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The conservation of the genetic integrity of Apis mellifera subspecies has emerged as an important objective. In 2019, the Emilia-Romagna region became the first Italian regional authority to issue a law specifically addressing the protection of the native Apis mellifera ligustica subspecies. In this study we analysed a highly informative portion of the mitochondrial DNA (mtDNA), widely used for assessing genetic diversity of honey bee populations. By analysing 1143 honey bees sampled after the introduction of this law, we provided a snapshot of the distribution of mtDNA haplotypes in this region. The two most frequent mtDNA haplotypes were C1 (characteristic of A. m. ligustica) and C2 (characteristic of A. m. carnica), reported in 86.5% and 11.0% of the analysed bees, respectively. About 1.3% and 1.1% of the analysed bees carried mtDNA haplotypes of the A and M lineages (haplotypes A1a, A1e, A4, A26, A65 and two novel ones, A2w and A6a; M3, M3a, M4 and M79). Continued genetic monitoring will be important to assess the impact of this regional law over the coming years. Based on the obtained results, we recommend a more stringent policy to prevent the erosion of the genetic integrity of the native subspecies A. m. ligustica. [ABSTRACT FROM AUTHOR]

Abstract The conservation of the genetic integrity of Apis mellifera subspecies has emerged as an important objective. In 2019, the Emilia-Romagna region became the first Italian regional authority to issue a law specifically addressing the protection of the native Apis mellifera ligustica subspecies. In this study we analysed a highly informative portion of the mitochondrial DNA (mtDNA), widely used for assessing genetic diversity of honey bee populations. By analysing 1143 honey bees sampled after the introduction of this law, we provided a snapshot of the distribution of mtDNA haplotypes in this region. The two most frequent mtDNA haplotypes were C1 (characteristic of A. m. ligustica) and C2 (characteristic of A. m. carnica), reported in 86.5% and 11.0% of the analysed bees, respectively. About 1.3% and 1.1% of the analysed bees carried mtDNA haplotypes of the A and M lineages (haplotypes A1a, A1e, A4, A26, A65 and two novel ones, A2w and A6a; M3, M3a, M4 and M79). Continued genetic monitoring will be important to assess the impact of this regional law over the coming years. Based on the obtained results, we recommend a more stringent policy to prevent the erosion of the genetic integrity of the native subspecies A. m. ligustica.

The problems of the relationship between the cultures of the Volga-Kama region in general and the formation of the Udmurt ethnos involve scientists of various specializations. The main task of this work is to make a comprehensive description of the burial from the Tolyonsky burial ground located on the territory of the Debessky district of Udmurtia on the right bank of the Cheptsy River. Excavations were carried out by V.A. Semenov in the 1980s. Archaeological and anthropological material is stored in the Ivanova Historical and Cultural Museum-Reserve Idnakar of the Udmurt Republic. The absolute radiocarbon dating of the sample was obtained: 1440 ± 69 years. An anthropological study of the remains revealed a predominant body type as thoracic or pectorally muscular; the living body length was 166 cm. The studied individual was distinguished by brachycephaly, large latitudinal dimensions of the face, and a protruding nose shape. A scientific restoration of the appearance by the method of M.M. Gerasimov was carried out; contour and graphic portraits (full face and profile) were presented. Taking into account the results of genetic analysis, a watercolor artistic portrait was made. For the first time, comprehensive paleogenetic studies have been carried out on a representative of the Polomskaya archaeological culture. The Y-chromosomal haplogroup N1a1a1a2b (B181) and the mitochondrial haplogroup U4 have been established with high reliability. The identified haplogroups in individual no. 93 from the Tolyonsky burial ground along the male and female lines are geographically confined to the Central Volga-Ural region, which is consistent with the results of anthropological and archaeological research. With a probability of more than 97%, the color of the iris of the eyes is predicted to be blue in this individual. [ABSTRACT FROM AUTHOR]

Abstract Background Allotetraploid oilseed rape (Brassica napus L.) is an important worldwide oil-producing crop. The origin of rapeseed is still undetermined due to the lack of wild resources. Despite certain genetic architecture and phylogenetic studies have been done focus on large group of Brassica nuclear genomes, the organelle genomes information under global pattern is largely unknown, which provide unique material for phylogenetic studies of B. napus. Here, based on de novo assemblies of 1,579 B. napus accessions collected globally, we constructed the chloroplast and mitochondrial pan-genomes of B. napus, and investigated the genetic diversity, phylogenetic relationships of B. napus, B. rapa and B. oleracea. Results Based on mitotype-specific markers and mitotype-variant ORFs, four main cytoplasmic haplotypes were identified in our groups corresponding the nap, pol, ole, and cam mitotypes, among which the structure of chloroplast genomes was more conserved without any rearrangement than mitochondrial genomes. A total of 2,092 variants were detected in chloroplast genomes, whereas only 326 in mitochondrial genomes, indicating that chloroplast genomes exhibited a higher level of single-base polymorphism than mitochondrial genomes. Based on whole-genome variants diversity analysis, eleven genetic difference regions among different cytoplasmic haplotypes were identified on chloroplast genomes. The phylogenetic tree incorporating accessions of the B. rapa, B. oleracea, natural and synthetic populations of B. napus revealed multiple origins of B. napus cytoplasm. The cam-type and pol-type were both derived from B. rapa, while the ole-type was originated from B. oleracea. Notably, the nap-type cytoplasm was identified in both the B. rapa population and the synthetic B. napus, suggesting that B. rapa might be the maternal ancestor of nap-type B. napus. Conclusions The phylogenetic results provide novel insights into the organelle genomic evolution of Brassica species. The natural rapeseeds contained at least four cytoplastic haplotypes, of which the predominant nap-type might be originated from B. rapa. Besides, the organelle pan-genomes and the overall variation data offered useful resources for analysis of cytoplasmic inheritance related agronomical important traits of rapeseed, which can substantially facilitate the cultivation and improvement of rapeseed varieties.

