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1.
Plant J ; 97(4): 730-748, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30422341

RESUMO

Over the last 40 years, new sunflower downy mildew isolates (Plasmopara halstedii) have overcome major gene resistances in sunflower, requiring the identification of additional and possibly more durable broad-spectrum resistances. Here, 354 RXLR effectors defined in silico from our new genomic data were classified in a network of 40 connected components sharing conserved protein domains. Among 205 RXLR effector genes encoding conserved proteins in 17 P. halstedii pathotypes of varying virulence, we selected 30 effectors that were expressed during plant infection as potentially essential genes to target broad-spectrum resistance in sunflower. The transient expression of the 30 core effectors in sunflower and in Nicotiana benthamiana leaves revealed a wide diversity of targeted subcellular compartments, including organelles not so far shown to be targeted by oomycete effectors such as chloroplasts and processing bodies. More than half of the 30 core effectors were able to suppress pattern-triggered immunity in N. benthamiana, and five of these induced hypersensitive responses (HR) in sunflower broad-spectrum resistant lines. HR triggered by PhRXLRC01 co-segregated with Pl22 resistance in F3 populations and both traits localized in 1.7 Mb on chromosome 13 of the sunflower genome. Pl22 resistance was physically mapped on the sunflower genome recently sequenced, unlike all the other downy mildew resistances published so far. PhRXLRC01 and Pl22 are proposed as an avirulence/resistance gene couple not previously described in sunflower. Core effector recognition is a successful strategy to accelerate broad-spectrum resistance gene identification in complex crop genomes such as sunflower.


Assuntos
Helianthus/metabolismo , Helianthus/microbiologia , Oomicetos/patogenicidade , Doenças das Plantas/microbiologia , Mapeamento Cromossômico , Resistência à Doença/genética , Resistência à Doença/fisiologia , Genótipo , Virulência/genética , Virulência/fisiologia
2.
Front Plant Sci ; 9: 1780, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30564260

RESUMO

Resistance to downy mildew (Plasmopara halstedii) in sunflower (Helianthus annuus L.) is conferred by major resistance genes, denoted Pl. Twenty-two Pl genes have been identified and genetically mapped so far. However, over the past 50 years, wide-scale presence of only a few of them in sunflower crops led to the appearance of new, more virulent pathotypes (races) so it is important for sunflower varieties to carry as wide a range of resistance genes as possible. We analyzed phenotypically 12 novel resistant sources discovered in breeding pools derived from two wild Helianthus species and in eight wild H. annuus ecotypes. All were effective against at least 16 downy mildew pathotypes. We mapped their resistance genes on the sunflower reference genome of 3,600 Mb, in intervals that varied from 75 Kb to 32 Mb using an AXIOM® genotyping array of 49,449 SNP. Ten probably new genes were identified according to resistance spectrum, map position, hypersensitive response to the transient expression of a P. halstedii RXLR effector, or the ecotype/species from which they originated. The resistance source HAS6 was found to carry the first downy mildew resistance gene mapped on chromosome 11, whereas the other resistances were positioned on chromosomes 1, 2, 4, and 13 carrying already published Pl genes that we also mapped physically on the same reference genome. The new genes were designated Pl23-Pl32 according to the current nomenclature. However, since sunflower downy mildew resistance genes have not yet been sequenced, rules for designation are discussed. This is the first large scale physical mapping of both 10 new and 10 already reported downy mildew resistance genes in sunflower.

