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1.
Biology (Basel) ; 10(7)2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-34356540

RESUMO

Eggplant is a solanaceous crop cultivated worldwide for its edible fruit. Eggplant breeding programs are mainly aimed to the generation of F1 hybrids by crossing two highly homozygous, pure lines, which are traditionally obtained upon several self crossing generations, which is an expensive and time consuming process. Alternatively, fully homozygous, doubled haploid (DH) individuals can be induced from haploid cells of the germ line in a single generation. Several attempts have been made to develop protocols to produce eggplant DHs principally using anther culture and isolated microspore culture. Eggplant could be considered a moderately recalcitrant species in terms of ability for DH production. Anther culture stands nowadays as the most valuable technology to obtain eggplant DHs. However, the theoretical possibility of having plants regenerated from somatic tissues of the anther walls cannot be ruled out. For this reason, the use of isolated microspores is recommended when possible. This approach still has room for improvement, but it is largely genotype-dependent. In this review, we compile the most relevant advances made in DH production in eggplant, their application to breeding programs, and the future perspectives for the development of other, less genotype-dependent, DH technologies.

2.
Methods Mol Biol ; 2287: 41-103, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270025

RESUMO

In this chapter, we present a list of species (and few interspecific hybrids) where haploids and/or doubled haploids have been published, including the method by which they were obtained and the corresponding references. This list is an update of the compilation work of Maluszynski et al. published in 2003, including new species for which protocols were not available at that time, and also novel methodologies developed during these years. The list includes 383 different backgrounds. In this book, we present full protocols to produce DHs in 43 of the species included in this list. In addition, this book includes a chapter for one species not included in the list. This makes a total of 384 species where haploids and/or DHs have been reported up to date.


Assuntos
Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/genética , Melhoramento Vegetal/métodos , Haploidia , Partenogênese , Pólen/genética , Pólen/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.
Plant Physiol ; 176(1): 418-431, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29146775

RESUMO

TANGLED1 (TAN1) and AUXIN-INDUCED-IN-ROOTS9 (AIR9) are microtubule-binding proteins that localize to the division site in plants. Their function in Arabidopsis (Arabidopsis thaliana) remained unclear because neither tan1 nor air9 single mutants have a strong phenotype. We show that tan1 air9 double mutants have a synthetic phenotype consisting of short, twisted roots with disordered cortical microtubule arrays that are hypersensitive to a microtubule-depolymerizing drug. The tan1 air9 double mutants have significant defects in division plane orientation due to failures in placing the new cell wall at the correct division site. Full-length TAN1 fused to yellow fluorescent protein, TAN1-YFP, and several deletion constructs were transformed into the double mutant to assess which regions of TAN1 are required for its function in root growth, root twisting, and division plane orientation. TAN1-YFP expressed in tan1 air9 significantly rescued the double mutant phenotype in all three respects. Interestingly, TAN1 missing the first 126 amino acids, TAN1-ΔI-YFP, failed to rescue the double mutant phenotype, while TAN1 missing a conserved middle region, TAN1-ΔII-YFP, significantly rescued the mutant phenotype in terms of root growth and division plane orientation but not root twisting. We use the tan1 air9 double mutant to discover new functions for TAN1 and AIR9 during phragmoplast guidance and root morphogenesis.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/citologia , Arabidopsis/genética , Proteínas de Ciclo Celular/genética , Divisão Celular , Proteínas Associadas aos Microtúbulos/genética , Mutação/genética , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Benzamidas/farmacologia , Padronização Corporal/efeitos dos fármacos , Proteínas de Ciclo Celular/metabolismo , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Proteínas Associadas aos Microtúbulos/metabolismo , Paclitaxel/farmacologia , Fenótipo , Raízes de Plantas/citologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Prófase/efeitos dos fármacos , Proteínas Recombinantes de Fusão/metabolismo
5.
Plant Physiol ; 173(1): 863-871, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27881728

RESUMO

A sensitive and dynamically responsive auxin signaling reporter based on the DII domain of the INDOLE-3-ACETIC ACID28 (IAA28, DII) protein from Arabidopsis (Arabidopsis thaliana) was modified for use in maize (Zea mays). The DII domain was fused to a yellow fluorescent protein and a nuclear localization sequence to simplify quantitative nuclear fluorescence signal. DII degradation dynamics provide an estimate of input signal into the auxin signaling pathway that is influenced by both auxin accumulation and F-box coreceptor concentration. In maize, the DII-based marker responded rapidly and in a dose-dependent manner to exogenous auxin via proteasome-mediated degradation. Low levels of DII-specific fluorescence corresponding to high endogenous auxin signaling occurred near vasculature tissue and the outer layer and glume primordia of spikelet pair meristems and floral meristems, respectively. In addition, high DII levels were observed in cells during telophase and early G1, suggesting that low auxin signaling at these stages may be important for cell cycle progression.


