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Orchids and arbuscular mycorrhiza (AM) plants evolved independently and have different structures and fungal partners, but they both facilitate nutrient uptake. Orchid mycorrhiza (OM) supports orchid seed germination, but unlike AM, its role in disease resistance of mature plants is largely unknown. Here, we examined whether OM induces systemic disease resistance against a necrotrophic pathogen in a similar fashion to AM. We investigated the priming effect of mycorrhizal fungi inoculation on resistance of a terrestrial orchid, Bletilla striata, to soft rot caused by Dickeya fangzhongdai. We found that root colonization by a compatible OM fungus primed B. striata seedlings and induced systemic resistance against the infection. Transcriptome analysis showed that priming was mediated by the downregulation of jasmonate and ethylene pathways and that these pathways are upregulated once infection occurs. Comparison with the reported transcriptome of AM fungus-colonized rice leaves revealed similar mechanisms in B. striata and in rice. These findings highlight a novel aspect of commonality between OM and AM plants in terms of induced systemic resistance. [ABSTRACT FROM AUTHOR]

The tiny terrestrial orchids (plant height less than 8 cm) in the genus Corybas dependent on mycorrhizal fungal (OMF) partners for seed germination and seedling development. The OMF community of the Corybas remains poorly understood, although the relevant knowledge is very important for in situ and ex situ conservation of these orchids. In this study, we characterized OMF richness and compositions of three helmet-orchid species, i.e., Corybas geminigibbus, C. himalaicus, and C. shanlinshiensis, from their natural habitats by using Illumina sequencing of the internal transcribed spacer 2 region. We found that fungal colonization was restricted in the rhizomes of the helmet-orchids instead of their tuberoids, and serendipitoid fungi were predominant, while tulasnelloid were absent in the three investigated Corybas species regardless of their altitude. The three Corybas species shared 27 serendipitoid operational taxonomic units that are different to those of their related orchids, the genera of Cyrtostylis and Stigmatodactylus. Corybas shanlinshiensis alone had a range of ectomycorrhizal fungi (mainly russuloid and thelephoroid) broader than C. himalaicus and C. geminigibbus. Our study provides new information about terrestrial orchid–fungi associations and may further contribute to orchid conservation. [ABSTRACT FROM AUTHOR]

Dendrobium officinale Kimura & Migo in the genus Dendrobium of Orchidaceae is an important medicinal plant that produces various bibenzyl and phenanthrene derivatives. In some orchids, these derivatives have been reported to increase with fungal infection. Bibenzyl biosynthesis is regulated by bibenzyl synthase (BBS). Although six genes of the BBS family have been registered from D. officinale, their gene regulation mechanisms are unclear. The infection of Dendrobium with mycorrhizal fungi also reportedly increases the expression of genes involved in biosynthesis; however, the effect of mycorrhizal fungi on bibenzyl production is unknown. The present study examined the effects of three mycorrhizal fungi isolated from D. officinale on BBS gene expression and bibenzyl production over time. One of the Tulasnellaceae operational taxonomic units induced BBS gene expression and increased two representative bibenzyls, gigantol and dendrophenol, at specific time points. Furthermore, 19 BBS sequences were cloned from 12 Dendrobium species, and a phylogenetic analysis was performed. The results indicated that repeated BBS gene duplication occurred during the evolution of the genus, and further duplication occurred after speciation. These results suggest that it is possible to optimize metabolite production by selecting suitable symbiotic fungi. [ABSTRACT FROM AUTHOR]

Orchids are crucial for the horticulture industry. Mycorrhizal fungi benefit crops by improving nutrition, plant growth, and disease resistance. However, the mycorrhizal association of horticultural hybrid orchids is poorly understood. To address this, we investigated mycorrhizal colonization in the entire root system and assessed the mycorrhizal community using a Dendrobium cultivar, D. Stardust 'Firebird', obtained from three nurseries. Additionally, we isolated and tested mycorrhizal fungi in symbiotic culture to assess their role in the seed germination and growth of Dendrobium species. All plants were colonized by mycorrhizal fungi, with a higher colonization rate in mature than in juvenile plants. Molecular identification of mycorrhizal fungi by Sanger and high-throughput sequencing revealed that the cultivar was associated with a phylogenetically diverse group of fungi, including mycorrhizal fungi from Tulasnellaceae, and several wood-decaying fungi. The Tulasnellaceae isolates significantly enhanced the seed germination of three Dendrobium species and increased the survival rate and growth of asymbiotic seedlings of D. moniliforme. This study is the first comprehensive examination of mycorrhizal associations in horticultural orchid hybrids, providing valuable insights for commercial production. [ABSTRACT FROM AUTHOR]

Multi-generational asexual reproduction of Gastrodia elata Bl. will cause seedling species degeneration. Sexual reproduction of Gastrodia elata Bl. seed is an effective method to solve the problem of degeneration. The development of Gastrodia elata Bl. seeds cannot be separated from the germination fungus. However, there are few strains of germination fungus in production, and there is also the problem of species degradation in application for many years. It is very important for the sexual reproduction of Gastrodia elata Bl. to isolate more new strains of excellent germination fungus from the origin. This study used the Gastrodia elata Bl. f. glauca S. chow seeds germination vegetative propagation corms capture method to isolate its symbiotic germination fungus, and comprehensively identified the species of germination fungus by colony morphology, ITS, sporocarps regeneration and germination function, and compared the growth characteristics and germination ability with other germination fungus (Mycena purpureofusca, Mycena dendrobii and Mycena osmundicola). The germination fungus was isolated from the vegetative propagation corms of Gastrodia elata Bl. f. glauca S. chow seeds and named GYGL-1. After comprehensive identification, GYGL-1 was Mycetinis scorodonius. Compared with other germination fungus, GYGL-1 has fast germination speed, vigorous growth, and high germination ability for Gastrodia elata Bl. f. glauca S. chow seeds. Innovated the isolation method of Gastrodia elata Bl. seeds germination fungus, obtained the regenerated sporocarps of the germination fungus, and discovered that Mycetinis scorodonius has a new function of germinating Gastrodia elata Bl. f. glauca S. chow seeds, enriching the resource library of Gastrodia elata Bl. germination fungus. [ABSTRACT FROM AUTHOR]

Basidiomycota is one of the major phyla in the fungal tree of life. The outline of Basidiomycota provides essential taxonomic information for researchers and workers in mycology. In this study, we present a time-framed phylogenomic tree with 487 species of Basidiomycota from 127 families, 47 orders, 14 classes and four subphyla; we update the outline of Basidiomycota based on the phylogenomic relationships and the taxonomic studies since 2019; and we provide notes for each order and discuss the history, defining characteristics, evolution, justification of orders, problems, significance, and plates. Our phylogenomic analysis suggests that the subphyla diverged in a time range of 443–490 Myr (million years), classes in a time range of 312–412 Myr, and orders in a time range of 102–361 Myr. Families diverged in a time range of 50–289 Myr, 76–224 Myr, and 62–156 Myr in Agaricomycotina, Pucciniomycotina, and Ustilaginomycotina, respectively. Based on the phylogenomic relationships and divergence times, we propose a new suborder Mycenineae in Agaricales to accommodate Mycenaceae. In the current outline of Basidiomycota, there are four subphyla, 20 classes, 77 orders, 297 families, and 2134 genera accepted. When building a robust taxonomy of Basidiomycota in the genomic era, the generation of molecular phylogenetic data has become relatively easier. Finding phenotypical characters, especially those that can be applied for identification and classification, however, has become increasingly challenging. [ABSTRACT FROM AUTHOR]

