Parasexual Recombination Research Articles

P335 Genome plasticity in Candida albicans : acutting-edge strategy for evolution, adaptation, and survival

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjectives Candida albicans is the most implicated fungal species that grows as a commensal or opportunistic pathogen in the human host. The genome plasticity is central to its adaptation and has long been of considerable interest. Understanding the factors that affect the genome is crucial for a proper understanding of species and rapid development and adjustment of therapeutic strategies to mitigate their spread.MethodsHere, we present an up-to-date overview of recent genomic studies involving C. albicans and discuss the accumulating evidence that shows how mitotic recombination events, ploidy dynamics, aneuploidy, and loss of heterozygosity (LOH) influence evolution, adaptation, and survival in C. albicans. This review was exclusively based on a thorough literature review on research papers investigating C. albicans and its genome. The Boolean search for relevant literatures was performed in Google scholar, Science direct, PubMed, and Web of science using a combination of the following precise keywords: Candida albicans, Genome plasticity, Parasexual events, Ploidy dynamics, Aneuploidy, LOH, and Mutations. All the papers that relate to the genome and genetics of C. albicans were included in the study.Results Candida albicans has a diploid heterozygous genome that is highly dynamic and can display variation from small to large-scale chromosomal rearrangement and aneuploidy, which have implications for drug resistance, virulence, and pathogenicity. The instability in the genome arising from stress or exposure to the host environment can be detrimental to survival. However, C. albicans can tactically exploit this instability in their genome as a survival strategy. Our investigation showed that there is a need for more studies to understand the regulation and the exact mechanisms that lead to chromosomal instability in C. albicans. It is important to understand how increasing ploidy affects genome stability. It is also important to unravel how tetraploids lose chromosomes to revert to diploids. Whether other ploidy states can be efficiently sustained also remains to be unraveled. Moreover, as the understanding of the pathogenicity of fungi continues to evolve, there is a continuing challenge in grasping the processes and factors that mediate the switching from commensal to the pathogen. It remains unclear whether parasexual processes also operate within natural populations. More studies are needed to appreciate better parasexual recombination events and mutational processes in C. albicans evolution and its link to persistence and infection establishment. The interrelationship between the host and the microbes can promote microbial fitness in the host and help the host acquire some beneficial features such as resistance against several pathogens. However, the long-term consequences on host fitness are largely unknown. This evidence provides an avenue to study evolutionary forces controlling C. albicans fitness level after host exposure.ConclusionThe variation in the genome of C. albicans greatly influences its biology, stress response approaches, and its virulence and resistant potentials. Understanding how C. albicans generate diversity is very crucial as this will help to design strategies mitigating their spread.

The Emergence of Fusarium oxysporum f. sp. apii Race 4 and Fusarium oxysporum f. sp. coriandrii Highlights Major Obstacles Facing Agricultural Production in Coastal California in a Warming Climate: A Case Study.

Currently, Fusarium oxysporum f. sp. apii (Foa) race 4 in celery and F. oxysporum f. sp. coriandrii (Foci) in coriander have the characteristics of emerging infectious plant diseases in coastal southern California: the pathogens are spreading, yield losses can be severe, and there are currently no economical solutions for their control. Celery, and possibly coriander, production in these regions is are likely to have more severe disease from projected warmer conditions in the historically cool, coastal regions. Experimental evidence shows that Foa race 4 causes much higher disease severity when temperatures exceed 21°C. A phylogenomic analysis indicated that Foa race 4, an older, less virulent, and uncommon Foa race 3, and two Foci are closely related in their conserved genomes. These closely related genotypes are somatically compatible. Foa race 4 can also cause disease in coriander and the two organisms readily form “hetero” conidial anastomosis tubes (CAT), further increasing the likelihood of parasexual recombination and the generation of novel pathotypes. A horizontal chromosome transfer event likely accounts for the difference in host range between Foci versus Foa races 4 and 3 because they differ primarily in one or two accessory chromosomes. How Foa race 4 evolved its hyper-virulence is unknown. Although the accessory chromosomes of Foa races 3 and 4 are highly similar, there is no evidence that Foa race 4 evolved directly from race 3, and races 3 and 4 probably only have a common ancestor. Foa race 2, which is in a different clade within the Fusarium oxysporum species complex (FOSC) than the other Foa, did not contribute to the evolution of race 4, and does not form CATs with Foa race 4; consequently, while inter-isolate CAT formation is genetically less restrictive than somatic compatibility, it might be more restricted between FOSC clades than currently known. Other relatively new F. oxysporum in coastal California include F. oxysporum f. sp. fragariae on strawberry (Fof). Curiously, Fof “yellows-fragariae” isolates also have similar core genomes to Foa races 4 and 3 and Foci, perhaps suggesting that there may be core genome factors in this lineage that favor establishment in these soils.

