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1.
Cytogenet Genome Res ; 161(5): 249-256, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34433167

RESUMO

B chromosomes occur in different species of the small characid fishes of the genus Moenkhausia. These supernumerary elements, that do not recombine with chromosomes of the standard A complement and follow their own evolutionary mechanism vary in number, morphology, and distribution. Here, we show karyotypic data of individuals of 2 populations of Moenkhausia oligolepis of the Brazilian Amazon (Pedro Correia and Taboquinha streams, Tocantins river basin), both with a diploid number of 50 chromosomes and karyotypic formula of 10m + 32sm + 8a. In addition to the normal complement, we also observed the occurrence of B chromosomes in the 2 populations with intra- and interindividual variation ranging from 0 to 10 Bs, independent of sex. The C-banding pattern evidenced heterochromatic blocks located mainly in the pericentromeric region of the chromosomes, while the B chromosomes appeared euchromatic. Silver-stained nucleolus organizer regions were identified in multiples sites, and some of these blocks were positive when stained with chromomycin A3. The karyotype analysis and the application of whole-chromosome painting in populations of M. oligolepis reinforce the conservation of the basal diploid number for the genus, as well as the evolutionary tendency in these fishes to carry B chromosomes. Both populations turned out to be in different stages of stability and expansion of their B chromosomes. We further suggest that the origin of these chromosomes is due to the formation of isochromosomes. Here, we identified a pair of complement A chromosomes involved in this process.


Assuntos
Characidae/genética , Instabilidade Cromossômica , Cromossomos/química , Cariotipagem/métodos , Animais , Brasil , Cromomicina A3/química , Bandeamento Cromossômico , Mapeamento Cromossômico , Feminino , Corantes Fluorescentes/química , Hibridização in Situ Fluorescente , Cariótipo , Masculino , Mitose , Ploidias
2.
Methods Mol Biol ; 2287: 105-125, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270026

RESUMO

Determination of the ploidy level is an essential step when trying to produce doubled haploids (DHs) in any species. Each species and method used to produce DHs has its own frequency of DH production, which means that the rest of plants produced stay haploid. Since haploids are of little use for breeding purposes, it is necessary to distinguish them from true DHs. For this, several methodologies are available, including flow cytometry, chromosome counting, chloroplast counting in stomatal guard cells, measurement of stomatal size and length, counting of nucleoli, evaluation of pollen formation and viability, analysis of cell size, and analysis of morphological markers. However, not all of them are equally easy to use, affordable, reliable, reproducible, and resolutive and therefore useful for a particular case. In this chapter, we revise these methods available to assess the ploidy level of plants, discussing their respective advantages and limitations, and provide some troubleshooting tips and hints to help decide which to choose in each case.


Assuntos
Melhoramento Vegetal/métodos , Zea mays/crescimento & desenvolvimento , Zea mays/genética , Cruzamentos Genéticos , Haploidia , Ploidias , Sementes/genética , Sementes/crescimento & desenvolvimento
3.
Methods Mol Biol ; 2288: 129-144, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270009

RESUMO

Rapeseed (Brassica napus) is one of the most important oilseed crops worldwide. It is also a model system to study the process of microspore embryogenesis, due to the high response of some B. napus lines, and to the refinements of the protocols. This chapter presents a protocol for the induction of haploid and DH embryos in B. napus through isolated microspore culture in two specific backgrounds widely used in DH research, the high response DH4079 line and the low response DH12075 line. We also present methods to identify the best phenological window to identify buds with microspores/pollen at the right developmental stage to induce this process. Methods to determine microspore/pollen viability and to check the ploidy by flow cytometry are also described.


Assuntos
Brassica napus/crescimento & desenvolvimento , Brassica napus/genética , Melhoramento Vegetal/métodos , Aclimatação/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Meios de Cultura/química , Diploide , Citometria de Fluxo , Genótipo , Germinação/genética , Haploidia , Homozigoto , Biologia Molecular/métodos , Ploidias , Pólen/genética , Pólen/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
4.
Methods Mol Biol ; 2288: 145-162, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270010

RESUMO

Culture of isolated microspores is a widely used method to obtain haploid and doubled haploid plants for many crop species. This protocol describes the steps necessary to obtain a large number of microspore derived embryos for pakchoi (Brassica rapa L. ssp. chinensis) and zicaitai (Brassica rapa L. ssp. сhinensis Hanelt var. purpuraria Kitam).


