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Biology subjects

Sanders, I. R.

Publications and source records attributed to Sanders, I. R..

10 recordsLinked to original sources

Intricate asymbiotic hyphal interactions of Rhizophagus irregularis revealed with single-plane observations in a new microfluidic device

Arbuscular mycorrhizal fungi (AMF) are essential symbionts to most land plants. There is great interest in using AMF inoculation treatments to enhance restoration or agricultural efforts. However, little is known about AMF asymbiotic interactions that could shape soil ecosystems and inoculation outcomes. Studying these early traits is challenging given the obligate nature of AMF. Microfluidic devices enable high-resolution, single-plane, time-resolved observation of AMF hyphal traits in controlled, plant-less environments. The AMF-AnastomosisChip, introduced in this work, additionally separates spores and hyphae into lanes for detailed observations prior to entering a shared interaction zone. Using this device with Rhizophagus irregularis, we phenotyped key hyphal traits from germination to anastomosis. We compared the effect of a fatty acid (FA) treatment to other environmental factors in shaping AMF growth and fusion (anastomosis). Among our results, we confirm known effects of the FA treatment in increasing hyphal branching and longevity but interestingly show that multiple hyphae in a lane can negate FA effects. We also newly reveal that anastomosis occurs for 100% of interactions among hyphae from different lanes, but only for 50% of in-lane interactions, potentially linked to spore origin. Our findings validate the AMF-AnastomosisChip as a versatile platform for increasing discovery in trait-based microbial ecology, including for asymbiotic AMF interactions with environmental factors. This work sets the foundation for future studies with the AMF-AnastomosisChip on the effects of nutrient content, plant-derived molecules, or microbial community members on the AMF traits characterized in this study.

microbiology↗

Diurnal plant and Rhizophagus irregularis transcriptional patterns are linked to shifts in cassava tissue partitioning in the field

O_LICassava is a globally-important crop whose yields can radically increase with arbuscular mycorrhizal fungi (AMF) inoculation. However, extensive background noise in field environments makes it very challenging to understand how the cassava-AMF symbiosis confers benefits, which is especially important for future applications of AMF treatments. C_LIO_LIIn two field experiments, we combined transcriptomics and allometric analyses to investigate functional variation in cassava-AMF interactions using sterile, single isolates of Rhizophagus irregularis. We developed a novel Index of Symbiotic Transcriptional Activity (ISTA) and accounted for sampling times to reduce transcriptomic noise and improve links to biomass traits. C_LIO_LIISTA significantly correlated with cassava shoot biomass in an isolate-dependent manner, and allometric analyses revealed that R. irregularis isolates can either reinforce or uncouple cassava shoot-root relationships to maximize root yields. Differential expression and co-expression network analyses uncovered isolate-specific plant and fungal gene module responses. Including ISTA and sampling time as random effects enhanced detection of gene candidates, including down-regulated genes linked to higher yield. C_LIO_LIOur study uses novel and translatable transcriptomic tools to readily dissect variably field data, allowing new links to be found between AMF symbiotic functions and cassava yields. C_LI

molecular biology↗

Dissecting mycorrhizal fungal trait variation, its genetic basis and trade-offs

Despite the importance of arbuscular mycorrhizal fungi (AMF) in worldwide ecosystems, there is relatively little controlled phenotyping of their intraspecific fungal trait diversity, which limits our determination of how AMF traits link to their genetic differences. Here, we monitor 33 in vitro fungal traits in a population of 48 genetically diverse isolates of Rhizophagus irregularis, over 18 weeks, across plate compartments with and without direct plant root influence. This study reveals that intraspecific AMF variability is extensive, and that R. irregularis traits are mostly positively correlated, especially in root-free environments. The few trait trade-offs (negative correlations) found existed in proximity to the host root. Given the large screening population and the variance decomposition method of repeatability, we find that the genetic individuality of isolates significantly explained variation in approximately half of the AMF traits measured, and especially those related to hyphal growth, branching structures and reproduction near plant roots. Especially, differences in the frequency and fineness of branching right after growth from the plant host, which did not correlate to nearly any other traits, were explained by isolate identity (i.e., genotype). The new information we reveal on R. irregularis trait relationships, over time and in different environments, as well as trait links to genetic differences among isolates, provides the foundation for future understanding of AMF functional diversity. Our genetic findings also provide critical groundwork for genome wide association studies on the non-plant-related traits of R. irregularis.

ecology↗

The model arbuscular mycorrhizal fungus Rhizophagus irregularis harbours endosymbiotic bacteria with a highly reduce genome

