Genome-wide transcriptome and gene family analysis reveal candidate genes associated with potassium uptake of maize colonized by arbuscular mycorrhizal fungi.

BACKGROUND: Potassium (K) is an essential nutrient for plant growth and development. Maize (Zea mays) is a widely planted crops in the world and requires a huge amount of K fertilizer. Arbuscular mycorrhizal fungi (AMF) are closely related to the K uptake of maize. Genetic improvement of maize K utilization efficiency will require elucidating the molecular mechanisms of maize K uptake through the mycorrhizal pathway. Here, we employed transcriptome and gene family analysis to elucidate the mechanism influencing the K uptake and utilization efficiency of mycorrhizal maize. METHODS AND RESULTS: The transcriptomes of maize were studied with and without AMF inoculation and under different K conditions. AM symbiosis increased the K concentration and dry weight of maize plants. RNA sequencing revealed that genes associated with the activity of the apoplast and nutrient reservoir were significantly enriched in mycorrhizal roots under low-K conditions but not under high-K conditions. Weighted gene correlation network analysis revealed that three modules were strongly correlated with K content. Twenty-one hub genes enriched in pathways associated with glycerophospholipid metabolism, glycerolipid metabolism, starch and sucrose metabolism, and anthocyanin biosynthesis were further identified. In general, these hub genes were upregulated in AMF-colonized roots under low-K conditions. Additionally, the members of 14 gene families associated with K obtain were identified (ARF: 38, ILK: 4, RBOH: 12, RUPO: 20, MAPKK: 89, CBL: 14, CIPK: 44, CPK: 40, PIN: 10, MYB: 174, NPF: 79, KT: 19, HAK/HKT/KUP: 38, and CPA: 8) from maize. The transcript levels of these genes showed that 92 genes (ARF:6, CBL:5, CIPK:13, CPK:2, HAK/HKT/KUP:7, PIN:2, MYB:26, NPF:16, RBOH:1, MAPKK:12 and RUPO:2) were upregulated with AM symbiosis under low-K conditions. CONCLUSIONS: This study indicated that AMF increase the resistance of maize to low-K stress by regulating K uptake at the gene transcription level. Our findings provide a genome-level resource for the functional assignment of genes regulated by K treatment and AM symbiosis in K uptake-related gene families in maize. This may contribute to elucidate the molecular mechanisms of maize response to low K stress with AMF inoculation, and provided a theoretical basis for AMF application in the crop field.

Bacillus subtilis and Rhizophagus intraradices Improve Vegetative Growth, Yield, and Fruit Quality of Fragaria × ananassa var. San Andreas.

Strawberry cultivation requires strategies that maintain or improve its yield within a scheme in which reducing fertilizers and other chemical products can make its consumption safer and more environmentally friendly. This study aims to evaluate the effect of Bacillus subtilis and Rhizophagus intraradices on strawberry growth, yield, and fruit quality. B. subtilis and R. intraradices were inoculated and co-inoculated under three fertilization levels of 225-100-250, 112-50-125, and 0-0-0 kg∙ha-1 of N, P2O5 and K2O. Vegetative growth was evaluated in plant height (cm), leaf area (cm2), aerial fresh weight (g), aerial dry weight (g), and plant coverage (%) variables. Fruit quality parameters such as total acidity (g∙100 mL-1), soluble solids (Brix°), and firmness (kg) were also determined, as well as the number of fruits per m2 and yield (t∙ha-1). The results showed that the pre-treatment of root immersion in a nutrient solution with B. subtilis and the fractionation of 6 L B. subtilis inoculation per plant at a concentration of 107 CFU∙mL-1, in combination with 225-100-250 kg∙ha-1 of N, P2O5, and K2O, achieved the highest accumulation of dry matter (12.9 ± 1.9 g∙plant-1), the highest number of fruits (28.2 ± 4.5 fruits∙m-2), and the highest yield (7.2 ± 1.4 t∙ha-1). In addition, this treatment increased the soluble sugar content by 34.78% and fruit firmness by 26.54% compared to the control without inoculation. This study highlights the synergistic effect of mineral nutrition and microbial inoculation with B. subtilis in increasing strawberry yield and fruit quality.