In 2017, two candidate species of Mycena were reported from Japan, with the Japanese names “Togari-sakura-take” and “Mitsuhida-sakura-take”. However, to date, no taxonomic study or formal description has been undertaken for these two species. In the present study, we conducted comprehensive morphological and molecular phylogenetic examinations of “Togari-sakura-take” and “Mitsuhida-sakura-take”, and compared them to known species within the genus Mycena. We performed phylogenetic analyses on a concatenated dataset, including the internal transcribed spacer region of ribosomal RNA, RNA polymerase II largest subunit, and translation elongation factor-1 alpha genes. “Togari-sakura-take” formed a clade with Mycena subulata, which was recently described from China, whereas “Mitsuhida-sakura-take” formed a distinct independent clade. We identified the former as M. subulata based on molecular phylogenetic analyses and morphological observations. However, the Japanese specimens displayed dextrinoid cheilocystidia and caulocystidia as well as the inamyloidity of basidiospores, which differed from the original description of M. subulata based on the materials from China. “Mitsuhida-sakura-take” was characterized by its remarkably dense lamellae and could be distinguished from known Mycena species by the combination of absent pleurocystidia and presence of bowling pin-shaped cheilocystidia. Here, we describe “Mitsuhida-sakura-take” as a new species, named Mycena densilamellata, in the section Calodontes. [ABSTRACT FROM AUTHOR]

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Epiphytic orchids comprise 68% of vascular epiphytes globally; nevertheless, many are endangered. One such epiphytic orchid is Phalaenopsis japonica, which is widely used in the floricultural industry. This study aimed to identify the mycorrhizal fungi of adult P. japonica and their roles in seed germination and seedling development. Root samples were collected from 32 adults across 4 sites in southern Japan, and mycorrhizal fungi were identified using Sanger and high-throughput sequencing (HTS). The results show phylogenetically diverse mycobionts, mainly Ceratobasidiaceae (CE) and Tulasnellaceae (TU), with dominant OTUs designated CE6 and CE22. Sanger sequencing found 9 OTUs, 4 CE, and 5 TU; HTS detected 22 OTUs, 4 CE, 16 TU, and 2 Serendipitaceae. Seeds inoculated with CE6 improved germination and protocorm development compared with other strains. In addition, asymbiotic seedlings inoculated with CE6, CE22, and TU18 displayed varying effects in growth, with CE6 being the most notable. While TU18 did not promote seed germination, it effectively promoted leaf development in seedlings. Overall, Ceratobasidiaceae was predominantly associated with seed germination, seedling growth, and the adult stages, with CE6 and CE22 becoming the primary partners throughout the life history of P. japonica. Our findings illuminate mycorrhizal symbiosis in epiphytic habitats, offering conservation and commercial production insights. [ABSTRACT FROM AUTHOR]

The orchid family (Orchidaceae) is one of the most diverse plant families in the world, but at the same time also contains one of the largest number of rare and endangered species. While conservation actions such as in situ and ex situ conservation and prohibition of international trade of wild orchids have achieved positive results to slow down the current decline of orchids, species with high medicinal or ornamental value may require more advanced measures. Recent pilot studies using novel cultivation techniques have successfully reintroduced endangered epiphytic orchids and facilitated the commercial cultivation of medicinal orchids. Because fungal partners play a key role in in situ symbiotic seed germination and industrial cultivation, we advocate for the development of fungus banks in laboratories engaging in orchid research, where fungi obtained from germinating seeds or seedlings can be studied and used to improve orchid germination under both in vitro and in situ conditions. Furthermore, these fungi could be shared nationally and internationally, enhancing orchid conservation efforts across the globe. Similar to seed banks, the development of fungus banks will reduce the possibility of fungi going extinct and ensure their availability for reintroduction programs and commercial cultivation. With the availability of both a fungus and seed bank, the conservation of threatened orchid species can be significantly enhanced by improving restoration programs and commercial cultivation of medicinal orchids and releasing the pressure on wild populations from harvesting. [ABSTRACT FROM AUTHOR]

The degree of specificity between fully myco-heterotrophic (MH) orchids and mycorrhizal fungi is regarded as high, but some species undergo a mycobiont shift as they transition from juvenile to adult plant. We investigated morphological and physiological aspects of the interaction between the fully MH Cyrtosia septentrionalis and its four known mycobionts to elucidate developmental consequences of variable, life-stage-dependent specificity. Of five randomly sampled germinated seeds co-cultured with each mycobiont, Physisporinus sp. 'TK-10' colonized all, Armillaria gallica and Desarmillaria tabescens colonized one, and A. mellea subsp. nipponica colonized none. Whereas 16.2% of aseptically germinated seeds exhibited onward growth when co-cultured with TK-10, just 1.5-2.6% did so with the other species. Even so, A. gallica colonized and formed internal rhizomorphs within rhizomes established with TK-10, suggesting that this mycobiont can replace and potentially can oust the latter. We infer that the orchid can associate with Armillaria and Desarmillaria mycobionts throughout its life, but that TK-10 enhances early growth. However, because TK-10 has a higher wood-rotting capacity than A. gallica, rapid resource exhaustion may cause nutrient supply shortages. We hypothesize that secondary colonization by Armillaria or Desarmillaria species triggers TK-10 displacement and that this mycobiont shift sustains orchid growth for longer. [ABSTRACT FROM AUTHOR]

Objective: To Identify the endophytic fungal community of the G. skinneri root in urban populations, showing differences in composition between velamen and cortex to better understand the fungus-root interaction in these orchids. Design/Methodology/Approach: We collected tissues from velamen and root from five specimens of G. skinneri growing on urban trees in Tapachula, Chiapas. We extracted DNA, PCR amplified the ITS marker, sequenced on the Illumina platform followed by diversity analyses and taxonomic assignment. Results: We detected 845 OTUs that were assigned to the Fungi kingdom. Velamen and cortex share 403 OTUs, 402 were found exclusively in velamen and 40 in cortex. Besides orchid mycorrhiza forming fungi, we detected other species (e.g Alternaria sp., Beauveria sp., Fusarium sp., Glomus sp. and Tricoderma sp.) that could be involved in root physiology during development and defense against pathogens and predators. Study Limitations/Implications: Metagenomic studies provide substantial amounts of data that go beyond conventional studies. However, the information generated is still limited regarding the role of each endophyte. Findings/Conclusions: Despite these limitations, our work fills a knowledge gap because we detected endophytes that were previously unknown for G. skinneri, leading to new research questions about rootendophyte relationships. [ABSTRACT FROM AUTHOR]

Many orchid species are endangered due to anthropogenic pressures such as habitat destruction and overharvesting, meanwhile, all orchids rely on orchid mycorrhizal fungi (OMF) for seed germination and seedling growth. Therefore, a better understanding of this intimate association is crucial for orchid conservation. Isolation and identification of OMF remain challenging as many fungi are unculturable. In our study, we tested the efficiency of both culture-dependent and culture-independent methods to describe OMF diversity in multiple temperate orchids and assessed any phylogenetic patterns in cultivability. The culture-dependent method involved the cultivation and identification of single pelotons (intracellular hyphal coils), while the culture-independent method used next-generation sequencing (NGS) to identify root-associated fungal communities. We found that most orchid species were associated with multiple fungi, and the orchid host had a greater impact than locality on the variability in fungal communities. The culture-independent method revealed greater fungal diversity than the culture-dependent one, but despite the lower detection, the isolated fungal strains were the most abundant OMF in adult roots. Additionally, the abundance of NGS reads of cultured OTUs was correlated with the extent of mycorrhizal root colonization in orchid plants. Finally, this limited-scale study tentatively suggests that the cultivability character of OMF may be randomly distributed along the phylogenetic trees of the rhizoctonian families. [ABSTRACT FROM AUTHOR]

