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1.
Theriogenology ; 173: 64-72, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34339905

RESUMO

The aims of this study were to analyze the effects of different concentrations of rutin on primordial follicle survival and development after in vitro culture of sheep ovarian tissue, and to verify the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway in the rutin actions. Ovarian fragments were fixed for histological analysis (fresh control) or cultured in α-minimum essential medium alone (α-MEM+: control medium) or in α-MEM+supplemented with different concentrations of rutin (0.1; 1 or 10 µg/mL) for 7 days. Inhibition of the PI3K activity was performed in fragments cultured with 50 µM LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3 (apoptosis) and Akt phosphorylation (p-Akt). The results showed that 1 µg/mL rutin has a greater percentage of normal follicles (P < 0.05) than those of α-MEM+ and other rutin treatments. In addition, 1 µg/mL rutin maintained the follicular apoptosis similar (P > 0.05) to that of the fresh control and lower than α-MEM+ and 10 µg/mL rutin. All rutin concentrations increased (P < 0.05) follicular activation compared to fresh control and α-MEM+. Furthermore, follicular and oocyte diameters increased (P < 0.05) only after culture with 1 µg/mL rutin. After PI3K inhibition, there was a reduction (P < 0.05) of rutin follicular effects. In conclusion, rutin at 1 µg/mL reduces apoptosis, promotes activation and growth of sheep primordial follicles through the modulation of the PI3K/Akt signaling pathway after in vitro culture of ovine ovarian tissue.


Assuntos
Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Animais , Apoptose , Feminino , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Rutina/farmacologia , Ovinos , Técnicas de Cultura de Tecidos/veterinária
2.
Methods Mol Biol ; 2287: 187-197, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270030

RESUMO

The production of doubled haploids (DHs) has proved to be a highly valuable tool to obtain new cultivars. Among the cereals, barley (Hordeum vulgare L.) is the most successful species in large-scale haploid production. Techniques employed for this purpose are based on either the gynogenetic or the androgenetic pathway. Interspecific cross with Hordeum bulbosum L., haploid gene inducer (the hap gene), ovary culture, anther culture (AC), and isolated microspore culture (IMC) are the most used methods. Among all of them, IMC is regarded as a particularly effective system owing to the great increase in green plant numbers per spike and also the higher induction of chromosome doubling when compared with other methods. Thus, IMC provides the best way to mass scale production of new varieties.


Assuntos
Cromossomos de Plantas , Gametogênese Vegetal , Hordeum/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos/métodos , Meios de Cultura , Haploidia , Hordeum/genética , Pólen/genética , Pólen/crescimento & desenvolvimento
3.
Methods Mol Biol ; 2287: 215-226, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270032

RESUMO

The generation of doubled haploid barley plants by means of the so-called "Bulbosum" method has been practiced for meanwhile five decades. It rests upon the pollination of barley by its wild relative Hordeum bulbosum. This can result in the formation of hybrid embryos whose further development is typically associated with the loss of the pollinator's chromosomes. In recent years, this principle has, however, only rarely been used owing to the availability of efficient methods of anther and microspore culture. On the other hand, immature pollen-derived embryogenesis is to some extent prone to segregation bias in the resultant populations of haploids, which is due to its genotype dependency. Therefore, the principle of uniparental genome elimination has more recently regained increasing interest within the plant research and breeding community. The development of the present protocol relied on the use of the spring-type barley cultivar Golden Promise. The protocol is the result of a series of comparative experiments, which have addressed various methodological facets. The most influential ones included the method of emasculation, the temperature at flowering and early embryo development, the method, point in time and concentration of auxin administration for the stimulation of caryopsis development, the developmental stage at embryo dissection, as well as the nutrient medium used for embryo rescue. The present protocol allows the production of haploid barley plants at an efficiency of ca. 25% of the pollinated florets.


