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1.
Trends Ecol Evol ; 35(1): 34-42, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31703819

RESUMO

Hybridization has broad evolutionary consequences, from fueling or counteracting speciation to facilitating adaptation to novel environments. Hybridization and subsequent introgression appear widespread along the tree of life. However, our understanding of how distinct evolutionary forces shape admixed genomes and the fate of introgressed genetic variants remains scarce. Most admixture research in animals has focused on diploid organisms. We propose that haplodiploid organisms can help resolve open questions about the genomic consequences of hybridization in natural populations. The ploidy difference between haploid males and diploid females, the availability of genome-wide male haplotypes, and ongoing cases of admixture make haplodiploid organisms promising models to improve our knowledge with regards to the evolution of hybrid genomes.


Assuntos
Diploide , Hibridização Genética , Animais , Feminino , Genômica , Haploidia , Masculino
2.
Chemosphere ; 242: 125145, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31678852

RESUMO

Neonicotinoid insecticides are currently of major concern for the health of wild and managed insects that provide key ecosystem services like pollination. Even though sublethal effects of neonicotinoids are well known, there is surprisingly little information on how they possibly impact developmental stability, and to what extent genetics are involved. This holds especially true for haploid individuals because they are hemizygous at detoxification loci and may be more susceptible. Here we take advantage of haplodiploidy in Western honey bees, Apis mellifera, to show for the first time that neonicotinoids affect developmental stability in diploid females (workers), and that haploid males (drones) are even more susceptible. Phenotypic fore wing venation abnormalities and fluctuating wing asymmetry, as measures of developmental instability, were significantly increased under field-realistic neonicotinoid-exposure of colonies. The higher susceptibility of haploid drones suggests that heterozygosity can play a key role in the ability to buffer the sublethal effects of neonicotinoids. Aiming to improve conservation efforts, our findings highlight the urgent need to better understand the role that genetics plays at enabling non-target organisms to cope with insecticide exposure.


Assuntos
Abelhas/fisiologia , Inseticidas/toxicidade , Neonicotinoides/toxicidade , Animais , Ecossistema , Feminino , Haploidia , Inseticidas/análise , Masculino , Neonicotinoides/análise , Nitrocompostos , Polinização , Asas de Animais
3.
J Appl Genet ; 60(3-4): 291-300, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31506777

RESUMO

Agriculture will benefit from a rigorous characterization of genes for adult plant resistance (APR) since this gene class was recognized to provide more durable protection from plant diseases. The present study reports the identification of APR loci to powdery mildew in German winter wheat cultivars Cortez and Atlantis. Cortez was previously shown to carry all-stage resistance gene Pm3e. To avoid interference of Pm3e in APR studies, line 6037 that lacked Pm3e but showed field resistance from doubled-haploid (DH) population Atlantis/Cortez was used in two backcrosses to Atlantis for the establishment of DH population 6037/Atlantis//Atlantis. APR was assessed in the greenhouse 10, 15, and 20 days after inoculation (dai) from the 4-leaf stage onwards and combined with single-nucleotide polymorphism data in a genome-wide association study (GWAS) and a linkage map-based quantitative trait loci (QTL) analysis. In GWAS, two QTL were detected: one on chromosome 1BL 10 dai, the other on chromosome 2BL 20 dai. In conventional QTL analysis, both QTL were detected with all three disease ratings: the QTL on chromosome 1BL explained a maximum of 35.2% of the phenotypic variation 10 dai, whereas the QTL on chromosome 2BL explained a maximum of 43.5% of the phenotypic variation 20 dai. Compared with GWAS, linkage map-based QTL analysis allowed following the dynamics of QTL action. The two large-effect QTL for APR to powdery mildew with dynamic gene action can be useful for the enhancement of wheat germplasm.