Parasexual recombination in Macrophomina pseudophaseolina and vegetative compatibility reactions in M. euphorbiicola

Parasexual recombination in Macrophomina pseudophaseolina and vegetative compatibility reactions in M. euphorbiicola

A Novel Spo11 Homologue Functions as a Positive Regulator in Cyst Differentiation in Giardia lamblia.

Giardia lamblia persists in a dormant state with a protective cyst wall for transmission. It is incompletely known how three cyst wall proteins (CWPs) are coordinately synthesized during encystation. Meiotic recombination is required for sexual reproduction in animals, fungi, and plants. It is initiated by formation of double-stranded breaks by a topoisomerase-like Spo11. It has been shown that exchange of genetic material in the fused nuclei occurs during Giardia encystation, suggesting parasexual recombination processes of this protozoan. Giardia possesses an evolutionarily conserved Spo11 with typical domains for cleavage reaction and an upregulated expression pattern during encystation. In this study, we asked whether Spo11 can activate encystation process, like other topoisomerases we previously characterized. We found that Spo11 was capable of binding to both single-stranded and double-stranded DNA in vitro and that it could also bind to the cwp promoters in vivo as accessed in chromatin immunoprecipitation assays. Spo11 interacted with WRKY and MYB2 (named from myeloblastosis), transcription factors that can activate cwp gene expression during encystation. Interestingly, overexpression of Spo11 resulted in increased expression of cwp1-3 and myb2 genes and cyst formation. Mutation of the Tyr residue for the active site or two conserved residues corresponding to key DNA-binding residues for Arabidopsis Spo11 reduced the levels of cwp1-3 and myb2 gene expression and cyst formation. Targeted disruption of spo11 gene with CRISPR/Cas9 system led to a significant decrease in cwp1-3 and myb2 gene expression and cyst number. Our results suggest that Spo11 acts as a positive regulator for Giardia differentiation into cyst.

Empirical Support for the Pattern of Competitive Exclusion between Insect Parasitic Fungi.

Fungal entomopathogens are largely facultative parasites and play an important role in controlling the density of insect populations in nature. A few species of these fungi have been used for biocontrol of insect pests. The pattern of the entomopathogen competition for insect individuals is still elusive. Here, we report the empirical competition for hosts or niches between the inter- and intra-species of the entomopathogens Metarhizium robertsii and Beauveria bassiana. It was found that the synergistic effect of coinfection on virulence increase was not evident, and the insects were largely killed and mycosed by M. robertsii independent of its initial co-inoculation dosage and infection order. For example, >90% dead insects were mycosed by M. robertsii even after immersion in a spore suspension with a mixture ratio of 9:1 for B. bassiana versus M. robertsii. The results thus support the pattern of competitive exclusion between insect pathogenic fungi that occurred from outside to inside the insect hosts. Even being inferior to compete for insects, B. bassiana could outcompete M. robertsii during co-culturing in liquid medium. It was also found that the one-sided mycosis of insects occurred during coinfection with different genotypic strains of either fungi. However, parasexual recombination was evident to take place between the compatible strains after coinfection. The data of this study can help explain the phenomena of the exclusive mycosis of insect individuals, but co-occurrence of entomopathogens in the fields, and suggest that the synergistic effect is questionable regarding the mixed use of fungal parasites for insect pest control.