Assuntos
Hordeum/crescimento & desenvolvimento , Hordeum/genética , Melhoramento Vegetal/métodos , Técnicas de Cultura de Tecidos/métodos , Genótipo , Haploidia , Hordeum/embriologia , Pólen/genética , Pólen/crescimento & desenvolvimento , Polinização
4.
Methods Mol Biol ; 2287: 227-244, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270033

RESUMO

The use of doubled haploid (DH) plants in plant breeding programmes is the fastest route to release new varieties (4-6 years), allowing for a rapid response to end-user needs. Microspore embryogenesis is one of the most efficient methods for DH plant production in bread wheat. In this process, microspores triggered by a stress treatment or by application of bioactive compounds are reprogrammed to follow an embryogenic pathway that leads to the production of haploid or DH plants. In this chapter, we describe a protocol for anther culture of bread wheat. This protocol is based on an osmotic and starvation treatment of the anthers followed by the application of a microtubule disrupting agent. Anthers are cultured in an ovary pre-conditioned medium with mature ovaries from cv. Caramba. This protocol has been applied to a wide range of genotypes and F1s from bread and spelt wheat.


Assuntos
Pão/análise , Flores/crescimento & desenvolvimento , Flores/genética , Melhoramento Vegetal/métodos , Técnicas de Cultura de Tecidos/métodos , Triticum/crescimento & desenvolvimento , Triticum/genética , Haploidia , Pólen/genética , Pólen/crescimento & desenvolvimento
5.
Methods Mol Biol ; 2287: 257-266, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270035

RESUMO

Doubled haploid (DH) plant production belongs to modern biotechnology methods of plant breeding. The main advantage of DH plant production methods is the development of genetically homozygous lines in one generation, whilst in conventional breeding programmes, the development of homozygous lines requires more generations. The present chapter describes an efficient protocol for DH plant production in spelt wheat genotypes using in vitro anther culture.


Assuntos
Flores/crescimento & desenvolvimento , Melhoramento Vegetal/métodos , Técnicas de Cultura de Tecidos/métodos , Triticum/crescimento & desenvolvimento , Flores/genética , Haploidia , Técnicas In Vitro/métodos , Pólen/genética , Pólen/crescimento & desenvolvimento , Triticum/genética
6.
Methods Mol Biol ; 2287: 281-293, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270037

RESUMO

Isolated microspore culture systems have been designed in maize by several groups, mainly from the late 1980s to early 2000s. However, even with optimized protocols, microspore embryogenesis induction has remained very dependent on the genotype in maize, with elite germplasm generally displaying no response or very low response. Yet, these last few years, significant progress has been accomplished in understanding and controlling microspore embryogenesis induction in model dicot and monocot species. This knowledge may be transferred to maize, and isolated microspore culture may gain new interest in this crop, at least for embryogenesis research. The methods we hereby present in detail permit the purification of 3-12 × 105 viable microspores per maize tassel, at the favorable stage for microspore embryogenesis. When cultured in appropriate liquid media, microspores from responsive genotypes give rise to androgenic embryos, which can then be regenerated into fertile doubled haploid plants.


Assuntos
Gametogênese Vegetal , Técnicas de Cultura de Tecidos/métodos , Zea mays/crescimento & desenvolvimento , Meios de Cultura , Haploidia , Pólen/embriologia , Pólen/genética , Pólen/crescimento & desenvolvimento , Zea mays/embriologia , Zea mays/genética
7.
Methods Mol Biol ; 2287: 295-312, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270038

RESUMO

Here, we describe a method of triticale isolated microspore culture for production of doubled haploid plants via androgenesis. We use this method routinely because it is highly efficient and works well on different triticale genotypes. To force microspores into becoming embryogenic, we apply a 21-day cold pretreatment. The shock of cold facilitates redirecting microspores from their predestined pollen developmental program into the androgenesis pathway. Ovaries are included in our culture methods to help with embryogenesis, and the histone deacytelase inhibitor Trichostatin A (TSA) is added to further improve androgenesis and increase our ability to recover green doubled haploid plants.