Assuntos
Ascomicetos/genética , Resistência à Doença/genética , Doenças das Plantas/genética , Locos de Características Quantitativas/genética , Triticum/genética , Ascomicetos/patogenicidade , Cruzamento , Cromossomos de Plantas/genética , Cruzamentos Genéticos , Ligação Genética/genética , Estudo de Associação Genômica Ampla , Genótipo , Haploidia , Fenótipo , Doenças das Plantas/microbiologia , Triticum/microbiologia
4.
Theor Appl Genet ; 132(12): 3227-3243, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31555890

RESUMO

KEY MESSAGE: Increased efficiencies achieved in different steps of DH line production offer greater benefits to maize breeding programs. Doubled haploid (DH) technology has become an integral part of many commercial maize breeding programs as DH lines offer several economic, logistic and genetic benefits over conventional inbred lines. Further, new advances in DH technology continue to improve the efficiency of DH line development and fuel its increased adoption in breeding programs worldwide. The established method for maize DH production covered in this review involves in vivo induction of maternal haploids by a male haploid inducer genotype, identification of haploids from diploids at the seed or seedling stage, chromosome doubling of haploid (D0) seedlings and finally, selfing of fertile D0 plants. Development of haploid inducers with high haploid induction rates and adaptation to different target environments have facilitated increased adoption of DH technology in the tropics. New marker systems for haploid identification, such as the red root marker and high oil marker, are being increasingly integrated into new haploid inducers and have the potential to make DH technology accessible in germplasm such as some Flint, landrace, or tropical material, where the standard R1-nj marker is inhibited. Automation holds great promise to further reduce the cost and time in haploid identification. Increasing success rates in chromosome doubling protocols and/or reducing environmental and human toxicity of chromosome doubling protocols, including research on genetic improvement in spontaneous chromosome doubling, have the potential to greatly reduce the production costs per DH line.


Assuntos
Haploidia , Melhoramento Vegetal/métodos , Zea mays/genética , Cromossomos de Plantas , Produtos Agrícolas/genética , Fenótipo
5.
Theor Appl Genet ; 132(12): 3333-3345, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31559526

RESUMO

KEY MESSAGE: Doubled-haploid libraries from landraces capture native genetic diversity for a multitude of quantitative traits and make it accessible for breeding and genome-based studies. Maize landraces comprise large allelic diversity. We created doubled-haploid (DH) libraries from three European flint maize landraces and characterized them with respect to their molecular diversity, population structure, trait means, variances, and trait correlations. In total, 899 DH lines were evaluated using high-quality genotypic and multi-environment phenotypic data from up to 11 environments. The DH lines covered 95% of the molecular variation present in 35 landraces of an earlier study and represent the original three landrace populations in an unbiased manner. A comprehensive analysis of the target trait plant development at early growth stages as well as other important agronomic traits revealed large genetic variation for line per se and testcross performance. The majority of the 378 DH lines evaluated as testcrosses outperformed the commercial hybrids for early development. For total biomass yield, we observed a yield gap of 15% between mean testcross yield of the commercial hybrids and mean testcross yield of the DH lines. The DH lines also exhibited genetic variation for undesirable traits like root lodging and tillering, but correlations with target traits early development and yield were low or nonsignificant. The presented diversity atlas is a valuable, publicly available resource for genome-based studies to identify novel trait variation and evaluate the prospects of genomic prediction in landrace-derived material.


Assuntos
Variação Genética , Genética Populacional , Melhoramento Vegetal , Zea mays/genética , Cruzamentos Genéticos , Europa (Continente) , Genótipo , Haploidia , Fenótipo
6.
Plant Sci ; 287: 110189, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31481211