Copulating cancer cells

Cancer cells are notoriously tricky; despite treatments designed to kill them, they can keep returning by quickly evolving or diversifying their genetics. Mutation is one way that cancer cells can switch up their genetics. Now, a group led by Andriy Marusyk of the University of South Florida has found another mechanism, one that looks a lot like sex. In in vivo and in vitro tests, Marusyk’s team found that a small number of cancer cells can combine their DNA by fusing together, taking on characteristics of both parent cells ( Nat. Ecol. Evol. 2021, DOI: 10.1038/s41559-020-01367-y ). Researchers have seen cancer cells fuse before, but it wasn’t clear if there was a benefit to the cells. Marusyk’s team wondered if parasexual recombination—combining two cells to create one new one—might be happening. To find out, the researchers made two populations of breast cancer cells, each modified to produce a different fluorescent

Spontaneous cell fusions as a mechanism of parasexual recombination in tumour cell populations.

The initiation and progression of cancers reflect the underlying process of somatic evolution, in which the diversification of heritable phenotypes provides a substrate for natural selection, resulting in the outgrowth of the most fit subpopulations. Although somatic evolution can tap into multiple sources of diversification, it is assumed to lack access to (para)sexual recombination-a key diversification mechanism throughout all strata of life. On the basis of observations of spontaneous fusions involving cancer cells, the reported genetic instability of polypoid cells and the precedence of fusion-mediated parasexual recombination in fungi, we asked whether cell fusions between genetically distinct cancer cells could produce parasexual recombination. Using differentially labelled tumour cells, we found evidence of low-frequency, spontaneous cell fusions between carcinoma cells in multiple cell line models of breast cancer both in vitro and in vivo. While some hybrids remained polyploid, many displayed partial ploidy reduction, generating diverse progeny with heterogeneous inheritance of parental alleles, indicative of partial recombination. Hybrid cells also displayed elevated levels of phenotypic plasticity, which may further amplify the impact of cell fusions on the diversification of phenotypic traits. Using mathematical modelling, we demonstrated that the observed rates of spontaneous somatic cell fusions may enable populations of tumour cells to amplify clonal heterogeneity, thus facilitating the exploration of larger areas of the adaptive landscape (relative to strictly asexual populations), which may substantially accelerate a tumour's ability to adapt to new selective pressures.

Parasexual recombination enablesAspergillus fumigatusto persist in cystic fibrosis

Aspergillus fumigatus is a saprobic fungus that causes a range of pulmonary diseases, some of which are characterised by fungal persistence such as is observed in cystic fibrosis (CF) patients. Creation of genetic variation is critical for A. fumigatus to adapt to the lung environment, but biofilm formation, especially in CF patients, may preclude mutational supply in A. fumigatus due to its confinement to the hyphal morphotype. We tested our hypothesis that genetic variation is created through parasexual recombination in chronic biofilms by phenotypic and genetic analysis of A. fumigatus isolates cultured from different origins.As diploids are the hallmark of parasex, we screened 799 A. fumigatus isolates obtained from patients with CF, chronic pulmonary lung disease and acute invasive aspergillosis, and from the environment for spore size. Benomyl sensitivity, nuclear content measurements through fluorescence-activated cell sorting and scanning electron microscopy were used to confirm the diploid state of large size spores. Whole genome sequencing was used to characterise diploid-associated genetic variation.We identified 11 diploids in isolates recovered from six of 11 (55%) CF patients and from one of 24 (4%) chronic aspergillosis patients, but not in 368 isolates from patients with acute Aspergillus infection and the environment. Diploid formation was associated with accumulation of mutations and variable haploid offspring including a voriconazole-resistant isolate.Parasexual recombination allows A. fumigatus to adapt and persist in CF patients, and plays a role in azole resistance development. Our findings are highly significant for understanding the genetics and biology of A. fumigatus in the human lung.

Genomic Instability in Fungal Plant Pathogens.