Full myco‐heterotrophic orchid Gastrodia elata Bl. is widely distributed in Northeast Asia, and previous research has not fully investigated the symbiotic fungal community of its early immature tubers. This study utilized Illumina sequencing to compare symbiotic fungal communities in natural G. elata immature tubers and their habitats. LEfSe (Linear Discriminant Analysis Effect Size) was used to screen for Biomarkers that could explain variations among different fungal communities, and correlation analyses were performed among Biomarkers and other common orchid mycorrhizal fungi. Our results illustrate that the symbiotic fungal communities of immature G. elata tubers cannot be simply interpreted as subsets of the environmental fungal communities because some key members cannot be traced back to the environment. The early growth of G. elata was related to a small group of fungi, such as Sebacina, Thelephora, and Inocybe, which were also common mycorrhizal fungi from other orchids. In addition, Mycena, Auricularia, and Cryptococcus were unique fungal partners of G. elata, and many new species have yet to be discovered. Possible symbiotic Mycena should be M. plumipes and its sibling species in this case. Our results provide insight into the symbiotic partner switch and trophic pattern change during the development and maturation of G. elata. [ABSTRACT FROM AUTHOR]

Plant–fungal interactions are ubiquitous across ecosystems and contribute significantly to plant ecology and evolution. All orchids form obligate symbiotic relationships with specific fungi for germination and early growth, and the distribution of terrestrial orchid species has been linked to occurrence and abundance of specific orchid mycorrhizal fungi (OMF) in the soil. The availability of OMF can therefore be a habitat requirement that is relevant to consider when establishing management and conservation strategies for threatened orchid species, but knowledge on the spatial distribution of OMF in soil is limited. We here studied the mycorrhizal associations of three terrestrial orchid species (Anacamptis pyramidalis, Orchis purpurea and Platanthera chlorantha) found in a local orchid diversity hotspot in eastern Denmark, and investigated the abundance of the identified mycorrhizal fungi in the surrounding soil. We applied ITS metabarcoding to samples of orchid roots, rhizosphere soil and bulk soil collected at three localities, supplemented with standard barcoding of root samples with OMF specific primers, and detected 22 Operational Taxonomic Units (OTUs) putatively identified as OMF. The three orchid species displayed different patterns of OMF associations, supporting the theory that association with specific fungi constitutes part of an orchid's ecological niche allowing co‐occurrence of many species in orchid‐rich habitats. The identified mycorrhizal partners in the basidiomycete families Tulasnellaceae and Ceratobasidiaceae (Cantharallales) were detected in low abundance in rhizosphere soil, and appeared almost absent from bulk soil at the localities. This finding highlights our limited knowledge of the ecology and trophic mode of OMF outside orchid tissues, as well as challenges in the detection of specific OMF with standard methods. Potential implications for management and conservation strategies are discussed. [ABSTRACT FROM AUTHOR]

Gastrodia elata Blume, a fully mycoheterotrophic perennial plant of the family Orchidaceae, is a traditional Chinese herb with medicinal and edible value. Interestingly, G. elata requires symbiotic relationships with Mycena and Armillaria strains for seed germination and plant growth, respectively. However, there is no comprehensive summary of the symbiotic mechanism between fungi and G. elata. Here, the colonization and digestion of hyphae, the bidirectional exchange of nutrients, the adaptation of fungi and G. elata to symbiosis, and the role of microorganisms and secondary metabolites in the symbiotic relationship between fungi and G. elata are summarized. We comprehensively and deeply analyzed the mechanism of symbiosis between G. elata and fungi from three perspectives: morphology, nutrition, and molecules. The aim of this review was to enrich the understanding of the mutualistic symbiosis mechanisms between plants and fungi and lay a theoretical foundation for the ecological cultivation of G. elata. [ABSTRACT FROM AUTHOR]

Purpose. Improvement of the gene pool of the cultivated form of eggplant (Solanum melongena L.) through interspecific hybridisation with Solanum aethiopicum L. and gametic breeding; analysis of the interspecific lines for a set of valuable quantitative traits determining the yield structure. Methods. Studies for the comprehensive evaluation of eggplant lines of interspecific origin were carried out during 2021-2023 in protected soil conditions at the experimental base of the Institute of Vegetable and Melon Growing of the National Academy of Agrarian Sciences of Ukraine. Phenological observations and biometric measurements of plants of four lines were carried out. The stability of the manifestation of seven quantitative traits during the research years was determined by the methods of variation statistics [calculation of the mean square deviation (σ), coefficient of variation (V), Pearson's paired linear correlation (rp)]. Results. The analysis identified eggplant lines of interspecific origin that were superior to the standard variety 'Almaz' in terms of the level and stability of quantitative traits. Thus, F6(Pavlotas-20 / 'Almaz')I1 and ВС1[F5 (Pavlotas-20 / 'Almaz')]I1 showed a statistically significant increase in fruit width (by 16.48% and 14.85%, respectively). Meanwhile, line BC2[F5 (Pavlotas-20 / 'Almaz')]I1 exhibited higher productivity (by 18.11%) compared to the standard variety. The correlation analysis shows that the trait "Productivity of one plant" has a strong positive relationship with the trait "Number of fruits per plant" (rp = 0.75) and a strong negative relationship with "Average fruit width" (rp = -0.70) and "Duration of the period from mass germination to technical fruit maturity" (rp = -0.72). The selected line BC2[F5 (Pavlotas-20 / 'Almaz')]I1 exhibited a four-day shorter period from mass germination to technical fruit maturity compared to the standard variety. Conclusions. The line BC2[F5 (Pavlotas-20 / 'Almaz')]I1 was isolated based on the complex of quantitative traits. It underwent an additional stage of gametophytic breeding, resulting in a high adaptive potential to growing conditions. [ABSTRACT FROM AUTHOR]

Huotanjun (Burnt Charcoal Mushroom) is a group of edible mushrooms widely hunted and eaten in southern China. Taxonomically, they belong to Russula sect. Nigricantinae. Confusion with the fatal Russula subnigricans cause many poisoning incidents in Asia. For a long time, European names were used in Asia but the Asian genotypes are not the same as European species. We studied specimens collected in the field or bought from markets in southern China. Analysis of four DNA loci (ITS, nc LSU, TEF1-α, RPB2) showed that, except for a specimen of Russula adusta from subalpine Abies forest in southwestern China, all the specimens represent species different from European and North American ones, suggesting prevalent continental endemism in this group. Four new species are here described. Among these, Russula sinoadusta and R. zhuzuijun are Asian counterparts of European R. adusta; R. brunneocystidiata and R. huotanjun are closely related and morphologically highly similar to European R. densifolia and R. fuliginosa. Using multi-locus data, we were able to establish the phylogenetic position of R. subnigricans. It is nested in R. sect. Nigricantinae and sister to R. latolamellata, a species also having distant lamellae and only reddening context. Except for R. sinoadusta which is known only from China, all of our other species are shared by China and Japan. We analyzed the presence/absence of the diagnostic toxin of R. subnigricans cycloprop-2-ene carboxylic acid for 12 species and did not detect the polymeric products in any of them but R. subnigricans. Nevertheless, considering the fact that several potentially edible species share the spaced lamellae and only reddening discoloration with R. subnigricans, we strongly recommend to only eat species with decidedly blackening context and gills. [ABSTRACT FROM AUTHOR]

Physisporinus, a genus in Polyporales, Basidiomycota, is a versatile fungus that lives as a wood decomposer, a potential pathogen of standing trees, and an orchid mycobiont. We previously reported that some Physisporinus species inhabiting wet wood in aquatic environments such as streams and waterfalls form synnema-like structures (SSs) bearing acanthophyses at their apices, and that they produce acanthophyses on vegetative hyphae when cultured on agar media. In this study, we investigated the acanthophysis-forming ability in Physisporinus and allied genera, and experimentally demonstrated the function of SSs. Phylogenetic analyses and observations of Meripilus, Physisporinus and Rigidoporus cultures showed that all of the strains forming acanthophyses belonged to Physisporinus, whereas strains of Meripilus and Rigidoporus did not produce acanthophyses. These findings suggest that SS/acanthophysis formation is a useful taxonomic character for members of Physisporinus. When Physisporinus strains were cultured under oxygen (O2) concentrations of 5, 10, 20 and 40%, most of those cultured under 20% O2 formed the most acanthophyses. According to these experimental data, the SSs/acanthophyses in Physisporinus were considered to have a respiratory function. Physisporinus probably acquired the SS/acanthophysis- forming ability to adapt to moist and/or aquatic habitats and to decay wet wood in which the O2 concentration is often low. [ABSTRACT FROM AUTHOR]

