, while four were found in Brassica oleracea L. (B. oleracea). The results show that the identified Whirly genes not only have homology but also share the same chromosomal positions. Phylogenetic analysis indicates that Whirly genes in monocots and dicots exhibit high conservation. In the evolutionary process, the Whirly gene family in B. napus experienced events of intron/exon loss. Collinearity insights point to intense purifying selection post-duplication. Promoter regions housed diverse cis-acting elements linked to photoresponse, anaerobic initiation, and methyl jasmonate responsiveness. Notably, elements tied to abscisic acid signaling and meristem expression were prominent in diploid ancestors but subdued in tetraploid B. napus. Tissue-specific expression unveiled analogous patterns within subfamily genes. Subsequent qRT-PCR analysis spotlighted BnAWHY1b's potential significance in abiotic stress response, particularly drought. These findings can be used as theoretical foundations to understand the functions and effects of the Whirly gene family in B. napus, providing references for the molecular mechanism of gene evolution between this species and its diploid ancestors.

GenoFig: a user-friendly application for the visualization and comparison of genomic regions.

Understanding the molecular evolutionary history of organisms usually requires visual comparison of genomic regions from related species or strains. Although several applications already exist to achieve this task, they are either too old, too limited, or too complex for most user's needs. GenoFig is a graphical application for the visualization of prokaryotic genomic regions, intended to be as easy to use as possible and flexible enough to adapt to a variety of needs. GenoFig allows the personalized representation of annotations extracted from GenBank files in a consistent way across sequences, using regular expressions. It also provides several unique options to optimize the display of homologous regions between sequences, as well as other more classical features such as sequence GC percent or GC-skew representations. In summary, GenoFig is a simple, free, and highly configurable tool to explore the evolution of specific genomic regions in prokaryotes and to produce publication-ready figures. Genofig is fully available at https://forgemia.inra.fr/public-pgba/genofig under a GPL 3.0 license.

Genome-wide identification and analyses of ZmAPY genes reveal their roles involved in maize development and abiotic stress responses

Apyrase is a class of enzyme that catalyzes the hydrolysis of nucleoside triphosphates/diphosphates (NTP/NDP), which widely involved in regulation of plant growth and stress responses. However, apyrase family genes in maize have not been identified, and their characteristics and functions are largely unknown. In this study, we identified 16 apyrases (named as ZmAPY1-ZmAPY16) in maize genome, and analyzed their phylogenetic relationships, gene structures, chromosomal distribution, upstream regulatory transcription factors and expression patterns. Analysis of the transcriptome database unveiled tissue-specific and abiotic stress-responsive expression of ZmAPY genes in maize. qPCR analysis further confirmed their responsiveness to drought, heat, and cold stresses. Association analyses indicated that variations of ZmAPY5 and ZmAPY16 may regulate maize agronomic traits and drought responses. Our findings shed light on the molecular characteristics and evolutionary history of maize apyrase genes, highlighting their roles in various biological processes and stress responses. This study forms a basis for further exploration of apyrase functions in maize.

Fortieth Anniversary Virtual Issues: Neutralist–Selectionist Debate

Fortieth Anniversary Virtual Issues: Neutralist–Selectionist Debate

Speciation in the Iranian plateau: Molecular phylogeny and evolutionary history of the Persian long‐tailed desert lizard

AbstractMesalina watsonana is a well‐known species of small lacertid lizards with an extensive species distribution that exhibits high genetic diversity. The species has a wide distribution range in Iran, some parts of Turkmenistan, Afghanistan, Pakistan, and northwest India. This study aims to generate phylogenetic and phylogeographic evidence to derive taxonomic recommendations supporting. Furthermore, this species can be used as a model for examining Iranian Platuea's historical biogeography. We conducted a very detailed sampling of its distribution and used genetic approaches. Phylogenetic analyses were done implementing two mitochondrial (Cytb and 16S) and one nuclear (C‐mos) gene fragments. Combination of these results indicated that seven well‐supported distinct clades exist within this species complex, i.e. Kerman clade, Esfarayen clade, Halil clade, Ardestan clade, M. watsonana clade, Bardaskan Clade, and Khuzestan Clade in Iran. Also, our results revealed that several distinct clades diverged due to geologic events when the Dasht‐e‐Kavir and the Zagros Mountains were formed. It seems that the ancestor of M. watsonana spread to Iranian Plateau before the formation of the Zagros Mountains. Orogenic activities of the Zagros Mountains and the formation of deserts have influenced the separation of these lineages from the Late Miocene by allopatric speciation. Generally, our findings suggest that each of the seven clades corresponding to distinct geographic regions deserves to be elevated to the species level.