RESUMO

Microspores exposed to some stress factors may display cell totipotency and could be reprogrammed towards embryogenic development. Plant breeding and genetic engineering widely use haploids/doubled haploids (DHs) derived from in vitro-cultured microspores, but the mechanism of this process remains poorly understood. Recently published data suggest that microspore embryogenesis (ME) is accompanied by changes in DNA methylation and chromatin reorganization. Here, we used two triticale DH lines (DH19 and DH28), significantly different with respect to embryogenic potential. To change DNA methylation levels, we applied two cytosine-analogs: 5-azacytidine (AC) and 2'-deoxy-5-azacytidine (DAC) treatments. We found that chemically-induced DNA demethylation caused chromatin relaxation and dysregulation of marker genes (TaTPD1-like, GSTF2, GSTA2, CHI3, Tad1, TaNF-YA7, SERK2, TaME1) related to ME. Both drugs showed significant cytotoxicity in a dose-dependent manner. We noticed that lines varied in terms of overall DNA methylation levels and responded in a different way to hypomethylation caused by the drugs. DH19 (low embryogenic) after inhibitors treatment, showed higher microspore viability, but its recalcitrancy was not overcome. For highly embryogenic DH28, we noted significantly higher effectiveness of embryo-like structure production and plant regeneration. In summary, our study provides new insight into the role of DNA methylation in ME initiation. They suggest potential benefits resulting from the utilization of epigenetic inhibitors to improve the process of DHs production.


Assuntos
Triticale/genética , Cruzamento , Metilação de DNA , DNA de Plantas/genética , Haploidia , Pólen/embriologia , Pólen/genética , Triticale/embriologia
8.
Plant Cell Rep ; 38(12): 1515-1525, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31473791

RESUMO

KEY MESSAGE: A single division meiosis mechanism of meiotic restitution is incompletely penetrant but significantly associated with restored fertility in triticale haploids (n = 21, genome formula ABR). Meiotic restitution, or failure of meiosis to produce gametes with a reduced chromosome number, can lead to the restoration of fertility in allohaploids. Meiotic restitution is of major interest for producing doubled haploids, as haploid plants undergoing meiotic restitution can often form seeds without the need to apply mitosis inhibitors to double chromosome number. We aimed to characterize meiotic restitution in a population of 183 haploids (n = 21, genome formula ABR) derived from an F1 wheat-rye hybrid where one parent was known to carry factors responsible for restoration of fertility in wide-cross haploids. Based on cytological analysis, approximately half of the plants analyzed were characterized by normal meiosis, while half showed at least some cytological evidence of meiotic restitution. However, this mechanism was incompletely penetrant in the population, with no individual plant showing 100% unreduced gamete formation: restitution occurred sectorially within each anther and was not observed in all the anthers of a given plant. Hence, the absence of meiotic restitution could not be confirmed conclusively for any individual plant, confounding this analysis. However, cytological observation of meiotic restitution was significantly associated with seed set, further confirming the role of meiotic restitution in fertility restoration. Our results provide insight into this mechanism of unreduced gamete formation, and provide a basis for future work identifying the genetic factors responsible for this trait.


Assuntos
Cromossomos de Plantas/genética , Meiose/fisiologia , Triticale/genética , Haploidia , Hibridização Genética/genética , Hibridização Genética/fisiologia , Meiose/genética , Poliploidia
9.
BMC Genomics ; 20(1): 641, 2019 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-31399045

RESUMO

BACKGROUND: Although the Y chromosome plays an important role in male sex determination and fertility, it is currently understudied due to its haploid and repetitive nature. Methods to isolate Y-specific contigs from a whole-genome assembly broadly fall into two categories. The first involves retrieving Y-contigs using proportion sharing with a female, but such a strategy is prone to false positives in the absence of a high-quality, complete female reference. A second strategy uses the ratio of depth of coverage from male and female reads to select Y-contigs, but such a method requires high-depth sequencing of a female and cannot utilize existing female references. RESULTS: We develop a k-mer based method called DiscoverY, which combines proportion sharing with female with depth of coverage from male reads to classify contigs as Y-chromosomal. We evaluate the performance of DiscoverY on human and gorilla genomes, across different sequencing platforms including Illumina, 10X, and PacBio. In the cases where the male and female data are of high quality, DiscoverY has a high precision and recall and outperforms existing methods. For cases when a high quality female reference is not available, we quantify the effect of using draft reference or even just raw sequencing reads from a female. CONCLUSION: DiscoverY is an effective method to isolate Y-specific contigs from a whole-genome assembly. However, regions homologous to the X chromosome remain difficult to detect.