Lipids Mediate Arbuscule Development and Senescence in Tomato Roots Colonized by Arbuscular Mycorrhizae Fungus under Drought Stress.

Arbuscular mycorrhizae (AM) symbiosis can enhance plant resistance to drought stress (DS). This study aimed to investigate the DS effects on lipids at different stages of symbiosis and to link lipid profiles to arbuscule dynamics in tomato roots colonized by AM fungi. DS increased mycorrhizal colonization and arbuscule abundance at an early stage but decreased them at a later stage, delayed arbuscule development, and accelerated arbuscule senescence at a later stage. DS decreased the contents of phospholipids (PLs) and saturated neutral lipids (NLs) at the early stage but increased the contents of saturated PLs and unsaturated NLs at the late stage. Specifically, DS inhibited AM-specific PL contents but increased AM-specific NL contents, which was supported by the expression of RAM2, STR/STR2. These data indicate the negative effect of DS on AM symbiosis and arbuscule dynamics with the effect size depending on the symbiosis stage, which highlights the importance of the symbiosis stage under abiotic stress.

Mycorrhizae in mine wasteland reclamation.

Mycorrhizae are found on about 70-80 % of the roots of all plant species; ectomycorrhizae (ECM) are mostly found on woody plants and gymnosperms, whereas arbuscular mycorrhizal fungi (AMF) are found on 80-90 % of all plant species. In abandoned mining sites, woody plants dominate, while non-woody species remain scarce. However, this pattern depends on the specific mine site and its ecological context. This review article explores the potential of using mycorrhizae-plant associations to enhance and facilitate the remediation of mine wastelands and metal-polluted sites. In this review, we employed reputable databases to collect articles and relevant information on mycorrhizae and their role in plant growth and soil fertility spanning from the 1990s up to 2024. Our review found that the abilities of plants selected for minewasteland reclamation can be harnessed effectively if their mycorrhizae utilization is known and considered. Our findings indicate that AMF facilitates plant cohabitation by influencing species richness, feedback effects, shared mycelial networks, and plant-AMF specificity. Several types of mycorrhizae have been isolated from mine wastelands, including Glomus mosseae, which reduces heavy metal accumulation in plants, and Rhizophagus irregularis, which enhances plant growth and survival in revegetated mine sites. Additionally, studies on ECM in surface mine spoil restoration stands highlight their role in enhancing fungal biodiversity and providing habitats for rare and specialized fungal species. Recent research shows that ECM and AMF fungi can interact synergistically to enhance plant growth, with ECM improving plant nitrogen absorption and AMF increasing nitrogen use efficiency. Our review also found that despite their critical role in improving plant growth and resilience, there remains limited knowledge about the specific mechanisms by which mycorrhizae communicate with each other and other microorganisms, such as bacteria, root-associated fungi, soil protozoa, actinomycetes, nematodes, and endophytes, within the soil matrix. This article highlights the connection between mycorrhizae and plants and other microorganisms in mine wastelands, their role in improving soil structure and nutrient cycling, and how mycorrhizae can help restore soil fertility and promote plant growth, thus improving the overall environmental quality of mine wasteland sites.

Effects of High Salinity and Water Stress on Wetland Grasses from the Spanish Mediterranean Coast.