Fungi and fungal-like organisms (oomycetes) that cause diseases in plants have impacted human communities for centuries and probably from the dawn of agriculture. In modern agriculture, there is a constant race between new strategies to manage fungal plant pathogens and their ability to adapt. An important component in this race is fungal genetic diversity. Mechanisms such as sexual and parasexual recombination that contribute to the creation of novel allele combinations in fungal plant pathogens are briefly discussed in the first part of this review. Advances in genomics have enabled the investigation of chromosomal aberrations of agriculturally important fungal isolates at the nucleotide level. Some of these cases are summarized in the second part of this review; it is claimed that the effect of chromosomal aberrations on pathogenicity should be studied mechanistically. More data on the effect of gene copy number variations on phenotypes that are relevant to agriculture are especially needed. Genome rearrangements through translocations have shaped the genome of fungal plant pathogens by creating lineage-specific chromosome territories encoding for genes participating in plant diseases. Pathogenicity chromosomes are unique cases of such lineage-specific genetic elements, interestingly these chromosomes can be transferred horizontally and thus transforming a non-pathogenic strain to a pathogenic one. The third part of this review describes our attempts to reveal mutators in fungal plant pathogens by identifying fungi that lack important DNA repair genes or respond to DNA damage in an unconventional way. We found that a group of fungal plant pathogens lack conserved genes that are needed for an important Holliday junction resolution pathway. In addition, in Fusarium oxysporum, the rate-limiting step in dNTP production is not induced under DNA replication stress. This is very different from organisms from bacteria to humans. It remains to be seen if these mechanisms promote genetic instability in fungal plant pathogens.

Evolution of Fluconazole-Resistant Candida albicans Strains by Drug-Induced Mating Competence and Parasexual Recombination.

The clonal population structure of Candida albicans suggests that (para)sexual recombination does not play an important role in the lifestyle of this opportunistic fungal pathogen, an assumption that is strengthened by the fact that most C. albicans strains are heterozygous at the mating type locus (MTL) and therefore mating-incompetent. On the other hand, mating might occur within clonal populations and allow the combination of advantageous traits that were acquired by individual cells to adapt to adverse conditions. We have investigated if parasexual recombination may be involved in the evolution of highly drug-resistant strains exhibiting multiple resistance mechanisms against fluconazole, an antifungal drug that is commonly used to treat infections by C. albicans Growth of strains that were heterozygous for MTL and different fluconazole resistance mutations in the presence of the drug resulted in the emergence of derivatives that had become homozygous for the mutated allele and the mating type locus and exhibited increased drug resistance. When MTLa/a and MTLα/α cells of these strains were mixed in all possible combinations, we could isolate mating products containing the genetic material from both parents. The initial mating products did not exhibit higher drug resistance than their parental strains, but further propagation under selective pressure resulted in the loss of the wild-type alleles and increased fluconazole resistance. Therefore, fluconazole treatment not only selects for resistance mutations but also promotes genomic alterations that confer mating competence, which allows cells in an originally clonal population to exchange individually acquired resistance mechanisms and generate highly drug-resistant progeny.IMPORTANCE Sexual reproduction is an important mechanism in the evolution of species, since it allows the combination of advantageous traits of individual members in a population. The pathogenic yeast Candida albicans is a diploid organism that normally propagates in a clonal fashion, because heterozygosity at the mating type locus (MTL) inhibits mating between cells. Here we show that C. albicans cells that have acquired drug resistance mutations during treatment with the commonly used antifungal agent fluconazole rapidly develop further increased resistance by genome rearrangements that result in simultaneous loss of heterozygosity for the mutated allele and the mating type locus. This enables the drug-resistant cells of a population to switch to the mating-competent opaque morphology and mate with each other to combine different individually acquired resistance mechanisms. The tetraploid mating products reassort their merged genomes and, under selective pressure by the drug, generate highly resistant progeny that have retained the advantageous mutated alleles. Parasexual propagation, promoted by stress-induced genome rearrangements that result in the acquisition of mating competence in cells with adaptive mutations, may therefore be an important mechanism in the evolution of C. albicans populations.

Study on Parasexual Recombination between <i>Pyricularia oryzae</i> and <i>Pyricularia grisea</i>

Rice blast fungus is notorious for changeability in pathogenicity, but it lacks sexual life cycle. It can be postulated that such an imperfect fungus has another mechanism for generating genetic variations. Recent studies concerning comparative genomics reveal that parasexual recombination may play important role in the evolution of rice blast fungus. To observe the parasexualism of rice blast (Pyricularia oryzae) and crabgrass blast (Pyricularia grisea) fungus double inoculation and punch method were applied in this experiment. A total of 520 isolates collected from the double inoculated lesions was subjected to PCR-RFLP analysis of the ITS region to identify subcultures of the inoculated rice blast isolates. As a result, four isolates from the three double inoculated lesions with SA13-1ME and TP106 were identified as subcultures of TP106. To access the recombination genotypes, a total of 17 isolates from the three lesions was subjected to MAGGY-DNA fingerprint analysis. However, recombinant DNA fingerprint patterns between TP106 and SA13-1ME were not detected among the 17 isolates. Although TP022 was not recovered from the double inoculated lesions, the fact that TP106 was recovered from the double inoculated lesion indicates that rice blast fungus can invade and colonized in blast lesion on crabgrass. The opportunistic infection on the double inoculated lesions observed in this study potentially provides new insight into the life cycle of rice blast pathogen.

