Sexual Crosses Research Articles

Fusarium mirum sp. nov, intertwining Fusarium madaense and Fusarium andiyazi, pathogens of tropical grasses

Many species in the Fusarium fujikuroi Species Complex (FFSC) have an affinity for grass species, with whom they live in an endophytic association or cause disease. We recovered isolates of Fusarium from agriculturally important grasses in Africa and Brazil, and characterized them with morphological markers, mating type, and Amplified Fragment Length Polymorphisms (AFLPs). We also conducted multi-locus phylogenetic analyses based on partial DNA sequences of translation elongation factor-1α (TEF1), β-tubulin (TUB), and the second largest subunit of RNA polymerase (RPB2) gene regions. Sexual cross fertility was used to test the biological species concept and the sexual stage of F. madaense is described. A novel species within the FFSC, Fusarium mirum, that is different from the other known species in the complex, was formally described. Fusarium mirum, F. madaense, and Fusarium andiyazi are a tightly intertwined species trio that are morphologically identical, but phylogenetically distinguishable, and amongst whom interspecific genetic exchange may still occur. These three species are so close that they cannot be reliably distinguished if only sequences of the TEF1 gene are used. In pathogenicity tests, all tested isolates of F. madaense from sugarcane, sorghum, maize, millet and Brachiaria could induce stalk rot in sorghum, maize and millet, and pokkah boeng in sugarcane. This study increases our understanding of the diversity of species within the FFSC that cause disease in tropical grasses or act as endophytes, and their geographic distributions. The genetically close relationship between F. mirum, F. madaense, and F. andiyazi provides an opportunity to study and identify factors underlying their limited inter-specific cross-fertility and sympatric speciation.

All Plant Breeding Technologies Are Equal, but Some Are More Equal Than Others: The Case of GM and Mutagenesis.

A pervasive opposition to genetically modified (GM) foods has developed from the notion that they pose a risk to human and environmental health. Other techniques for the genetic modification of plants, such as sexual crossing and mutagenesis breeding, have mostly remained unchallenged. This research aims to investigate public perception of plant breeding technologies. Specifically, sexual crossing, mutagenesis, transgenics (GM) and gene editing. It was expected that attitudes and intentions would be most positive and the perception of risk lowest for plant genetic modification through sexual crosses. Scores on these variables were expected to be similar between mutagenesis, GM and gene editing. It was also expected that attitudes, intentions and risk perception would change (becoming more positive) once participants learned about foods developed through these technologies. Participants reported their attitudes, intentions and risk perception at two points in time. At Time 2, they were presented with pictures of food items developed through sexual crossing, GM and mutagenesis. The results showed that mutagenesis stood out as the most negatively perceived technology, whereas genetic development via sexual crosses was generally perceived as positive. The results highlight the importance of messaging, framing in consumer attitudes.

Take-All Disease: New Insights into an Important Wheat Root Pathogen.

Take-all disease, caused by the fungal root pathogen Gaeumannomyces tritici, is considered to be the most important root disease of wheat worldwide. Here we review the advances in take-all research over the last 15 years, focusing on the identification of new sources of genetic resistance in wheat relatives and the role of the microbiome in disease development. We also highlight recent breakthroughs in the molecular interactions between G. tritici and wheat, including genome and transcriptome analyses. These new findings will aid the development of novel control strategies against take-all disease. In light of this growing understanding, the G. tritici-wheat interaction could provide a model study system for root-infecting fungal pathogens of cereals.

The History of Botrytis Taxonomy, the Rise of Phylogenetics, and Implications for Species Recognition.