The impacts of climate change are reaching unprecedented levels, heightening the risk of species loss and ecosystem service degradation. Wetlands, highly threatened ecosystems, serve vital ecological functions by capturing carbon, filtering water, and harbouring diverse wildlife. Coastal wetlands encounter many challenges, such as increased drought periods and escalating salinity levels, severely impacting plant biodiversity. Assessing how plants respond to various environmental stress factors is imperative for devising successful conservation strategies. In the present study, we examined three representative grass species found in various habitats within the Albufera Natural Park, close to the city of Valencia on the Spanish Mediterranean coast: Imperata cylindrica, Phragmites australis, and Saccharum ravennae. High salinity and water stress conditions were induced by subjecting the plants to irrigation with solutions containing 200, 400, 600, and 800 mM NaCl or withholding irrigation altogether to mimic coastal flooding and drought scenarios. The treatments were maintained until noticeable wilting of the plants occurred, at which point a range of stress biomarkers were determined, including photosynthetic pigments, ions, osmolytes, oxidative stress markers, and antioxidant metabolites, as well as antioxidant enzyme activities. Saccharum ravennae displayed the highest sensitivity to salt stress, whereas I. cylindrica appeared to be the most tolerant. The primary salinity tolerance mechanism observed in I. cylindrica and P. australis was a blockage of ion transport from the root zone to the aerial part, together with the salt-induced accumulation of proline and soluble sugars to high concentrations in the former. No significant effects of the water deficit treatment on the growth or biochemical parameters were observed for any of the analysed species. These findings offer valuable information for the effective management and conservation of coastal wetlands facing the challenges posed by climate change.

Biostimulant and Arbuscular Mycorrhizae Application on Four Major Biomass Crops as the Base of Phytomanagement Strategies in Metal-Contaminated Soils.

Using contaminated land to grow lignocellulosic crops can deliver biomass and, in the long term, improve soil quality. Biostimulants and microorganisms are nowadays an innovative approach to define appropriate phytomanagement strategies to promote plant growth and metal uptake. This study evaluated biostimulants and mycorrhizae application on biomass production and phytoextraction potential of four lignocellulosic crops grown under two metal-contaminated soils. Two greenhouse pot trials were setup to evaluate two annual species (sorghum, hemp) in Italy and two perennial ones (miscanthus, switchgrass) in China, under mycorrhizae (M), root (B2) and foliar (B1) biostimulants treatments, based on humic substances and protein hydrolysates, respectively, applied both alone and in combination (MB1, MB2). MB2 increased the shoot dry weight (DW) yield in hemp (1.9 times more), sorghum (3.6 times more) and miscanthus (tripled) with additional positive effects on sorghum and miscanthus Zn and Cd accumulation, respectively, but no effects on hemp metal accumulation. No treatment promoted switchgrass shoot DW, but M enhanced Cd and Cr shoot concentrations (+84%, 1.6 times more, respectively) and the phytoextraction efficiency. Root biostimulants and mycorrhizae were demonstrated to be more efficient inputs than foliar biostimulants to enhance plant development and productivity in order to design effective phytomanagement strategies in metal-contaminated soil.

Unraveling plant-microbe symbioses using single-cell and spatial transcriptomics.

Plant-microbe symbioses require intense interaction and genetic coordination to successfully establish in specific cell types of the host and symbiont. Traditional RNA-seq methodologies lack the cellular resolution to fully capture these complexities, but single-cell and spatial transcriptomics (ST) are now allowing scientists to probe symbiotic interactions at an unprecedented level of detail. Here, we discuss the advantages that novel spatial and single-cell transcriptomic technologies provide in studying plant-microbe endosymbioses and highlight key recent studies. Finally, we consider the remaining limitations of applying these approaches to symbiosis research, which are mainly related to the simultaneous capture of both plant and microbial transcripts within the same cells.

A transition from arbuscular to ectomycorrhizal forests halts soil carbon sequestration during subtropical forest rewilding.