3.
Front Plant Sci ; 8: 1633, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28983306

RESUMO

Prediction of hybrid performance using incomplete factorial mating designs is widely used in breeding programs including different heterotic groups. Based on the general combining ability (GCA) of the parents, predictions are accurate only if the genetic variance resulting from the specific combining ability is small and both parents have phenotyped descendants. Genomic selection (GS) can predict performance using a model trained on both phenotyped and genotyped hybrids that do not necessarily include all hybrid parents. Therefore, GS could overcome the issue of unknown parent GCA. Here, we compared the accuracy of classical GCA-based and genomic predictions for oil content of sunflower seeds using several GS models. Our study involved 452 sunflower hybrids from an incomplete factorial design of 36 female and 36 male lines. Re-sequencing of parental lines allowed to identify 468,194 non-redundant SNPs and to infer the hybrid genotypes. Oil content was observed in a multi-environment trial (MET) over 3 years, leading to nine different environments. We compared GCA-based model to different GS models including female and male genomic kinships with the addition of the female-by-male interaction genomic kinship, the use of functional knowledge as SNPs in genes of oil metabolic pathways, and with epistasis modeling. When both parents have descendants in the training set, the predictive ability was high even for GCA-based prediction, with an average MET value of 0.782. GS performed slightly better (+0.2%). Neither the inclusion of the female-by-male interaction, nor functional knowledge of oil metabolism, nor epistasis modeling improved the GS accuracy. GS greatly improved predictive ability when one or both parents were untested in the training set, increasing GCA-based predictive ability by 10.4% from 0.575 to 0.635 in the MET. In this scenario, performing GS only considering SNPs in oil metabolic pathways did not improve whole genome GS prediction but increased GCA-based prediction ability by 6.4%. Our results show that GS is a major improvement to breeding efficiency compared to the classical GCA modeling when either one or both parents are not well-characterized. This finding could therefore accelerate breeding through reducing phenotyping efforts and more effectively targeting for the most promising crosses.

4.
Theor Appl Genet ; 130(6): 1099-1112, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28255669

RESUMO

KEY MESSAGE: SNP genotyping of 114 cultivated sunflower populations showed that the multiplication process and the main traits selected during breeding of sunflower cultivars drove molecular diversity of the populations. The molecular diversity in a set of 114 cultivated sunflower populations was studied by single-nucleotide polymorphism genotyping. These populations were chosen as representative of the 400 entries in the INRA collection received or developed between 1962 and 2011 and made up of land races, open-pollinated varieties, and breeding pools. Mean allele number varied from 1.07 to 1.90. Intra-population variability was slightly reduced according to the number of multiplications since entry but some entries were probably largely homozygous when received. A principal component analysis was used to study inter-population variability. The first 3 axes accounted for 17% of total intra-population variability. The first axis was significantly correlated with seed oil content, more closely than just the distinction between oil and confectionary types. The second axis was related to the presence or absence of restorer genes and the third axis to flowering date and possibly to adaptation to different climates. Our results provide arguments highlighting the effect of the maintenance process on the within population genetic variability as well as on the impact of breeding for major agronomic traits on the between population variability of the collection. Propositions are made to improve sunflower population maintenance procedures to keep maximum genetic variability for future breeding.


Assuntos
Genética Populacional , Helianthus/genética , Melhoramento Vegetal , Polimorfismo de Nucleotídeo Único , Alelos , DNA de Plantas/genética , Ligação Genética , Genótipo
5.
Front Plant Sci ; 7: 1887, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28066456

RESUMO

Plasmopara halstedii is an obligate biotrophic oomycete causing downy mildew disease on sunflower, Helianthus annuus, an economically important oil crop. Severe symptoms of the disease (e.g., plant dwarfism, leaf bleaching, sporulation and production of infertile flower) strongly impair seed yield. Pl resistance genes conferring resistance to specific P. halstedii pathotypes were located on sunflower genetic map but yet not cloned. They are present in cultivated lines to protect them against downy mildew disease. Among the 16 different P. halstedii pathotypes recorded in France, pathotype 710 is frequently found, and therefore continuously controlled in sunflower by different Pl genes. High-throughput sequencing of cDNA from P. halstedii led us to identify potential effectors with the characteristic RXLR or CRN motifs described in other oomycetes. Expression of six P. halstedii putative effectors, five RXLR and one CRN, was analyzed by qRT-PCR in pathogen spores and in the pathogen infecting sunflower leaves and selected for functional analyses. We developed a new method for transient expression in sunflower plant leaves and showed for the first time subcellular localization of P. halstedii effectors fused to a fluorescent protein in sunflower leaf cells. Overexpression of the CRN and of 3 RXLR effectors induced hypersensitive-like cell death reactions in some sunflower near-isogenic lines resistant to pathotype 710 and not in susceptible corresponding lines, suggesting they could be involved in Pl loci-mediated resistances.