Assuntos
Cromossomos Humanos Y/genética , Análise de Sequência de DNA/métodos , Feminino , Haploidia , Humanos , Masculino , Análise de Sequência de DNA/economia , Fatores de Tempo
10.
Theor Appl Genet ; 132(11): 3035-3045, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31377817

RESUMO

KEY MESSAGE: Novel large-effect consistent QTL for anther extrusion (AE) to improve cross-pollination were mapped in doubled haploid populations derived from IPK gene bank spring wheat accessions. TaAP2-D, an ortholog of Cleistogamy1 in barley, is a likely candidate gene for AE in wheat. To establish a robust hybrid wheat breeding system, male lines harboring alleles that promote outcrossing should be developed. In this study, we developed two doubled haploid (DH) populations of hexaploid spring wheat (Triticum aestivum L.) by crossing accessions taken from IPK gene bank. In both populations, the phenotypic data of anther extrusion (AE) based on three years of field trials showed a wide variation and approximated a normal distribution. Both populations were genotyped with a 15 k Infinium single nucleotide polymorphism (SNP) array resulting in 3567 and 3457 polymorphic SNP markers for DH population-1 and DH population-2, respectively. Composite interval mapping identified quantitative trait loci (QTL) on chromosomes 1D, 2D, 4A, 4B, 5A, 5D, 6A, and 6B; with consistent QTL (that are identified in all the years) on chromosome 4A in DH population-1, and on chromosomes 2D and 6B in DH population-2. The consistent QTL explained 17.2%, 32.9%, and 12.3% of the phenotypic variances, respectively. Genic scan of the chromosome 2D-QTL showed that the wheat gene TaAP2-D, an ortholog of Cleistogamy1 which promotes AE via swelling of the lodicules in barley, lies within the QTL region. A diagnostic marker was developed for TaAP2-D that showed co-segregation with the AE phenotype. This study shows the use of gene bank diversity reservoir to find alleles which are otherwise difficult to detect in elite populations. The identification of large-effect consistent QTL for AE is expected to help form efficient male parental lines suitable for hybrid wheat seed production and serve as a source for map-based cloning.


Assuntos
Flores/crescimento & desenvolvimento , Variação Genética , Locos de Características Quantitativas , Triticum/genética , Mapeamento Cromossômico , Genética Populacional , Haploidia , Fenótipo
11.
Theor Appl Genet ; 132(11): 3023-3033, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31410494

RESUMO

KEY MESSAGE: Based on their consistency over environments, two QTL identified in Lillian on chromosomes 5A and 7A could be useful targets for marker assisted breeding of common bunt resistance. Common bunt of wheat (Triticum aestivum L.) caused by Tilletia tritici and T. laevis is an economically important disease because of losses in grain yield and reduced grain quality. Resistance can be quantitative, under the control of multiple small effect genes. The Canada Western Red Spring wheat variety Lillian is moderately resistant to common bunt races found on the Canadian prairies. This study was conducted to identify and map quantitative trait loci (QTL) conferring resistance against common bunt in Lillian. A doubled haploid population comprising 280 lines was developed from F1 plants of the cross of Lillian by Vesper. The lines were inoculated at seeding with the two races L16 (T. laevis) and T19 (T. tritici), grown in field near Swift Current, SK, in 2014, 2015 and 2016 and assessed for disease incidence. The lines were genotyped with the 90 K iSelect SNP genotyping assay, and a high-density genetic map was constructed. Quantitative trait locus analysis was performed with MapQTL.6® software. Two relatively stable common bunt resistance QTL, detected in two of the 3 years, were identified on chromosomes 5A and 7A from Lillian. In addition, three less stable QTL, appearing in one out of 3 years, were identified: one was contributed by Lillian on chromosome 3D and two were contributed by Vesper on chromosomes 1D and 2A. Epistatic interaction was identified for the bunt incidence between 3D and 7A resulting in greater bunt resistance. Future bunt resistance breeding will benefit from combining these QTL through gene pyramiding.