Ecological succession and restoration rapidly promote multiple dimensions of ecosystem functions and mitigate global climate change. However, the factors governing the limited capacity to sequester soil organic carbon (SOC) in old forests are poorly understood. Ecological theory predicts that plants and microorganisms jointly evolve into a more mutualistic relationship to accelerate detritus decomposition and nutrient regeneration in old than young forests, likely explaining the changes in C sinks across forest succession or rewilding. To test this hypothesis, we conducted a field experiment of root-mycorrhizal exclusion in successional subtropical forests to investigate plant-decomposer interactions and their effects on SOC sequestration. Our results showed that SOC accrual rate at the 0-10 cm soil layer was 1.26 mg g-1 yr-1 in early-successional arbuscular mycorrhizal (AM) forests, which was higher than that in the late-successional ectomycorrhizal (EcM) forests with non-significant change. A transition from early-successional AM to late-successional EcM forests increase fungal diversity, especially EcM fungi. In the late-successional forests, the presence of ectomycorrhizal hyphae promotes SOC decomposition and nutrient cycle by increasing soil nitrogen and phosphorus degrading enzyme activity as well as saprotrophic microbial richness. Across early- to late-successional forests, mycorrhizal priming effects on SOC decomposition explain a slow-down in the capacity of older forests to sequester soil C. Our findings suggest that a transition from AM to EcM forests supporting greater C decomposition can halt the capacity of forests to provide nature-based global climate change solutions.

The trade-off regulation of arbuscular mycorrhiza on alfalfa growth dose-dependent on gradient Mo exposure.

Molybdenum (Mo) is an essential nutrient for leguminous plants, but the effects of Mo exposure on plant growth, especially in relation to soil microorganisms, are not fully understood. This study employed alfalfa (Medicago sativa L.) to evaluate the physiochemical responses to gradient soil Mo variations and explore the potential regulatory role of rhizosphere microorganism - arbuscular mycorrhizal fungi (AMF) in modulating Mo's impact on plant physiology, with a focus on metabolic pathways. The results showed that Mo exerted hormetic effect (facilitation at low doses; inhibition at high doses) on alfalfa growth, promoting biomass (below 90.94 mg/kg, with a 63.98 % maximum increase), root length (below 657.11 mg/kg, with a 39.29 % maximum increase), and plant height (below 304.03 mg/kg, with an 18.4 % maximum increase). Excess Mo (1000 mg/kg) resulted in a reduction in photosynthesis and biomass growth due to increased oxidative stress (p < 0.05). Within the stimulatory zones, AMF enhanced Mo accumulation in alfalfa, augmenting its phytological effects. Exceed the stimulatory zones, AMF enhanced alfalfa Fe uptake and reduced the generation of reactive oxygen species (ROS) induced by excess Mo by shifting the redox homeostasis-controlled enzyme from peroxidase (POD) to superoxide dismutase (SOD), thereby improving alfalfa's tolerance to Mo. Metabolomic analysis further revealed that AMF promoted the biosynthesis of indole acetic acid (IAA) and various amino acids in Mo-stressed alfalfa (p < 0.05), which accelerated alfalfa growth and mitigated Mo-induced phytotoxicity. These insights provide a foundation for developing sustainable management strategies for Mo-exposed soils using AMF inoculants, such as minimizing Mo fertilizer application in Mo-deficient soils and facilitating the reclamation of Mo-contaminated soils.

The biological controls of soil carbon accumulation following wildfire and harvest in boreal forests: A review.

Boreal forests are frequently subjected to disturbances, including wildfire and clear-cutting. While these disturbances can cause soil carbon (C) losses, the long-term accumulation dynamics of soil C stocks during subsequent stand development is controlled by biological processes related to the balance of net primary production (NPP) and outputs via heterotrophic respiration and leaching, many of which remain poorly understood. We review the biological processes suggested to influence soil C accumulation in boreal forests. Our review indicates that median C accumulation rates following wildfire and clear-cutting are similar (0.15 and 0.20 Mg ha-1 year-1, respectively), however, variation between studies is extremely high. Further, while many individual studies show linear increases in soil C stocks through time after disturbance, there are indications that C stock recovery is fastest early to mid-succession (e.g. 15-80 years) and then slows as forests mature (e.g. >100 years). We indicate that the rapid build-up of soil C in younger stands appears not only driven by higher plant production, but also by a high rate of mycorrhizal hyphal production, and mycorrhizal suppression of saprotrophs. As stands mature, the balance between reductions in plant and mycorrhizal production, increasing plant litter recalcitrance, and ectomycorrhizal decomposers and saprotrophs have been highlighted as key controls on soil C accumulation rates. While some of these controls appear well understood (e.g. temporal patterns in NPP, changes in aboveground litter quality), many others remain research frontiers. Notably, very little data exists describing and comparing successional patterns of root production, mycorrhizal functional traits, mycorrhizal-saprotroph interactions, or C outputs via heterotrophic respiration and dissolved organic C following different disturbances. We argue that these less frequently described controls require attention, as they will be key not only for understanding ecosystem C balances, but also for representing these dynamics more accurately in soil organic C and Earth system models.