Assuntos
Brassica rapa/crescimento & desenvolvimento , Brassica rapa/genética , Melhoramento Vegetal/métodos , Brassica rapa/ultraestrutura , Cloroplastos/ultraestrutura , Cromossomos de Plantas/ultraestrutura , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Meios de Cultura/química , Diploide , Germinação/genética , Haploidia , Homozigoto , Microscopia de Fluorescência , Biologia Molecular/métodos , Ploidias , Pólen/genética , Pólen/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
5.
Methods Mol Biol ; 2288: 163-180, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270011

RESUMO

Brassica carinata, also known as Ethiopian or Abyssinian mustard, is a drought- and heat-tolerant oilseed with great potential as a dedicated industrial feedstock crop for use in biofuel and other bio-based applications. Doubled haploid technology, a system that allows for the rapid development of doubled haploid, completely homozygous plants through microspore embryogenesis, has been applied routinely in both B. carinata breeding and basic research. Here, we present a comprehensive isolated microspore culture protocol detailing the various steps involved in doubled haploid plant production for this species, from growing donor plants over harvesting flower buds and isolating, culturing and inducing microspores to regenerating doubled haploid embryos and plantlets.


Assuntos
Mostardeira/crescimento & desenvolvimento , Mostardeira/genética , Melhoramento Vegetal/métodos , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Meios de Cultura/química , Diploide , Haploidia , Homozigoto , Biologia Molecular/métodos , Ploidias , Pólen/genética , Pólen/crescimento & desenvolvimento , Pólen/ultraestrutura , Técnicas de Cultura de Tecidos
6.
Methods Mol Biol ; 2288: 201-216, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270013

RESUMO

Broccoli (Brassica olearecea var. italica) is a cole crop grown for its floral heads and stalks. It is rich in bioactive chemicals good for human health. Broccoli has been consumed as a vegetable since Roman times, but its production and consumption have increased significantly over the past few decades. Breeders try to develop new broccoli varieties with high yield, improved quality, and resistance to biotic and abiotic stresses. Almost all new broccoli varieties are F1 hybrids. Development of inbred broccoli lines that can be used as parents in hybrid production is a time-consuming and difficult process. Haploidization techniques can be utilized as a valuable support in broccoli breeding programs to speed up the production of genetically pure genotypes. Haploid plants of broccoli can be produced from immature male gametophytes via anther and microspore cultures with similar success rates. The most important parameters affecting the success of haploidization in broccoli are the genetic background (genotype) and the developmental stage of the microspores. Broccoli genotypes differ in their responses to androgenesis induction. The highest androgenesis response could be induced from microspores in late uninucleate and early binucleate stages. Recovery of diploid broccoli plants from haploids is possible via spontaneous and induced doubling. Doubled haploid (DH) broccoli lines are considered to be fully homozygous. Therefore, the production of DH lines is an alternative way to obtain pure inbred lines that can be utilized as parents in the development of new F1 hybrid varieties showing high levels of heterosis, high-quality heads, and uniform harvestable crop. We are using an anther culture-based haploid plant production system to develop DH broccoli lines in our broccoli breeding program. DH broccoli lines are produced from different genetic backgrounds within a year and handed to broccoli breeders.


Assuntos
Brassica/crescimento & desenvolvimento , Brassica/genética , Melhoramento Vegetal/métodos , Aclimatação/genética , Brassica/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Meios de Cultura/química , Diploide , Citometria de Fluxo , Flores/genética , Flores/crescimento & desenvolvimento , Haploidia , Homozigoto , Vigor Híbrido/genética , Biologia Molecular/métodos , Ploidias , Pólen/genética , Pólen/crescimento & desenvolvimento , Regeneração/genética , Técnicas de Cultura de Tecidos
7.
Methods Mol Biol ; 2288: 217-232, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270014

RESUMO

Here, we describe the first protocol of European radish (Raphanus sativus L. subsp. sativus convar. radicula) for obtaining doubled haploid plants through in vitro microspore culture, in which the full cycle of doubled haploid formation was successfully achieved. Using this protocol, a yield of up to eight embryoids per Petri dish can be obtained. Effectiveness of this protocol was confirmed for several genotypes of European radish.