Botrytis is one of the oldest, most well studied, and most economically important fungal taxa. Nonetheless, many species in this genus have remained obscured for nearly 300 years because of the difficulty in distinguishing these species by conventional mycological methods. Aided by the use of phylogenetic tools, the genus is currently undergoing a taxonomic revolution. The number of putative species in the genus has nearly doubled over the last 10 years and more species are likely to be discovered in the future. The implementation of phylogenetic species recognition concepts in Botrytis is providing for more resolution on the relatedness among species than ever before, and this has helped to overcome issues in historical species recognition using morphology, sexual crosses, and pathogenicity tests. Meanwhile, the use of genetic tools is helping to reveal surprising insight into this archetypal necrotroph's behavior, making these approaches increasingly important in species recognition and identification. As Botrytis taxonomy continues to evolve at a rapid pace, researchers should be encouraged to continue to employ the powerful tool of phylogenetics while considering how it fits into a larger framework of classical Botrytis species recognition. Starting points for discussion on how to move forward with Botrytis species recognition are included herein, with an emphasis on the implications and utility of new species descriptions.

Review of Horseradish Breeding for Improved Root Quality and Yield in Illinois, USA

Horseradish cultivars are highly heterozygous clones and are maintained through asexual propagation, using root cuttings. For many years, horseradish was believed to be sterile and therefore impossible to improve by traditional sexual crosses. Prior to the 20th century, the only way to improve horseradish was to select and plant root cuttings from the most desirable plants. Moreover, the development of new improved horseradish cultivars has also been somewhat limited by the lack of viable seed resulting from low fertility of horseradish flowers. However, in Illinois, USA, a horseradish breeding program was initiated in the 1950s to develop additional cultivars to widen the genetic base of the few cultivars being grown at the time. In more recent years, the proven cross breeding technique has been primarily used to obtain new genotypes of horseradish, since it is more efficient in producing new improved cultivars, compared to the polycross method that had been previously used for decades to obtain new genetic combinations. Since horseradish is a minor specialty crop with very little available information regarding breeding procedures, this review was developed to provide a better understanding of pollination barriers and methods for fertility improvement, traditional breeding procedures and cultivar development, and traditional breeding achievements and limitations. The development of new horseradish cultivars is essential for the sustained success of the Illinois, USA industry since it provides growers with a continuous supply of new selections that have increased vigor, outstanding root quality, and high yields.

Marker-Free Transplastomic Plants by Excision of Plastid Marker Genes Using Directly Repeated DNA Sequences.

Excision of marker genes using DNA direct repeats makes use of the efficient native homologous recombination pathway present in the plastids of algae and plants. The method is simple, efficient, and widely applicable to plants and green algae. Marker excision frequency is dependent on the length and number of directly repeated sequences. When two repeats are used a repeat size of greater than 600bp promotes efficient excision of the marker gene. A wide variety of sequences can be used to make the direct repeats. Only a single round of transformation is required and there is no requirement to introduce site-specific recombinases by retransformation or sexual crosses. Selection is used to maintain the marker and ensure homoplasmy of transgenic plastid genomes (plastomes). Release of selection allows the accumulation of marker-free plastomes generated by marker excision, which is a spontaneous and unidirectional process. Cytoplasmic sorting allows the segregation of cells with marker-free transgenic plastids. The marker-free shoots resulting from direct repeat mediated excision of marker genes have been isolated by vegetative propagation of shoots in the T0 generation. Alternatively, accumulation of marker-free plastomes during growth, development and flowering of T0 plants allows for the collection of seeds that give rise to a high proportion of marker-free T1 seedlings. The procedure enables precise plastome engineering involving insertion of transgenes, point mutations and deletion of genes without the inclusion of any extraneous DNA. The simplicity and convenience of direct repeat excision facilitates its widespread use to isolate marker-free crops.

Transposon activation is a major driver in the genome evolution of cultivated olive trees (Olea europaea L.).

The primary domestication of olive (Olea europaea L.) in the Levant dates back to the Neolithic period, around 6,000-5,500 BC, as some archeological remains attest. Cultivated olive trees are reproduced clonally, with sexual crosses being the sporadic events that drive the development of new varieties. In order to determine the genomic changes which have occurred in a modern olive cultivar, the genome of the Picual cultivar, one of the most popular olive varieties, was sequenced. Additional 40 cultivated and 10 wild accessions were re-sequenced to elucidate the evolution of the olive genome during the domestication process. It was found that the genome of the 'Picual' cultivar contains 79,667 gene models, of which 78,079 were protein-coding genes and 1,588 were tRNA. Population analyses support two independent events in olive domestication, including an early possible genetic bottleneck. Despite genetic bottlenecks, cultivated accessions showed a high genetic diversity driven by the activation of transposable elements (TE). A high TE gene expression was observed in presently cultivated olives, which suggests a current activity of TEs in domesticated olives. Several TEs families were expanded in the last 5,000 or 6,000 years and produced insertions near genes that may have been involved in selected traits during domestication as reproduction, photosynthesis, seed development, and oil production. Therefore, a great genetic variability has been found in cultivated olive as a result of a significant activation of TEs during the domestication process.