Assuntos
Resistência à Doença/genética , Doenças das Plantas/genética , Locos de Características Quantitativas , Triticum/genética , Basidiomycota/patogenicidade , Mapeamento Cromossômico , Cromossomos de Plantas , Genes de Plantas , Genótipo , Haploidia , Fenótipo , Doenças das Plantas/microbiologia , Triticum/microbiologia
12.
PLoS One ; 14(7): e0218564, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31291290

RESUMO

Owing to the hierarchical organization of biology, from genomes over transcriptomes and proteomes down to metabolomes, there is continuous debate about the extent to which data and interpretations derived from one level, e.g. the transcriptome, are in agreement with other levels, e.g. the metabolome. Here, we tested the effect of ocean acidification (OA; 400 vs. 1000 µatm CO2) and its modulation by light intensity (50 vs. 300 µmol photons m-2 s-1) on the biomass composition (represented by 75 key metabolites) of diploid and haploid life-cycle stages of the coccolithophore Emiliania huxleyi (RCC1216 and RCC1217) and compared these data with interpretations from previous physiological and gene expression screenings. The metabolite patterns showed minor responses to OA in both life-cycle stages. Whereas previous gene expression analyses suggested that the observed increased biomass buildup derived from lipid and carbohydrate storage, this dataset suggests that OA slightly increases overall biomass of cells, but does not significantly alter their metabolite composition. Generally, light was shown to be a more dominant driver of metabolite composition than OA, increasing the relative abundances of amino acids, mannitol and storage lipids, and shifting pigment contents to accommodate increased irradiance levels. The diploid stage was shown to contain vastly more osmolytes and mannitol than the haploid stage, which in turn had a higher relative content of amino acids, especially aromatic ones. Besides the differences between the investigated cell types and the general effects on biomass buildup, our analyses indicate that OA imposes only negligible effects on E. huxleyi´s biomass composition.


Assuntos
Haptófitas/crescimento & desenvolvimento , Água do Mar/química , Biomassa , Dióxido de Carbono/análise , Diploide , Haploidia , Haptófitas/genética , Haptófitas/metabolismo , Concentração de Íons de Hidrogênio , Estágios do Ciclo de Vida , Metaboloma , Oceanos e Mares , Transcriptoma
13.
PLoS Biol ; 17(7): e3000350, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31265461

RESUMO

Mutagenic screening is powerful for identifying key genes involved in developmental processes. However, such screens are successful only in lower organisms. Here, we develop a targeted genetic screening approach in mice through combining androgenetic haploid embryonic stem cells (AG-haESCs) and clustered regularly interspaced palindromic repeats/CRISPR-associated protein 9 (CRISPR-Cas9) technology. We produced a mutant semi-cloned (SC) mice pool by oocyte injection of AG-haESCs carrying constitutively expressed Cas9 and an single guide RNA (sgRNA) library targeting 72 preselected genes in one step and screened for bone-development-related genes through skeletal analysis at birth. This yielded 4 genes: Zic1 and Clec11a, which are required for bone development, and Rln1 and Irx5, which had not been previously considered. Whereas Rln1-/- mice exhibited small skeletal size only at birth, Irx5-/- mice showed skeletal abnormalities both in postnatal and adult phases due to decreased bone mass and increased bone marrow adipogenesis. Mechanistically, iroquois homeobox 5 (IRX5) promotes osteoblastogenesis and inhibits adipogenesis by suppressing peroxisome proliferator activated receptor γ (PPARγ) activation. Thus, AG-haESC-mediated functional mutagenic screening opens new avenues for genetic interrogation of developmental processes in mice.