Analyzing genetic diversity of chloroplast genomes in Liliales

AbstractLiliales is a monocotyledonous order and contains both photosynthetic and mycoheterotrophic species that distribute locally or worldwide. In this study, thegenetic diversity of chloroplast genomes in Liliales was explored regarding theirnucleotide diversity and repeated composition. The analysis of nucleotide diversity revealed various hotspots in large and small single-copy regions whereas the IR regions had low sequence divergence. Although each family has specific hotspots, the rps15-ycf1 region was commonly found as a highly variable area in the cpDNA of observed taxa. In the cpDNA of Liliales, mononucleotide simple sequence repeat (SSR) is the most common type. The majority of SSRs are located in non-coding regions. Similarly, more long repeats were found in non-coding areas than in coding sequences. Additionally, the complement repeat exceeds forward type in the cpDNA of Liliales. The highest number of long repeats was found in Corsia dispar whereas that of SSRswas detected in Smilax china. The results of nucleotide diversity and repeat analyses provided fundamental information for further studies on population genetics, molecular marker development and evolutionary history of Liliales.® 2022 Journal of Science and Technology - NTTU

Molecular characterization and evolutionary history analysis of a novel Kudzu mosaic virus (KMV) infecting the Kudzu (Pueraria montana)

ObjectivesPlant viruses are a significant contributor to the economic losses experienced in numerous plant species. Begomoviruses exhibit a wide range of host species, encompassing both cultivated and non-cultivated plants. The kudzu plant, scientifically known as Pueraria montana, is indigenous to the Southeast Asian region. Moreover, it exhibits varied attributes with regards to its utilization in the fields of medicine, food, cosmetics, and the herbal industry. MethodsDuring a routine survey, some P. montana plants having typical symptoms of yellowing, mosaic, and mottle were observed in areas including Fort-Minro (FM) and Anari of district Dera Ghazi Khan and Maari (MA) of district Rajanpur. The genomic DNA was extracted from the collected samples. The polymerase chain reaction (PCR) and rolling circular amplification (RCA) was carried out to isolate the DNA and DNA- β from the collected plant specimens. An evolutionary research was conducted to assess the lineage of this virus inside a novel geographical region. ResultsA 500-bp amplicon size was obtained from the samples of the FM and MA, while the approximately 2.7-kb amplicon size of its full genome was obtained from the samples of both locations. The results of the sequence analysis demonstrated conclusively that the plants in question are infected with Kudzu mosaic virus (KMV). The evolutionary history analysis has confirmed the virus's lineage in a new region, and the nucleotide frequencies of obtained sequences were variable. ConclusionThe presence of a novel bipartite begomovirus in the tropical region has been substantiated using a phylogenetic and evolutionary investigation. The observation of a white fly population on infected plants has also suggested that the transmission of this virus through the migration of this insect vector can be a viable mechanism.

In silico epitope prediction and evolutionary analysis reveals capsid mutation patterns for enterovirus B.

Enterovirus B (EVB) is a common species of enterovirus, mainly consisting of Echovirus (Echo) and Coxsackievirus B (CVB). The population is generally susceptible to EVB, especially among children. Since the 21st century, EVB has been widely prevalent worldwide, and can cause serious diseases, such as viral meningitis, myocarditis, and neonatal sepsis. By using cryo-electron microscopy, the three-dimensional (3D) structures of EVB and their uncoating receptors (FcRn and CAR) have been determined, laying the foundation for the study of viral pathogenesis and therapeutic antibodies. A limited number of epitopes bound to neutralizing antibodies have also been determined. It is unclear whether additional epitopes are present or whether epitope mutations play a key role in molecular evolutionary history and epidemics, as in influenza and SARS-CoV-2. In the current study, the conformational epitopes of six representative EVB serotypes (E6, E11, E30, CVB1, CVB3 and CVB5) were systematically predicted by bioinformatics-based epitope prediction algorithm. We found that their epitopes were distributed into three clusters, where the VP1 BC loop, C-terminus and VP2 EF loop were the main regions of EVB epitopes. Among them, the VP1 BC loop and VP2 EF loop may be the key epitope regions that determined the use of the uncoating receptors. Further molecular evolution analysis based on the VP1 and genome sequences showed that the VP1 C-terminus and VP2 EF loop, as well as a potential "breathing epitope" VP1 N-terminus, were common mutation hotspot regions, suggesting that the emergence of evolutionary clades was driven by epitope mutations. Finally, footprints showed mutations were located on or near epitopes, while mutations on the receptor binding sites were rare. This suggested that EVB promotes viral epidemics by breaking the immune barrier through epitope mutations, but the mutations avoided the receptor binding sites. The bioinformatics study of EVB epitopes may provide important information for the monitoring and early warning of EVB epidemics and developing therapeutic antibodies.