Gastrodia elata Bl. f. glauca is an important traditional Chinese medicinal plant. The yield and quality of Gastrodia elata Bl. have significantly decreased due to multigenerational asexual reproduction. Therefore, it is necessary to have sexual reproduction of Gastrodia elata Bl. to supplement the market supply. Seeds of G. elata Bl. have no endosperm, and their sexual reproduction depends on the nutrients provided by the embryo cells infected by Mycena fungi to complete seed germination. However, Mycena fungi are small and have many species, and not all Mycena fungi can promote the germination of G. elata Bl. seeds. Therefore, it is of great significance to isolate and identify suitable germination fungi and explore the mechanism for improving the production performance and yield, and quality of G. elata Bl. Six closely related Mycena isolates, JFGL-01, JFGL-02, JFGL-03, JFGL-04, JFGL-05, and JFGL-06, were isolated from the leaves and protocorms of G. elata Bl. f. glauca and were identified as Mycena purpureofusca. The mycelial state and number of germinating protocorms were used as indicators to preferentially select Mycena fungi, and it was concluded that JFGL-06 had the best mycelial state and ability to germinate G. elata Bl. seeds. Finally, a mechanism to increase the yield of G. elata Bl. was explored by comparing the changes in nutrient elements and microbial diversity in the soil around G. elata Bl. with different strains. JFGL-06 proved to be an excellent Mycena fungal strain suitable for G. elata Bl. f. glauca. Compared with the commercial strain, JFGL-06 significantly increased the C, N, Na, Mg, S, Cl, K, Ca, and Fe contents of the soil surrounding the protocorms of G. elata Bl. f. glauca. JFGL-06 improved the composition, diversity, and metabolic function of the surrounding soil microbial community of G. elata Bl. f. glauca protocorms at the phylum, class, and genus levels, significantly increased the relative abundance of bacteria such as Acidobacteria and fungi such as Trichoderma among the dominant groups, and increased the abundance of functional genes in metabolic pathways such as nucleotide metabolism and energy metabolism. There was a significant reduction in the relative abundance of bacteria, such as Actinomycetes, and fungi, such as Fusarium, in the dominant flora, and a reduced abundance of functional genes, such as amino acid metabolism and xenobiotic biodegradation and metabolism. This is the main reason why the JFGL-06 strain promoted high-quality and high-yield G. elata Bl. f. glauca in Changbai Mountain. [ABSTRACT FROM AUTHOR]

Background and aims The introduction of crassulacean acid metabolism (CAM) into C3 crops has been considered as a means of improving water-use efficiency. In this study, we investigated photosynthetic and leaf structural traits in F 1 hybrids between Cymbidium ensifolium (female C3 parent) and C. bicolor subsp. pubescens (male CAM parent) of the Orchidaceae. Methods Seven F 1 hybrids produced through artificial pollination and in vitro culture were grown in a greenhouse with the parent plants. Structural, biochemical and physiological traits involved in CAM in their leaves were investigated. Key results Cymbidium ensifolium accumulated very low levels of malate without diel fluctuation, whereas C. bicolor subsp. pubescens showed nocturnal accumulation and diurnal consumption of malate. The F 1s also accumulated malate at night, but much less than C. bicolor subsp. pubescens. This feature was consistent with low nocturnal fixation of atmospheric CO2 in the F 1s. The δ13C values of the F 1s were intermediate between those of the parents. Leaf thickness was thicker in C. bicolor subsp. pubescens than in C. ensifolium , and those of the F 1s were more similar to that of C. ensifolium. This was due to the difference in mesophyll cell size. The chloroplast coverage of mesophyll cell perimeter adjacent to intercellular air spaces of C. bicolor subsp. pubescens was lower than that of C. ensifolium , and that of the F 1s was intermediate between them. Interestingly, one F 1 had structural and physiological traits more similar to those of C. bicolor subsp. pubescens than the other F 1s. Nevertheless, all F 1s contained intermediate levels of phosphoenolpyruvate carboxylase but as much pyruvate, Pi dikinase as C. bicolor subsp. pubescens. Conclusions CAM traits were intricately inherited in the F 1 hybrids, the level of CAM expression varied widely among F 1 plants, and the CAM traits examined were not necessarily co-ordinately transmitted to the F 1s. [ABSTRACT FROM AUTHOR]

Background: Mycorrhiza is a ubiquitous form of symbiosis based on the mutual, beneficial exchange of resources between roots of autotrophic (AT) plants and heterotrophic soil fungi throughout a complex network of fungal mycelium. Mycoheterotrophic (MH) and mixotrophic (MX) plants can parasitise this system, gaining all or some (respectively) required nutrients without known reciprocity to the fungus. We applied, for the first time, an ecological stoichiometry framework to test whether trophic mode of plants influences their elemental carbon (C), nitrogen (N), and phosphorus (P) composition and may provide clues about their biology and evolution within the framework of mycorrhizal network functioning. Results: We analysed C:N:P stoichiometry of 24 temperate orchid species and P concentration of 135 species from 45 plant families sampled throughout temperate and intertropical zones representing the three trophic modes (AT, MX and MH). Welch's one-way ANOVA and PERMANOVA were used to compare mean nutrient values and their proportions among trophic modes, phylogeny, and climate zones. Nutrient concentration and stoichiometry significantly differentiate trophic modes in orchids. Mean foliar C:N:P stoichiometry showed a gradual increase of N and P concentration and a decrease of C: nutrients ratio along the trophic gradient AT < MX < MH, with surprisingly high P requirements of MH orchids. Although P concentration in orchids showed the trophy-dependent pattern regardless of climatic zone, P concentration was not a universal indicator of trophic modes, as shown by ericaceous MH and MX plants. Conclusion: The results imply that there are different evolutionary pathways of adaptation to mycoheterotrophic nutrient acquisition, and that the high nutrient requirements of MH orchids compared to MH plants from other families may represent a higher cost to the fungal partner and consequently lead to the high fungal specificity observed in MH orchids. [ABSTRACT FROM AUTHOR]

Context: Giving up photosynthetic function is a bold evolutionary step for a plant, yet the evolutionary transition from autotrophy to mycoheterotrophy has occurred dozens of times. Comparing plastome sequences of mycoheterotrophs and autotrophs has identified recurring patterns of gene loss throughout a range of land plants, though more comparisons are required to see how broadly these patterns apply across the diversity of terrestrial plants. Mycoheterotrophy is especially common in Orchidaceae, with more than 40 transitions from autotrophy to mycoheterotrophy in the family. Aims: We sought to test generalised patterns of plastome degradation seen in other mycoheterotrophs by comparing two species in the genus Dipodium (Orchidaceae); one species is photosynthetic and the other appears to be a facultative mycoheterotroph species. Methods: We sequenced and assembled the plastomes of two Dipodium species and compared the two genomes to identify gene degradation or loss. Results: The two plastomes were nearly identical, with no degradation of photosynthesis genes in the putative mycoheterotroph, and both species have undergone loss or pseudogenisation of all plastid ndh (nicotinamide adenine dinucleotide + hydrogen specific dehydrogenase) genes. Conclusions: These results contrast with most other comparisons between photosynthetic and likely mycoheterotrophic relatives, where rapid degradation in mycoheterotroph plastome genes is common, and may suggest the leafless Dipodium species are capable of photosynthesis and may be in the early stages of transitioning to a fully heterotrophic lifestyle. Implications: Further investigation of trophic transitions in Dipodium , including sequencing more plastomes and measuring photosynthetic capability of the putative heterotrophs, will yield insights into the evolution of plant lineages that lose the ability to photosynthesise. Some plant species rely on fungal symbioses for energy, rather than the sun; such plants typically follow a general pattern of gene loss as they transition away from being photosynthetic. We found that an Australian orchid species typically regarded as being fully reliant on the fungal symbiont, retains the genes required for photosynthesis. This species may be able to utilise both energy sources, or may rather be only in the early stages of transitioning to fungal reliance. [ABSTRACT FROM AUTHOR]