Arbuscular mycorrhizal fungi (AMF; Glomeromycotina) are symbionts of most plant species that are known to possess unique intracytoplasmic endosymbiotic bacteria with an enigmatic role. Candidatus Moeniiplasma glomeromycotorum (CaMg) was shown to be widespread along the AMF phylogeny and present in most AMF species and isolates of those species. The model AMF species, Rhizophagus irregularis, that can be cultivated in vitro and for which a lot of genomic information now exists, would be the ideal model to study the true nature of the CaMg-AMF symbiosis. However, R. irregularis was never found to host endobacteria. Here we show by DNA sequencing that R. irregularis can, indeed, host CaMg (Ri-CaMg). However, this appears rare as only one R. irregularis isolate out of 58 hosted CaMg. In that isolate, the endosymbiotic bacterial population was genetically homogenous. By sequencing the complete genome of the bacteria, we found that its genome is among the smallest of all known CaMg and Mycoplasma-like genomes, with a highly reduced gene repertoire, suggesting a strong adaptation to the intracellular life. We discuss our findings in the light of previous literature on CaMg and on the same AMF isolates and suggest that these endosymbionts are more likely parasites than non-obligatory mutualists.

microbiology↗

Recombinant haplotypes reflect sexual reproduction in symbiotic arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi (AMF) are part of the most widespread fungal-plant symbiosis. They colonize at least 80% of plant species, promote plant growth and plant diversity. These fungi are multinucleated and contain either one or two haploid nuclear genotypes (monokaryon and dikaryon) identified by the alleles at a putative mating-type locus. This taxon has been considered as an ancient asexual scandal because of the lack of observable sexual structures. Despite identification of a putative mating-type locus and functional activation of genes related to mating when two isolates co-exist, it remains unknown if AMF take part in a mainly sexual life cycle. We used publicly available genome sequences to test if dikaryon genomes display signatures of sexual reproduction in the form of reciprocal recombination patterns, or if they display signatures of parasexual reproduction involving gene conversion. We used short-read and long-read sequence data to identify nucleus genotype-specific haplotypes within dikaryons and then compared them to orthologous gene sequences from related monokaryon isolates displaying the same putative MAT-types. We observed that these genotype-specific haplotypes display reciprocal recombination and not gene conversion. These results are consistent with a sexual origin of the dikaryon rather than a parasexual origin and provides an important step to understand the life cycle of these globally important symbiotic fungi.

evolutionary biology↗

Inoculation with highly-related mycorrhizal fungal siblings, and their interaction with plant genoptypes, strongly shapes tropical mycorrhizal fungal community structure

Arbuscular mycorrhizal fungi (AMF) have the potential to increase crop yields and all globally important crops form the mycorrhizal symbiosis. Only a few studies have investigated the impact of introduced AMF on local AMF communities and most studies have only investigated effects of one isolate. We studied the impact on AMF community structure of inoculating roots of the globally important crop cassava with highly genetically-related clonal siblings of two genetically different Rhizophagus irregularis isolates. We hypothesized that inoculation with R. irregularis siblings differentially influences the structure and the diversity of the pre-existing AMF community colonizing cassava. Alpha and beta taxonomic and phylogenetic AMF diversity were strongly and significantly altered differentially following inoculation with sibling AMF progeny. In most cases, the effects were also cassava-genotype specific. Although biomass production and AMF colonization were also both differentially affected by inoculation with sibling R. irregularis progeny these variables were not correlated with changes in the AMF community structure. The results highlight that investigations on the impact of an introduced AMF species, that use only one isolate, are unlikely to be representative of the overall effects of that AMF species and that the genetic identity of the host must be considered. The amount of inoculum added was very small and effects were observed 12 months following inoculation. That such a small amount of almost genetically identical fungal inoculum can strongly differentially influence AMF community structure 12 months following inoculation, indicates that AMF communities in tropical soils are not very resistant to perturbation.

ecology↗

Using variation in arbuscular mycorrhizal fungi to drive the productivity of the food security crop cassava

The unprecedented challenge to feed the rapidly growing human population can only be achieved with major changes in how we combine technology with agronomy1. Despite their potential few beneficial microbes have truly been demonstrated to significantly increase productivity of globally important crops in real farming conditions2,3. The way microbes are employed has largely ignored the successes of crop breeding where naturally occurring intraspecific variation of plants has been used to increase yields. Doing this with microbes requires establishing a link between variation in the microbes and quantitative traits of crop growth along with a clear demonstration that intraspecific microbial variation can potentially lead to large differences in crop productivity in real farming conditions. Arbuscular mycorrhizal fungi (AMF), form symbioses with globally important crops and show great potential to improve crop yields2. Here we demonstrate the first link between patterns of genome-wide intraspecific AMF variation and productivity of the globally important food crop cassava. Cassava, one of the most important food security crops, feeds approximately 800 million people daily4. In subsequent field trials, inoculation with genetically different isolates of the AMF Rhizophagus irregularis altered cassava root productivity by up to 1.46-fold in conventional cultivation in Colombia. In independent field trials in Colombia, Kenya and Tanzania, clonal sibling progeny of homokaryon and dikaryon parental AMF enormously altered cassava root productivity by up to 3 kg per plant and up to a 3.69-fold productivity difference. Siblings were clonal and, thus, qualitatively genetically identical. Heterokaryon siblings can vary quantitatively but monokaryon siblings are identical. Very large among-AMF sibling effects were observed at each location although which sibling AMF was most effective depended strongly on location and cassava variety. We demonstrate the enormous potential of genetic, and possibly epigenetic variation, in AMF to greatly alter productivity of a globally important crop that should not be ignored. A microbial improvement program to accelerate crop yield increases over that possible by plant breeding or GMO technology alone is feasible. However, such a paradigm shift can only be realised if researchers address how plant genetics and local environments affect mycorrhizal responsiveness of crops to predict which fungal variant will be effective in a given location.