Assuntos
Proteínas de Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Telófase/fisiologia , Fatores de Transcrição/metabolismo , Zea mays/citologia , Proteínas de Arabidopsis/genética , Fase G1/fisiologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Ácidos Indolacéticos/farmacologia , Meristema/genética , Meristema/metabolismo , Plantas Geneticamente Modificadas , Domínios Proteicos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transdução de Sinais , Imagem com Lapso de Tempo , Fatores de Transcrição/genética , Zea mays/efeitos dos fármacos , Zea mays/genética , Zea mays/metabolismo
6.
PLoS One ; 9(1): e87216, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24475254

RESUMO

The Pathogen and Circadian Controlled 1 (PCC1) gene, previously identified and further characterized as involved in defense to pathogens and stress-induced flowering, codes for an 81-amino acid protein with a cysteine-rich C-terminal domain. This domain is essential for homodimerization and anchoring to the plasma membrane. Transgenic plants with the ß-glucuronidase (GUS) reporter gene under the control of 1.1 kb promoter sequence of PCC1 gene display a dual pattern of expression. At early post-germination, PCC1 is expressed only in the root vasculature and in the stomata guard cells of cotyledons. During the transition from vegetative to reproductive development, PCC1 is strongly expressed in the vascular tissue of petioles and basal part of the leaf, and it further spreads to the whole limb in fully expanded leaves. This developmental pattern of expression together with the late flowering phenotype of long-day grown RNA interference (iPCC1) plants with reduced PCC1 expression pointed to a regulatory role of PCC1 in the photoperiod-dependent flowering pathway. iPCC1 plants are defective in light perception and signaling but are not impaired in the function of the core CO-FT module of the photoperiod-dependent pathway. The regulatory effect exerted by PCC1 on the transition to flowering as well as on other reported phenotypes might be explained by a mechanism involving the interaction with the subunit 5 of the COP9 signalosome (CSN).


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Membrana Celular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Arabidopsis/metabolismo , Complexo do Signalossomo COP9 , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica de Plantas/genética , Oligonucleotídeos/genética , Interferência de RNA , Reação em Cadeia da Polimerase em Tempo Real , Técnicas do Sistema de Duplo-Híbrido
7.
J Exp Bot ; 65(4): 907-21, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24371253

RESUMO

The extensive support for abscisic acid (ABA) involvement in the complex regulatory networks controlling stress responses and development in plants contrasts with the relatively recent role assigned to nitric oxide (NO). Because treatment with exogenous ABA leads to enhanced production of NO, it has been widely considered that NO participates downstream of ABA in controlling processes such as stomata movement, seed dormancy, and germination. However, data on leaf senescence and responses to stress suggest that the functional interaction between ABA and NO is more complex than previously thought, including not only cooperation but also antagonism. The functional relationship is probably determined by several factors including the time- and place-dependent pattern of accumulation of both molecules, the threshold levels, and the regulatory factors important for perception. These factors will determine the actions exerted by each regulator. Here, several examples of well-documented functional interactions between NO and ABA are analysed in light of the most recent reported data on seed dormancy and germination, stomata movements, leaf senescence, and responses to abiotic and biotic stresses.