Caracterização de Isolados de Fusarium oxysporum f. sp. cubense (Foc) Quanto à Compatibilidade Vegetativa e à Patogenicidade em Cultivares de Bananeira Diferenciadoras de Raças no Brasil

Fusarium wilt is caused by Fusarium oxysporum f. sp. cubense (Foc) a highly variable pathogen. However, the effect of heterokaryosis on the taxonomic classification and the importance of the formation of a heterokaryon between wild isolates of Fusarium species is still unclear and remains widely debated. Just as parasexual recombination has been reported in F. oxysporum f. sp. cubense , to explain the genetic relationships among isolates belonging to the same race or formae specialis . Asexual and genetically isolated groups identified in these studies are called vegetative compatibility groups (VCG). However, another way of determining races of this pathogen has been through indicator varieties, using the Gros Michel variety for Race 1; Bluggoe for Race 2; Great Naine for Race 4 The present study aimed to determine the genetic diversity among isolates of Foc, through these two techniques in order to identify the races of the pathogen, which do exist in Brazil. The genetics analysis was based on the technique of UPGMA, using the Jaccard similarity coefficient. The technique allowed VCG group the isolates from the Northeast and Southeast regions of Brazil in the VCG 1 and the Northern Region, the VCG 2. While the pathogenicity test allowed us to determine the presence of race 1, through its pathogenicity in Gros Michel and race 4 in Grand Naine. Besides, group them within the same VCG. Keywords: Fusarium wilt ; parasexual recombination; asexual reproduction.

Phylogenetic analyses on the diversity of Aspergillus fumigatus sensu lato based on five orthologous loci.

One hundred isolates of Aspergillus fumigatus sensu lato mainly from China, as well as from Australia, France, India, Indonesia, Ireland, UK, and USA were analyzed to infer their sequence types (STs) and population diversity based on partial calmodulin, calcineurin regulatory subunit B, beta-tubulin, cytochrome C and calcineurin catalytic subunit A genes as well as their mating types, using ClonalFrame, Structure and MEGA software. Our results inferred 48 STs and showed that most of the STs or lineages evolved independently and without clear population structure among them. Whereas one lineage was recognized that could be a true population and in which one clade might diverge into another distinct lineage, namely, a cryptic species, A. neoellipticus. In addition, we found that mutation, parasexual, and sexual recombination could, respectively, play specific roles in the evolution of these fungi. Our results also showed that MAT1-1/MAT1-2 mating type ratios of A. fumigatus sensu lato was biased to nearly 1:1.4 (20/28) when clone-corrected, but when not clone-corrected, the ratio of MAT1-1/MAT1-2 was so biased as near 1:2 (35/65), which might mean that isolates with MAT1-2 are in the process of losing sexual ability preceding those with MAT1-1.

Vegetative compatibility groups and sexual reproduction among Spanish Monilinia fructicola isolates obtained from peach and nectarine orchards, but not Monilinia laxa

The frequency of occurrence of Monilinia fructicola in the Ebro Valley, Spain has increased since its first appearance in 2006, and M. fructicola has displaced Monilinia laxa, the native species which is the main cause of brown rot in peaches in this valley. In order to determine the characteristics that may be related to the displacement, we studied the capacity to generate new genotypic combinations of M. fructicola under laboratory conditions. The morphology and parasitic ability from ten field isolates of M. fructicola and M. laxa collected from three different orchards in the valley, and sampling from five different lesions were studied. Nitrate-non-utilising (nit) mutants were generated in order to test the isolates for vegetative compatibility which was done by assessing their colony growth when cultured singly or in pairs on media that contained different nitrogen sources. For the M. fructicola isolates, five vegetative compatibility groups (VCGs) were identified using the nit mutants and six VCGs were identified when they were grown on potato dextrose agar dishes. In all instances, the vegetatively compatible M. fructicola isolates came mainly from the same orchard. Only one VCG displays the same morphological and competition characteristics. No VCGs were identified among the M. laxa isolates. We did not find any apothecia of M. laxa and M. fructicola isolates in the soil of the three orchards, but we were able to produce apothecia of M. fructicola in the laboratory. Our finding of sexual reproduction and VCGs in the M. fructicola isolates suggests that the genetic variability of M. fructicola could be maintained by sexual and/or parasexual recombination.