Obtaining monokaryotic and dikaryotic mycelial cultures of two Amazonian strains of Geastrum (Geastraceae, Basidiomycota)

ABSTRACT The high diversity of the genus Geastrum and the difficulty of obtaining mycelial cultures impairs the study of the ecophysiology and the exploration of the biotechnological potential of the taxon. In this study, different culture media were tested to obtain mycelial cultures for G. lloydianum and G. subiculosum collected in the Brazilian Amazon. Data on spore germination, and isolation of monokaryotic cultures and in vitro sexual reproduction are presented, as well as a brief morphological description of the cultures obtained. For both species, Potato Dextrose Agar (PDA) was the most promising of the tested culture media. The highest growth in agar culture ever recorded for this genus is reported (4.9 mm per week for G. lloydianum and 7.5 mm for G. subiculosum). In the PDA culture medium, spores germinated after 35-40 days of incubation and the isolation of monokaryotic cultures of the two species, as well as in vitro sexual crosses, were successfully performed.

Sexual attractions, behaviors, and boundary crossings between sport psychology professionals and their athlete-clients: Prevalence, attitudes, and supervision

Participants were 181 sport performance professionals (SPPs); 92 reported being sexually attracted to their athlete-clients (ACs), though few SPPs sought supervision regarding such attractions. In regards to specific behaviors, approximately half reported discussing personal matters unrelated to their work, whereas far fewer had engaged in sexual behaviors with their ACs, such as discussing sexual matters unrelated to their work, and caressing or intimately touching an AC. Common nonsexual boundary crossings (NSBCs) included consulting with an AC in public places, working with an AC at practice, and working with an AC at a competition. Sexual attractions exist and NSBCs occur, thus SPPs need to be trained in these issues to be able to successfully navigate them. Lay summary: About half of the sport psychology professionals (SPPs) reported being sexually attracted to an athlete-client (AC). Typical boundary crossings included: consulting with an AC in public and private places and travelling with ACs. Therefore SPPs’ should be ethically trained and seek supervision to effectively work with such attractions.

Development of an effective protocol for in vitro multiplication of peppermint (Mentha piperita)

Mentha piperita is a completely sterile hybrid due to the fact that male organs of flower fail to develop. The stamens remain abnormally short, sometimes degenerate and wither from bud. As a result it is not amenable to improvement by sexual crosses. Ruthless exploitation of peppermint due to growing interest in the commercialization of plant based medicines has lead to drastic decrease of natural resource base which will result in extinction. In order to meet pharmaceutical needs and to provent the plants from extinction, development of rapid large scale propagation systems is a necessity. In vitro multiplication was carried out for mass production of disease free planting material without seasonal constraints ensuring germplasm conservation and sustainable use of the medicinal crop.

European Court of Justice delivers no justice to Europe on genome-edited crops.

European Court of Justice delivers no justice to Europe on genome-edited crops.

Infectious particle identity determines dissemination and disease outcome for the inhaled human fungal pathogen Cryptococcus.