Background: Allotetraploid oilseed rape (Brassica napus L.) is an important worldwide oil-producing crop. The origin of rapeseed is still undetermined due to the lack of wild resources. Despite certain genetic architecture and phylogenetic studies have been done focus on large group of Brassica nuclear genomes, the organelle genomes information under global pattern is largely unknown, which provide unique material for phylogenetic studies of B. napus. Here, based on de novo assemblies of 1,579 B. napus accessions collected globally, we constructed the chloroplast and mitochondrial pan-genomes of B. napus, and investigated the genetic diversity, phylogenetic relationships of B. napus, B. rapa and B. oleracea. Results: Based on mitotype-specific markers and mitotype-variant ORFs, four main cytoplasmic haplotypes were identified in our groups corresponding the nap, pol, ole, and cam mitotypes, among which the structure of chloroplast genomes was more conserved without any rearrangement than mitochondrial genomes. A total of 2,092 variants were detected in chloroplast genomes, whereas only 326 in mitochondrial genomes, indicating that chloroplast genomes exhibited a higher level of single-base polymorphism than mitochondrial genomes. Based on whole-genome variants diversity analysis, eleven genetic difference regions among different cytoplasmic haplotypes were identified on chloroplast genomes. The phylogenetic tree incorporating accessions of the B. rapa, B. oleracea, natural and synthetic populations of B. napus revealed multiple origins of B. napus cytoplasm. The cam-type and pol-type were both derived from B. rapa, while the ole-type was originated from B. oleracea. Notably, the nap-type cytoplasm was identified in both the B. rapa population and the synthetic B. napus, suggesting that B. rapa might be the maternal ancestor of nap-type B. napus. Conclusions: The phylogenetic results provide novel insights into the organelle genomic evolution of Brassica species. The natural rapeseeds contained at least four cytoplastic haplotypes, of which the predominant nap-type might be originated from B. rapa. Besides, the organelle pan-genomes and the overall variation data offered useful resources for analysis of cytoplasmic inheritance related agronomical important traits of rapeseed, which can substantially facilitate the cultivation and improvement of rapeseed varieties. [ABSTRACT FROM AUTHOR]

Pet hair has a demonstrated value in resolving legal issues. Cat hair is chronically shed and it is difficult to leave a home with cats without some level of secondary transfer. The power of cat hair as an evidentiary resource may be underused because representative genetic databases are not available for exclusionary purposes. Mitochondrial control region databases are highly valuable for hair analyses and have been developed for the cat. In a representative worldwide data set, 83% of domestic cat mitotypes belong to one of twelve major types. Of the remaining 17%, 7.5% are unique within the published 1394 sample database. The current research evaluates the sample size necessary to establish a representative population for forensic comparison of the mitochondrial control region for the domestic cat. For most worldwide populations, randomly sampling 50 unrelated local individuals will achieve saturation at 95%. The 99% saturation is achieved by randomly sampling 60-170 cats, depending on the numbers of mitotypes available in the population at large. Likely due to the recent domestication of the cat and minimal localized population substructure, fewer cats are needed to meet mitochondria DNA control region database practical saturation than for humans or dogs. Coupled with the available worldwide feline control region database of nearly 1400 cats, minimal local sampling will be required to establish an appropriate comparative representative database and achieve significant exclusionary power.

Several different mitochondrial clades have been found in natural populations of Drosophila melanogaster. Most often, the difference is in single nucleotide substitutions, some of which are conservative. Some clades are rare, and others dominate. It has been reported that clade III dominates over clades V and VI in seven populations of D. melanogaster. We compared D. melanogaster strains with different mitotypes by locomotor activity (using TriKinetics Drosophila Activity Monitor), energy expenditure (by indirect calorimetry, based on measuring oxygen consumption) and life span (under extreme conditions at 29 °C). The nuclear genomes of these strains were aligned for several generations by backcrosses. According to our data, individuals with the mitotype from clade III had a higher level of locomotor activity and longer life span. In terms of energy expenditure, the strains studied did not differ. However, the same level of energy expenditure may be differently distributed between the state of activity and the state of rest or sleep. If the energy expenditure during the sleep in flies with different locomotor activity is the same, then an individual with the same overall energy expenditure can move a greater distance or be active longer. This can be interpreted as an advantage of the strain with the mitotype from clade III compared to the other two mitotypes studied. If individuals have different energy expenditure values at rest, the strains with lower energy expenditure at rest spend less energy during forced inactivity. In this case, the mitotype from clade III should also be advantageous. What nucleotide substitutions in the mitotype from clade III can provide an adaptive advantage is not clear yet. We assume that individuals with widespread clade М(III) may have adaptive advantages compared to other mitotypes due to their greater locomotor activity even with the same energy expenditure. Further studies are required, for mitotypes are polymorphic for single nucleotide polymorphism not only between but also within the clades.

Eukaryotic species are reproductively isolated by sterility barriers that prevent interspecies fertilization (prezygotic sterility barrier) or the fertilization results in infertile offspring (postzygotic sterility barrier). The Saccharomyces species are isolated by postzygotic sterility barriers. Their allodiploid hybrids form no viable gametes (ascospores) and the viable ascospores of the allotetraploids cannot fertilize (conjugate). Our previous work revealed that this mechanism of reproductive isolation differs from those operating in plants and animals and we designated it double sterility barrier (the failure of homeologous chromosomes to pair and the repression of mating by mating-type heterozygosity). Other studies implicated nucleo-mitochondrial incompatibilities in the sterility of the Saccharomyces hybrids, a mechanism assumed to play a central role in the reproductive isolation of animal species. In this project the mitochondrial genomes of 50 cevarum (S. cerevisiae × S. uvarum) hybrids were analyzed. 62% had S. cerevisiae mitotypes, 4% had S. uvarum mitotypes, and 34% had recombinant mitotypes. All but one hybrid formed viable spores indicating that they had genomes larger than allodiploid. Most of these spores were sterile (no sporulation in the clone of vegetative descendants; a feature characteristic of allodiploids). But regardless of their mitotypes, most hybrids could also form fertile alloaneuploid spore clones at low frequencies upon the loss of the MAT-carrying chromosome of the S. uvarum subgenome during meiosis. Hence, the cevarum alloploid nuclear genome is compatible with both parental mitochondrial genomes as well as with their recombinants, and the sterility of the hybrids is maintained by the double sterility barrier (determined in the nuclear genome) rather than by nucleo-mitochondrial incompatibilities. During allotetraploid sporulation both the nuclear and the mitochondrial genomes of the hybrids could segregate but no correlation was observed between the sterility or the fertility of the spore clones and their mitotypes. Nucleo-mitochondrial incompatibility was manifested as respiration deficiency in certain meiotic segregants. As respiration is required for meiosis-sporulation but not for fertilization (conjugation), these segregants were deficient only in sporulation. Thus, the nucleo-mitochondrial incompatibility affects the sexual processes only indirectly through the inactivation of respiration and causes only partial sterility in certain segregant spore clones.