Fern mycorrhizae do not respond to fertilization in a tropical montane forest.

Ferns are known to have a lower incidence of mycorrhization than angiosperms. It has been suggested that this results from carbon being more limiting to fern growth than nutrient availability, but this assertion has not been tested yet. In the present study, we took advantage of a fertilization experiment with nitrogen and phosphorus on cloud forest plots of the Ecuadorean Andes for 15 years. A previous analysis revealed changes in the abundances of fern species in the fertilized plots compared to the control plots and hypothesized that this might be related to the responses of the mycorrhizal relationships to nutrient availability. We revisited the plots to assess the root-associated fungal communities of two epiphytic and two terrestrial fern species that showed shifts in abundance. We sampled and analyzed the roots of 125 individuals following a metabarcoding approach. We recovered 1382 fungal ASVs, with a dominance of members of Tremellales (Basidiomycota) and Heliotales (Ascomycota). The fungal diversity was highly partitioned with little overlap between individuals. We found marked differences between terrestrial and epiphytic species, with the latter fundamentally missing arbuscular mycorrhizal fungi (AMF). We found no effect of fertilization on the diversity or relative abundance of the fungal assemblages. Still, we observed a direct impact of phosphorus fertilization on its concentration in the fern leaves. We conclude that fern-fungi relationships in the study site are not restricted by nutrient availability and suggest the existence of little specificity on the fungal partners relative to the host fern species.

Distinct interactions of ericoid mycorrhizae and plant growth-promoting bacteria: impacts on blueberry growth and heat resilience.

Blueberries confront substantial challenges from climate change, such as rising temperatures and extreme heat, necessitating urgent solutions to ensure productivity. We hypothesized that ericoid mycorrhizal fungi (ErM) and plant growth-promoting bacteria (PGPB) would establish symbiotic relationships and increase heat stress tolerance in blueberries. A growth chamber study was designed with low (25/20°C) and high temperature (35/30°C) conditions with micropropagated blueberry plantlets inoculated with ErM, PGPB, and both. Gas exchange and chlorophyll fluorescence properties of the leaves were monitored throughout the growth. At harvest, biochemical assays and biomass analysis were performed to evaluate potential oxidative stress induced by elevated temperatures. ErM application boosted root biomass under 25/20°C conditions but did not impact photosynthetic efficiency. In contrast, PGPB demonstrated a dual role: enhancing photosynthetic capacity and reducing stomatal conductance notably under 35/30°C conditions. Moreover, PGPB showcased conflicting effects, reducing oxidative damage under 25/20°C conditions while intensifying it during 47°C heat shock. A significant highlight lies in the opposing effects of ErM and PGPB on root growth and stomatal conductance, signifying their reciprocal influence on blueberry plant behavior, which may lead to increased water uptake or reduced water use. Understanding these complex interactions holds promise for refining sustainable strategies to overcome climate challenges.

Draft genome sequences of Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 isolated from superabsorbent polymer granules inoculated with an arbuscular mycorrhizal fungus.

Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 were isolated from superabsorbent polymer granules cultured with an arbuscular mycorrhizal fungus. Whole-genome sequencing of these three strains revealed genome sizes of 4.57 Mb, 7.13 Mb, and 5.49 Mb with G + C contents of 36.9%, 62.5%, and 58.2%, respectively.