The majority of invasive human fungal pathogens gain access to their human hosts via the inhalation of spores from the environment into the lung, but relatively little is known about this infectious process. Among human fungal pathogens the most frequent cause of inhaled fatal fungal disease is Cryptococcus, which can disseminate from the lungs to other tissues, including the brain, where it causes meningoencephalitis. To determine the mechanisms by which distinct infectious particles of Cryptococcus cause disseminated disease, we evaluated two developmental cell types (spores and yeast) in mouse models of infection. We discovered that while both yeast and spores from several strains cause fatal disease, there was a consistently higher fungal burden in the brains of spore-infected mice. To determine the basis for this difference, we compared the pathogenesis of avirulent yeast strains with their spore progeny derived from sexual crosses. Strikingly, we discovered that spores produced by avirulent yeast caused uniformly fatal disease in the murine inhalation model of infection. We determined that this difference in outcome is associated with the preferential dissemination of spores to the lymph system. Specifically, mice infected with spores harbored Cryptococcus in their lung draining lymph nodes as early as one day after infection, whereas mice infected with yeast did not. Furthermore, phagocyte depletion experiments revealed this dissemination to the lymph nodes to be dependent on CD11c+ phagocytes, indicating a critical role for host immune cells in preferential spore trafficking. Taken together, these data support a model in which spores capitalize on phagocytosis by immune cells to escape the lung and gain access to other tissues, such as the central nervous system, to cause fatal disease. These previously unrealized insights into early interactions between pathogenic fungal spores and lung phagocytes provide new opportunities for understanding cryptococcosis and other spore-mediated fungal diseases.

Transcriptome sequence resource for the cucurbit powdery mildew pathogen Podosphaera xanthii

Podosphaera xanthii is the main causal agent of cucurbit powdery mildew in Southern Italy. Illumina sequencing of mRNA from two P. xanthii isolates of opposite mating types (MAT1-1 and MAT1-2) and their sexual cross was used to obtain a detailed de novo Trinity-based assembly of the transcriptome of the fungus. Over 60 million of high-quality paired-end reads were obtained and assembled into 71,095 contigs corresponding to putative transcripts that were functionally annotated. More than 55% of the assembled transcripts (40,221 contigs) had a significant hit in BLASTx search and included sequences related to sexual compatibility and reproduction, as well as several classes of transposable elements and putative mycoviruses. The availability of these new transcriptomic data and investigations on potential source of genetic variation in P. xanthii will promote new insights on the pathogen and its interactions with host plants and associated microbiome.

Genetic improvement of naked-tufted seed mutants in upland cotton (Gossypium hirsutum L.)

This research evaluated the potential for using sexual crosses for genetic improvement of fiber yield, lint percent, AFIS fiber quality, and fiber initiation of two naked-tufted mutants of cotton (Gossypium hirsutum L.). The naked tufted lines were crossed with ten fuzzy breeding lines to generate 10 F2 populations. The F2 distributions were tested with Chi Square for Goodness of Fit to two monogenic and two digenetic segregation ratios. Five of the six F2 populations made with Atlas-NS-129 segregated as either a single dominant gene (3 Naked-tufted: 1 fuzzy) or as two dominant genes (9 Naked-tufted: 7 fuzzy). Three of the four F2 populations made with SC 9023-ns-57 segregated as either a single recessive gene (3 Fuzzy: 1 naked-tufted) or as duplicate recessive genes (9 Fuzzy: 7 naked-tufted). However, the F2 populations when crossed with the Fuzzy line of Tejas 48 showed a reversal of dominance. The F2 population of Atlas-NS-129 X Tejas 48 fit a 9 Fuzzy: 7 naked tufted ratio expected of two recessive genes. The F2 population of the cross of SC 9023-ns X Tejas 48 fit a 3 Naked-tufted: 1 fuzzy ratio expected of a single dominant gene. Additional studies will be necessary to fully determine the number of gene(s) and the expected phenotypic ratios of individual naked-tufted mutants in crosses with fuzzy seeded breeding lines. Eight of the F2 and F3 populations were also evaluated for lint yield, lint percent and AFIS fiber quality traits. The F2 and F3 populations of the cross of Atlas-NS-129 X Holland 338 produced the highest average lint yields (1089 kg ha−1) as well as excellent fiber quality. These data indicate that it may be possible to develop relatively high yielding naked tufted cultivars by crossing these mutant lines with elite higher yielding breeding lines.