Summary: Stable isotope signatures of fungal sporocarps have been instrumental in identifying carbon gains of chlorophyllous orchids from a fungal source. Yet, not all mycorrhizal fungi produce macroscopic sporocarps and frequently fungi of different taxa occur in parallel in orchid roots.To overcome this obstacle, we investigated stable isotope signatures of fungal pelotons extracted from orchid roots and compared these data to the respective orchid and reference plant tissues. Anoectochilus sandvicensis and Epipactis palustris represented specialized or unspecialized rhizoctonia‐associated orchids. Epipactis atrorubens and Epipactis leptochila are orchids considered ectomycorrhiza‐associated with different preferences for Basidio‐ and Ascomycota.13C enrichment of rhizoctonia pelotons was minor compared with plant tissues and significantly lower than enrichments of pelotons from ectomycorrhizal Epipactis species. 15N values of pelotons from E. leptochila and E. atrorubens showed similar patterns as known for respective sporocarps of ectomycorrhizal Ascomycota and Basidiomycota, however, with an offset towards lower 15N enrichments and nitrogen concentrations.Our results suggest an explicit fungal nutrition source of orchids associated with ectomycorrhizal fungi, whereas the low 13C enrichment in rhizoctonia‐associated orchids and fungal pelotons hamper the detection of carbon gains from fungal partners. 15N isotopic pattern of orchids further suggests a selective transfer of 15N‐enriched protein‐nitrogen into orchids. [ABSTRACT FROM AUTHOR]

These signatures have been shown to reflect the ecophysiology of fungi and demonstrate that fungi that can utilize organic nitrogen exhibit higher SP 15 sp N than those fungi restricted to mineral nitrogen sources (Gebauer & Taylor, [4]; Lilleskov I et al i ., [14]). Keywords: arbuscular mycorrhizal fungi; carbon; DNA sequencing; ITS2; mycoheterotrophy; nitrogen; rhizoctonia EN arbuscular mycorrhizal fungi carbon DNA sequencing ITS2 mycoheterotrophy nitrogen rhizoctonia 1166 1172 7 07/17/23 20230815 NES 230815 Data availability Isotope abundance data are available in the Supporting Information. For the orchid-associated fungi, hyphae are only slightly enriched in SP 13 sp C in relation to both reference and orchid plants, remaining unclear whether these orchids gain C from the associated fungi based on these results. Yet, values of SP 13 sp C and SP 15 sp N are poorly known for arbuscular mycorrhizal fungi (but see e.g. Courty I et al i ., [1]; Suetsugu I et al i ., [24], for isotope values of fungal spores), and the orchid-associated fungi known as "rhizoctonia" in natural conditions. [Extracted from the article]

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Some plant species took an alternative evolutionary pathway in which they lost their photosynthetic capacity to depend exclusively on carbon supplied by arbuscular mycorrhizal fungi (AMF) in an association called mycoheterotrophy. Among them is Voyriella parviflora, a species of the family Gentianaceae, which is found in tropical regions such as the Amazon basin. Here, we assessed the identity of AMF symbionts associated with this species. DNA was isolated from eight Gentianaceae specimens and from litter and surrounding roots of photosynthetic plants. The atp1 gene was amplified by Sanger sequencing to determine the taxonomic affiliation of the mycoheterotrophic plants. A 280 bp region of the 18S rRNA gene of AMF was amplified with primers NS31/AML2 by high-throughput sequencing. The mycoheterotrophic specimens were assigned to V. parviflora with a bootstrap support of 72%. Glomus was the most abundant AMF genus, both in the mycoheterotrophic plants and in the litter and roots of photosynthetic plants. In addition, a few Glomus genotypes were abundantly enriched in the mycoheterotrophic plants, with only a few specimens colonized by Gigaspora, Acaulospora, and Scutellospora in a low proportion. These genotypes formed a cluster within a larger clade, suggesting that V. parviflora shows a preferential association with a narrow Glomus lineage which is not phylogenetically close to a previously identified V. parviflora's associated lineage. Furthermore, detecting fungi from other families suggests that V. parviflora is colonized by other genera, although with low frequency. These findings provide new insights into the association between AMF and mycoheterotrophic species and highlight the importance of considering trap culture-independent approaches in understanding this symbiosis. [ABSTRACT FROM AUTHOR]

The first genome sequenced of a eukaryotic organism was for Saccharomyces cerevisiae, as reported in 1996, but it was more than 10 years before any of the zygomycete fungi, which are the early-diverging terrestrial fungi currently placed in the phyla Mucoromycota and Zoopagomycota, were sequenced. The genome for Rhizopus delemar was completed in 2008; currently, more than 1000 zygomycete genomes have been sequenced. Genomic data from these early-diverging terrestrial fungi revealed deep phylogenetic separation of the two major clades—primarily plant—associated saprotrophic and mycorrhizal Mucoromycota versus the primarily mycoparasitic or animal-associated parasites and commensals in the Zoopagomycota. Genomic studies provide many valuable insights into how these fungi evolved in response to the challenges of living on land, including adaptations to sensing light and gravity, development of hyphal growth, and co-existence with the first terrestrial plants. Genome sequence data have facilitated studies of genome architecture, including a history of genome duplications and horizontal gene transfer events, distribution and organization of mating type loci, rDNA genes and transposable elements, methylation processes, and genes useful for various industrial applications. Pathogenicity genes and specialized secondary metabolites have also been detected in soil saprobes and pathogenic fungi. Novel endosymbiotic bacteria and viruses have been discovered during several zygomycete genome projects. Overall, genomic information has helped to resolve a plethora of research questions, from the placement of zygomycetes on the evolutionary tree of life and in natural ecosystems, to the applied biotechnological and medical questions. [ABSTRACT FROM AUTHOR]

During reclamation organisms like fungi directly interact at the soil–plant interface and can potentially impact plant growth, health, and overall revegetation success. Therefore, a study was conducted to determine how topsoil amendments (mycorrhizae, biochar, compost, manure) influence root‐associated fungal communities and growth of fast growing Populus balsamifera trees on a phosphogypsum (PG) reclamation site in Alberta, Canada. Tree height, diameter, biomass, and diversity of Ascomycota and Basidiomycota were assessed using Illumina MiSeq sequencing targeting the fungal internal transcribed spacer region 1 (ITS1) and ITS2. Topsoil amendments and root location (topsoil vs. PG within a treatment), greatly affected the root fungal community. Basidiomycota preferentially grew in PG, and had a high treatment specific root fungal community with highest diversity of ectomycorrhizal fungi and basidiomycete yeast in biochar PG and compost PG. Populus balsamifera roots were associated with a high diversity of Ascomycota, found equally in roots in topsoil and PG, with a trend towards higher diversity and abundance in PG. The core genus found across all samples was Fusarium, a potential plant pathogen. All topsoil amendments strongly reduced abundance of Fusarium and other Ascomycota in poplar roots. Biochar and compost reduced number and abundance of potential fungal pathogens and increased number of potential plant growth promoting Basidiomycota, indicating favorable growth conditions for P. balsamifera on a phosphygypsum reclamation site by application of these amendments. This study indicates compost and biochar can be effective soil amendments to support P. balsamifera establishment and growth on a PG reclamation site. [ABSTRACT FROM AUTHOR]