Medaka (comprising two species Oryzias latipes and Oryzias sakaizumii) is a small freshwater fish that is globally used as an experimental model organism. Wild Japanese medaka populations have been declining and their genetic diversity has been affected by extensive artificial introduction of non-native populations and 'himedaka', an orange-red variety derived from O. latipes, which is a commercially popular in Japan. To detect artificially caused genetic introgression in wild medaka populations, we developed a novel and effective nuclear markers. Sequencing analysis of a single-copy nuclear DNA (scnDNA) belonging to linkage group 21 (LG21b) was performed to identify specific haplotype in O. sakaizumii, nine genetically localized groups of O. latipes, and himedaka populations. A total of 28 haplotypes were identified from the 16 tested wild populations. Of the 28 haplotypes, 27 were considered specific to O. sakaizumii and to each genetically localized group of O. latipes; thus, these haplotypes could be used as diagnostic markers to detect artificially caused genetic introgression. Eight haplotypes were confirmed in himedaka, and four were considered specific to himedaka. Our tests involved direct sequencing of the 3′-sequence of the LG21b region as a diagnostic marker, which effectively detected artificially caused genetic introgression in wild populations. Analysis of the actual level of genetic introgression will be improved by incorporating the new scnDNA marker in combination with conventional DNA markers of medaka. [ABSTRACT FROM AUTHOR]

Eukaryotic species are reproductively isolated by sterility barriers that prevent interspecies fertilization (prezygotic sterility barrier) or the fertilization results in infertile offspring (postzygotic sterility barrier). The Saccharomyces species are isolated by postzygotic sterility barriers. Their allodiploid hybrids form no viable gametes (ascospores) and the viable ascospores of the allotetraploids cannot fertilize (conjugate). Our previous work revealed that this mechanism of reproductive isolation differs from those operating in plants and animals and we designated it double sterility barrier (the failure of homeologous chromosomes to pair and the repression of mating by mating-type heterozygosity). Other studies implicated nucleo-mitochondrial incompatibilities in the sterility of the Saccharomyces hybrids, a mechanism assumed to play a central role in the reproductive isolation of animal species. In this project the mitochondrial genomes of 50 cevarum (S. cerevisiae × S. uvarum) hybrids were analyzed. 62% had S. cerevisiae mitotypes, 4% had S. uvarum mitotypes, and 34% had recombinant mitotypes. All but one hybrid formed viable spores indicating that they had genomes larger than allodiploid. Most of these spores were sterile (no sporulation in the clone of vegetative descendants; a feature characteristic of allodiploids). But regardless of their mitotypes, most hybrids could also form fertile alloaneuploid spore clones at low frequencies upon the loss of the MAT -carrying chromosome of the S. uvarum subgenome during meiosis. Hence, the cevarum alloploid nuclear genome is compatible with both parental mitochondrial genomes as well as with their recombinants, and the sterility of the hybrids is maintained by the double sterility barrier (determined in the nuclear genome) rather than by nucleo-mitochondrial incompatibilities. During allotetraploid sporulation both the nuclear and the mitochondrial genomes of the hybrids could segregate but no correlation was observed between the sterility or the fertility of the spore clones and their mitotypes. Nucleo-mitochondrial incompatibility was manifested as respiration deficiency in certain meiotic segregants. As respiration is required for meiosis-sporulation but not for fertilization (conjugation), these segregants were deficient only in sporulation. Thus, the nucleo-mitochondrial incompatibility affects the sexual processes only indirectly through the inactivation of respiration and causes only partial sterility in certain segregant spore clones. [ABSTRACT FROM AUTHOR]

China is the largest producer and exporter of royal jelly in the world. The high production of royal jelly in China is mainly attributed to a high royal jelly-producing lineage of honeybees (Apis mellifera) (HRJB). However, few studies have been conducted on the genetic characterization of HRJB. In this study, the mitochondrial DNA intergenic region between cytochrome oxidase I and II (COI-COII) and the mitochondrial NADH dehydrogenase subunit 2 sequences (ND2) were determined for 90 HRJB colonies, collected from the regions of China where HRJB originated, and 25 unimproved A. m. ligustica colonies from China. COI-COII sequence analysis revealed two mitotypes (C1 and C2d) in HRJB colonies and one mitotype (C1) in unimproved A. m. ligustica colonies. The main mitotype (C1) in HRJB accounted for 93% of the colonies. Based on ND2 sequences, four and two mitotypes were found in HRJB and unimproved A. m. ligustica colonies, respectively. Sequence alignment showed that nucleotides in three positions of the ND2 sequence were different between the main mitotype of HRJB and that of unimproved A. m. ligustica. Our study suggested that HRJB was bred from A. m. ligustica and possibly had genetic characteristics different from unimproved A. m. ligustica.

Growing interest has been emerging on the need to monitor the genetic integrity of the European Apis mellifera subspecies that could be threatened by the human-mediated dispersion of non-native populations and lines. Mitochondrial DNA (mtDNA) lineages can provide useful information for this purpose. In this study, we took advantage of the environmental DNA (eDNA) contained in the honey, which can be analyzed to detect the main groups of mitotypes of the honey bees that produced it. In this study, we applied this eDNA to produce a distribution map all over the Italian peninsula and the two major islands (Sicily and Sardinia) of the following three honey bee mtDNA lineages: A, C and M. A total of 607 georeferenced honey samples, produced in all Italian regions, was analyzed to detect these lineages. The A lineage was widespread in Sicily, as expected, considering that A. m. siciliana carries the African lineage. Surprisingly, this lineage was also reported in about 14% of all other samples produced in almost all continental regions, and in Sardinia. The applied method obtained an updated distribution map of honey bee mtDNA lineages that could be useful to design policies for the conservation of Italian honey bee genetic resources.

