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1.
Elife ; 102021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34596043

RESUMO

Evolutionary adaptation to a constant environment is driven by the accumulation of mutations which can have a range of unrealized pleiotropic effects in other environments. These pleiotropic consequences of adaptation can influence the emergence of specialists or generalists, and are critical for evolution in temporally or spatially fluctuating environments. While many experiments have examined the pleiotropic effects of adaptation at a snapshot in time, very few have observed the dynamics by which these effects emerge and evolve. Here, we propagated hundreds of diploid and haploid laboratory budding yeast populations in each of three environments, and then assayed their fitness in multiple environments over 1000 generations of evolution. We find that replicate populations evolved in the same condition share common patterns of pleiotropic effects across other environments, which emerge within the first several hundred generations of evolution. However, we also find dynamic and environment-specific variability within these trends: variability in pleiotropic effects tends to increase over time, with the extent of variability depending on the evolution environment. These results suggest shifting and overlapping contributions of chance and contingency to the pleiotropic effects of adaptation, which could influence evolutionary trajectories in complex environments that fluctuate across space and time.


Assuntos
Adaptação Biológica , Aptidão Genética , Pleiotropia Genética/fisiologia , Saccharomyces cerevisiae/fisiologia , Aclimatação , Diploide , Meio Ambiente , Haploidia , Saccharomyces cerevisiae/genética
2.
BMC Genomics ; 22(1): 699, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-34579651

RESUMO

BACKGROUND: Nutrition and cell size play an important role in the determination of caste differentiation in queen and worker of honeybees (Apis mellifera), whereas the haploid genome dominates the differentiation of drones. However, the effects of female developmental environment on the development of males remain unclear. In this study, young drone larvae were transferred into worker cells (WCs) or remained in drone cells (DCs) to rear drones. The drone larvae were also grafted into queen cells (QCs) for 48 h and then transplanted into drone cells until emerging. Morphological indexes and reproductive organs of these three types of newly emerged drones were measured. Newly emerged drones and third instar drone larvae from WCs, DCs and QCs were sequenced by RNA sequencing (RNA-Seq). RESULTS: The amount of food remaining in cells of the QC and WC groups was significantly different to that in the DC group at the early larval stage. Morphological results showed that newly emerged DC drones had bigger body sizes and more well-developed reproductive tissues than WC and QC drones, whereas the reproductive tissues of QC drones were larger than those of WC drones. Additionally, whole body gene expression results showed a clear difference among three groups. At larval stage there were 889, 1761 and 1927 significantly differentially expressed genes (DEGs) in WC/DC, QC/DC and WC/QC comparisons, respectively. The number of DEGs decreased in adult drones of these three comparisons [678 (WC/DC), 338 (QC/DC) and 518 (WC/QC)]. A high number of DEGs were involved in sex differentiation, growth, olfaction, vision, mammalian target of rapamycin (mTOR), Wnt signaling pathways, and other processes. CONCLUSIONS: This study demonstrated that the developmental environment of honeybee females can delay male development, which may serve as a model for understanding the regulation of sex differentiation and male development in social insects by environmental factors.


Assuntos
Diferenciação Sexual , Olfato , Animais , Abelhas/genética , Feminino , Haploidia , Larva/genética , Masculino , Análise de Sequência de RNA
3.
Elife ; 102021 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-34533136

RESUMO

Eukaryotic life cycles alternate between haploid and diploid phases and in phylogenetically diverse unicellular eukaryotes, expression of paralogous homeodomain genes in gametes primes the haploid-to-diploid transition. In the unicellular chlorophyte alga Chlamydomonas, KNOX and BELL TALE-homeodomain genes mediate this transition. We demonstrate that in the liverwort Marchantia polymorpha, paternal (sperm) expression of three of five phylogenetically diverse BELL genes, MpBELL234, and maternal (egg) expression of both MpKNOX1 and MpBELL34 mediate the haploid-to-diploid transition. Loss-of-function alleles of MpKNOX1 result in zygotic arrest, whereas a loss of either maternal or paternal MpBELL234 results in variable zygotic and early embryonic arrest. Expression of MpKNOX1 and MpBELL34 during diploid sporophyte development is consistent with a later role for these genes in patterning the sporophyte. These results indicate that the ancestral mechanism to activate diploid gene expression was retained in early diverging land plants and subsequently co-opted during evolution of the diploid sporophyte body.