Multiple transitions between realms shape relict lineages of Proteus cave salamanders.

In comparison to biodiversity on Earth's surface, subterranean biodiversity has largely remained concealed. The olm (Proteus anguinus) is one of the most enigmatic extant cave inhabitants, and until now little was known regarding its genetic structure and evolutionary history. Olms inhabit subterranean waters throughout the Dinaric Karst of the western Balkans, with a seemingly uniform phenotypic appearance of cave-specialized traits: an elongate body, snout and limbs, degenerated eyes and loss of pigmentation ("white olm"). Only a single small region in southeastern Slovenia harbours olms with a phenotype typical of surface animals: pigmented skin, eyes, a blunt snout and short limbs ("black olm"). We used a combination of mitochondrial DNA and genome-wide single nucleotide polymorphism data to investigate the molecular diversity, evolutionary history and biogeography of olms along the Dinaric Karst. We found nine deeply divergent species-level lineages that separated between 17 and 4 million years ago, while molecular diversity within lineages was low. We detected no signal of recent admixture between lineages and only limited historical gene flow. Biogeographically, the contemporaneous distribution of lineages mostly mirrors hydrologically separated subterranean environments, while the historical separation of olm lineages follows microtectonic and climatic changes in the area. The reconstructed phylogeny suggests at least four independent transitions to the cave phenotype. Two of the species-level lineages have miniscule ranges and may represent Europe's rarest amphibians. Their rarity and the decline in other lineages call for protection of their subterranean habitats.

The deep-rooted origin of disulfide-rich spider venom toxins.

Spider venoms are a complex concoction of enzymes, polyamines, inorganic salts and disulfide-rich peptides (DRPs). Although DRPs are widely distributed and abundant, their evolutionary origin has remained elusive. This knowledge gap stems from the extensive molecular divergence of DRPs and a lack of sequence and structural data from diverse lineages. By evaluating DRPs under a comprehensive phylogenetic, structural and evolutionary framework, we have not only identified 78 novel spider toxin superfamilies but also provided the first evidence for their common origin. We trace the origin of these toxin superfamilies to a primordial knot - which we name 'Adi Shakti', after the creator of the Universe according to Hindu mythology - 375 MYA in the common ancestor of Araneomorphae and Mygalomorphae. As the lineages under evaluation constitute nearly 60% of extant spiders, our findings provide fascinating insights into the early evolution and diversification of the spider venom arsenal. Reliance on a single molecular toxin scaffold by nearly all spiders is in complete contrast to most other venomous animals that have recruited into their venoms diverse toxins with independent origins. By comparatively evaluating the molecular evolutionary histories of araneomorph and mygalomorph spider venom toxins, we highlight their contrasting evolutionary diversification rates. Our results also suggest that venom deployment (e.g., prey capture or self-defense) influences evolutionary diversification of DRP toxin superfamilies.

Molecular evolution and population genetics of glutamate decarboxylase acid resistance pathway in lactic acid bacteria.