Assuntos
Melhoramento Vegetal/métodos , Raphanus/crescimento & desenvolvimento , Raphanus/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/fisiologia , Meios de Cultura/química , Diploide , Corantes Fluorescentes , Genótipo , Haploidia , Homozigoto , Indóis , Biologia Molecular/métodos , Ploidias , Pólen/genética , Pólen/crescimento & desenvolvimento , Raphanus/fisiologia , Regeneração/genética , Coloração e Rotulagem , Técnicas de Cultura de Tecidos
8.
Methods Mol Biol ; 2288: 235-250, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270015

RESUMO

Eggplant is one of the five important, worldwide-distributed solanaceous crops. The use of anther culture technology to produce pure, 100% homozygous doubled haploid lines for hybrid seed production is possible since 1982, where the first protocol of wide application to different eggplant materials was published. From then on, different improvements and adaptations to different materials have been made. In parallel, protocols to implement isolated microspore culture technology in eggplant have been developed principally in the last decade, which opens the door for a more efficient DH production in this species. In this chapter, two protocols, one for anther and other for isolated microspore culture in eggplant, are described. Some steps and materials are common to both approaches. A detailed description of each step from is provided.


Assuntos
Melhoramento Vegetal/métodos , Solanum melongena/crescimento & desenvolvimento , Solanum melongena/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/fisiologia , Meios de Cultura/química , Diploide , Flores/genética , Flores/crescimento & desenvolvimento , Corantes Fluorescentes , Haploidia , Homozigoto , Indóis , Biologia Molecular/métodos , Ploidias , Pólen/genética , Pólen/crescimento & desenvolvimento , Regeneração/genética , Solanum melongena/fisiologia , Coloração e Rotulagem , Técnicas de Cultura de Tecidos
9.
Methods Mol Biol ; 2288: 251-266, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270016

RESUMO

The shed-microspore culture technique is an alternative sub-method combining anther and isolated microspore culture to induce microspore embryogenesis. Recently, its effective use in different types of peppers has drawn attention, because it has a higher embryo yield potential compared to anther culture and is more practical than isolated microspore culture. In this chapter, a stepwise protocol for shed-microspore culture of ornamental pepper is described. This protocol includes the steps of donor plant growth conditions, the choice of suitable flower buds based on DAPI staining of microspores, application of a cold pretreatment to flower buds, surface sterilization of the buds, shed-microspore culture of anthers, stress treatments, regeneration of androgenic in vitro plantlets, their acclimatization and ploidy analysis, and in vivo chromosome doubling of the haploid plants.


Assuntos
Capsicum/crescimento & desenvolvimento , Capsicum/genética , Melhoramento Vegetal/métodos , Pólen/crescimento & desenvolvimento , Pólen/genética , Capsicum/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/fisiologia , Meios de Cultura/química , Diploide , Flores/genética , Flores/crescimento & desenvolvimento , Corantes Fluorescentes , Haploidia , Homozigoto , Indóis , Biologia Molecular/métodos , Ploidias , Regeneração/genética , Coloração e Rotulagem , Técnicas de Cultura de Tecidos
10.
BMC Plant Biol ; 21(1): 325, 2021 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-34229602

RESUMO

BACKGROUND: Plant phylogeographic studies of species in subtropical China have mainly focused on rare and endangered species, whereas few studies have been conducted on taxa with relatively wide distribution, especially polyploid species. We investigated the cytotype and haplotype distribution pattern of the Actinidia chinensis complex, a widespread geographically woody liana with variable ploidy in subtropical China comprising two varieties, with three chloroplast fragments DNA (ndhF-rpl132, rps16-trnQ and trnE-trnT). Macroevolutionary, microevolutionary and niche modeling tools were also combined to disentangle the origin and the demographic history of the species or cytotypes. RESULTS: The ploidy levels of 3338 individuals from 128 populations sampled throughout the species distribution range were estimated with flow cytometry. The widespread cytotypes were diploids followed by tetraploids and hexaploids, whereas triploids and octoploids occurred in a few populations. Thirty-one chloroplast haplotypes were detected. The genetic diversity and genetic structure were found to be high between varieties (or ploidy races) chinensis and deliciosa. Our results revealed that these two varieties inhabit significantly different climatic niche spaces. Ecological niche models (ENMs) indicate that all varieties' ranges contracted during the Last Inter Glacial (LIG), and expanded eastward or northward during the Last Glacial Maximum (LGM). CONCLUSIONS: Pliocene and Plio-Pleistocene climatic fluctuations and vicariance appear to have played key roles in shaping current population structure and historical demography in the A. chinensis complex. The polyploidization process also appears to have played an important role in the historical demography of the complex through improving their adaptability to environmental changes.