Assuntos
Diploide , Células Germinativas Vegetais , Marchantia/genética , Genes de Plantas , Haploidia , Filogenia
4.
Elife ; 102021 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-34523420

RESUMO

Although studies of Saccharomyces cerevisiae have provided many insights into mutagenesis and DNA repair, most of this work has focused on a few laboratory strains. Much less is known about the phenotypic effects of natural variation within S. cerevisiae's DNA repair pathways. Here, we use natural polymorphisms to detect historical mutation spectrum differences among several wild and domesticated S. cerevisiae strains. To determine whether these differences are likely caused by genetic mutation rate modifiers, we use a modified fluctuation assay with a CAN1 reporter to measure de novo mutation rates and spectra in 16 of the analyzed strains. We measure a 10-fold range of mutation rates and identify two strains with distinctive mutation spectra. These strains, known as AEQ and AAR, come from the panel's 'Mosaic beer' clade and share an enrichment for C > A mutations that is also observed in rare variation segregating throughout the genomes of several Mosaic beer and Mixed origin strains. Both AEQ and AAR are haploid derivatives of the diploid natural isolate CBS 1782, whose rare polymorphisms are enriched for C > A as well, suggesting that the underlying mutator allele is likely active in nature. We use a plasmid complementation test to show that AAR and AEQ share a mutator allele in the DNA repair gene OGG1, which excises 8-oxoguanine lesions that can cause C > A mutations if left unrepaired.


Assuntos
Variação Genética , Mutação Puntual , Saccharomyces cerevisiae/genética , Alelos , Sistemas de Transporte de Aminoácidos Básicos/genética , Reparo do DNA , Diploide , Teste de Complementação Genética , Haploidia , Taxa de Mutação , Fenótipo , Saccharomyces cerevisiae/classificação , Proteínas de Saccharomyces cerevisiae/genética
5.
PLoS One ; 16(8): e0249505, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34343170

RESUMO

Assessment of genetic purity of parental inbred lines and their resultant F1 hybrids is an essential quality control check in maize hybrid breeding, variety release and seed production. In this study, genetic purity, parent-offspring relationship and diversity among the inbred lines were assessed using 92 single-nucleotide polymorphism (SNP) markers. A total of 188 maize genotypes, comprising of 26 inbred lines, four doubled haploid (DH) lines and 158 single-cross maize hybrids were investigated in this study using Kompetitive Allele Specific Polymerase Chain Reaction (KASP) genotyping assays. The bi-allelic data was analyzed for genetic purity and diversity parameters using GenAlex software. The SNP markers were highly polymorphic and 90% had polymorphic information content (PIC) values of > 0.3. Pairwise genetic distances among the lines ranged from 0.05 to 0.56, indicating a high level of dissimilarity among the inbred lines. A maximum genetic distance of (0.56) was observed between inbred lines CKDHL0089 and CML443 while the lowest (0.05) was between I-42 and I-40. The majority (67%) of the inbred lines studied were genetically pure with residual heterozygosity of <5%, while only 33% had heterozygosity levels of >5%. Inbred lines, which were not pure, require purification through further inbreeding. Cluster analysis partitioned the lines into three distinct genetic clusters with the potential to contribute new beneficial alleles to the maize breeding program. Out of the 68 hybrids (43%) that passed the parent-offspring test, seven hybrids namely; SCHP29, SCHP95, SCHP94, SCHP134, SCHP44, SCHP114 and SCHP126, were selected as potential candidates for further evaluation and release due to their outstanding yield performance.