Mechanism underlying 2n male and female gamete formation in lemon via cytological and molecular marker analysis

The unreduced (2n) gametes have long been used in triploid breeding of citrus. In lemon, the previously reported mechanisms for 2n megagametophyte formation were controversial, whereas that for 2n pollen production is still unknown. Herein, the frequency of and mechanism underlying 2n megagametophyte and 2n pollen formation in ‘Eureka’ lemon were investigated based on cytological observation and genotyping of the triploid hybrids between ‘Eureka’ lemon and ‘Early gold’ sweet orange. As a result, 4.79% of the viable pollens of ‘Eureka’ lemon were identified as the 2n pollen with a larger diameter (70.16 ± 3.92 µm). The 2n pollen might be resulted from the formation of parallel spindles at meiosis stage II. Among the 204 plantlets regenerated from embryo rescue following the sexual cross, 12 were triploids as identified by flow cytometry. According to the analysis of heterozygosity transmission using 13 pericentromeric single nucleotide polymorphism (SNP) markers and 20 randomly distributed simple sequence repeat (SSR) markers, 11 triploids were identified to be originated from the fertilization of 2n megagametophytes of ‘Eureka’ lemon, with a frequency of 5.39%. Among them, nine 2n megagametophytes were supposed to be arisen from the second division restitution (SDR), whereas the other two were from post-meiotic genome doubling (PMD). These results to understand the mechanism underlying 2n gamete formation in lemon are valuable for its efficient polyploid breeding.

Morphology, phylogeny, and sexual stage of Fusarium caatingaense and Fusarium pernambucanum, new species of the Fusarium incarnatum-equiseti species complex associated with insects in Brazil

ABSTRACTBased on morphological and molecular phylogenetic markers and the fertility of sexual crosses, two novel species of Fusarium associated with Dactylopius opuntiae (Hemiptera: Dactylopiidae) and Aleurocanthus woglumi (Hemiptera: Aleyrodidae) from northeastern Brazil are described as Fusarium caatingaense and F. pernambucanum. Partial sequences of five loci were generated for 29 entomopathogenic Fusarium isolates. Multilocus phylogenetic analyses demonstrated that F. caatingaense and F. pernambucanum belong to the Incarnatum clade of the Fusarium incarnatum-equiseti species complex (FIESC). These species displayed common morphological characters such as the production of various types of aerial conidia formed on monophialides and polyphialides and differ from each other mainly in the dimensions and morphology of their sporodochial conidia. Mating type polymerase chain reaction (PCR) revealed 17 MAT1-1 isolates and 12 MAT1-2 isolates, all of them heterothallic. Fertile perithecia were produced in 4.2% of infraspecific crosses of F. caatingaense and in 13.3% of infraspecific crosses of F. pernambucanum after 2–3 wk. Crosses between F. caatingaense and F. pernambucanum did not result in fertile perithecia. We demonstrate the existence of a sexual stage in species of the Incarnatum clade and describe the morphological characters of these sexual morphs for the first time. These results suggest that previously unknown sexual cycles contribute to the high genetic diversity within FIESC.

Fusarium udum revisited: a common, but poorly understood member of the Fusarium fujikuroi species complex

Fusarium udum is the causal agent of a wilt disease on pigeon pea (Cajanus cajan) in tropical regions. This species shares morphological characteristics with F. oxysporum, leading to misidentification when the diagnosis is solely based on morphological markers. The sexual stage of this fungus was observed on stems of Cajanus in India and was formally described as Gibberella indica. In Brazil, a wilt disease is reported on Crotalaria, but the etiological agent has not been identified correctly so far. In this study, we tested the hypothesis that the causal agent of a wilt on Crotalaria belongs to the same species pathogenic to C. cajan. Strains obtained from diseased Crotalaria spp. were characterized through molecular phylogeny of TEF, TUB and RPB2, laboratory crosses, morphological markers, and pathogenicity tests. Strains from Crotalaria from Brazil formed a well-supported clade with F. udum strains from Crotalaria and Cajanus from other countries. Strains from Brazil intercrossed among themselves and also with others from all geographic regions, and formed fertile perithecia, defining a distinct mating population inside the Fusarium fujikuroi species complex. One strain obtained from Cajanus cajan in India is indicated as epitype, and female-fertile tester strains of both mating types were selected for the identification of field isolates through sexual crosses. These results confirm that the species associated with wilt disease on Crotalaria and Cajanus is F. udum. Wilt symptoms caused by F. udum in Cr. ochroleuca are described and illustrated.