Plant mitochondrial genomes characteristically contain extensive structural variation that can be used to define and classify cytoplasm types. We developed markers based on structural variation in the mitochondrial genomes of fertile and cytoplasmic male sterility (CMS) pepper lines and applied them to a panel of Capsicum accessions. We designed a total of 20 sequence characterized amplified region (SCAR) markers based on DNA rearrangement junctions or cytoplasm-specific segments that did not show high similarity to any nuclear mitochondrial DNA segments. We used those markers to classify the mitotypes of 96 C. annuum accessions into 15 groups. Precise genotyping of other Capsicum species (C. frutescens, C. chinense, and C. baccatum) was hampered because of various stoichiometric levels of marker amplicons. We developed a multiplex PCR system based on four of the markers that efficiently classified the C. annuum accessions into five mitotype groups. Close relationships between specific mitotypes and morphological phenotypes implied that diversification or domestication of C. annuum germplasm might have been accompanied by structural rearrangements of mitochondrial DNA or the selection of germplasms with specific mitotypes. Meanwhile, CMS lines shared the same amplification profile of markers with another mitotype. Further analysis using mitochondrial DNA (mtDNA) markers based on single-nucleotide polymorphisms (SNPs) or insertions and deletions (InDels) and CMS-specific open reading frames (orfs) provided new information about the origin of the CMS-specific mitotype and evaluation of candidates for CMS-associated genes, respectively.

Plant mitochondrial genomes characteristically contain extensive structural variation that can be used to define and classify cytoplasm types. We developed markers based on structural variation in the mitochondrial genomes of fertile and cytoplasmic male sterility (CMS) pepper lines and applied them to a panel of Capsicum accessions. We designed a total of 20 sequence characterized amplified region (SCAR) markers based on DNA rearrangement junctions or cytoplasm-specific segments that did not show high similarity to any nuclear mitochondrial DNA segments. We used those markers to classify the mitotypes of 96 C. annuum accessions into 15 groups. Precise genotyping of other Capsicum species (C. frutescens , C. chinense , and C. baccatum) was hampered because of various stoichiometric levels of marker amplicons. We developed a multiplex PCR system based on four of the markers that efficiently classified the C. annuum accessions into five mitotype groups. Close relationships between specific mitotypes and morphological phenotypes implied that diversification or domestication of C. annuum germplasm might have been accompanied by structural rearrangements of mitochondrial DNA or the selection of germplasms with specific mitotypes. Meanwhile, CMS lines shared the same amplification profile of markers with another mitotype. Further analysis using mitochondrial DNA (mtDNA) markers based on single-nucleotide polymorphisms (SNPs) or insertions and deletions (InDels) and CMS-specific open reading frames (orf s) provided new information about the origin of the CMS-specific mitotype and evaluation of candidates for CMS-associated genes, respectively. [ABSTRACT FROM AUTHOR]

Gracilaria spp. are economically important for the production of agar. The distribution of ecotypes of Gracilaria caudata all along the Brazilian coast raises questions regarding whether they are undergoing speciation and are possibly reproductively incompatible. Therefore, in the present work, we selected different female and male gametophytes of G. caudata from three populations along an extended Brazilian coastline (from 3° to 23° S) to perform crossing tests and observe possible reproductive barriers. In addition, we tested post-zygotic isolation, by following meiosis, the production of haploids, and the reproduction of haploids, in the progeny of these inter-population crosses. DNA sequences of the mitochondrial cytochrome c oxidase subunit 1 gene (COI) were used to determine relationships among all gametophytes employed in the crosses and to understand inheritance of mitochondria. The crosses showed interfertility between all populations tested. Neither pre- nor postzygotic isolation was seen in G. caudata from Brazil. Individuals with haplotypes that differ by 1–5 bp (0.15–0.79% divergences) in COI were reproductively compatible. In conclusion, the exchange of genetic material among populations from a wide geographic distribution allows consideration of G. caudata as a biological species. [ABSTRACT FROM AUTHOR]

Mitochondrial DNA (mtDNA) and the dietary macronutrient ratio are known to influence a wide range of phenotypic traits including longevity, fitness and energy production. Commonly mtDNA mutations are posited to be selectively neutral or reduce fitness and, to date, no selectively advantageous mtDNA mutations have been experimentally demonstrated in adult female Drosophila. Here we propose that a ND V161L mutation interacted with diets differing in their macronutrient ratios to influence organismal physiology and mitochondrial traits, but further studies are required to definitively show no linked mtDNA mutations are functionally significant. We utilized two mtDNA types (mitotypes) fed either a 1:2 Protein: Carbohydrate (P:C) or 1:16 P:C diet. When fed the former diet, Dahomey females harboring the V161L mitotype lived longer than those with the Alstonville mitotype and had higher climbing, basal reactive oxygen species (ROS) and elevated glutathione S-transferase E1 expression. The short lived Alstonville females ate more, had higher walking speed and elevated mitochondrial functions as suggested by respiratory control ratio (RCR), mtDNA copy number and expression of mitochondrial transcription termination factor 3. In contrast, Dahomey females fed 1:16 P:C were shorter lived, had higher fecundity, walking speed and mitochondrial functions. They had reduced climbing. This result suggests that mtDNA cannot be assumed to be a strictly neutral evolutionary marker when the dietary macronutrient ratio of a species varies over time and space and supports the hypothesis that mtDNA diversity may reflect the amount of time since the last selective sweep rather than strictly demographic processes.

Wolbachia is a genus of maternally inherited bacteria that is widespread in field populations of Drosophila melanogaster. However, there are no sufficient data on Wolbachia infection among laboratory mutant stocks. We show the wide prevalence of Wolbachia among 353 mutant stocks from the collection of the Genetics of Populations Laboratory, Institute of Cytology and Genetics (ICG), Novosibirsk, Russia. The endosymbiont has been stably inherited in laboratory stocks for a long period of time. Two uninfected stocks from the collection are considered as a result of bacteria loss during maintaining them in the laboratory. There are three Wolbachia genotypes: wMel, wMelCS, and wMelCS2 in the collection. As endosymbiont is coinherited with mytochondria the definite cytotypes are formed from Wolbachia genotypes and mytotypes. We have revealed four cytotypes: M-MEL, M-w-, S-CS, and S-w– in the collection that had been described earlier for field populations of D. melanogaster. The cytotype and genotype frequency patterns differ significantly from those encountered in the wild, that is accounted for genealogy of each stock.