6.
Mol Plant Pathol ; 16(2): 109-22, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25476405

RESUMO

Downy mildew of sunflower is caused by Plasmopara halstedii (Farlow) Berlese & de Toni. Plasmopara halstedii is an obligate biotrophic oomycete pathogen that attacks annual Helianthus species and cultivated sunflower, Helianthus annuus. Depending on the sunflower developmental stage at which infection occurs, the characteristic symptoms range from young seedling death, plant dwarfing, leaf bleaching and sporulation to the production of infertile flowers. Downy mildew attacks can have a great economic impact on sunflower crops, and several Pl resistance genes are present in cultivars to protect them against the disease. Nevertheless, some of these resistances have been overcome by the occurrence of novel isolates of the pathogen showing increased virulence. A better characterization of P. halstedii infection and dissemination mechanisms, and the identification of the molecular basis of the interaction with sunflower, is a prerequisite to efficiently fight this pathogen. This review summarizes what is currently known about P. halstedii, provides new insights into its infection cycle on resistant and susceptible sunflower lines using scanning electron and light microscopy imaging, and sheds light on the pathogenicity factors of P. halstedii obtained from recent molecular data. TAXONOMY: Kingdom Stramenopila; Phylum Oomycota; Class Oomycetes; Order Peronosporales; Family Peronosporaceae; Genus Plasmopara; Species Plasmopara halstedii. DISEASE SYMPTOMS: Sunflower seedling damping off, dwarfing of the plant, bleaching of leaves, starting from veins, and visible white sporulation, initially on the lower side of cotyledons and leaves. Plasmopara halstedii infection may severely impact sunflower seed yield. INFECTION PROCESS: In spring, germination of overwintered sexual oospores leads to sunflower root infection. Intercellular hyphae are responsible for systemic plant colonization and the induction of disease symptoms. Under humid and fresh conditions, dissemination structures are produced by the pathogen on all plant organs to release asexual zoosporangia. These zoosporangia play an important role in pathogen dissemination, as they release motile zoospores that are responsible for leaf infections on neighbouring plants. DISEASE CONTROL: Disease control is obtained by both chemical seed treatment (mefenoxam) and the deployment of dominant major resistance genes, denoted Pl. However, the pathogen has developed fungicide resistance and has overcome some plant resistance genes. Research for more sustainable strategies based on the identification of the molecular basis of the interaction are in progress. USEFUL WEBSITES: http://www.heliagene.org/HP, http://lipm-helianthus.toulouse.inra.fr/dokuwiki/doku.php?id=start, https://www.heliagene.org/PlasmoparaSpecies (soon available).


Assuntos
Helianthus/microbiologia , Oomicetos/patogenicidade , Fungicidas Industriais/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Virulência
7.
Theor Appl Genet ; 126(5): 1337-56, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23435733

RESUMO

Association mapping and linkage mapping were used to identify quantitative trait loci (QTL) and/or causative mutations involved in the control of flowering time in cultivated sunflower Helianthus annuus. A panel of 384 inbred lines was phenotyped through testcrosses with two tester inbred lines across 15 location × year combinations. A recombinant inbred line (RIL) population comprising 273 lines was phenotyped both per se and through testcrosses with one or two testers in 16 location × year combinations. In the association mapping approach, kinship estimation using 5,923 single nucleotide polymorphisms was found to be the best covariate to correct for effects of panel structure. Linkage disequilibrium decay ranged from 0.08 to 0.26 cM for a threshold of 0.20, after correcting for structure effects, depending on the linkage group (LG) and the ancestry of inbred lines. A possible hitchhiking effect is hypothesized for LG10 and LG08. A total of 11 regions across 10 LGs were found to be associated with flowering time, and QTLs were mapped on 11 LGs in the RIL population. Whereas eight regions were demonstrated to be common between the two approaches, the linkage disequilibrium approach did not detect a documented QTL that was confirmed using the linkage mapping approach.