Assuntos
Desenvolvimento Ósseo/genética , Regulação da Expressão Gênica no Desenvolvimento , Marcação de Genes/métodos , Testes Genéticos/métodos , Células-Tronco Embrionárias Murinas/metabolismo , Animais , Sistemas CRISPR-Cas , Células Cultivadas , Haploidia , Fatores de Crescimento de Células Hematopoéticas/genética , Fatores de Crescimento de Células Hematopoéticas/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Camundongos , Camundongos Knockout , Relaxina/genética , Relaxina/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
14.
Genome Res ; 29(7): 1115-1122, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31221725

RESUMO

New genes are a major source of novelties, and a disproportionate amount of them are known to show testis expression in later phases of male gametogenesis in different groups such as mammals and plants. Here, we propose that this enhanced expression is a consequence of haploid selection during the latter stages of male gametogenesis. Because emerging adaptive mutations will be fixed faster if their phenotypes are expressed by haploid rather than diploid genotypes, new genes with advantageous functions arising during this unique stage of development have a better chance to become fixed. To test this hypothesis, expression levels of genes of differing evolutionary age were examined at various stages of Drosophila spermatogenesis. We found, consistent with a model based on haploid selection, that new Drosophila genes are both expressed in later haploid phases of spermatogenesis and harbor a significant enrichment of adaptive mutations. Additionally, the observed overexpression of new genes in the latter phases of spermatogenesis was limited to the autosomes. Because all male cells exhibit hemizygous expression for X-linked genes (and therefore effectively haploid), there is no expectation that selection acting on late spermatogenesis will have a different effect on X-linked genes in comparison to initial diploid phases. Together, our proposed hypothesis and the analyzed data suggest that natural selection in haploid cells elucidates several aspects of the origin of new genes by explaining the general prevalence of their testis expression, and a parsimonious solution for new alleles to avoid being lost by genetic drift or pseudogenization.


Assuntos
Evolução Molecular , Haploidia , Seleção Genética , Espermatogênese/genética , Animais , Drosophila , Mutação em Linhagem Germinativa , Masculino , Modelos Genéticos , Testículo/metabolismo , Cromossomo X
15.
Nat Plants ; 5(6): 575-580, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31182848

RESUMO

Doubled haploid (DH) breeding based on in vivo haploid induction has led to a new approach for maize breeding1. All modern haploid inducers used in DH breeding are derived from the haploid inducer line Stock6. Two key quantitative trait loci, qhir1 and qhir8, lead to high-frequency haploid induction2. Mutation of the gene MTL/ZmPLA1/NLD in qhir1 could generate a ~2% haploid induction rate (HIR)3-5; nevertheless, this mutation is insufficient for modern haploid inducers whose average HIR is ~10%6. Therefore, cloning of the gene underlying qhir8 is important for illuminating the genetic basis of haploid induction. Here, we present the discovery that mutation of a non-Stock6-originating gene in qhir8, namely, ZmDMP, enhances and triggers haploid induction. ZmDMP was identified by map-based cloning and further verified by CRISPR-Cas9-mediated knockout experiments. A single-nucleotide change in ZmDMP leads to a 2-3-fold increase in the HIR. ZmDMP knockout triggered haploid induction with a HIR of 0.1-0.3% and exhibited a greater ability to increase the HIR by 5-6-fold in the presence of mtl/zmpla1/nld. ZmDMP was highly expressed during the late stage of pollen development and localized to the plasma membrane. These findings provide important approaches for studying the molecular mechanism of haploid induction and improving DH breeding efficiency in maize.


Assuntos
Haploidia , Proteínas de Membrana/genética , Proteínas de Plantas/genética , Zea mays/genética , Mapeamento Cromossômico , Cromossomos de Plantas , Técnicas de Inativação de Genes , Mutação
16.
PLoS One ; 14(6): e0218538, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31242252

RESUMO

The diploid genotypes of males are widely thought to determine sperm phenotypes, yet recent work shows that the haploid genetics of the individual sperm cell also contributes significantly. We tested seven sperm phenotypes, flagellar length and six behaviors, looking for correlations between genetic and phenotypic variability. While flagellar length appears to be controlled by the diploid genotype of the source, variation in three of the behavioral phenotypes, linearity, wobble, and progression are significantly correlated with the heterozygosity of the male producer. Because males that are more genetically variable produce a sperm set that is more diverse in its haploid genotypes, we suggest that the correlations may reflect significant haploid genetic control of sperm swimming behaviors.