Mitochondrial DNA (mtDNA) and the dietary macronutrient ratio are known to influence a wide range of phenotypic traits including longevity, fitness and energy production. Commonly mtDNA mutations are posited to be selectively neutral or reduce fitness and, to date, no selectively advantageous mtDNA mutations have been experimentally demonstrated in adult female Drosophila. Here we propose that a ND V161L mutation interacted with diets differing in their macronutrient ratios to influence organismal physiology and mitochondrial traits, but further studies are required to definitively show no linked mtDNA mutations are functionally significant. We utilized two mtDNA types (mitotypes) fed either a 1:2 Protein: Carbohydrate (P:C) or 1:16 P:C diet. When fed the former diet, Dahomey females harboring the V161L mitotype lived longer than those with the Alstonville mitotype and had higher climbing, basal reactive oxygen species (ROS) and elevated glutathione S-transferase E1 expression. The short lived Alstonville females ate more, had higher walking speed and elevated mitochondrial functions as suggested by respiratory control ratio (RCR), mtDNA copy number and expression of mitochondrial transcription termination factor 3. In contrast, Dahomey females fed 1:16 P:C were shorter lived, had higher fecundity, walking speed and mitochondrial functions. They had reduced climbing. This result suggests that mtDNA cannot be assumed to be a strictly neutral evolutionary marker when the dietary macronutrient ratio of a species varies over time and space and supports the hypothesis that mtDNA diversity may reflect the amount of time since the last selective sweep rather than strictly demographic processes. [ABSTRACT FROM AUTHOR]

In this study, we tested the Ion Torrent next generation semiconductor based sequencing technology for meat species identification in several highly processed and complex meat products and meat derived broths (a döner kebab, a beef/pork paté, a meat based filling of tortellini, one instantaneous granular preparation of broth stock made by meat and two ready to use meat broths from different producers). The detection protocol included the sequencing of targeted mitochondrial DNA (mtDNA) regions amplified with universal primer pairs and a bioinformatic pipeline designed to interpret sequencing results. Six libraries were sequenced producing a total of 1,363,351 filtered reads. Data mining detected expected and unexpected meat species in the analysed products. Pork was identified in the kebab and Bubalus bubalis DNA was identified in the beef/pork paté. For products for which the precise meat species ingredient information was not available, it was possible to obtain it. Human contamination based on human detected reads could be useful to evaluate the hygienic level of highly processed products. Mitochondrial haplotypes (mitotypes) were identified for several mtDNA-species combinations providing another level of information useful for the authentication of meat derived products. This work defined a methodological framework to establish assays using this sequencing platform for routine species identification in complex and highly processed food. [ABSTRACT FROM AUTHOR]

In Japan, Globodera pallida was first reported in Abashiri city, Hokkaido in 2015. Sequences of ribosomal DNA (rDNA) and mitochondrial cytochrome b gene (cytb) from the Japanese population of G. pallida were compared with those of overseas populations. The representative sequences for rDNA and for cytb from the Japanese population had 100% identity with the sequences from G. pallida populations of North America and Europe in GenBank. Phylogenetic analysis also showed that they clustered in the clade that included populations from Europe and North America. From these results, we concluded that the Japanese population of G. pallida is close to populations that have been reported in Europe and North America. Recent studies suggested that a few populations of G. pallida were introduced into Europe from South America and have spread to various places around the world, and according to our results, into Japan also. [ABSTRACT FROM AUTHOR]

During Quaternary glaciations, the ranges of Northern Eurasia forest species periodically experienced contraction followed by subsequent re-colonizations in the interglacial intervals. However, unlike the broadleaf trees of temperate forests, taiga species seem not to have retreated fully to southern regions in unfavorable periods and possibly survived at mid-latitudes in multiple refugia. Here, we report a study of genetic variation of three mitochondrial DNA markers in 90 populations of Scots pine ( Pinus sylvestris) located from Eastern Europe to Eastern Siberia. The geographic distribution of seven mitotypes demonstrated the split between western and eastern populations approximately along the 38th meridian. Genetic diversity in the western part was significantly higher than in the eastern one. Five mitotypes were western- and one eastern-specific. One mitotype was common in both regions, but in the eastern part it occurred only in the South Urals and adjacent areas. The geographic structure in the mitotype distribution supports a hypothesis of post-glacial re-colonization of the studied territory from the European and Ural refugia. [ABSTRACT FROM AUTHOR]

China is the largest producer and exporter of royal jelly in the world. The high production of royal jelly in China is mainly attributed to a high royal jelly-producing lineage of honeybees (Apis mellifera) (HRJB). However, few studies have been conducted on the genetic characterization of HRJB. In this study, the mitochondrial DNA intergenic region between cytochrome oxidase I and II (COI-COII) and the mitochondrial NADH dehydrogenase subunit 2 sequences (ND2) were determined for 90 HRJB colonies, collected from the regions of China where HRJB originated, and 25 unimproved A. m. ligustica colonies from China. COI-COII sequence analysis revealed two mitotypes (C1 and C2d) in HRJB colonies and one mitotype (C1) in unimproved A. m. ligustica colonies. The main mitotype (C1) in HRJB accounted for 93% of the colonies. Based on ND2 sequences, four and two mitotypes were found in HRJB and unimproved A. m. ligustica colonies, respectively. Sequence alignment showed that nucleotides in three positions of the ND2 sequence were different between the main mitotype of HRJB and that of unimproved A. m. ligustica. Our study suggested that HRJB was bred from A. m. ligustica and possibly had genetic characteristics different from unimproved A. m. ligustica. [ABSTRACT FROM AUTHOR]

Three chloroplast and one mitochondrial DNA markers were developed and used for genotyping of 60 trees in two populations of the Siberian stone pine (Pinus sibirica Du Tour). Two chloroplast loci were monomorphic in both populations, and one polymorphic with two alleles. Therefore, four chloroplast haplotypes were revealed totally. A mitochondrial DNA marker had two alleles or haplotypes (mitotypes).