Assuntos
Genes de Plantas/genética , Melhoramento Vegetal/métodos , Polimorfismo de Nucleotídeo Único/genética , Zea mays/genética , Alelos , Cromossomos de Plantas/genética , Variação Genética/genética , Haploidia , Hormônio do Crescimento Humano , Hibridização Genética/genética
6.
Nat Commun ; 12(1): 4369, 2021 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-34272403

RESUMO

There is a strong demand for methods that can efficiently reconstruct valid super-resolution intact genome 3D structures from sparse and noise single-cell Hi-C data. Here, we develop Single-Cell Chromosome Conformation Calculator (Si-C) within the Bayesian theory framework and apply this approach to reconstruct intact genome 3D structures from single-cell Hi-C data of eight G1-phase haploid mouse ES cells. The inferred 100-kb and 10-kb structures consistently reproduce the known conserved features of chromatin organization revealed by independent imaging experiments. The analysis of the 10-kb resolution 3D structures reveals cell-to-cell varying domain structures in individual cells and hyperfine structures in domains, such as loops. An average of 0.2 contact reads per divided bin is sufficient for Si-C to obtain reliable structures. The valid super-resolution structures constructed by Si-C demonstrate the potential for visualizing and investigating interactions between all chromatin loci at the genome scale in individual cells.


Assuntos
Cromatina/metabolismo , Cromossomos/metabolismo , Células-Tronco Embrionárias/metabolismo , Genoma , Análise de Célula Única/métodos , Animais , Teorema de Bayes , Cromatina/química , Cromatina/genética , Cromossomos/química , Cromossomos/genética , Fase G1 , Haploidia , Hibridização in Situ Fluorescente , Camundongos , Conformação Molecular
7.
BMC Plant Biol ; 21(1): 353, 2021 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-34311707

RESUMO

BACKGROUND: In breeding programs, the selection of cultivars with the highest yield potential consisted in the selection of the yield per se, which resulted in cultivars with higher grains per spike (GN) and occasionally increased grain weight (GW) (main numerical components of the yield). In this study, quantitative trait loci (QTL) for GW, GN and spike fertility traits related to GN determination were mapped using two doubled haploid (DH) populations (Baguette Premium 11 × BioINTA 2002 and Baguette 19 × BioINTA 2002). RESULTS: In total 305 QTL were identified for 14 traits, out of which 12 QTL were identified in more than three environments and explained more than 10% of the phenotypic variation in at least one environment. Eight hotspot regions were detected on chromosomes 1A, 2B, 3A, 5A, 5B, 7A and 7B in which at least two major and stable QTL sheared confidence intervals. QTL on two of these regions (R5A.1 and R5A.2) have previously been described, but the other six regions are novel. CONCLUSIONS: Based on the pleiotropic analysis within a robust physiological model we conclude that two hotspot genomic regions (R5A.1 and R5A.2) together with the QGW.perg-6B are of high relevance to be used in marker assisted selection in order to improve the spike yield potential. All the QTL identified for the spike related traits are the first step to search for their candidate genes, which will allow their better manipulation in the future.


Assuntos
Locos de Características Quantitativas/genética , Triticum/genética , Cromossomos de Plantas/genética , Haploidia , Fenótipo , Melhoramento Vegetal
8.
Methods Mol Biol ; 2289: 23-45, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270061

RESUMO

Anther and microspore culture for producing haploid plants are very complex systems and include general effects where the specific effects must be identified and optimized to develop culture systems capable of producing the large numbers of haploids required by breeding programs. These general effects include genotype, physiological state of the source plant, age of the anthers and microspores, preculture treatments, culture conditions, and culture media. Design of experiments (DoE) is an experimental approach specifically designed to identify and optimize the multiple factors that make up complex systems, and is ideally suited for developing in vitro systems to produce haploids. The basic DoE strategy starts by screening multiple factors thought to affect the responses being measured. Screening identifies factors with large and small effects. Factors with large effects are used to manipulate the system, and are moved to the DoE optimization phase such as response surface methodology. Factors with small or trivial effects are eliminated from further consideration, and this simplifies the system. The basic concepts of fractional factorial designs and how to use them are explained. Fractional factorials are the most important DoE screening tool and are the first experiments run before DoE optimization experiments. To illustrate the unique properties of fractional factorials, a detailed example is provided that includes all of the calculations so that no statistical software is required.