Deletion of the sex-determining gene SXI1\u03b1 enhances the spread of mitochondrial introns in Cryptococcus neoformans

BackgroundHoming endonuclease genes (HEGs) are widely distributed genetic elements in the mitochondrial genomes of a diversity of eukaryotes. Due to their ability to self-propagate within and between genomes, these elements can spread rapidly in populations. Whether and how such elements are controlled in genomes remains largely unknown.ResultsHere we report that the HEG-containing introns in the mitochondrial COX1 gene in Cryptococcus neoformans are mobile and that their spread in sexual crosses is influenced by mating type (MAT) α-specific homeodomain gene SXI1α. C. neoformans has two mating types, MATa and MATα. In typical crosses between strains of the two mating types, only a small portion (< 7%) of diploid fusants inherited the HEGs from the MATα parent. However, disruption of the SXI1α gene resulted in the majority (> 95%) of the diploid fusants inheriting the HEG-containing introns from the MATα parent, a frequency significantly higher than those of intronless mitochondrial genes.ConclusionsOur results suggest that SXI1α not only determines uniparental mitochondrial inheritance but also inhibits the spread of HEG-containing introns in the mitochondrial genome in C. neoformans.

Mejoramiento genético acelerado de angiospermas perennes vía inducción floral por sobre-expresión del gen FT

Los bosques y selvas enfrentan el reto de satisfacer la demanda por recursos de una población en crecimiento, así como la amenaza del rápido cambio climático que exacerba la magnitud y frecuencia de estreses bióticos y abióticos. Para ello, es urgente acelerar el mejoramiento genético de especies forestales. Sin embargo, sus largas etapas juveniles y asincronía floral retrasan peligrosamente este proceso. El presente ensayo explora los adelantos biotecnológicos en inducción floral y su potencial aplicación en especies forestales. Entre los genes identificados y caracterizados que participan en la ruta de señalización de la floración, especial atención se destina al gen FLOWERING LOCUS T, considerado un integrador de rutas de señalización altamente conservado entre las angiospermas, que, al sobre-expresarse por ingeniería genética, es capaz de inducir la floración de forma eficiente. Esta novedosa estrategia biotecnológica se ha utilizado, recientemente, para segregar genes de resistencia a enfermedades, en un menor tiempo, en germoplasma comercial de manzana y ciruela. Permite soslayar barreras naturales que por mucho tiempo han restringido a las especies forestales al mejoramiento por selección, principalmente. Entre sus ventajas está la de poder restringirla al proceso y no al producto, para acelerar las cruzas sexuales sin modificar genéticamente la progenie; se aleja así de la controversia alrededor de la liberación y consumo de organismos genéticamente modificados, y de los costos y trámites obligatorios para los OGM para monitoreo de posibles riesgos. Se proyecta como una tecnología que puede acelerar, significativamente, el mejoramiento de especies forestales.

Persistence of &amp;lt;i&amp;gt;Venturia inaequalis&amp;lt;/i&amp;gt; Populations Resistant to Strobilurins in the Field and in the Glasshouse

The research was focused on the persistence of resistance of V. inaequalis to strobilurins from resistant populations occurring in the field. It was studied with two types of experiments: the first in apple orchards where resistance occurred in 2005 and employ of these fungicides was suspended from 2006 to 2011, realizing every year sensitivity/resistance assays on strobilurins. In the second was verified the sensitivity/resistance of the same 2015 populations, repeatedly multiplied (ten inoculations) in glasshouse on apple seedlings. Results showed that the resistance survived in orchards all the years of tests (six years), despite sensitivity fluctuations occurred on infected leaves due to sexual crosses, competition with sensible strains and soil management. In glasshouse, the level of resistance of all populations increased quickly in first inoculations and values remained high (EC50 > 10 mg·L-1) in all the following inoculations, until the last, the 10th. V. inaequalis resistant strains seemed to show high fitness: relevant persistence in the field, and a capacity of prevail on sensible ones in absence of factors of field variability.