Population structure of Fusarium oxysporum f. sp. radicis-lycopersici in Florida inferred from vegetative compatibility groups and microsatellites

Fusarium crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL) is a serious soilborne disease reducing tomato yields in Florida, a leading state in fresh market tomato production in the United States. One hundred and twenty five isolates of FORL were collected from the three main tomato-growing counties in Florida between 2006 and 2008. Vegetative compatibility groups (VCGs) and 10 microsatellite loci were used to infer the population structure of FORL. Up to 69.8 % of the isolates could be assigned to one of three VCGs, 0094, 0098 or 0099, with frequencies of 38.6, 24.4, and 6.8 % respectively. A medium level of population differentiation (Φst = 0.159) was detected among the three counties, and three optimal clusters (populations) were supported by discriminant analysis of principal components. In addition, each population had some individuals that likely migrated from other populations. Migration probably played an important role in shaping the population structure of FORL since repeated VCGs and multilocus genotypes were observed in the three counties. Considerable migrants (> 1.33 migrants per generation) were also detected between the three counties, resulting in an increase in the effective population size and genetic diversity of F. oxysporum f. sp. radicis-lycopersici. Although migration is an important evolutionary force, mutation and parasexual recombination could not be completely ruled out as contributing causes to the genetic diversity of FORL. Management strategies that limit the movement of F. oxysporum f. sp. radicis-lycopersici are necessary to reduce the evolutionary potential of the pathogen.

Heterokaryon Incompatibility Is Suppressed Following Conidial Anastomosis Tube Fusion in a Fungal Plant Pathogen

It has been hypothesized that horizontal gene/chromosome transfer and parasexual recombination following hyphal fusion between different strains may contribute to the emergence of wide genetic variability in plant pathogenic and other fungi. However, the significance of vegetative (heterokaryon) incompatibility responses, which commonly result in cell death, in preventing these processes is not known. In this study, we have assessed this issue following different types of hyphal fusion during colony initiation and in the mature colony. We used vegetatively compatible and incompatible strains of the common bean pathogen Colletotrichum lindemuthianum in which nuclei were labelled with either a green or red fluorescent protein in order to microscopically monitor the fates of nuclei and heterokaryotic cells following hyphal fusion. As opposed to fusion of hyphae in mature colonies that resulted in cell death within 3 h, fusions by conidial anastomosis tubes (CAT) between two incompatible strains during colony initiation did not induce the vegetative incompatibility response. Instead, fused conidia and germlings survived and formed heterokaryotic colonies that in turn produced uninucleate conidia that germinated to form colonies with phenotypic features different to those of either parental strain. Our results demonstrate that the vegetative incompatibility response is suppressed during colony initiation in C. lindemuthianum. Thus, CAT fusion may allow asexual fungi to increase their genetic diversity, and to acquire new pathogenic traits.

Evidence of resistance gene markers parasexual recombination in Magnaporthe grisea

A selectable marker gene conferring resistance to bialaphos (Bar) was introduced into rice blast isolate 95-8-36 and another conferring resistance to hygromycin (Hph) into isolate P32-1 of Magnaporthe grisea by Agrobacterium tumefaciens- mediated transformation (ATMT) to demonstrate exchange of DNA. Many of the transformants of 95-8-36 and P32-1 were mixed by grinding and cultured on oat agar media. Spores were collected with sterile water and inoculated in PSA selection medium with hygromycin and glufosinate in order to screen the recombinants. Some isolates were obtained from the medium and confirmed to have Bar and Hph genes by PCR technique, indicating that M. grisea made parasexual recombination under the artificial conditions. The recombinants could inherit the double resistance stably such that their growth rate and sporulation ability also resemble their original isolates. Key words: Parasexual recombination, Magnaporthe grisea, resistant gene mark,Agrobacterium tumefaciens-mediated transformation (ATMT).