Glutamate decarboxylase (GAD) pathway (GDP) is a major acid resistance mechanism enabling microorganisms' survival in low pH environments. We aimed to study the molecular evolution and population genetics of GDP in Lactic Acid Bacteria (LAB) to understand evolutionary processes shaping adaptation to acidic environments comparing species where the GDP genes are organized in an operon structure (Levilactobacillus brevis) versus lack of an operon structure (Lactiplantibacillus plantarum). Within species molecular population genetic analyses of GDP genes in L. brevis and L. plantarum sampled from diverse fermented food and other environments showed abundant synonymous and non-synonymous nucleotide diversity, mostly driven by low frequency changes, distributed throughout the coding regions for all genes in both species. GAD genes showed higher level of replacement polymorphism compared to transporter genes (gadC and YjeM) for both species, and GAD genes that are outside of an operon structure showed even higher level of replacement polymorphism. Population genetic tests suggest negative selection against replacement changes in all genes. Molecular structure and amino acid characteristics analyses showed that in none of the GDP genes replacement changes alter 3D structure or charge distribution supporting negative selection against non-conservative amino acid changes. Phylogenetic and between species divergence analyses suggested adaptive protein evolution on GDP genes comparing phylogenetically distant species, but conservative evolution comparing closely related species. GDP genes within an operon structure showed slower molecular evolution and higher conservation. All GAD and transporter genes showed high codon usage bias in examined LAB species suggesting high expression and utilization of acid resistance genes. Substantial discordances between species, GAD, and transporter gene tree topologies were observed suggesting molecular evolution of GDP genes do not follow speciation events. Distribution of operon structure on the species tree suggested multiple independent gain or loss of operon structure in LABs. In conclusion, GDP genes in LABs exhibit a dynamic molecular evolutionary history shaped by gene loss, gene transfer, negative and positive selection to maintain its active role in acid resistance mechanism, and enable organisms to thrive in acidic environments.

Transcriptome Analysis of Air Space-Type Variegation Formation in Trifolium pratense.

Air space-type variegation is the most diverse among the species of known variegated leaf plants and is caused by conspicuous intercellular spaces between the epidermal and palisade cells and among the palisade cells at non-green areas. Trifolium pratense, a species in Fabaceae with V-shaped air space-type variegation, was selected to explore the application potential of variegated leaf plants and accumulate basic data on the molecular regulatory mechanism and evolutionary history of leaf variegation. We performed comparative transcriptome analysis on young and adult leaflets of variegated and green plants and identified 43 candidate genes related to air space-type variegation formation. Most of the genes were related to cell-wall structure modification (CESA, CSL, EXP, FLA, PG, PGIP, PLL, PME, RGP, SKS, and XTH family genes), followed by photosynthesis (LHCB subfamily, RBCS, GOX, and AGT family genes), redox (2OG and GSH family genes), and nitrogen metabolism (NodGS family genes). Other genes were related to photooxidation, protein interaction, and protease degradation systems. The downregulated expression of light-responsive LHCB subfamily genes and the upregulated expression of the genes involved in cell-wall structure modification were important conditions for air space-type variegation formation in T. pratense. The upregulated expression of the ubiquitin-protein ligase enzyme (E3)-related genes in the protease degradation systems were conducive to air space-type variegation formation. Because these family genes are necessary for plant growth and development, the mechanism of the leaf variegation formation in T. pratense might be a widely existing regulation in air space-type variegation in nature.

Diet evolution of carnivorous and herbivorous mammals in Laurasiatheria

BackgroundLaurasiatheria contains taxa with diverse diets, while the molecular basis and evolutionary history underlying their dietary diversification are less clear.ResultsIn this study, we used the recently developed molecular phyloecological approach to examine the adaptive evolution of digestive system-related genes across both carnivorous and herbivorous mammals within Laurasiatheria. Our results show an intensified selection of fat and/or protein utilization across all examined carnivorous lineages, which is consistent with their high-protein and high-fat diets. Intriguingly, for herbivorous lineages (ungulates), which have a high-carbohydrate diet, they show a similar selection pattern as that of carnivorous lineages. Our results suggest that for the ungulates, which have a specialized digestive system, the selection intensity of their digestive system-related genes does not necessarily reflect loads of the nutrient components in their diets but appears to be positively related to the loads of the nutrient components that are capable of being directly utilized by the herbivores themselves. Based on these findings, we reconstructed the dietary evolution within Laurasiatheria, and our results reveal the dominant carnivory during the early diversification of Laurasiatheria. In particular, our results suggest that the ancestral bats and the common ancestor of ruminants and cetaceans may be carnivorous as well. We also found evidence of the convergent evolution of one fat utilization-related gene, APOB, across carnivorous taxa.ConclusionsOur molecular phyloecological results suggest that digestive system-related genes can be used to determine the molecular basis of diet differentiations and to reconstruct ancestral diets.