The ant genus Linepithema is widely known, thanks to the pest species Linepithema humile (Mayr), which is easily mistaken for Linepithema micans (Forel) due to their morphological similarity. Like L. humile, L. micans is associated to the main grapevine pest in Brazil, Eurhizococcus brasiliensis (Wille), also known as ground pearl. Therefore, the present study uses mtDNA fragments to expand the knowledge of haplotype diversity and distribution of L. micans in the state of Rio Grande do Sul (Brazil), to understand the genetic differences of the populations identified in this study. We identified 15 haplotypes of L. micans spread across different localities. Twelve of these haplotypes were new for the species. The high haplotype diversity uncovered in Rio Grande do Sul (Brazil) for this species was predictable, as L. micans is in its native environment. Additional studies that take gene flow into account may reveal interesting aspects of diversity in these populations. [ABSTRACT FROM AUTHOR]

DNA variation in 402 bp of the mitochondrial control region flanked by repeat sequences RS2 and RS3 was evaluated by Sanger sequencing in 152 English domestic cats, in order to determine the significance of matching DNA sequences between hairs found with a victim’s body and the suspect’s pet cat. Whilst 95% of English cats possessed one of the twelve globally widespread mitotypes, four new variants were observed, the most common of which (2% frequency) was shared with the evidential samples. No significant difference in mitotype frequency was seen between 32 individuals from the locality of the crime and 120 additional cats from the rest of England, suggesting a lack of local population structure. However, significant differences were observed in comparison with frequencies in other countries, including the closely neighbouring Netherlands, highlighting the importance of appropriate genetic databases when determining the evidential significance of mitochondrial DNA evidence. [ABSTRACT FROM AUTHOR]

Distinct populations of the potato cyst nematode ( PCN) Globodera pallida exist in the UK that differ in their ability to overcome various sources of resistance. An efficient method for distinguishing between populations would enable pathogen-informed cultivar choice in the field. Science and Advice for Scottish Agriculture ( SASA) annually undertake national DNA diagnostic tests to determine the presence of PCN in potato seed and ware land by extracting DNA from soil floats. These DNA samples provide a unique resource for monitoring the distribution of PCN and further interrogation of the diversity within species. We identify a region of mitochondrial DNA descriptive of three main groups of G. pallida present in the UK and adopt a metagenetic approach to the sequencing and analysis of all SASA samples simultaneously. Using this approach, we describe the distribution of G. pallida mitotypes across Scotland with field-scale resolution. Most fields contain a single mitotype, one-fifth contain a mix of mitotypes, and less than 3% contain all three mitotypes. Within mixed fields, we were able to quantify the relative abundance of each mitotype across an order of magnitude. Local areas within mixed fields are dominated by certain mitotypes and indicate towards a complex underlying 'pathoscape'. Finally, we assess mitotype distribution at the level of the individual cyst and provide evidence of 'hybrids'. This study provides a method for accurate, quantitative and high-throughput typing of up to one thousand fields simultaneously, while revealing novel insights into the national genetic variability of an economically important plant parasite. [ABSTRACT FROM AUTHOR]

In response to the challenge of food security in the new century, heterosis exploitation is being intensified as an effective way to increase grain yield of rice. Thus, breeding of new cytoplasmic male sterility (CMS) is necessary so as to prevent homogenization of hybrid rice in commercial breeding programs. As the rapid development of molecular biology, molecular marker based selection can not only increase the efficiency and accuracy for breeding, but also provide us the possibility to create new CMS by genotypic identification. In this study, we developed a novel CMS rice by screening rice male sterility-inducing cytoplasm and maintainer candidates using mitotype-specific and nuclear molecular markers, respectively. Field trials reveal that the newly bred CMS line has good general combining ability and special combining ability for generation of hybrid rice, which means that the molecular marker system provides us an efficient and practicable approach for development of new CMS in rice breeding programs. [ABSTRACT FROM AUTHOR]

This study was conducted to analyze the process of Africanization of managed honey bee (Apis mellifera) colonies by determining the frequency of African and European morphotypes and mitotypes (mtDNA) in three different ecological environments of northern Mexico. Colonies (n= 151) were sampled in 1) temperate semi-dry; 2) semi-warm semi-dry; and 3) temperate sub-humid regions in the state of Zacatecas. The mtDNA type was determined by PCR-RFLP and the morphotype by the Fast Africanized Bee Identification System (FABIS). Out of all the colonies sampled in all areas, the mtDNA analysis showed a significantly higher frequency of European maternal lineage (77.5%) than of African maternal lineage (22.5%; P <0.0001). The morphometric analysis classified 47% of the colonies as European and 42.4% of them as Africanized. The frequency of colonies with African or European mitotypes and morphotypes varied significantly between regions (P <0.05) with results indicating a higher degree of Africanization in the semi-warm semi-dry region. Conversely, the highest frequency of colonies with the European morphotype and mitotype occurred in the temperate semi-dry region. These results suggest that the environment affects the degree of Africanization of honey bee colonies in northern Mexico. Colonies established at higher altitudes and in more temperate climates have more European genotypes than colonies established in tropical regions. Several hypotheses are discussed to explain these results. [ABSTRACT FROM AUTHOR]

Pet hair has a demonstrated value in resolving legal issues. Cat hair is chronically shed and it is difficult to leave a home with cats without some level of secondary transfer. The power of cat hair as an evidentiary resource may be underused because representative genetic databases are not available for exclusionary purposes. Mitochondrial control region databases are highly valuable for hair analyses and have been developed for the cat. In a representative worldwide data set, 83% of domestic cat mitotypes belong to one of twelve major types. Of the remaining 17%, 7.5% are unique within the published 1394 sample database. The current research evaluates the sample size necessary to establish a representative population for forensic comparison of the mitochondrial control region for the domestic cat. For most worldwide populations, randomly sampling 50 unrelated local individuals will achieve saturation at 95%. The 99% saturation is achieved by randomly sampling 60–170 cats, depending on the numbers of mitotypes available in the population at large. Likely due to the recent domestication of the cat and minimal localized population substructure, fewer cats are needed to meet mitochondria DNA control region database practical saturation than for humans or dogs. Coupled with the available worldwide feline control region database of nearly 1400 cats, minimal local sampling will be required to establish an appropriate comparative representative database and achieve significant exclusionary power. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome sequences of Brassica species have provided insight into inter- and intraspecific variation of plant mitochondrial genomes. However, the size of mitochondrial genome sequenced for Brassica oleracea hitherto does not match to its physical mapping data. This fact led us to investigate B. oleracea mitochondrial genome in detail. Here we report novel B. oleracea mitochondrial genome, derived from var. capitata, a cabbage cultivar ''Fujiwase''. The genome was assembled into a 219,952-bp circular sequence that is comparable to the mitochondrial genomes of other Brassica species (ca. 220-232 kb). This genome contained 34 protein-coding genes, 3 rRNA genes and 17 tRNA genes. Due to absence of a large repeat (140 kb), the mitochondrial genome of ''Fujiwase'' is clearly smaller than the previously reported mitochondrial genome of B. oleracea accession ''08C717'' (360 kb). In both mitotypes, all genes were identical, except cox2- 2, which was present only in the Fujiwase type. At least two rearrangement events via large and small repeat sequences have contributed to the structural differences between the two mitotypes. PCR-based marker analysis revealed that the Fujiwase type is predominant, whereas the 08C717 type coexists at low frequency in all B. oleracea cultivars examined. Intraspecific variations in the mitochondrial genome in B. oleracea may occur because of heteroplasmy, coexistence of different mitotypes within an individual, and substoichiometric shifting. Our data indicate that the Fujiwase-type genome should be used as the representative genome of B. oleracea. [ABSTRACT FROM AUTHOR]