Assuntos
Melhoramento Vegetal/métodos , Plantas/genética , Meios de Cultura/metabolismo , Genótipo , Haploidia , Software
9.
Methods Mol Biol ; 2289: 47-67, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270062

RESUMO

Efficient doubled haploid (DH) plant production is of great interest in the plant breeding industry and research because homozygous lines are obtained within a single generation shortening the breeding cycle substantially. DH protocol development can be a time- and resource-consuming process due to numerous factors affecting its success and efficiency. Here we present concepts and examples about how critical success factors can be identified throughout a DH protocol and an early microspore response monitored by simple impedance flow cytometry (IFC) measurements, which will help to optimize each step of an androgenesis-based DH protocol.


Assuntos
Citometria de Fluxo/métodos , Tecnologia/métodos , Impedância Elétrica , Haploidia , Melhoramento Vegetal/métodos , Plantas/genética
10.
Methods Mol Biol ; 2289: 71-85, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270063

RESUMO

As in any other economically important crop, the possibility of producing fully homozygous, doubled haploid lines in cucumber allows for faster and cheaper breeding. At present, the fastest way to doubled haploidy is the production of cucumber haploid plants and duplication of their chromosomes to make them doubled haploid. In this chapter, we describe a complete protocol to successfully produce cucumber doubled haploid plants, including the evaluation of their ploidy level by flow cytometry. Briefly, this protocol involves a first step of anther culture to induce microspores to divide and proliferate forming calli. The calli produced are isolated from anthers and transferred first to a liquid medium and then to a solid medium to induce organogenesis. Organogenic shoots will eventually give rise to entire DH plants.


Assuntos
Cucumis sativus/genética , Melhoramento Vegetal/métodos , Técnicas de Cultura de Tecidos/métodos , Meios de Cultura/metabolismo , Flores/genética , Haploidia
11.
Methods Mol Biol ; 2289: 3-22, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270060

RESUMO

Haploid production is of great importance in plant breeding programs. Doubled haploid technology accelerates the generation of inbred lines with homozygosity in all loci in a single year. Haploids can be induced in vitro via cultivating the haploid gametes or in vivo through inter- and intraspecific hybridization. Haploid induction through centromere engineering is a novel system that is theoretically applicable to many plant species. The present review chapter discusses the proposed molecular mechanisms of selective chromosome elimination in early embryogenesis and the effects of kinetochore component modifications on proper chromosome segregation. Finally, the advantages and limitations of the CENH3-mediated haploidization approach and its applications are highlighted.


Assuntos
Centrômero/genética , Plantas/genética , Animais , Produtos Agrícolas/genética , Genoma de Planta/genética , Haploidia , Humanos , Hibridização Genética/genética , Cinetocoros/fisiologia , Melhoramento Vegetal/métodos
12.
Methods Mol Biol ; 2289: 97-110, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270065

RESUMO

Doubled haploid (DH) technology is very advantageous in plant breeding. This technique is beneficial for reducing the time required to obtain pure lines and contributes to the selection efficiency. Using this technique, 100% homozygosity can be achieved in a single generation, while the development of stable lines using the traditional self-pollination method takes from 6 to 8 years. It has long been used in diverse crops including cucurbits. DHs can be obtained via parthenogenesis (pollination mostly with irradiated pollen), gynogenesis (in vitro culture of ovules and ovaries), and androgenesis (in vitro culture of microspores and anthers). All these methods have been used for over 30 years to develop haploid and DH lines in cucurbit crops. Nowadays, many researchers benefit from these techniques routinely. However, there are still many limits for using DH technology in watermelon breeding programmes. The number of developed DH lines is still very low.In this chapter, we present a protocol based on the different studies on haploids and DHs induced in watermelon through irradiated pollen technique, unfertilized ovule/ovary culture and anther/microspore culture. According to the results of all these studies, it is crucial to develop an efficient protocol for haploid embryo induction to enhance the frequency of obtaining haploid embryos in watermelon.