Assuntos
Actinidia/classificação , Actinidia/citologia , Cloroplastos/classificação , Filogeografia , Teorema de Bayes , China , DNA de Cloroplastos/genética , Ecossistema , Variação Genética , Genética Populacional , Haplótipos/genética , Método de Monte Carlo , Ploidias
11.
Nat Commun ; 12(1): 4202, 2021 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-34244507

RESUMO

Biochemical reactions typically depend on the concentrations of the molecules involved, and cell survival therefore critically depends on the concentration of proteins. To maintain constant protein concentrations during cell growth, global mRNA and protein synthesis rates are tightly linked to cell volume. While such regulation is appropriate for most proteins, certain cellular structures do not scale with cell volume. The most striking example of this is the genomic DNA, which doubles during the cell cycle and increases with ploidy, but is independent of cell volume. Here, we show that the amount of histone proteins is coupled to the DNA content, even though mRNA and protein synthesis globally increase with cell volume. As a consequence, and in contrast to the global trend, histone concentrations decrease with cell volume but increase with ploidy. We find that this distinct coordination of histone homeostasis and genome content is already achieved at the transcript level, and is an intrinsic property of histone promoters that does not require direct feedback mechanisms. Mathematical modeling and histone promoter truncations reveal a simple and generalizable mechanism to control the cell volume- and ploidy-dependence of a given gene through the balance of the initiation and elongation rates.


Assuntos
Histonas/biossíntese , Modelos Genéticos , Biossíntese de Proteínas/genética , RNA Mensageiro/biossíntese , Transcrição Genética , DNA Fúngico/genética , Genoma Fúngico , Histonas/genética , Ploidias , Regiões Promotoras Genéticas/genética , RNA Fúngico/biossíntese , RNA Fúngico/genética , RNA Mensageiro/genética , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/biossíntese , Proteínas de Saccharomyces cerevisiae/genética
12.
Nat Commun ; 12(1): 4264, 2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34253736

RESUMO

Single-cell RNA-seq reveals the role of pathogenic cell populations in development and progression of chronic diseases. In order to expand our knowledge on cellular heterogeneity, we have developed a single-nucleus RNA-seq2 method tailored for the comprehensive analysis of the nuclear transcriptome from frozen tissues, allowing the dissection of all cell types present in the liver, regardless of cell size or cellular fragility. We use this approach to characterize the transcriptional profile of individual hepatocytes with different levels of ploidy, and have discovered that ploidy states are associated with different metabolic potential, and gene expression in tetraploid mononucleated hepatocytes is conditioned by their position within the hepatic lobule. Our work reveals a remarkable crosstalk between gene dosage and spatial distribution of hepatocytes.


Assuntos
Fígado/metabolismo , Ploidias , Análise de Sequência de RNA , Análise de Célula Única , Animais , Biomarcadores/metabolismo , Núcleo Celular/metabolismo , Doença Crônica , Secções Congeladas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ontologia Genética , Hepatócitos/metabolismo , Fígado/patologia , Hepatopatias/patologia , Camundongos Endogâmicos C57BL , Regeneração , Células-Tronco/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Genética
13.
Int J Mol Sci ; 22(9)2021 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-34067074

RESUMO

The recently discovered interleukin (IL)- 32 isoform IL-32θ exerts anti-metastatic effects in the breast tumor microenvironment. However, the involvement of IL-32θ in breast cancer cell proliferation is not yet fully understood; therefore, the current study aimed to determine how IL-32θ affects cancer cell growth and evaluated the responses of IL-32θ-expressing cells to other cancer therapy. We compared the functions of IL-32θ in triple-negative breast cancer MDA-MB-231 cells that stably express IL-32θ, with MDA-MB-231 cells transfected with a mock vector. Slower growth was observed in cells expressing IL-32θ than in control cells, and changes were noted in nuclear morphology, mitotic division, and nucleolar size between the two groups of cells. Interleukin-32θ significantly reduced the colony-forming ability of MDA-MB-231 cells and induced permanent cell cycle arrest at the G1 phase. Long-term IL-32θ accumulation triggered permanent senescence and chromosomal instability in MDA-MB-231 cells. Genotoxic drug doxorubicin (DR) reduced the viability of MDA-MB-231 cells not expressing IL-32θ more than in cells expressing IL-32θ. Overall, these findings suggest that IL-32θ exerts antiproliferative effects in breast cancer cells and initiates senescence, which may cause DR resistance. Therefore, targeting IL-32θ in combination with DR treatment may not be suitable for treating metastatic breast cancer.