Background Cytoplasmic male sterility (CMS) is not only important for exploiting heterosis in crop plants, but also as a model for investigating nuclear-cytoplasmic interaction. CMS may be caused by mutations, rearrangement or recombination in the mitochondrial genome. Understanding the mitochondrial genome is often the first and key step in unraveling the molecular and genetic basis of CMS in plants. Comparative analysis of the mitochondrial genome of the hau CMS line and its maintainer line in B. juneca (Brassica juncea) may help show the origin of the CMS-associated gene orf288. Results Through next-generation sequencing, the B. juncea hau CMS mitochondrial genome was assembled into a single, circular-mapping molecule that is 247,903 bp in size and 45.08% in GC content. In addition to the CMS associated gene orf288, the genome contains 35 proteinencoding genes, 3 rRNAs, 25 tRNA genes and 29 ORFs of unknown function. The mitochondrial genome sizes of the maintainer line and another normal type line "J163-4" are both 219,863 bp and with GC content at 45.23%. The maintainer line has 36 genes with protein products, 3 rRNAs, 22 tRNA genes and 31 unidentified ORFs. Comparative analysis the mitochondrial genomes of the hau CMS line and its maintainer line allowed us to develop specific markers to separate the two lines at the seedling stage. We also confirmed that different mitotypes coexist substoichiometrically in hau CMS lines and its maintainer lines in B. juncea. The number of repeats larger than 100 bp in the hau CMS line (16 repeats) are nearly twice of those found in the maintainer line (9 repeats). Phylogenetic analysis of the CMS-associated gene orf288 and four other homologous sequences in Brassicaceae show that orf288 was clearly different from orf263 in Brassica tournefortii despite of strong similarity. Conclusion The hau CMS mitochondrial genome was highly rearranged when compared with its isonuclear maintainer line mitochondrial genome. This study may be useful for studying the mechanism of natural CMS in B. juncea, performing comparative analysis on sequenced mitochondrial genomes in Brassicas, and uncovering the origin of the hau CMS mitotype and structural and evolutionary differences between different mitotypes. [ABSTRACT FROM AUTHOR]

Genetic variation in the major histocompatibility complex (MHC) is known to affect disease resistance in many species. Investigations of MHC diversity in populations of wild species have focused on the antigen presenting class IIβ molecules due to the known polymorphic nature of these genes and the role these molecules play in pathogen recognition. Studies of MHC haplotype variation in the turkey (Meleagris gallopavo) are limited. This study was designed to examine MHC diversity in a group of Eastern wild turkeys (Meleagris gallopavo silvestris) collected during population expansion following reintroduction of the species in southern Wisconsin, USA. Southern blotting with BG and class IIβ probes and single nucleotide polymorphism (SNP) genotyping was used to measure MHC variation. SNP analysis focused on single copy MHC genes flanking the highly polymorphic class IIβ genes. Southern blotting identified 27 class IIβ phenotypes, whereas SNP analysis identified 13 SNP haplotypes occurring in 28 combined genotypes. Results show that genetic diversity estimates based on RFLP (Southern blot) analysis underestimate the level of variation detected by SNP analysis. Sequence analysis of the mitochondrial D-loop identified 7 mitochondrial haplotypes (mitotypes) in the sampled birds. Results show that wild turkeys located in southern Wisconsin have a genetically diverse MHC and originate from several maternal lineages. [ABSTRACT FROM AUTHOR]

Brassica rapa L. is an important vegetable and oilseed crop. Cytoplasmic diversity of 36 B. rapa accessions was analyzed using the mitochondria-specific markers. Twelve representative materials including five additional Brassica species and one Eruca sativa Mill. were used as references. A modified multiplex PCR amplification using four pairs of primers was performed to test the mitochondrial types (mitotypes) of the tested materials. Ten accessions were detected with Cam-I mitotype which could amplify 500 and 800 bp bands, twenty-two accessions with Cam-II mitotype which could amplify 500, 800 and 906 bp bands, one accession with Pol mitotype. Interestingly, three B. rapa accessions were revealed with nap mitotype, two of them were local landraces in northern Shaanxi, the third one was a variety from Gansu province which was developed using one local landrace from Northern Shaanxi as female parent. The considerable cytoplasmic diversity in B. rapa provides useful information on studying the possible origin and evolution of B. rapa accessions, and conservation of the germplasm. [ABSTRACT FROM AUTHOR]

Primers were developed that allow for rapid, reliable and inexpensive screening of cytochrome b by analysis of single-stranded conformational polymorphisms. Twenty different haplotypes were identified from six species of Iberian Barbus. These primers proved useful for population and species level studies, and could also be valuable in population genetic and phylogenetic studies of other cyprinin fishes. [ABSTRACT FROM AUTHOR]

The complete mitochondrial DNA sequence of the frigate tuna Auxis thazard and two divergent mitotypes (Mitotype I and Mitotype II) of the bullet tuna Auxis rochei have been determined. The total length of the mitogenomes was 16,506, 16,501 and 16,503 bp, respectively. All mitogenomes had a gene content (13 protein-coding, 2 rRNAs and 22 tRNAs) and organization similar to those observed in most other vertebrates. The major non-coding region (control region) ranged between 843 and 847 bp in length, and showed the typical conserved blocks. Phylogenetic analyses revealed a monophyletic origin of Auxis with respect to other tuna fish. Molecular data here presented provide a useful tool for evolutionary as well as population genetic studies. [ABSTRACT FROM AUTHOR]