microbiology↗

Investigating unexplained genetic variation and its expression in the arbuscular mycorrhizal fungus Rhizophagus irregularis

Arbuscular mycorrhizal fungi (AMF) are important symbionts of plants. Recently, studies of the AMF Rhizophagus irregularis recorded within-isolate genetic variation that does not completely match the proposed homokaryon or heterokaryon state (where heterokaryons comprise a population of two distinct nucleus genotypes). We re-analysed published data showing that bi-allelic sites (and their frequencies), detected in proposed homo- and heterokaryote R. irregularis isolates, were similar across independent studies using different techniques. This indicated that observed within-fungus genetic variation was not an artefact of sequencing and that such within-fungus genetic variation possibly exists. We looked to see if bi-allelic transcripts from three R. irregularis isolates matched those observed in the genome as this would give a strong indication of whether bi-allelic sites recorded in the genome were reliable variants. In putative homokaryon isolates, very few bi-allelic transcripts matched those in the genome. In a putative heterokaryon, a large number of bi-allelic transcripts matched those in the genome. Bi-allelic transcripts also occurred in the same frequency in the putative heterokaryon as predicted from allele frequency in the genome. Our results indicate that while within-fungus genome variation in putative homokaryon and heterokaryon AMF was highly similar in 2 independent studies, there was little support that this variation is transcribed in homokaryons. In contrast, within-fungus variation thought to be segregated among two nucleus genotypes in a heterokaryon isolate was indeed transcribed in a way that is proportional to that seen in the genome.

microbiology↗

Dual RNA-seq reveals large-scale non-conserved genotype x genotype specific genetic reprograming and molecular crosstalk in the mycorrhizal symbiosis

Arbuscular mycorrhizal fungi (AMF) impact plant growth and are a major driver of plant diversity and productivity. We quantified the contribution of intra-specific genetic variability in cassava (Manihot esculenta) and Rhizophagus irregularis to gene reprogramming in symbioses using dual RNA-sequencing. A large number of cassava genes exhibited altered transcriptional responses to the fungus but transcription of most of these plant genes (72%) responded in a different direction or magnitude depending on the plant genotype. Two AMF isolates displayed large differences in their transcription, but the direction and magnitude of the transcriptional responses for a large number of these genes was also strongly influenced by the genotype of the plant host. This indicates that unlike the highly conserved plant genes necessary for the symbiosis establishment, plant and fungal gene transcriptional responses are not conserved and are greatly influenced by plant and fungal genetic differences, even at the within-species level. The transcriptional variability detected allowed us to identify an extensive gene network showing the interplay in plant-fungal reprogramming in the symbiosis. Key genes illustrated that the two organisms jointly program their cytoskeleton organisation during growth of the fungus inside roots. Our study reveals that plant and fungal genetic variation plays a strong role in shaping the genetic reprograming in response to symbiosis, indicating considerable genotype x genotype interactions in the mycorrhizal symbiosis. Such variation needs to be considered in order to understand the molecular mechanisms between AMF and their plant hosts in natural communities.

plant biology↗

Within-species phylogenetic relatedness of a common mycorrhizal fungus affects evenness in plant communities through effects on dominant species

Arbuscular mycorrhizal fungi (AMF) have been shown to influence plant community structure and diversity. Studies based on single plant - single AMF isolate experiments show that within AMF species variation leads to large differential growth responses of different plant species. Because of these differential effects, genetic differences among isolates of an AMF species could potentially have strong effects on the structure of plant communities.\n\nWe tested the hypothesis that within species variation in the AMF R. irregularis significantly affects plant community structure and plant co-existence. We took advantage of a recent genetic characterization of several isolates using double-digest restriction-site associated DNA sequencing (ddRADseq). This allowed us to test not only for the impact of within AMF species variation on plant community structure but also for the role of the R. irregularis phylogeny on plant community metrics. Nine isolates of R. irregularis, belonging to three different genetic groups (Gp1, Gp3 and Gp4), were used as either single inoculum or as mixed diversity inoculum. Plants in a mesocosm representing common species that naturally co-exist in European grasslands were inoculated with the different AMF treatments.\n\nWe found that within-species differences in R. irregularis did not strongly influence the performance of individual plants or the structure of the overall plant community. However, the evenness of the plant community was affected by the phylogeny of the fungal isolates, where more closely-related AMF isolates were more likely to affect plant community evenness in a similar way compared to more genetically distant isolates.\n\nThis study underlines the effect of within AMF species variability on plant community structure. While differential effects of the AMF isolates were not strong, a single AMF species had enough functional variability to change the equilibrium of a plant community in a way that is associated with the evolutionary history of the fungus.

ecology↗