Assuntos
Gametogênese Vegetal , Técnicas de Cultura de Tecidos/métodos , Triticale/crescimento & desenvolvimento , Meios de Cultura , Haploidia , Pólen/embriologia , Pólen/genética , Pólen/crescimento & desenvolvimento , Triticale/embriologia , Triticale/genética
8.
Int J Mol Sci ; 22(14)2021 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-34299165

RESUMO

In vitro tissue culture plant regeneration is a complicated process that requires stressful conditions affecting the cell functioning at multiple levels, including signaling pathways, transcriptome functioning, the interaction between cellular organelles (retro-, anterograde), compounds methylation, biochemical cycles, and DNA mutations. Unfortunately, the network linking all these aspects is not well understood, and the available knowledge is not systemized. Moreover, some aspects of the phenomenon are poorly studied. The present review attempts to present a broad range of aspects involved in the tissue culture-induced variation and hopefully would stimulate further investigations allowing a better understanding of the phenomenon and the cell functioning.


Assuntos
Regulação da Expressão Gênica de Plantas , Fenômenos Fisiológicos Vegetais , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Técnicas de Cultura de Tecidos/métodos , Técnicas de Cultura de Tecidos/normas , Metilação de DNA , Epigênese Genética , Proteínas de Plantas/genética
9.
Aging (Albany NY) ; 13(13): 16938-16956, 2021 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-34292877

RESUMO

Macrophage accumulation and nitrosative stress are known mechanisms underlying age-related cardiovascular pathology and functional decline. The cardiac muscle microenvironment is known to change with age, yet the direct effects of these changes have yet to be studied in-depth. The present study sought to better elucidate the role that biochemical and biomechanical alterations in cardiac tissue have in the altered phenotype and functionality of cardiac resident macrophages observed with increasing age. To accomplish this, naïve bone marrow derived macrophages from young mice were seeded onto either functionalized poly-dimethyl-siloxane hydrogels ranging in stiffness from 2kPA to 64kPA or onto tissue culture plastic, both of which were coated with either young or aged solubilized mouse cardiac extracellular matrix (cECM). Both biomechanical and biochemical alterations were found to have a significant effect on macrophage polarization and function. Increased substrate stiffness was found to promote macrophage morphologies associated with pro-inflammatory macrophage activation, increased expression of pro-inflammatory inducible nitric oxide synthase protein with increased nitric oxide secretion, and attenuated arginase activity and protein expression. Additionally, exposure to aged cECM promoted attenuated responsivity to both canonical pro-inflammatory and anti-inflammatory cytokine signaling cues when compared to young cECM treated cells. These results suggest that both biomechanical and biochemical changes in the cardiovascular system play a role in promoting the age-related shift towards pro-inflammatory macrophage populations associated with cardiovascular disease development.


Assuntos
Microambiente Celular/fisiologia , Coração/fisiologia , Macrófagos/fisiologia , Macrófagos/ultraestrutura , Envelhecimento/patologia , Envelhecimento/fisiologia , Animais , Arginase/metabolismo , Fenômenos Biomecânicos , Células da Medula Óssea , Citocinas/metabolismo , DNA/biossíntese , Ativação de Macrófagos , Camundongos , Camundongos Endogâmicos C57BL , Miocárdio/patologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/biossíntese , Fenótipo , Transdução de Sinais , Técnicas de Cultura de Tecidos
10.
Int J Mol Sci ; 22(12)2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-34203791

RESUMO

For in vitro modeling of human joints, osteochondral explants represent an acceptable compromise between conventional cell culture and animal models. However, the scarcity of native human joint tissue poses a challenge for experiments requiring high numbers of samples and makes the method rather unsuitable for toxicity analyses and dosing studies. To scale their application, we developed a novel method that allows the preparation of up to 100 explant cultures from a single human sample with a simple setup. Explants were cultured for 21 days, stimulated with TNF-α or TGF-ß3, and analyzed for cell viability, gene expression and histological changes. Tissue cell viability remained stable at >90% for three weeks. Proteoglycan levels and gene expression of COL2A1, ACAN and COMP were maintained for 14 days before decreasing. TNF-α and TGF-ß3 caused dose-dependent changes in cartilage marker gene expression as early as 7 days. Histologically, cultures under TNF-α stimulation showed a 32% reduction in proteoglycans, detachment of collagen fibers and cell swelling after 7 days. In conclusion, thin osteochondral slice cultures behaved analogously to conventional punch explants despite cell stress exerted during fabrication. In pharmacological testing, both the shorter diffusion distance and the lack of need for serum in the culture suggest a positive effect on sensitivity. The ease of fabrication and the scalability of the sample number make this manufacturing method a promising platform for large-scale preclinical testing in joint research.