Inoculation of black turtle beans (Phaseolus vulgaris) with mycorrhizal fungi increases the nutritional quality of seeds.

The use of arbuscular mycorrhizal fungi (AMF) as biofertilizers has proven successful in boosting the yield and nutritional quality of a variety of crops. AMF associate with plant roots and exchange soil nutrients for photosynthetically derived C in the form of sugars and lipids. Past research has shown that not all AMF species are equal in their benefit to nutrient uptake and crop health, and that the most beneficial AMF species appear to vary by host species. Although an important human food staple, especially in developing regions where nutrient deficiency is a prevalent threat to public health, little work has been done to test the effectiveness of AMF in enhancing the nutritional quality of common bean (Phaseolus vulgaris L.). Therefore, our objective was to determine the most beneficial AMF species for inoculation of this important crop. We inoculated black beans (Phaseolus vulgaris black turtle beans) with eight individual AMF species and one mixed species inoculum in an outdoor pot trial over 3 months and assessed the extent to which they altered yield, mineral nutrient and anthocyanin concentration of seeds and leaf tissues. Despite seeing no yield effects from inoculation, we found that across treatments percent root length colonized by AMF was positively correlated with plant tissue P, Cu, and Zn concentration. Underlying these broad benefits, seeds from plants inoculated with three AMF species, Claroideoglomus claroideum (+15%), Funneliformis mosseae (+13%), and Gigaspora rosea (+11%) had higher P concentration than non-mycorrhizal plants. C. claroideum also increased seed potassium (K) and copper (Cu), as well as leaf aluminum (Al) concentration making it a promising candidate to further test the benefit of individual AMF species on black bean growth in field trials.

IPD3, a master regulator of arbuscular mycorrhizal symbiosis, affects genes for immunity and metabolism of non-host Arabidopsis when restored long after its evolutionary loss.

Arbuscular mycorrhizal symbiosis (AM) is a beneficial trait originating with the first land plants, which has subsequently been lost by species scattered throughout the radiation of plant diversity to the present day, including the model Arabidopsis thaliana. To explore if elements of this apparently beneficial trait are still present and could be reactivated we generated Arabidopsis plants expressing a constitutively active form of Interacting Protein of DMI3, a key transcription factor that enables AM within the Common Symbiosis Pathway, which was lost from Arabidopsis along with the AM host trait. We characterize the transcriptomic effect of expressing IPD3 in Arabidopsis with and without exposure to the AM fungus (AMF) Rhizophagus irregularis, and compare these results to the AM model Lotus japonicus and its ipd3 knockout mutant cyclops-4. Despite its long history as a non-AM species, restoring IPD3 in the form of its constitutively active DNA-binding domain to Arabidopsis altered expression of specific gene networks. Surprisingly, the effect of expressing IPD3 in Arabidopsis and knocking it out in Lotus was strongest in plants not exposed to AMF, which is revealed to be due to changes in IPD3 genotype causing a transcriptional state, which partially mimics AMF exposure in non-inoculated plants. Our results indicate that molecular connections to symbiosis machinery remain in place in this nonAM species, with implications for both basic science and the prospect of engineering this trait for agriculture.

The Role of CLE Peptides in the Suppression of Mycorrhizal Colonization of Tomato.