Background: Root and soil microbial communities constitute the below-ground plant microbiome, are drivers of nutrient cycling, and affect plant productivity. However, our understanding of their spatiotemporal patterns is confounded by exogenous factors that covary spatially, such as changes in host plant species, climate, and edaphic factors. These spatiotemporal patterns likely differ across microbiome domains (bacteria and fungi) and niches (root vs. soil). Results: To capture spatial patterns at a regional scale, we sampled the below-ground microbiome of switchgrass monocultures of five sites spanning > 3 degrees of latitude within the Great Lakes region. To capture temporal patterns, we sampled the below-ground microbiome across the growing season within a single site. We compared the strength of spatiotemporal factors to nitrogen addition determining the major drivers in our perennial cropping system. All microbial communities were most strongly structured by sampling site, though collection date also had strong effects; in contrast, nitrogen addition had little to no effect on communities. Though all microbial communities were found to have significant spatiotemporal patterns, sampling site and collection date better explained bacterial than fungal community structure, which appeared more defined by stochastic processes. Root communities, especially bacterial, were more temporally structured than soil communities which were more spatially structured, both across and within sampling sites. Finally, we characterized a core set of taxa in the switchgrass microbiome that persists across space and time. These core taxa represented < 6% of total species richness but > 27% of relative abundance, with potential nitrogen fixing bacteria and fungal mutualists dominating the root community and saprotrophs dominating the soil community. Conclusions: Our results highlight the dynamic variability of plant microbiome composition and assembly across space and time, even within a single variety of a plant species. Root and soil fungal community compositions appeared spatiotemporally paired, while root and soil bacterial communities showed a temporal lag in compositional similarity suggesting active recruitment of soil bacteria into the root niche throughout the growing season. A better understanding of the drivers of these differential responses to space and time may improve our ability to predict microbial community structure and function under novel conditions. [ABSTRACT FROM AUTHOR]

Two new species of Scytinostroma viz. S. acystidiatum and S. macrospermum, are described from southwest China. Phylogeny based on ITS + nLSU dataset demonstrates that samples of the two species form two independent lineages and are different in morphology from the existing species of Scytinostroma. Scytinostroma acystidiatum is characterized by resupinate, coriaceous basidiomata with cream to pale yellow hymenophore, a dimitic hyphal structure with generative hyphae bearing simple septa, the absence of cystidia, and amyloid, broadly ellipsoid basidiospores measuring 4.7-7 x 3.5-4.7 mm. Scytinostroma macrospermum is characterized by resupinate, coriaceous basidiomata with cream to straw yellow hymenophore, a dimitic hyphal structure with generative hyphae bearing simple septa, numerous cystidia embedded or projecting from hymenium, and inamyloid, ellipsoid basidiospores measuring 9-11 x 4.5-5.5 mm. The differences between the new species and morphologically similar and phylogenetically related species are discussed. [ABSTRACT FROM AUTHOR]

Background: Mycoheterotrophs, acquiring organic carbon and other nutrients from mycorrhizal fungi, have evolved repeatedly with substantial plastid genome (plastome) variations. To date, the fine-scale evolution of mycoheterotrophic plastomes at the intraspecific level is not well-characterized. A few studies have revealed unexpected plastome divergence among species complex members, possibly driven by various biotic/abiotic factors. To illustrate evolutionary mechanisms underlying such divergence, we analyzed plastome features and molecular evolution of 15 plastomes of Neottia listeroides complex from different forest habitats. Results: These 15 samples of Neottia listeroides complex split into three clades according to their habitats approximately 6 million years ago: Pine Clade, including ten samples from pine-broadleaf mixed forests, Fir Clade, including four samples from alpine fir forests and Fir-willow Clade with one sample. Compared with those of Pine Clade members, plastomes of Fir Clade members show smaller size and higher substitution rates. Plastome size, substitution rates, loss and retention of plastid-encoded genes are clade-specific. We propose to recognized six species in N. listeroides complex and slightly modify the path of plastome degradation. Conclusions: Our results provide insight into the evolutionary dynamics and discrepancy of closely related mycoheterotrophic orchid lineages at a high phylogenetic resolution. [ABSTRACT FROM AUTHOR]

The autotrophic lifestyle of photosynthetic plants has profoundly shaped their body plan, physiology, and gene repertoire. Shifts to parasitism and heterotrophy have evolved at least 12 times in more than 4000 species, and this transition has consequently left major evolutionary footprints among these parasitic lineages. Features that are otherwise rare at the molecular level and beyond have evolved repetitively, including reduced vegetative bodies, carrion‐mimicking during reproduction, and the incorporation of alien genetic material. Here, I propose an integrated conceptual model, referred to as the funnel model, to define the general evolutionary trajectory of parasitic plants and provide a mechanistic explanation for their convergent evolution. This model connects our empirical understanding of gene regulatory networks in flowering plants with classical theories of molecular and population genetics. It emphasizes that the cascading effects brought about by the loss of photosynthesis may be a major force constraining the physiological capacity of parasitic plants and shaping their genomic landscapes. Here I review recent studies on the anatomy, physiology, and genetics of parasitic plants that lend support to this photosynthesis‐centered funnel model. Focusing on nonphotosynthetic holoparasites, I elucidate how they may inevitably reach an evolutionary terminal status (i.e., extinction) and highlight the utility of a general, explicitly described and falsifiable model for future studies of parasitic plants. [ABSTRACT FROM AUTHOR]

Campylocentrum pachyrrhizum Rchb.f. Rolfe, also known as the ribbon orchid, is native to southern Florida, the West Indies, and northern South America. In Florida, it is restricted to the Fakahatchee Strand and is currently state-listed as endangered, but virtually nothing is known about the orchid's biology, ecology, and current status. The purpose of this study was to document ribbon orchid populations within the northern portion of the Fakahatchee Strand (Florida Panther NWR) and to identify some of the biotic and abiotic factors critical to its survival. During 2016–2022, a total of 118 ribbon orchids were recorded at five sites on 21 host trees comprising two species (Annona glabra, Fraxinus caroliniana). The majority (80%) were rooted on pop ash, and over half (54%) were rooted at the base of trees 100–150 cm above the high water mark. The number of orchids decreased from year to year, with the largest drop (2017–2018) coinciding with damage from Hurricane Irma. At the end of the 2022 survey, only 44 orchids remained. Mycorrhizal fungi isolated from two plants were identified as a Ceratobasidium species. We urge land managers to take immediate action to safeguard this species throughout southern Florida. [ABSTRACT FROM AUTHOR]

Plant mitochondrial genomes (mitogenomes) exhibit fluid genome architectures, which could lead to the rapid erosion of genome synteny over a short evolutionary time scale. Among the species-rich orchid family, the leafy Cymbidium lancifolium and leafless Cymbidium macrorhizon are sister species with remarkable differences in morphology and nutritional physiology. Although our understanding of the evolution of mitochondria is incomplete, these sister taxa are ideal for examining this subject. In this study, the complete mitogenomes of C. lancifolium and C. macrorhizon, totaling 704,244 bp and 650,751 bp, respectively, were assembled. In the 2 mitogenomes, 38 protein-coding genes, 18 cis- and 6 trans-spliced introns, and approximately 611 Kb of homologous sequences are identical; overall, they have 99.4% genome-wide similarity. Slight variations in the mitogenomes of C. lancifolium and C. macrorhizon in repeat content (21.0 Kb and 21.6 Kb, respectively) and mitochondrial DNA of plastid origin (MIPT; 38.2 Kb and 37.5 Kb, respectively) were observed. The mitogenome architectures of C. lancifolium and C. macrorhizon are complex and comprise 23 and 22 mini-circular chromosomes, respectively. Pairwise comparisons indicate that the two mitogenomes are largely syntenic, and the disparity in chromosome numbers is likely due to repeat-mediated rearrangements among different chromosomes. Notably, approximately 93.2 Kb C. lancifolium mitochondrial sequences lack any homology in the C. macrorhizon mitogenome, indicating frequent DNA gains and losses, which accounts mainly for the size variation. Our findings provide unique insights into mitogenome evolution in leafy and leafless plants of sister species and shed light on mitogenome dynamics during the transition from mixotrophy to mycoheterotrophy. [ABSTRACT FROM AUTHOR]