Assuntos
Mapeamento Cromossômico , Cromossomos de Plantas/genética , Flores/fisiologia , Genes de Plantas/genética , Ligação Genética , Helianthus/genética , DNA de Plantas/genética , Marcadores Genéticos , Helianthus/crescimento & desenvolvimento , Desequilíbrio de Ligação , Fenótipo , Locos de Características Quantitativas
8.
Mol Ecol Resour ; 13(1): 10-20, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23039950

RESUMO

Long a major focus of genetic research and breeding, sunflowers (Helianthus) are emerging as an increasingly important experimental system for ecological and evolutionary studies. Here, we review the various attributes of wild and domesticated sunflowers that make them valuable for ecological experimentation and describe the numerous publicly available resources that have enabled rapid advances in ecological and evolutionary genetics. Resources include seed collections available from germplasm centres at the USDA and INRA, genomic and EST sequences, mapping populations, genetic markers, genetic and physical maps and other forward- and reverse-genetic tools. We also discuss some of the key evolutionary, genetic and ecological questions being addressed in sunflowers, as well as gaps in our knowledge and promising areas for future research.


Assuntos
Evolução Biológica , Bases de Dados Genéticas , Ecologia/métodos , Genoma de Planta/genética , Helianthus/genética , Sementes , Manejo de Espécimes , Marcadores Genéticos/genética , Mapeamento Físico do Cromossomo
9.
Theor Appl Genet ; 126(2): 359-67, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23052021

RESUMO

The resistance of sunflower to Plasmopara halstedii is conferred by major resistance genes denoted Pl. Previous genetic studies indicated that the majority of these genes are clustered on linkage groups 8 and 13. The Pl6 locus is one of the main clusters to have been identified, and confers resistance to several P. halstedii races. In this study, a map-based cloning strategy was implemented using a large segregating F2 population to establish a fine physical map of this cluster. A marker derived from a bacterial artificial chromosome (BAC) clone was found to be very tightly linked to the gene conferring resistance to race 300, and the corresponding BAC clone was sequenced and annotated. It contains several putative genes including three toll-interleukin receptor-nucleotide binding site-leucine rich repeats (TIR-NBS-LRR) genes. However, only one TIR-NBS-LRR appeared to be expressed, and thus constitutes a candidate gene for resistance to P. halstedii race 300.


Assuntos
Resistência à Doença/genética , Genes de Plantas/genética , Helianthus/genética , Oomicetos/fisiologia , Doenças das Plantas/genética , Locos de Características Quantitativas , Sequência de Aminoácidos , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Clonagem Molecular , Cruzamentos Genéticos , DNA de Plantas/genética , Helianthus/imunologia , Helianthus/microbiologia , Imunidade Inata , Dados de Sequência Molecular , Oomicetos/patogenicidade , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , RNA de Plantas/genética , Homologia de Sequência de Aminoácidos
10.
Theor Appl Genet ; 125(8): 1589-601, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22824968

RESUMO

Interest in phytosterol contents due to their potential benefits for human health has been largely documented in several crop species. Studies were focused mainly on total sterol content and their concentration or distribution in seed. This study aimed at providing new insight into the genetic control of total and individual sterol contents in sunflower seed through QTL analyses in a RIL population characterized over 2 years showing contrasted rainfall during seed filling. Results indicated that 13 regions on 9 linkage groups were involved in different phytosterol traits. Most of the QTL mapped were stable across years in spite of contrasted growing conditions. Some of them explained up to 30 % of phenotypic variation. Two QTL, located on LG10, near b1, and on LG14, were found to co-localize with QTL for oil content, indicating that likely, a part of the genetic variation for sterol content is only the result of genetic variation for oil content. However, three other QTL, stable over the 2 years, were found on LG1, LG4 and LG7 each associated with a particular class of sterols, suggesting that some enzymes known to be involved in the sterol metabolic pathway may determine the specificity of sterol profiles in sunflower seeds. These results suggest that it may be possible to introduce these traits as criteria in breeding programmes for quality in sunflower. The molecular markers linked to genetic factors controlling phytosterol contents could help selection during breeding programs.