Symbioses with beneficial microbes are widespread in plants, but these relationships must balance the energy invested by the plants with the nutrients acquired. Symbiosis with arbuscular mycorrhizal (AM) fungi occurs throughout land plants, but our understanding of the genes and signals that regulate colonization levels is limited, especially in non-legumes. Here, we demonstrate that in tomato, two CLV3/EMBRYO-SURROUNDING REGION (CLE) peptides, SlCLE10 and SlCLE11, act to suppress AM colonization of roots. Mutant studies and overexpression via hairy transformation indicate that SlCLE11 acts locally in the root to limit AM colonization. Indeed, SlCLE11 expression is strongly induced in AM-colonized roots, but SlCLE11 is not required for phosphate suppression of AM colonization. SlCLE11 requires the FIN gene that encodes an enzyme required for CLE peptide arabinosylation to suppress mycorrhizal colonization. However, SlCLE11 suppression of AM does not require two CLE receptors with roles in regulating AM colonization, SlFAB (CLAVATA1 ortholog) or SlCLV2. Indeed, multiple parallel pathways appear to suppress mycorrhizal colonization in tomato, as double mutant studies indicate that SlCLV2 and FIN have an additive influence on mycorrhizal colonization. SlCLE10 appears to play a more minor or redundant role, as cle10 mutants did not influence intraradical AM colonization. However, the fact that cle10 mutants had an elevated number of hyphopodia and that ectopic overexpression of SlCLE10 did suppress mycorrhizal colonization suggests that SlCLE10 may also play a role in suppressing AM colonization. Our findings show that CLE peptides regulate AM colonization in tomato and at least SlCLE11 likely requires arabinosylation for activity.

Unveiling the complexity of strigolactones: exploring structural diversity, biosynthesis pathways, and signaling mechanisms.

Strigolactone is the collective name for compounds containing a butenolide as a part of their structure, first discovered as compounds that induce seed germination of root parasitic plants. They were later found to be rhizosphere signaling molecules that induce hyphal branching of arbuscular mycorrhizal fungi, and, finally, they emerged as a class of plant hormones. Strigolactones are found in root exudates, where they display a great variability in their chemical structure. Their structure varies among plant species, and multiple strigolactones can exist in one species. Over 30 strigolactones have been identified, yet the chemical structure of the strigolactone that functions as an endogenous hormone and is found in the above-ground parts of plants remains unknown. We discuss our current knowledge of the synthetic pathways of diverse strigolactones and their regulation, as well as recent progress in identifying strigolactones as plant hormones. Strigolactone is perceived by the DWARF14 (D14), receptor, an α/ß hydrolase which originated by gene duplication of KARRIKIN INSENSITIVE 2 (KAI2). D14 and KAI2 signaling pathways are partially overlapping paralogous pathways. Progress in understanding the signaling mechanisms mediated by two α/ß hydrolase receptors as well as remaining challenges in the field of strigolactone research are reviewed.

Culture age of Tulasnella affects symbiotic germination of the critically endangered Wyong sun orchid Thelymitra adorata (Orchidaceae).

Orchids (Orchidaceae) are dependent on mycorrhizal fungi for germination and to a varying extent as adult plants. We isolated fungi from wild plants of the critically endangered terrestrial orchid Thelymitra adorata and identified them using a multi-region barcoding approach as two undescribed Tulasnella species, one in each of phylogenetic group II and III (OTU1) of the Tulasnellaceae. Using symbiotic propagation methods, we investigated the role of Tulasnella identity (species and isolate) and age post isolation, on the fungus's ability and efficacy in germinating T. adorata. The group II isolate did not support germination. Seed germination experiments were conducted using either (i) three different isolates of OTU1, (ii) 4- and 12-week-old fungal cultures (post isolation) of a single isolate of OTU1, and (iii) T. subasymmetrica which is widespread and known to associate with other species of Thelymitra. Culture age and fungal species significantly (P < 0.05) affected the time to germination and percentage of seed germination, with greater and faster germination with 4-week-old cultures. Tulasnella subasymmetrica was able to germinate T. adorata to leaf stage, although at slightly lower germination percentages than OTU1. The ability of T. adorata to germinate with T. subasymmetrica may allow for translocation sites to be considered outside of its native range. Our findings on the age of Tulasnella culture affecting germination may have applications for improving the symbiotic germination success of other orchids. Furthermore, storage of Tulasnella may need to take account of the culture age post-isolation, with storage at - 80 °C as soon as possible recommended, post isolation.