Analyzing genetic diversity of chloroplast genomes in Liliales

AbstractLiliales is a monocotyledonous order and contains both photosynthetic and mycoheterotrophic species that distribute locally or worldwide. In this study, thegenetic diversity of chloroplast genomes in Liliales was explored regarding theirnucleotide diversity and repeated composition. The analysis of nucleotide diversity revealed various hotspots in large and small single-copy regions whereas the IR regions had low sequence divergence. Although each family has specific hotspots, the rps15-ycf1 region was commonly found as a highly variable area in the cpDNA of observed taxa. In the cpDNA of Liliales, mononucleotide simple sequence repeat (SSR) is the most common type. The majority of SSRs are located in non-coding regions. Similarly, more long repeats were found in non-coding areas than in coding sequences. Additionally, the complement repeat exceeds forward type in the cpDNA of Liliales. The highest number of long repeats was found in Corsia dispar whereas that of SSRswas detected in Smilax china. The results of nucleotide diversity and repeat analyses provided fundamental information for further studies on population genetics, molecular marker development and evolutionary history of Liliales.® 2022 Journal of Science and Technology - NTTU

Characterization of blaNDM-positive Enterobacteriaceae reveals the clonal dissemination of Enterobacter hormaechei coharboring blaNDM and tet(X4) along the pork production chain

The convergence of blaNDM and tet(X4) in a single strain has been sporadically detected in diverse pathogens from different sources, causing widespread concerns. However, the genomic characterization of blaNDM-positive or blaNDM- and tet(X4)-coharboring Enterobacteriaceae in the pork production chain, as well as how they emerged, remains mostly unclear. In this study, forty blaNDM-positive Enterobacteriaceae, including eleven tet(X4)- and blaNDM-coharboring isolates, were isolated from a pig slaughterhouse and a retail pork market in Yangzhou, China. This is the first report presenting the coexistence of blaNDM and tet(X4) in Enterobacter hormaechei, and it was demonstrated that they could spread clonally from the slaughterhouse to the retail market. Furthermore, we also discovered that the slaughterhouse served as a possible reservoir for blaNDM-positive ST48 E. coli strains. The blaNDM-bearing IncX3 plasmid was found in twenty blaNDM-positive strains as well as all blaNDM-5- and tet(X4)-coharboring strains, and most of them (30/31) were conjugative. In Klebsiella aerogenes NTT31XS, blaNDM was found on a nonconjugative ~17 kb IncX3 plasmid. When compared to the typical IncX3 plasmid, it deleted large-scale regions, which might imply a low fitness cost dissemination strategy for the IncX3 plasmid. Despite being from different species, tet(X4)-bearing plasmids carried by all blaNDM-5- and tet(X4)-coharboring strains were nonconjugative and had identical architectures. Phylogenetic analysis suggested that the formation of K. aerogenes NTT31XS may have been mediated by translocation of tet(X4)-positive circular intermediates and recombination events of regions in typical blaNDM-bearing IncX3 plasmids. Overall, this study expounded the prevalence and transmission characteristics of blaNDM-positive Enterobacteriaceae strains in the two links of pork production chain before they were exposed to humans, and expanded our knowledge of the molecular features and evolutionary history of tet(X4)- and blaNDM-5-coharboring strains. Strict control measures should be implemented to prevent the spread of such pathogens along food production chains.

Molecular mechanisms and evolutionary history of phytomelatonin in flowering

Flowering is a critical stage in plant life history, which is coordinated by environmental signals and endogenous cues. Phytomelatonin is a widely distributed indoleamine present in all living organisms and plays pleiotropic roles in plant growth and development. Recent evidence has established that phytomelatonin could modulate flowering in many species, probably in a concentration-dependent manner. Phytomelatonin seems to associate with floral meristem identification and floral organ formation, and the fluctuation of phytomelatonin might be important for flowering. Regarding the underlying mechanisms, phytomelatonin interacts with the central components of floral gene regulatory networks directly or indirectly, including the MADS-box gene family, phytohormones, and reactive oxygen species (ROS). From an evolutionary point of view, the actions of phytomelatonin in flowering probably evolved during the period of the diversification of flowering plants and could be regarded as a functional extension of its primary activities. The presumed evolutionary history of phytomelatonin-modulated flowering is proposed, presented in the chronological order of the appearance of phytomelatonin and core flowering regulators, namely DELLA proteins, ROS, and phytohormones. Further efforts are needed to address some intriguing aspects, such as the exploration of the association between phytomelatonin and photoperiodic flowering, phytomelatonin-related floral MADS-box genes, the crosstalk between phytomelatonin and phytohormones, as well as its potential applications in agriculture.