Assuntos
Characidae/genética , Characidae/fisiologia , Espermatozoides/fisiologia , Animais , Characidae/classificação , Estudos de Associação Genética , Variação Genética , Haploidia , Heterozigoto , Hibridização Genética , Masculino , Microscopia de Vídeo , Fenótipo , Motilidade Espermática/genética , Motilidade Espermática/fisiologia , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/citologia
17.
Zhongguo Dang Dai Er Ke Za Zhi ; 21(6): 552-557, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31208508

RESUMO

OBJECTIVE: To investigate the clinical efficacy of haploid hematopoietic stem cells (haplo-HSC) combined with third-party umbilical cord blood (tpCB) transplantation in the treatment of X-linked chronic granulomatous disease (X-CGD). METHODS: The clinical data of 26 boys with X-CGD were retrospectively analyzed who were admitted to the Sixth Medical Center of PLA General Hospital between April 2014 and March 2018. All the patients were treated with haplo-HSC combined with tpCB transplantation. The median age of the patients was 3.5 years. The donor was the father in 25 cases and an aunt in 1 case. Transplantation was 5/6 HLA-matched in 9 cases, 4/6 in 12 cases, and 3/6 in 5 cases. The patients received busulfan, cyclophosphamide, fludarabine, or anti-thymocyte globulin for myeloablative preconditioning. Cyclosporine A and mycophenolate mofetil were used for prevention of acute graft-versus-host disease (aGVHD). Then the patients were treated with haploid bone marrow hematopoietic stem cells combined with tpCB transplantation on day 1 and haploid peripheral hematopoietic stem cells on day 2. The counts of median donor total nucleated cells, CD34+ cells, and CD3+ cells were 14.6×108/kg, 5.86×106/kg, and 2.13×108/kg respectively. RESULTS: The median time to neutrophil and platelet engraftment was 12 and 23 days after transplantation respectively. Full donor hematopoietic chimerism was observed on day 30. Twenty-five cases were from haplo-HSC and 1 was from cord blood. No primary implant failure and implant dysfunction occurred, and secondary implant failure occurred in one case. The NADPH oxidase activity returned to normal one month after transplantation. The incidence of grade I-II aGVHD and grade III-IV aGVHD was 35% and 15% respectively. Chronic GVHD (cGVHD) of the skin occurred in one case, and no progression was observed after steroid administration. During the follow-up period of 6-51 months, 25 patients survived, of whom 24 were disease-free (23 patients without cGVHD and 1 with cGVHD of the skin) and NADPH oxidase activity returned to normal; one patient developed secondary implant failure but survived; one patient died of viral interstitial pneumonia 16 months after transplantation. The 5-year event-free survival rate and overall survival rate were 81%±12% and 89%±10% respectively. CONCLUSIONS: Haplo-HSC combined with tpCB transplantation is one of the effective methods for the treatment of X-CGD in children.


Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Doença Enxerto-Hospedeiro , Doença Granulomatosa Crônica , Transplante de Células-Tronco Hematopoéticas , Pré-Escolar , Haploidia , Células-Tronco Hematopoéticas , Humanos , Masculino , Estudos Retrospectivos , Condicionamento Pré-Transplante
18.
Genome Biol Evol ; 11(7): 1939-1951, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31209485