Gentiana zollingeri (Gentianaceae) is an initial mycoheterotrophic plant that depends on a specific group of arbuscular mycorrhizal (AM) fungi for carbon source during underground growth after seed germination. In this study, a mycorrhizal fungus dominant in mycoheterotrophic seedlings of G. zollingeri was successfully isolated from a soil core collected from a point close to a flowering G. zollingeri. The AM fungal isolate was identified as conspecific or closely related to Dominikia aurea (Glomeraceae) by spore morphology and molecular phylogeny. Basic Local Alignment Search Tool (BLAST) searches against the MaarjAM database showed that the nuclear small subunit ribosomal DNA sequences of the isolate matched the AM fungal sequences obtained from a wide range of plants in various ecosystems, including several mycoheterotrophs. Thus, it is suggested that the AM fungal isolate is one of the cheating susceptible AM fungi. Furthermore, the sequences corresponded to those of a group of AM fungi dominantly detected in Japanese temperate forests. Accordingly, there is a possibility that mycoheterotrophic plants, including seedlings of G. zollingeri, may target AM fungi with a wide host range and ubiquitous distribution. [ABSTRACT FROM AUTHOR]

Most of our knowledge on the ericoid mycorrhizal (ErM) symbiosis comes from temperate heathlands characterized by acidic peaty soils and many experiments with a few ascomycetous fungi. However, ericaceous plants thrive in many other ecosystems and in temperate coniferous forests, their seedlings often prosper on decomposing wood. While wood is typically exploited by basidiomycetous ectomycorrhizal (EcM) and saprobic fungi, the role of ErM fungi (ErMF) is much less clear. We explored the cultivable mycobiota of surface sterilized hair roots of Vaccinium spp. growing on decomposing wood in two coniferous forests in Mid-Norway (Scandinavia) and Northern Bohemia (Central Europe). Obtained isolates were identified using molecular tools and their symbiotic potential was tested in vitro. While the detected community lacked the archetypal ErMF Hyaloscypha hepaticicola and the incidence of dark septate endophytes and EcM fungi was negligible, it comprised other frequent asexual ascomycetous ErMF, namely H. variabilis and Oidiodendron maius, together with several isolates displaying affinities to sexual saprobic H. daedaleae and H. fuckelii. Ascomycete-suppressing media revealed representatives of the saprobic basidiomycetous genera Coprinellus, Gymnopilus, Mycena (Agaricales), and Hypochnicium (Polyporales). In the resyntheses, the tested basidiomycetes occasionally penetrated the rhizodermal cells of their hosts but never formed ericoid mycorrhizae and in many cases overgrew and killed the inoculated seedlings. In contrast, a representative of the H. daedaleae/H. fuckelii-related isolates repeatedly formed what morphologically appears as the ErM symbiosis and supported host's growth. In conclusion, while basidiomycetous saprobic fungi have a potential to colonize healthy-looking ericaceous hair roots, the mode(-s) of their functioning remain obscure. For the first time, a lineage in Hyaloscypha s. str. (corresponding to the former Hymenoscyphus ericae aggregate) where sexual saprobes are intermingled with root symbionts has been revealed, shedding new light on the ecology and evolution of these prominent ascomycetous ErMF. [ABSTRACT FROM AUTHOR]

Orchids commonly rely on mycorrhizal fungi to obtain the necessary resources for seed germination and growth. Whereas most photosynthetic orchids typically associate with so-called rhizoctonia fungi to complete their life cycle, there is increasing evidence that other fungi may be involved as well and that the mycorrhizal communities associated with orchids may be more diverse. Coexisting orchid species also tend to associate with different fungi to reduce competition for similar resources and to increase long-term population viability. However, few studies have related the mycorrhizal communities in the rhizosphere to communities found in the roots of closely related coexisting orchid species. In this study, we used high-throughput sequencing to investigate the diversity and community composition of orchid mycorrhizal fungi in the roots and the rhizosphere of four Cypripedium species growing in forests in Northeast China. The results showed that the investigated Cypripedium species associated with a wide variety of fungi including members of Tulasnellaceae, Psathyrellaceae, and Herpotrichiellaceae, whereas members of Russulaceae, Cortinariaceae, Thelephoraceae, and Herpotrichiellaceae showed high abundance in rhizosphere soils. The diversity of fungi detected in the rhizosphere soil was much higher than that in the roots. The observed variation in fungal communities in Cypripedium roots was not related to forest site or orchid species. On the other hand, variation in mycorrhizal communities of rhizosphere soil was significantly related to sampling site. These results indicate that orchid mycorrhizal communities in the rhizosphere display considerable variation among sites and that orchids use only a subset of the locally available fungi. Future studies focusing on the fine-scale spatial distribution of orchid mycorrhizal fungi and more detailed assessments of local environmental conditions will provide novel insights into the mechanisms explaining variation of fungal communities in both orchid roots and the rhizosphere. [ABSTRACT FROM AUTHOR]

Copyright of Guihaia is the property of Guihaia Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Orchids have symbiotic relationships with ectomycorrhizal or saprotrophic ("rhizoctonia") fungi, which provide plants with nutrients. The extent of plant dependency on fungi for carbon supply is reflected by plant tissue δ13C abundance values. Little is known about how wide-spread partial mycoheterotrophy (PMH) is among orchids in nature and it is debated whether rhizoctonia-associated orchids possess mycoheterotrophy. Using orchids with ectomycorrhizal fungi and rhizoctonia-associated orchid species, we tested one hypothesis: terrestrial green orchids decrease their δ13C values in response to shade less than neighboring non-orchid vascular plants (evidence for PMH). We examined eight pairs of plant species (orchid paired with reference autotrophic plant growing close-by) in well-lit versus shady field conditions in replicated sites across the European part of Russia. In each site illumination level was recorded and plant material was examined for δ13C. We found that all tested orchids showed similar weaker response of tissue δ13C signature to relative light exposure in contrast to the stronger response in control plants, suggesting wide-spread PMH among orchids. There was no significant difference in orchids with ectomycorrhizal fungi and rhizoctonia-associated orchids in response of δ13C to shade, supporting mycoheterotrophic mechanisms of carbon transfer to plants in rhizoctonia-associated orchids. Proportions of fungi-derived carbon for individual orchid species ranged between 3 and 50%. Orchids reduced stomatal density in response to shade less or similarly to autotrophic plants, suggesting that mycoheterotrophic plants might be somewhat less water limited than autotrophic species due to fungal water supply. Our data provide support for broad prevalence of PMH among orchids and of the existence of PMH in rhizoctonia-associated orchids. These findings extend the ecological importance of mycorrhizal associations in belowground food webs. [ABSTRACT FROM AUTHOR]

The presence and diversity of endophytic fungi associated with host plants are important not just for host plant growth and defense, but also impact the production of medicinal secondary metabolites. However, the correlation between endophytic fungi and crocin production in Crocus sativus (CS) remains underexplored. Here, we explore the relationship between endophytic fungal diversity and crocin content among different CS tissues and field sites. Specifically, we isolated endophytic fungi from five different field sites (Shanghai, Jiande, Huzhou, Anhui, and Hebei) and five different tissues (corm, scape, leaf, petal, and stigma) and analyzed fungal community diversity, richness, and evenness. We identified a total of 32 endophytic fungal taxa, assigned to 7 orders within 4 classes (Eurotiomycetes, Agaricomycetes, Dothideomycetes, and Sordariomycetes). The most dominant order was Eurotiales, and the most dominant genera were Penicillium and Talaromyces. Species richness tended to be highest in belowgrown tissues, such as corm and scape. Additionally, several fungal taxa were found to be either site- or tissue-specific. Three genera in particular were correlated with crocin content: Penicillium, Sistotrema, and Bjerkandera. Given the fact that endophytic microorganisms can both promote the production of secondary metabolites in host plants and potentially produce secondary metabolites themselves, further study is required to understand the mechanistic relationship between these and other fungal genera and crocin production. [ABSTRACT FROM AUTHOR]