Assuntos
Senescência Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Interleucinas/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Neoplasias da Mama/patologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Forma do Núcleo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Instabilidade Genômica , Humanos , Fenótipo , Ploidias
14.
Int J Mol Sci ; 22(11)2021 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-34070394

RESUMO

The genetic control of host response to the fungal necrotrophic disease Septoria nodorum blotch (SNB) in bread wheat is complex, involving many minor genes. Quantitative trait loci (QTL) controlling SNB response were previously identified on chromosomes 1BS and 5BL. The aim of this study, therefore, was to align and compare the genetic map representing QTL interval on 1BS and 5BS with the reference sequence of wheat and identify resistance genes (R-genes) associated with SNB response. Alignment of QTL intervals identified significant genome rearrangements on 1BS between parents of the DH population EGA Blanco, Millewa and the reference sequence of Chinese Spring with subtle rearrangements on 5BL. Nevertheless, annotation of genomic intervals in the reference sequence were able to identify and map 13 and 12 R-genes on 1BS and 5BL, respectively. R-genes discriminated co-located QTL on 1BS into two distinct but linked loci. NRC1a and TFIID mapped in one QTL on 1BS whereas RGA and Snn1 mapped in the linked locus and all were associated with SNB resistance but in one environment only. Similarly, Tsn1 and WK35 were mapped in one QTL on 5BL with NETWORKED 1A and RGA genes mapped in the linked QTL interval. This study provided new insights on possible biochemical, cellular and molecular mechanisms responding to SNB infection in different environments and also addressed limitations of using the reference sequence to identify the full complement of functional R-genes in modern varieties.


Assuntos
Ascomicetos/crescimento & desenvolvimento , Resistência à Doença , Genes de Plantas , Proteínas de Plantas , Ploidias , Triticum , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Triticum/genética , Triticum/microbiologia
15.
Fertil Steril ; 116(2): 388-395, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33827765

RESUMO

OBJECTIVE: To assess the association between body mass index (BMI) and embryo aneuploidy and mosaicism in a cohort of patients undergoing in vitro fertilization (IVF) with trophectoderm biopsy for preimplantation genetic testing for aneuploidy (PGT-A) using next-generation sequencing technology. DESIGN: Retrospective cohort study. SETTING: Academic center. PATIENTS: Patients undergoing their first IVF cycle with trophectoderm biopsy and PGT-A at our center between January 1, 2017, and August 31, 2020. Patients classified as underweight on the basis of BMI (BMI <18.5 kg/m2) and patients who underwent fresh embryo transfers were excluded. INTERVENTION: None. MAIN OUTCOME MEASURES: Number and proportion of aneuploid, mosaic, and euploid embryos. RESULTS: The patients were stratified according to the World Health Organization's BMI classification: normal weight (18.5-24.9 kg/m2, n = 1,254), overweight (25-29.9 kg/m2, n = 351), and obese (≥30 kg/m2, n = 145). Age-adjusted regression models showed no relationship between BMI classification and the number or proportion of aneuploid embryos. There were no statistically significant associations between BMI classifications and the number or proportion of mosaic or euploid embryos. A subgroup analysis of patients classified into age groups of <35, 35-40, and >40 years similarly showed no relationships between BMI and embryo ploidy outcomes. CONCLUSION: Body mass index was not associated with the number or proportion of aneuploid, mosaic, or euploid embryos in this large cohort of patients undergoing IVF with PGT-A, suggesting that the negative effect of excess weight on reproductive outcomes was independent of the ploidy status of the embryo cohort.