Assuntos
Citrullus/genética , Produtos Agrícolas/genética , Haploidia , Óvulo Vegetal/genética , Partenogênese/genética , Melhoramento Vegetal/métodos , Pólen/genética , Polinização/genética
13.
Methods Mol Biol ; 2289: 87-95, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270064

RESUMO

Parthenogenesis is the main technique to produce doubled haploid lines in melon. Although parthenogenesis is a genotype-dependent process and melon has a huge genotypic diversity, we developed a successful protocol for haploid embryo production via pollination with irradiated pollen and a protocol for chromosome doubling of haploid plants of 'Piel de Sapo' genotypes. 'Piel de sapo' genotype has lower efficiencies during the process in comparison with other genotypes, for instance, of the agrestis subspecies. Nevertheless, the doubled haploid lines produced have a great potential as pure parentals for hybrid seed production.


Assuntos
Cucurbitaceae/genética , Partenogênese/genética , Genótipo , Haploidia , Polinização/genética , Sementes/genética
14.
Methods Mol Biol ; 2289: 111-121, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270066

RESUMO

Production of homozygous pure parental lines is the first stage of hybrid vegetable breeding. Unfortunately, producing pure lines takes a long time by classical breeding methods, especially in open-pollinated vegetable species, and this period can be up to 8-10 years. Recently, doubled haploid (DH) technology, as a set of biotechnological methods, has emerged as an alternative to classical breeding methods and allows for the generation of 100% homozygous pure double haploid lines in 1 or 2 years. Although haploid plants were successfully produced via irradiated pollen technique and gynogenesis in some Cucurbita species, haploid plants have not been obtained from some lines due to genotype dependency, and haploidy frequency is still not sufficient for use in a breeding program. Thus, anther culture technique has emerged as an alternative technique in the DH process. The main objective of this chapter is to provide explanatory information on anther culture technique applied in the Cucurbita genus. For this purpose , key points and details of methods and protocols of the anther culture technique are described in summer squash (Cucurbita pepo L.), pumpkin (Cucurbita moschata Duch.), and winter squash (Cucurbita maxima Duch.).


Assuntos
Cucurbita/genética , Técnicas de Cultura/métodos , Melhoramento Vegetal/métodos , Genótipo , Haploidia , Pólen/genética , Polinização/genética
15.
Methods Mol Biol ; 2289: 123-133, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270067

RESUMO

The development of F1 hybrid vegetable varieties emerges as a result of a great effort, long time, investment, knowledge, and advanced technology. The first stage of hybrid vegetable breeding is obtaining pure lines. It is possible to obtain homozygous parent lines used in the production of hybrid varieties with traditional breeding methods. This period takes 8-10 years, especially in some vegetables which are highly open-pollinated, such as Cucurbita spp. Androgenetic- and/or gynogenetic-based dihaploidization methods provide 100% homozygous pure haploid lines in 1-2 years and save time and effort.The DH frequency by irradiated pollen technique and anther culture strongly depends on the genotypic response, whereby their practical use in a breeding program is still limited. As a possible alternative technique, gynogenesis (unfertilized ovule/ovarium cultures) switches on to produce haploid plants in some Cucurbita species. In the Cucurbita genus, gynogenesis has been one of the most studied and popular DH techniques and presented remarkable results in recent years.


Assuntos
Cucurbita/genética , Haploidia , Óvulo Vegetal/genética , Melhoramento Vegetal/métodos , Pólen/genética , Polinização/genética
16.
Methods Mol Biol ; 2289: 135-145, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270068

RESUMO

Due to their many superior agronomic traits (high yield and fruit quality, resistance/tolerance to biotic and abiotic stress factors, etc.), hybrid vegetable cultivars are widely used in vegetable production all over the world. The first stage of hybrid vegetable breeding is to obtain homozygous pure parental lines. Unfortunately, producing pure lines takes a long time by classical breeding methods, especially in open-pollinated vegetable species, and this period can be up to 8-10 years. Recently, doubled haploid (DH) technology, as a biotechnological method, has emerged as an alternative to classical breeding methods and allows for the generation of pure (100% homozygous) DH lines in one or two years.However, the DH technique needs labor-intensive efforts and experiences as well as the use of appropriate production technologies. The main objective of this chapter is to provide explanatory information on the technique of induction of parthenogenesis by irradiated pollen applied to several species of the Cucurbita genus. For this purpose , key points and details of methods and protocols of this technique are described in summer squash (Cucurbita pepo L.), pumpkin (Cucurbita moschata Duch.), and winter squash (Cucurbita maxima Duch.).