Layered evolution of gene expression in “superfast” muscles for courtship

Identifying the molecular process of complex trait evolution is a core goal of biology. However, pinpointing the specific context and timing of trait-associated changes within the molecular evolutionary history of an organism remains an elusive goal. We study this topic by exploring the molecular basis of elaborate courtship evolution, which represents an extraordinary example of trait innovation. Within the behaviorally diverse radiation of Central and South American manakin birds, species from two separate lineages beat their wings together using specialized “superfast” muscles to generate a “snap” that helps attract mates. Here, we develop an empirical approach to analyze phylogenetic lineage-specific shifts in gene expression in the key snap-performing muscle and then integrate these findings with comparative transcriptomic sequence analysis. We find that rapid wing displays are associated with changes to a wide range of molecular processes that underlie extreme muscle performance, including changes to calcium trafficking, myocyte homeostasis and metabolism, and hormone action. We furthermore show that these changes occur gradually in a layered manner across the species history, wherein which ancestral genetic changes to many of these molecular systems are built upon by later species-specific shifts that likely finalized the process of display performance adaptation. Our study demonstrates the potential for combining phylogenetic modeling of tissue-specific gene expression shifts with phylogenetic analysis of lineage-specific sequence changes to reveal holistic evolutionary histories of complex traits.

A New Gene Family Diagnostic for Intracellular Biomineralization of Amorphous Ca Carbonates by Cyanobacteria.

Cyanobacteria have massively contributed to carbonate deposition over the geological history. They are traditionally thought to biomineralize CaCO3 extracellularly as an indirect byproduct of photosynthesis. However, the recent discovery of freshwater cyanobacteria-forming intracellular amorphous calcium carbonates (iACC) challenges this view. Despite the geochemical interest of such a biomineralization process, its molecular mechanisms and evolutionary history remain elusive. Here, using comparative genomics, we identify a new gene (ccyA) and protein family (calcyanin) possibly associated with cyanobacterial iACC biomineralization. Proteins of the calcyanin family are composed of a conserved C-terminal domain, which likely adopts an original fold, and a variable N-terminal domain whose structure allows differentiating four major types among the 35 known calcyanin homologs. Calcyanin lacks detectable full-length homologs with known function. The overexpression of ccyA in iACC-lacking cyanobacteria resulted in an increased intracellular Ca content. Moreover, ccyA presence was correlated and/or colocalized with genes involved in Ca or transport and homeostasis, supporting the hypothesis of a functional role of calcyanin in iACC biomineralization. Whatever its function, ccyA appears as diagnostic of intracellular calcification in cyanobacteria. By searching for ccyA in publicly available genomes, we identified 13 additional cyanobacterial strains forming iACC, as confirmed by microscopy. This extends our knowledge about the phylogenetic and environmental distribution of cyanobacterial iACC biomineralization, especially with the detection of multicellular genera as well as a marine species. Moreover, ccyA was probably present in ancient cyanobacteria, with independent losses in various lineages that resulted in a broad but patchy distribution across modern cyanobacteria.

Molecular phylogeography and evolutionary history of the pink rice borer (Lepidoptera: Noctuidae): Implications for refugia identification and pest management

AbstractUnderstanding phylogeography, demographic dynamics as well as evolutionary forces of agricultural pests can enhance our understanding of their distribution dynamics, which is critical for developing effective control strategies. The pink rice borer (PRB), Sesamia inferens (Walker), is a destructive pest of rice that is widely distributed in Asia; however, little is known regarding its phylogeography and evolutionary history. In this study, we investigated the genetic diversity, population genetic structure and demographic history of the PRB at the scale of its whole distribution in China. Both mitochondrial and microsatellite datasets indicated that PRB individuals exhibit high levels of genetic diversity. Two large phylogeographic clusters were identified, corresponding to Northern and Southern China regions, respectively. The glacial events during the penultimate glacial period likely led to the separation of these two clusters, and parts of Northern and Southern China areas acted as independent refugia for PRB during glacial period. Two microrefugia in the Sichuan Basin and Yunnan‐Guizhou Plateau were also identified, and the PRB from Sichuan Basin could be one of the ancestral sources for PRB individuals from the Northern China lineage. Our study not only provides information that has implications for refugia identification, but also provides an example on how to reveal the phylogeography, demography history and evolutionary forces of an agriculture pest.