Assuntos
Índice de Massa Corporal , Fertilização In Vitro , Ploidias , Diagnóstico Pré-Implantação , Aborto Espontâneo/epidemiologia , Adulto , Aneuploidia , Feminino , Humanos , Idade Materna , Pessoa de Meia-Idade , Mosaicismo , Obesidade/complicações , Gravidez , Estudos Retrospectivos
16.
Fertil Steril ; 116(2): 583-596, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33926715

RESUMO

OBJECTIVE: To quantify the percentage of monopronuclear-derived blastocysts (MNBs) that are potentially useful for reproductive purposes using classic and state-of-the-art chromosome analysis approaches, and to study chromosomal distribution in the inner cell mass (ICM) and trophectoderm (TE) for intertissue/intratissue concordance comparison. DESIGN: Prospective experimental study. SETTING: Single-center in vitro fertilization clinic and reproductive genetics laboratory. PATIENT(S): A total of 1,128 monopronuclear zygotes were obtained between June 2016 and December 2018. INTERVENTION(S): MNBs were whole-fixed or biopsied to obtain a portion of ICM and 2 TE portions (TE1 and TE2) and were subsequently analyzed by fluorescence in situ hybridization, new whole-genome sequencing, and fingerprinting by single-nucleotide polymorphism array-based techniques (a-SNP). MAIN OUTCOME MEASURE(S): We assessed MNB rate, ploidy rate, and chromosomal constitution by new whole-genome sequencing, and parental composition by comparative a-SNP, performed in a "trio"-format (embryo/parents). The 24-chromosome distribution was compared between the TE and the ICM and within the TE. RESULT(S): A total of 18.4% of monopronuclear zygotes progressed to blastocysts; 77.6% of MNBs were diploid; 20% of MNBs were male and euploid, which might be reproductively useful. Seventy-five percent of MNBs were biparental and half of them were euploid, indicating that 40% might be reproductively useful. Intratissue concordance (TE1/TE2) was established for 93.3% and 73.3% for chromosome matching. Intertissue concordance (TE/ICM) was established for 78.8%, but 57.6% for chromosome matching. When segmental aneuploidy was not considered, intratissue concordance and chromosome matching increased to 100% and 80%, respectively, and intertissue concordance and chromosome matching increased to 84.8% and 75.8%, respectively. CONCLUSION(S): The a-SNP-trio strategy provides information about ploidy, euploidy, and parental origin in a single biopsy. This approach enabled us to identify 40% of MNBs with reproductive potential, which can have a significant effect in the clinical setting. Additionally, segmental aneuploidy is relevant for mismatched preimplantation genetic testing of aneuploidies, both within and between MNB tissues. Repeat biopsy might clarify whether segmental aneuploidy is a prone genetic character.


Assuntos
Blastocisto/ultraestrutura , Cromossomos/ultraestrutura , Ploidias , Polimorfismo de Nucleotídeo Único , Biópsia , Blastocisto/patologia , Massa Celular Interna do Blastocisto/ultraestrutura , Impressões Digitais de DNA , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Hibridização in Situ Fluorescente , Estudos Prospectivos
17.
Genes (Basel) ; 12(4)2021 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-33810423

RESUMO

Consumers' choices are mainly based on fruit external characteristics such as the final size, weight, and shape. The majority of edible fruit are by tree fruit species, among which peach is the genomic and genetic reference for Prunus. In this research, we used a peach with a slow ripening (SR) phenotype, identified in the Fantasia (FAN) nectarine, associated with misregulation of genes involved in mesocarp identity and showing a reduction of final fruit size. By investigating the ploidy level, we observed a progressive increase in endoreduplication in mesocarp, which occurred in the late phases of FAN fruit development, but not in SR fruit. During fruit growth, we also detected that genes involved in endoreduplication were differentially modulated in FAN compared to SR. The differential transcriptional outputs were consistent with different chromatin states at loci of endoreduplication genes. The impaired expression of genes controlling cell cycle and endocycle as well as those claimed to play a role in fruit tissue identity result in the small final size of SR fruit.