Assuntos
Osso e Ossos/fisiologia , Custos e Análise de Custo , Técnicas de Cultura de Tecidos/economia , Técnicas de Cultura de Tecidos/métodos , Idoso , Idoso de 80 Anos ou mais , Agrecanas/genética , Agrecanas/metabolismo , Biomarcadores/metabolismo , Cartilagem Articular/metabolismo , Proliferação de Células , Sobrevivência Celular , Condrócitos/citologia , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Matriz Extracelular/metabolismo , Feminino , Humanos , Antígeno Ki-67/metabolismo , Masculino , Microscopia Confocal , Pessoa de Meia-Idade , Esclerose , Sobrevivência de Tecidos , Transcrição Genética , Fator de Necrose Tumoral alfa/metabolismo
11.
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
12.
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
13.
Methods Mol Biol ; 2288: 267-278, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270017

RESUMO

An efficient anther culture on double-layered media to produce doubled haploid (DH) plants in pepper (Capsicum annuum) was clearly shown to outperformed other techniques such as anther culture on solid medium and direct isolated microspore culture on liquid medium. It was even used for DH production in a cayenne type of hot pepper which was previously known as less responsive or even more recalcitrant to androgenesis than sweet bell pepper. Indeed, anther culture on double-layered media has been routinely used to produce DH plants on broad genotypes of C. annuum as parental candidates to develop hybrid varieties. The step-by-step protocol of pepper anther culture on double-layered media, we hereby present in detail, includes the growth of donor plants, the use of flower buds as anther source, flower bud disinfection, anther isolation, anther culture and incubation processes, embryo germination and plant acclimatization process, and transplanting of plants to the soil-compost medium in pots.


Assuntos
Capsicum/crescimento & desenvolvimento , Flores/crescimento & desenvolvimento , Germinação , Técnicas de Embriogênese Somática de Plantas/métodos , Técnicas de Cultura de Tecidos/métodos , Capsicum/genética , Meios de Cultura , Flores/genética , Haploidia
14.
Commun Biol ; 4(1): 861, 2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34253821

RESUMO

Mucosal exposure to infected semen accounts for the majority of HIV-1 transmission events, with rectal intercourse being the route with the highest estimated risk of transmission. Yet, the impact of semen inflammation on colorectal HIV-1 transmission has never been addressed. Here we use cynomolgus macaques colorectal tissue explants to explore the effect of leukocytospermia, indicative of male genital tract inflammation, on SIVmac251 infection. We show that leukocytospermic seminal plasma (LSP) has significantly higher concentration of a number of pro-inflammatory molecules compared to normal seminal plasma (NSP). In virus-exposed explants, LSP enhance SIV infection more efficiently than NSP, being the increased viral replication linked to the level of inflammatory and immunomodulatory cytokines. Moreover, LSP induce leukocyte accumulation on the apical side of the colorectal lamina propria and the recruitment of a higher number of intraepithelial dendritic cells than with NSP. These results suggest that the outcome of mucosal HIV-1 infection is influenced by the inflammatory state of the semen donor, and provide further insights into mucosal SIV/HIV-1 pathogenesis.


Assuntos
Colo/virologia , Células Dendríticas/virologia , Reto/virologia , Sêmen/virologia , Vírus da Imunodeficiência Símia/fisiologia , Replicação Viral/fisiologia , Animais , Colo/metabolismo , Citocinas/metabolismo , Infecções por HIV/metabolismo , Infecções por HIV/transmissão , Infecções por HIV/virologia , HIV-1/fisiologia , Leucócitos/metabolismo , Leucócitos/patologia , Leucócitos/virologia , Macaca mulatta , Masculino , Reto/metabolismo , Síndrome de Imunodeficiência Adquirida dos Símios/metabolismo , Síndrome de Imunodeficiência Adquirida dos Símios/transmissão , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Técnicas de Cultura de Tecidos
15.
Cells ; 10(7)2021 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-34206722