Assuntos
Ácido Abscísico/metabolismo , Regulação da Expressão Gênica de Plantas , Óxido Nítrico/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Plantas/metabolismo , Estresse Fisiológico , Germinação , Modelos Biológicos , Desenvolvimento Vegetal , Dormência de Plantas , Imunidade Vegetal , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/imunologia , Folhas de Planta/fisiologia , Fenômenos Fisiológicos Vegetais , Estômatos de Plantas/crescimento & desenvolvimento , Estômatos de Plantas/imunologia , Estômatos de Plantas/fisiologia , Plantas/imunologia , Sementes/crescimento & desenvolvimento , Sementes/imunologia , Sementes/fisiologia , Fatores de Tempo
8.
J Exp Bot ; 64(11): 3385-95, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23833195

RESUMO

Pathogen and Circadian Controlled 1 (PCC1) was previously characterized as a regulator of defence against pathogens and stress-activated transition to flowering. Plants expressing an RNA interference construct for the PCC1 gene (iPCC1 plants) showed a pleiotropic phenotype. They were hypersensitive to abscisic acid (ABA) as shown by reduced germination potential and seedling establishment, as well as reduced stomatal aperture and main root length in ABA-supplemented media. In addition, iPCC1 plants displayed alterations in polar lipid contents and their corresponding fatty acids. Importantly, a significant reduction in the content of phosphatidylinositol (PI) was observed in iPCC1 leaves when compared with wild-type plants. A trend in reduced levels of 18:0 and increased levels of 18:2 and particularly 18:3 was also detected in several classes of polar lipids. The enhanced ABA-mediated responses and the reduced content of PI might be responsible for iPCC1 plants displaying a complex pattern of defence against pathogens of different lifestyles. iPCC1 plants were more susceptible to the hemi-biotrophic oomycete pathogen Phytophthora brassicae and more resistant to the necrotrophic fungal pathogen Botrytis cinerea compared with wild-type plants.


Assuntos
Ácido Abscísico/farmacologia , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/genética , Resistência à Doença/efeitos dos fármacos , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Fosfatidilinositóis/metabolismo , Doenças das Plantas
9.
PLoS Pathog ; 7(7): e1002148, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21829351

RESUMO

Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS), including H(2)O(2) and O(2) (-), are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI) or catalase. H(2)O(2) was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA). Accordingly, ABA biosynthesis mutants (aba2 and aba3) were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending innate defenses.


Assuntos
Arabidopsis , Botrytis/imunologia , Peróxido de Hidrogênio/imunologia , Doenças das Plantas , Imunidade Vegetal/fisiologia , Folhas de Planta , Superóxidos/imunologia , Ácido Abscísico/genética , Ácido Abscísico/imunologia , Arabidopsis/imunologia , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/imunologia , Carboxiliases/genética , Carboxiliases/imunologia , Coenzima A Ligases/genética , Coenzima A Ligases/imunologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Lipídeos de Membrana/genética , Lipídeos de Membrana/imunologia , Mutação/imunologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/microbiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/imunologia , Plantas Geneticamente Modificadas/microbiologia , Trametes/genética
10.
Plant Cell Environ ; 33(1): 11-22, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19781011

RESUMO

Salicylic acid (SA) has been characterized as an activator of pathogen-triggered resistance of plants. SA also regulates developmental processes such as thermogenesis in floral organs and stress-induced flowering. To deepen our knowledge of the mechanism underlying SA regulation of flowering time in Arabidopsis, we compared the transcriptomes of SA-deficient late flowering genotypes with wild-type plants. Down- or up-regulated genes in SA-deficient plants were screened for responsiveness to ultraviolet (UV)-C light, which accelerates flowering in Arabidopsis. Among them, only Pathogen and Circadian Controlled 1 (PCC1) was up-regulated by UV-C light through a SA-dependent process. Moreover, UV-C light-activated expression of PCC1 was also dependent on the flowering activator CONSTANS (CO). PCC1 gene has a circadian-regulated developmental pattern of expression with low transcript levels after germination that increased abruptly by day 10. RNAi plants with very low expression of PCC1 gene were late flowering, defective in UV-C light acceleration of flowering and contained FLOWERING LOCUS T (FT) transcript levels below 5% of that detected in wild-type plants. Although PCC1 seems to function between CO and FT in the photoperiod-dependent flowering pathway, transgenic plants overexpressing a Glucocorticoid Receptor (GR)-fused version of CO strongly activated FT but not PCC1 after dexamethasone treatment.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Ácido Salicílico/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Ligação a DNA/metabolismo , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Análise de Sequência com Séries de Oligonucleotídeos , Fotoperíodo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo , Interferência de RNA , RNA de Plantas/genética , Ácido Salicílico/farmacologia , Fatores de Transcrição/metabolismo , Raios Ultravioleta
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