Assuntos
Perfilação da Expressão Gênica/métodos , Prunus persica/fisiologia , Locos de Características Quantitativas , Ciclo Celular , Endorreduplicação , Regulação da Expressão Gênica de Plantas , Fenótipo , Proteínas de Plantas/genética , Ploidias , Prunus persica/genética , Análise de Sequência de RNA
18.
Methods Mol Biol ; 2250: 257-270, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33900611

RESUMO

Bamboo, a fast-growing non-timber forest plant with many uses, is a valuable species for green development. However, bamboo flowering is very infrequent, extending, in general, for up to 120 years. Ecologically, bamboo species are generally better adapted to various environments than other grasses. Therefore, the species deserves a special status in what could be called Ecological Bioeconomy. An understanding of the genetic processes of bamboo can help us sustainably develop and manage bamboo forests. Transposable elements (TEs), jumping genes or transposons, are major genetic elements in plant genomes. The rapid development of the bamboo reference genome, at the chromosome level, reveals that TEs occupy over 63.24% of the genome. This is higher than found in rice, Brachypodium, and sorghum. The bamboo genome contains diverse families of TEs, which play a significant role in bamboo's biological processes including growth and development. TEs provide important clues for understanding the evolution of the bamboo genome. In this chapter, we briefly describe the current status of research on TEs in the bamboo genome, their regulation, and transposition mechanisms. Perspectives for future research are also provided.


Assuntos
Elementos de DNA Transponíveis/genética , Genoma de Planta/genética , Genômica/métodos , Sasa/genética , Bases de Dados Genéticas , Regulação da Expressão Gênica de Plantas , Variação Genética , Tamanho do Genoma/genética , Internet , Melhoramento Vegetal/economia , Melhoramento Vegetal/métodos , Ploidias , Sasa/classificação , Especificidade da Espécie
19.
Plant Sci ; 307: 110879, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33902847

RESUMO

Glume hairiness or pubescence that occurs in hexaploid common wheat and its relatives at different ploidy levels is a distinct morphological marker. Current knowledge about the genetic control of wheat glume hairiness is based on study of Hg1 (formerly Hg) on chromosome 1AS. Here, we report characterization of a new locus for hairy glume Hg2 in synthetic hexaploid wheat line CIGM86.944. Hg2 was inherited a dominant allele. Bulked segregant analysis and RNA-sequencing (BSR-Seq) was performed on an F2:3 population from cross CIGM86.944 × Shannong 29 (glabrous glume), which localized Hg2 in a 2.02 cM genetic interval corresponding to ∼1.08 Mb (754,001,564-755,082,433 Mb) on chromosome 2BL in the Chinese Spring reference genome. Gene annotation and expression identified TraesCS2B02 G562300.1 encoding diacylglycerol kinase 5 protein and TraesCS2B02 G561400.1 encoding a wound-responsive family protein as possible candidate genes regulating development of glume hairiness. The identification of Hg2 provides new insights into the genetic control of glume hairiness in wheat.


Assuntos
Mapeamento Cromossômico , Genes de Plantas , Marcadores Genéticos , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Triticum/anatomia & histologia , Triticum/genética , Fenótipo , Ploidias
20.
Plant Physiol Biochem ; 163: 296-307, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33892228

RESUMO

It has been described in rice that Si only plays a physical barrier that does not allow Fe to enter cell apoplast, causing Fe deficiency responses even under Fe sufficiency growth conditions. Most of the conclusions were attained at acidic pH, but rice is also grown at calcareous conditions, which especially induce Fe deficiency in the plants. In this study, we assay the effect of Si in rice suffering both Fe deficiency and sufficiency in hydroponics at two pHs (5.5 and 7.5). Plant biometric parameters, ROS concentration, enzymatic activities, and total phenolic compounds, as well as ploidy levels, have been determined. In general, both pHs promoted similar rice responses under Fe sufficiency and deficiency status, but at pH 7.5, stress was favored. Flow cytometry studies revealed that Fe deficiency increased the percentage of cells in higher ploidy levels. Moreover, under this Fe status, Si addition enhanced this effect. This increase contributed to maintaining chloroplast structure which may have preserved antioxidant activities, and fortified cell walls, diminishing Fe uptake. The first is considered a beneficial effect as plants presented acceptable SPAD values, well chloroplast structure, and qualitatively high fluorescence observed by confocal microscopy, even under Fe deficiency. But contributes to intensify the Fe shortage, by decreasing apoplast Fe pools. In summary, Si addition to rice plants may not only behave as an apoplastic barrier but may also protect plant chloroplast and alter the plant endoreplication cycle, giving a memory effect to cope with present and future stresses.


Assuntos
Oryza , Hidroponia , Ferro , Oryza/genética , Raízes de Plantas , Ploidias , Silício/farmacologia
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