Descendents of Apis mellifera scutellata Lepeletier (Hymenoptera: Apidae) (the Africanized honey bee) arrived in the United States in 1990. Whether this was the first introduction is uncertain. A survey of feral honey bees from non-Africanized areas of the southern United States revealed three colonies (from Georgia, Texas, and New Mexico) with a diagnostic African mitochondrial DNA cytochrome b/BglII fragment pattern. To assess maternal origin of these colonies, we developed a primer pair for amplification of a cytochrome b fragment and sequenced using internal sequencing primers. Samples of the three reported honey bee colonies plus another 42 representing the 10 subspecies known to have been introduced in the United States were sequenced. Of the three colonies, the colonies from Texas and New Mexico matched subspecies of European maternal ancestry, whereas the colony from Georgia was of African ancestry. Contrary to expectations, the mitotype of the latter colony was more similar to that exhibited by sub-Saharan A. m. scutellata than to the mitotypes common in north African A. m. intermissa Maa or Portuguese and Spanish A. m. iberiensis Engel. This finding was consistent with anecdotal evidence that A. m. scutellata has been sporadically introduced into the United States before the arrival of the Africanized honey bee from South America. [ABSTRACT FROM AUTHOR]

The invasion of Africanized honeybees (Apis meilifera L.) in the Americas provides a window of opportunity to study the dynamics of secondary contact of subspecies of bees that evolved in allopatry in ecologically distinctive habitats of the Old World. We report here the results of an 11-year mitochondrial DNA survey of a feral honeybee population from southern United States (Texas). The mitochondrial haplotype (mitotype) frequencies changed radically during the 11-year study period. Prior to immigration of Africanized honeybees, the resident population was essentially of eastern and western European maternal ancestry. Three years after detection of the first Africanized swarm there was a mitotype turnover in the population from predominantly eastern European to predominantly A. m. scutellata (ancestor of Africanized honeybees). This remarkable change in the mitotype composition coincided with arrival of the parasitic mite Varroa destructor, which was likely responsible for severe losses experienced by colonies of European ancestry. From 1997 onward the population stabilized with most colonies of A. m. scutellata maternal origin. [ABSTRACT FROM AUTHOR]

Abstract Previous morphological, allozyme and chloroplast DNA data have suggested that Pinus densata originated through hybridization between P . tabuliformi s and P . yunnanensis . In the present study, sequence and restriction site analyses of maternally inherited mitochondrial nad 1 intron were used to detect variation patterns in 19 populations of P . tabuliformis , P . yunnanensis and P . densata . A total of three mitotypes (A, B, C) were detected. All but one of the populations of P . yunnanensis possessed mitotype B while all populations of P . tabuliformis had mitotype A. Pinus densata populations, on the other hand, harboured both mitotypes A and B, which are characteristic of P . tabuliformis and P . yunnanensis , respectively. This result gives strong additional evidence supporting the hybrid origin of this diploid pine. The distribution of mitotypes indicated very different mating compositions and evolutionary history among P . densata populations. It seems that local founder populations and backcrosses may have played important roles in the early establishment of P . densata populations. The uplift of the Tibetan Plateau had a significant impact on the distribution of maternal lineages of P . densata populations. [ABSTRACT FROM AUTHOR]

One hundred and two olive RAPD profiles were sampled from all around the Mediterranean Basin. Twenty four clusters of RAPD profiles were shown in the dendrogram based on the Ward’s minimum variance algorithm using chi-square distances. Factorial discriminant analyses showed that RAPD profiles were correlated with the use of the fruits and the country or region of origin of the cultivars. This suggests that cultivar selection has occurred in different genetic pools and in different areas. Mitochondrial DNA RFLP analyses were also performed. These mitotypes supported the conclusion also that multilocal olive selection has occurred. This prediction for the use of cultivars will help olive growers to choose new foreign cultivars for testing them before an eventual introduction if they are well adapted to local conditions. [ABSTRACT FROM AUTHOR]

Simple Summary: The conservation of the genetic diversity of the native honey bee subspecies is a hot topic in many European countries. Mitochondrial DNA (mtDNA) analyses can provide some information that is useful to monitor the genetic integrity of Apis mellifera populations. A preliminary distribution of the main honey bee mitotypes in Italy was obtained more than 20 years ago. In this study we obtained an updated and more detailed distribution map of the main groups of honey bee mitotypes using an unconventional method that exploits the information derived from the environmental DNA contained in the honey. The results were quite different from the picture taken two decades ago. The African mtDNA lineage was spread all over Italy and not only in Sicily, where it is mainly attached to the A. m. siciliana subspecies, and where it was identified in the previous investigation. A reduction in the frequency of the M lineage was also evident, and, on the other hand, a counterbalanced increase in the C mitotypes was observed in regions all over. The obtained results provided an updated distribution map of the A, C and M groups of mitotypes in Italy, which could be a starting point to design appropriate conservation programs for native honey bee subspecies. Growing interest has been emerging on the need to monitor the genetic integrity of the European Apis mellifera subspecies that could be threatened by the human-mediated dispersion of non-native populations and lines. Mitochondrial DNA (mtDNA) lineages can provide useful information for this purpose. In this study, we took advantage of the environmental DNA (eDNA) contained in the honey, which can be analyzed to detect the main groups of mitotypes of the honey bees that produced it. In this study, we applied this eDNA to produce a distribution map all over the Italian peninsula and the two major islands (Sicily and Sardinia) of the following three honey bee mtDNA lineages: A, C and M. A total of 607 georeferenced honey samples, produced in all Italian regions, was analyzed to detect these lineages. The A lineage was widespread in Sicily, as expected, considering that A. m. siciliana carries the African lineage. Surprisingly, this lineage was also reported in about 14% of all other samples produced in almost all continental regions, and in Sardinia. The applied method obtained an updated distribution map of honey bee mtDNA lineages that could be useful to design policies for the conservation of Italian honey bee genetic resources. [ABSTRACT FROM AUTHOR]