Assuntos
Cucurbita/genética , Partenogênese/genética , Frutas/genética , Haploidia , Fenótipo , Melhoramento Vegetal/métodos , Pólen/genética
17.
Methods Mol Biol ; 2289: 149-166, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270069

RESUMO

This chapter deals with microspore embryogenesis in Citrus. Microspore embryogenesis allows to induce immature gametes (microspores) and to deviate them, in this case, the male one, from the normal gametophytic developmental route in the direction of the sporophytic one, yielding homozygous organisms (embryos and plants).


Assuntos
Citrus/genética , Desenvolvimento Embrionário/genética , Pólen/genética , Regulação da Expressão Gênica de Plantas/genética , Haploidia , Hordeum/genética
18.
Methods Mol Biol ; 2289: 167-178, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270070

RESUMO

Doubled haploids have a high impact on the improvement of heterozygous crops through hybridization. Anther culture is a doubled haploid technique for producing homozygous lines. In coconut, a tree species reported to be recalcitrant for tissue culture, a successful doubled haploid protocol was established through anther culture. All the factors affecting androgenesis induction have been optimized. In this chapter, a stepwise protocol, from doubled haploid induction including palm selection, anther isolation, pretreatment, and culture initiation, up to plant regeneration and thereafter acclimatization of the regenerated plants, is described. Furthermore, the protocol for testing the anther-derived plants for the ploidy level is also presented.


Assuntos
Cocos/genética , Flores/genética , Haploidia , Hibridização Genética/genética
19.
Methods Mol Biol ; 2289: 179-198, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270071

RESUMO

This chapter deals with induction of haploidy via parthenogenesis in Persian walnut and via microspore embryogenesis in almond and hazelnut. Haploid induction through in situ parthenogenesis using pollination with irradiated pollen to stimulate the embryogenic development of the egg cell, followed by in vitro culture of the immature haploid embryos. Microspore embryogenesis allows the induction of immature pollen grains (microspores), to move away from the normal gametophytic developmental route in the direction of the sporophytic one, yielding homozygous organisms (embryos in this case). Unlike other fruit crops (such as Citrus), regeneration of entire plants has not yet been obtained in our studied nut crops; however, it gives the methodology should be used to continue the roadmap.


Assuntos
Corylus/genética , Juglans/genética , Nozes/genética , Prunus dulcis/genética , Produtos Agrícolas/genética , Haploidia , Partenogênese/genética , Pólen/genética , Polinização/genética , Técnicas de Cultura de Tecidos/métodos
20.
Methods Mol Biol ; 2289: 199-219, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270072

RESUMO

Cork oak (Quercus suber L.) is a forest tree species of the family Fagaceae. It is characterized by long life cycles which hamper doubled haploid plant production to obtain homozygotes and pure lines. The time-consuming method of repeated backcrossings by conventional breeding techniques to produce pure lines is impractical in woody species. Nevertheless, biotechnology has offered new tools to make it possible. A doubled haploid plant or embryo is one that is developed by the doubling of a haploid set of chromosomes. A protocol to produce doubled haploids of cork oak has been developed through microspore embryogenesis. By a heat stress treatment, the microspores inside the anther leave the gametophytic pathway and react shifting their development to the sporophytic pathway by means of which haploid embryos are obtained. Chromosome duplication of haploids from cork oak anther cultures occurs either spontaneously or may be induced by the application of antimitotic agents (e.g., colchicine, oryzalin, amiprophos-methyl). Furthermore, a genetic test is designed through microsatellite markers to elucidate whether the diploid embryos obtained are originally haploids which spontaneously duplicated their genome, or alternatively those embryos are generated from the diploid tissue of the anther wall. Here we describe a detailed protocol to produce doubled haploid individuals from cork oak anther cultures by using temperature stress and antimitotic agents.


Assuntos
Flores/genética , Resposta ao Choque Térmico/genética , Quercus/genética , Diploide , Haploidia , Repetições de Microssatélites/genética , Melhoramento Vegetal/métodos , Sementes/genética , Temperatura
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