Genetic and pathogenic diversity of Neofusicoccum parvum in New Zealand vineyards

Genetic diversity of 50 isolates of Neofusicoccum parvum, the predominant species of the Botryosphaeriaceae recovered from grapevines displaying symptoms of dieback and decline in New Zealand, was compared to that of isolates from Australia, South Africa, and California. The eight universally primed polymerase chain reaction (UP-PCR) primers distinguished 56 genotypes, with only four clonal pairs found. Seven main groups were identified in a neighbour-joining (NJ) tree with isolates from different regions and vineyards of New Zealand, Australia, and California distributed in different groups, indicating a high level of intra and intervineyard genetic variation. All of the South African isolates were positioned in a separate UP-PCR group, indicating that these isolates were less related to the other N. parvum isolates. When compared to fungi that reproduce sexually the genetic diversity and Shannon diversity indices were low (0.076–0.249; 0.109–0.367, respectively), genetic identity levels were high (0.76–0.95), and genetic distance levels were low (0.04–0.27). The large number of genotypes and the low number of clones in the New Zealand N. parvum populations may be explained by parasexual recombination as anastomosis was observed between nonself pairings. Pathogenicity tests using isolates from different UP-PCR groups inoculated onto either green shoots or 1-y-old grapevines detected virulence diversity, indicating intra and intervineyard variation between isolates, however, no correlation was detected between UP-PCR group and virulence.

Parasexual Recombination in Magnaporthe oryzae

Parasexual recombination is thought to be one of the causes of variation in the pathogenicity of rice blast fungus, Magnaporthe oryzae, which breaks down resistant rice varieties. In this study, the virulence and fitness of parasexual recombinants of M. oryzae were examined in vitro and in the field. According to the results of the genetic analysis of the pathogenicity of parasexual recombinants of the fungus, the segregation ratios of avirulence and virulence among recombinants on rice cultivar Hattan 3 and line K59-1 were consistent with those of the sexual progeny of the fungus. This indicated that the avirulent genes of the parasexual recombinants were segregated in a manner similar to that in sexual reproduction. The fitness of parasexual recombinants derived from co-cultures of parental isolates was also investigated. The recombinants produced intermediate-sized lesions and intermediate numbers of spores between the parents on the host. Plants inoculated with the recombinant also showed intermediate disease severities between the parents in the field. Parasexual recombinants can attack a rice multiline system because their virulence and fitness is inherited from the parents.

A Single Mating-Type Locus Composed of Homeodomain Genes Promotes Nuclear Migration and Heterokaryosis in the White-Rot Fungus Phanerochaete chrysosporium

The white-rot basidiomycete fungus Phanerochaete chrysosporium (Agaricomycetes) is a model species that produces potent wood-degrading enzymes. The mating system of the species has been difficult to characterize due to its cryptic fruiting habit and lack of clamp connections in the heterokaryotic phase. By exploiting the draft genome sequence, we reevaluated the mating system of P. chrysosporium by studying the inheritance and segregation of putative mating-type gene homologues, the homeodomain transcription factor genes (MAT-A) and the pheromone receptors (MAT-B). A pattern of mating incompatibility and fructification consistent with a bipolar system with a single MAT locus was observed, but the rejection response was much weaker than that seen in other agaricomycete species, leading to stable heterokaryons with identical MAT alleles. The homeodomain genes appear to comprise the single MAT locus because they are heterozygous in wild strains and hyperpolymorphic at the DNA sequence level and promote aspects of sexual reproduction, such as nuclear migration, heterokaryon stability, and basidiospore formation. The pheromone receptor loci that might constitute a MAT-B locus, as in many other Agaricomycetes, are not linked to the MAT-A locus and display low levels of polymorphism. This observation is inconsistent with a bipolar mating system that includes pheromones and pheromone receptors as mating-type determinants. The partial uncoupling of nuclear migration and mating incompatibility in this species may be predicted to lead to parasexual recombination and may have contributed to the homothallic behavior observed in previous studies.