Assuntos
Helianthus/genética , Fitosteróis/metabolismo , Sementes/genética , Sementes/metabolismo , Vias Biossintéticas/genética , Clima , Cruzamentos Genéticos , França , Hibridização Genética , Endogamia , Fitosteróis/biossíntese , Óleos de Plantas/química , Locos de Características Quantitativas/genética , Óleo de Girassol
11.
Theor Appl Genet ; 125(5): 909-20, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22576236

RESUMO

Major gene resistance to sunflower downy mildew (Plasmopara halstedii) races 304 and 314 was found to segregate independently from the resistance to races 334, 307 and 304 determined by the gene Pl2, already positioned on Linkage Group (LG) 8 of sunflower molecular maps. Using a consensus SSR-SNP map constructed from the INEDI RIL population and a new RIL population FU × PAZ2, the positions of Pl2 and Pl5 were confirmed and the new gene, denoted Pl21, was mapped on LG13, at 8 cM from Pl5. The two RIL populations were observed for their quantitative resistance to downy mildew in the field and both indicated the existence of a QTL on LG8 at 20-40 cM from the major resistance gene cluster. In addition, for the INEDI population, a strong QTL on LG10, reported previously, was confirmed and a third QTL was mapped on LG7. A growth chamber test methodology, significantly correlated with field results, also revealed the major QTL on LG10, explaining 65 % of variability. This QTL mapped in the same area as a gene involved in stomatal opening and root growth, which may be suggested as a possible candidate to explain the control of this character. These results indicate that it should be possible to combine major genes and other resistance mechanisms, a strategy that could help to improve durability of sunflower resistance to downy mildew.


Assuntos
Mapeamento Cromossômico , Genes de Plantas/genética , Helianthus/genética , Helianthus/microbiologia , Imunidade Inata/genética , Oomicetos/patogenicidade , Doenças das Plantas/genética , Locos de Características Quantitativas , Cromossomos de Plantas/genética , DNA de Plantas/genética , Marcadores Genéticos , Genótipo , Helianthus/imunologia , Desequilíbrio de Ligação , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase , Polimorfismo Genético/genética
12.
Funct Plant Biol ; 38(3): 246-259, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32480881

RESUMO

Present work focussed on improving the description of organogenesis, morphogenesis and metabolism in a biophysical plant model (SUNFLO) applied to sunflower (Helianthus annuus L.). This first version of the model is designed for potential growth conditions without any abiotic or biotic stresses. A greenhouse experiment was conducted to identify and estimate the phenotypic traits involved in plant productivity variability of 26 sunflower genotypes. The ability of SUNFLO to discriminate the genotypes was tested on previous results of a field survey aimed at evaluating the genetic progress since 1960. Plants were phenotyped in four directions; phenology, architecture, photosynthesis and biomass allocation. Twelve genotypic parameters were chosen to account for the phenotypic variability. SUNFLO was built to evaluate their respective contribution to the variability of yield potential. A large phenotypic variability was found for all genotypic parameters. SUNFLO was able to account for 80% of observed variability in yield potential and to analyse the phenotypic variability of complex plant traits such as light interception efficiency or seed yield. It suggested that several ways are possible to reach high yields in sunflower. Unlike classical statistical analysis, this modelling approach highlights some efficient parameter combinations used by the most productive genotypes. The next steps will be to evaluate the genetic determinisms of the genotypic parameters.

13.
Infect Genet Evol ; 8(5): 534-40, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18450523

RESUMO

Plasmopara halstedii, the causal agent of sunflower downy mildew, displays a gene-for-gene interaction with its host plant, Helianthus annuus and other species of the genus. Monitoring of the evolution of virulent races in France over a 19-year period led to the identification of 14 different races (or pathotypes). Twelve expressed sequence tag (EST)-derived markers displaying SNPs and insertion-deletion variations have recently been identified in P. halstedii. We used these markers to study the genetic structure and the evolution of sunflower downy mildew races. Bayesian assignment analysis identified three genetically differentiated groups of isolates organized around the first three races described in France. Strong genetic substructuring according to geographic origin of races was observed, confirming that these three groups corresponded to three separate introductions into France of isolates with different genetic and phenotypic backgrounds. Our results suggest that multiple introductions of P. halstedii isolates may have provided the raw material for more complex processes in the evolution of races, such as recombination between races or clonal evolution through mitotic instability.


Assuntos
Helianthus/microbiologia , Oomicetos/genética , Oomicetos/isolamento & purificação , Doenças das Plantas/microbiologia , Polimorfismo de Nucleotídeo Único/genética , Evolução Biológica , França , Genótipo , Oomicetos/patogenicidade , Dinâmica Populacional
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