RESUMO

The lungs are affected by illnesses including asthma, chronic obstructive pulmonary disease, and infections such as influenza and SARS-CoV-2. Physiologically relevant models for respiratory conditions will be essential for new drug development. The composition and structure of the lung extracellular matrix (ECM) plays a major role in the function of the lung tissue and cells. Lung-on-chip models have been developed to address some of the limitations of current two-dimensional in vitro models. In this review, we describe various ECM substitutes utilized for modeling the respiratory system. We explore the application of lung-on-chip models to the study of cigarette smoke and electronic cigarette vapor. We discuss the challenges and opportunities related to model characterization with an emphasis on in situ characterization methods, both established and emerging. We discuss how further advancements in the field, through the incorporation of interstitial cells and ECM, have the potential to provide an effective tool for interrogating lung biology and disease, especially the mechanisms that involve the interstitial elements.


Assuntos
Dispositivos Lab-On-A-Chip , Pneumopatias/patologia , Pulmão/fisiologia , Regeneração/fisiologia , Mucosa Respiratória/citologia , COVID-19/patologia , COVID-19/terapia , COVID-19/virologia , Células Cultivadas , Matriz Extracelular/fisiologia , Humanos , Pulmão/citologia , Pulmão/patologia , Pneumopatias/fisiopatologia , Pneumopatias/terapia , Modelos Biológicos , Mucosa Respiratória/patologia , Mucosa Respiratória/fisiologia , SARS-CoV-2/patogenicidade , Técnicas de Cultura de Tecidos/instrumentação , Técnicas de Cultura de Tecidos/métodos
16.
Methods Mol Biol ; 2288: 91-102, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270006

RESUMO

We describe the production of doubled haploids through anther culture in caraway. Induction conditions for the cultivation of donor plants, anther collection, composition of culture media, and physical induction conditions for embryogenesis have been described. As a result, responsive lines with numerous haploid embryo production were obtained, which after colchicine treatment became fertile. From a practical point of view, two doubled haploid populations are tested under field conditions.


Assuntos
Carum/crescimento & desenvolvimento , Carum/genética , Melhoramento Vegetal/métodos , Carum/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Meios de Cultura/química , Diploide , Esterases/análise , Flores/genética , Flores/crescimento & desenvolvimento , Haploidia , Homozigoto , Isoenzimas/análise , Biologia Molecular/métodos , Pólen/genética , Pólen/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
17.
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
18.
Methods Mol Biol ; 2288: 113-126, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270008

RESUMO

Carrot is a vegetable of increasing economic importance. New hybrid cultivars are constantly required to meet the changing market needs. The application of anther culture significantly shortens the difficult and long-lasting breeding of carrot. We examined all the stages of the process of generating androgenic plants: induction of embryos in anther cultures, regeneration and acclimatization of produced plants, their evaluation, ploidy and homozygosity, and many other factors affecting their effectiveness. Every factor has been optimized by experimentally selecting the optimal level. As a result, a full protocol of producing homozygous plants using anther cultures was developed, which is presented in this chapter.


Assuntos
Daucus carota/crescimento & desenvolvimento , Daucus carota/genética , Melhoramento Vegetal/métodos , Aclimatação/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/fisiologia , Meios de Cultura/química , Daucus carota/fisiologia , Flores/genética , Flores/crescimento & desenvolvimento , Heterozigoto , Homozigoto , Isoenzimas/análise , Biologia Molecular/métodos , Regeneração/genética , Técnicas de Cultura de Tecidos
19.
Methods Mol Biol ; 2288: 103-111, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270007

RESUMO

Doubled haploidy technology is a powerful tool to accelerate the breeding of new crop varieties. Protocols are not universal, as even species within the same family require a specific process. Here we describe methods for developing doubled haploids for fennel and dill, both Apiaceae species which are used for food, flavorings, and medicine.


Assuntos
Anethum graveolens/crescimento & desenvolvimento , Anethum graveolens/genética , Foeniculum/crescimento & desenvolvimento , Foeniculum/genética , Melhoramento Vegetal/métodos , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Meios de Cultura/química , Diploide , Haploidia , Homozigoto , Biologia Molecular/métodos , Pólen/genética , Pólen/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
20.
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
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