RESUMO

In many studies, sex-related genes have been found to evolve rapidly. We therefore expect plant pollen genes to evolve faster than sporophytic genes. In addition, pollen genes are expressed as haploids which can itself facilitate rapid evolution because recessive advantageous and deleterious alleles are not masked by dominant alleles. However, this mechanism is less straightforward to apply in the model plant species Arabidopsis thaliana. For 1 Myr, A. thaliana has been self-compatible, a life history switch that has caused: a reduction in pollen competition, increased homozygosity, and a dilution of masking in diploid expressed, sporophytic genes. In this study, we have investigated the relative strength of selection on pollen genes compared with sporophytic genes in A. thaliana. We present two major findings: 1) before becoming self-compatible, positive selection was stronger on pollen genes than sporophytic genes for A. thaliana and 2) current polymorphism data indicate that selection is weaker on pollen genes compared with sporophytic genes. This weaker selection on pollen genes can in part be explained by their higher tissue specificity, which in outbreeding plants can be outweighed by the effects of haploid expression and pollen competition. These results indicate that since A. thaliana has become self-compatible, selection on pollen genes has become more relaxed. This has led to higher polymorphism levels and a higher build-up of deleterious mutations in pollen genes compared with sporophytic genes.


Assuntos
Arabidopsis/genética , Acúmulo de Mutações , Pólen/metabolismo , Arabidopsis/fisiologia , Diploide , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Genes de Plantas/genética , Genes de Plantas/fisiologia , Haploidia , Ploidias , Pólen/genética , Seleção Genética/genética
19.
PLoS Genet ; 15(6): e1008211, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31194744

RESUMO

Although evolutionary transitions from sexual to asexual reproduction are frequent in eukaryotes, the genetic bases of these shifts remain largely elusive. Here, we used classic quantitative trait analysis, combined with genomic and transcriptomic information to dissect the genetic basis of asexual, parthenogenetic reproduction in the brown alga Ectocarpus. We found that parthenogenesis is controlled by the sex locus, together with two additional autosomal loci, highlighting the key role of the sex chromosome as a major regulator of asexual reproduction. We identify several negative effects of parthenogenesis on male fitness, and different fitness effects of parthenogenetic capacity depending on the life cycle generation. Although allele frequencies in natural populations are currently unknown, we discuss the possibility that parthenogenesis may be under both sex-specific selection and generation/ploidally-antagonistic selection, and/or that the action of fluctuating selection on this trait may contribute to the maintenance of polymorphisms in populations. Importantly, our data provide the first empirical illustration, to our knowledge, of a trade-off between the haploid and diploid stages of the life cycle, where distinct parthenogenesis alleles have opposing effects on sexual and asexual reproduction and may help maintain genetic variation. These types of fitness trade-offs have profound evolutionary implications in natural populations and may structure life history evolution in organisms with haploid-diploid life cycles.


Assuntos
Partenogênese/genética , Feófitas/genética , Reprodução Assexuada/genética , Cromossomos Sexuais/genética , Evolução Biológica , Genoma/genética , Haploidia , Estágios do Ciclo de Vida/genética , Feófitas/crescimento & desenvolvimento , Polimorfismo Genético , Locos de Características Quantitativas/genética
20.
Nat Protoc ; 14(7): 1991-2014, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31160788

RESUMO

Ploidy represents the number of chromosome sets in a cell. Although gametes have a haploid genome (n), most mammalian cells have diploid genomes (2n). The diploid status of most cells correlates with the number of probable alleles for each autosomal gene and makes it difficult to target these genes via mutagenesis techniques. Here, we describe a 7-week protocol for the derivation of mouse haploid embryonic stem cells (hESCs) from female gametes that also outlines how to maintain the cells once derived. We detail additional procedures that can be used with cell lines obtained from the mouse Haplobank, a biobank of >100,000 individual mouse hESC lines with targeted mutations in 16,970 genes. hESCs can spontaneously diploidize and can be maintained in both haploid and diploid states. Mouse hESCs are genomically and karyotypically stable, are innately immortal and isogenic, and can be derived in an array of differentiated cell types; they are thus highly amenable to genetic screens and to defining molecular connectivity pathways.


Assuntos
Técnicas de Cultura de Células/métodos , Haploidia , Células-Tronco Embrionárias Murinas/citologia , Células-Tronco Embrionárias Murinas/fisiologia , Animais , Blastocisto/citologia , Linhagem Celular , Separação Celular/métodos , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Fluxo de Trabalho
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