Gastrodia elata is an achlorophyllous and fully mycoheterotrophic orchid which obtains carbon and other nutrients from Armillaria species in its life cycle. Many researchers suggested that plant hormones, as signing molecules, play a central role in the plant–fungi interaction. In the process of Armillaria gallica 012 m cultivation, both exogenous indole‐3‐acetic acid (IAA) and indole‐3‐butyric acid (IBA) distinctly stimulated the growth of mycelia in solid media. The differential expression genes (DEGs) of A. gallica 012 m with IAA versus blank control (BK) and IBA versus BK were investigated. The results showed that more than 80% of DEGs of the IAA group were coincident with the DEGs of the IBA group, and more than half of upregulated DEGs and most of the downregulated DEGs of the IAA group coincided with those DEGs of the IBA group. Above research implied that A. gallica 012 m could perceive IAA and IBA, and possess similar responses and signaling pathways to IAA and IBA. The overlapping differential genes of the IAA group and IBA group were analyzed by GO term, and the results showed that several DEGs identified were related to biological processes including positive regulation of the biological process and biological process. The downregulated NmrA‐like and FKBP_C genes might be benefit to the growth of mycelia. Those results can explain that exiguous IAA and IBA improved the growth of A. gallica to some extent. We speculate that IAA and IBA are signaling molecules, and regulate the expression of growth‐related genes of A. gallica 012 m by the same signaling pathway. [ABSTRACT FROM AUTHOR]

Summary: Cleistogamy, in which plants can reproduce via self‐fertilization within permanently closed flowers, has evolved in > 30 angiosperm lineages; however, consistent with Darwin's doubts about its existence, complete cleistogamy – the production of only cleistogamous flowers – has rarely been recognized. Thus far, the achlorophyllous orchid genus, Gastrodia, is the only known genus with several plausible completely cleistogamous species.Here, we analyzed the floral developmental transcriptomes of two recently evolved, completely cleistogamous Gastrodia species and their chasmogamous sister species to elucidate the possible changes involved in producing common cleistogamous traits.The ABBA‐BABA test did not support introgression and protein sequence convergence as evolutionary mechanisms leading to cleistogamy, leaving convergence in gene expression as a plausible mechanism. Regarding transcriptomic differentiation, the two cleistogamous species had common modifications in the expression of developmental regulators, exhibiting a gene family‐wide signature of convergent expression changes in MADS‐box genes. Our transcriptomic pseudotime analysis revealed a prolonged juvenile state and eventual maturation, a heterochronic pattern consistent with partial neoteny, in cleistogamous flower development.These findings indicate that transcriptomic partial neoteny, arising from changes in the expression of conserved developmental regulators, might have contributed to the rapid and repeated evolution of cleistogamous flowers in Gastrodia. [ABSTRACT FROM AUTHOR]

The study of congruency between phylogenies of interacting species can provide a powerful approach for understanding the evolutionary history of symbiotic associations. Orchid mycorrhizal fungi can survive independently of orchids making cospeciation unlikely, leading us to predict that any congruence would arise from host‐switches to closely related fungal species. The Australasian orchid subtribe Drakaeinae is an iconic group of sexually deceptive orchids that consists of approximately 66 species. In this study, we investigated the evolutionary relationships between representatives of all six Drakaeinae orchid genera (39 species) and their mycorrhizal fungi. We used an exome capture dataset to generate the first well‐resolved phylogeny of the Drakaeinae genera. A total of 10 closely related Tulasnella Operational Taxonomic Units (OTUs) and previously described species were associated with the Drakaeinae orchids. Three of them were shared among orchid genera, with each genus associating with 1–6 Tulasnella lineages. Cophylogenetic analyses show Drakaeinae orchids and their Tulasnella associates exhibit significant congruence (p < 0.001) in the topology of their phylogenetic trees. An event‐based method also revealed significant congruence in Drakaeinae–Tulasnella relationships, with duplications (35), losses (25), and failure to diverge (9) the most frequent events, with minimal evidence for cospeciation (1) and host‐switches (2). The high number of duplications suggests that the orchids speciate independently from the fungi, and the fungal species association of the ancestral orchid species is typically maintained in the daughter species. For the Drakaeinae–Tulasnella interaction, a pattern of phylogenetic niche conservatism rather than coevolution likely explains the observed phylogenetic congruency in orchid and fungal phylogenies. Given that many orchid genera are characterized by sharing of fungal species between closely related orchid species, we predict that these findings may apply to a wide range of orchid lineages. [ABSTRACT FROM AUTHOR]

Cypripedium japonicum Thunb. (Orchidaceae), once a common perennial herb, is now designated as endangered throughout most of its distribution due to habitat destruction and fragmentation, and the impacts of horticultural collection. We investigated the genetic characteristics of this species for conservation purposes, using microsatellite markers to examine the genetic diversity and structure of 15 native and 5 ex situ populations in Japan. The results imply that although allelic variation is low in Japanese C. japonicum, sexual reproduction by seed, as well as clonal propagation, may occur in some populations. Both native and ex situ populations were found to be genetically differentiated, indicating that some populations may have experienced recent population declines, genetic fragmentation, or bottlenecks. The degree of genetic drift from the putative ancestral population, inferred through STRUCTURE analysis, was more pronounced in northern populations than in southern populations. Some of the ex situ conserved populations exhibited a low degree of differentiation from ancestral native populations. Our results imply that conservation of C. japonicum in Japan is best supported by maintaining individual populations and their unique genetic characteristics. [ABSTRACT FROM AUTHOR]

References 1 Smallwood P.A., Trapnell D.W. Species Distribution Modeling Reveals Recent Shifts in Suitable Habitat for Six North American Cypripedium spp. (Orchidaceae). The presence of Orchids is declining worldwide, mostly due to habitat loss and other factors like climate change, which are likely to increase in severity in the coming decades. Szlachetko et al. [[4]] revised the orchid genus I Andinia i into multiple genera, based on the outcomes of morphological and molecular studies. [Extracted from the article]

Triploid is considered a reproductive barrier and also a bridge in the formation of polyploids. However, few reports are available in Cymbidium. In this study, diploid ‘Xiaofeng’, sexual triploid ‘Yuchan’ and ‘Huanghe’ of Cymbidium were used to evaluate hybridization compatibility of the triploids. Results showed that the sexual triploids were fertile whether they were used as male or female parents. ‘Yuchan’ produced male gametes of 1x, 1x~2x, 2x, 2x~3x, and 3x at frequencies of 8.89%, 77.78%, 6.67%, 3.33%, and 3.33%, respectively; while ‘Huanghe’ produced 3.33% 1x, 80.00% 1x~2x, 8.89% 2x, 5.56% 2x~3x, and 2.22% 3x male gametes. The cross of ‘Xiaofeng’ with ‘Yuchan’ produced progenies with a wide range of ploidy levels, including one diploid, 34 2×~3× aneuploids, 12 triploids, and one tetraploid, indicating that male gametes produced by sexual triploid were fertile and could be transmitted and fused with egg cells. On the other hand, 10 progenies obtained from the cross of ‘Yuchan’ × ‘Xiaofeng’ were all aneuploids. The cross of ‘Yuchan’ with ‘Huanghe’ produced 40 progenies including three 2×~3× aneuploids, nine 3×~4× aneuploids, 21 tetraploids, six 4×~5× aneuploids, and one pentaploid, suggesting that 2x gametes, instead of the unreduced ones played a more important role in the formation of tetraploids. The survival rates of the hybrids were all above 80.00%, with the tetraploids at 96.67%. Cytological analysis revealed that during meiosis of sexual polyploids, two chromosome sets of the 2n gamete were inclined to enter into the same daughter cell, resulting in the production of 2x gametes. Our results indicate that the triploid cymbidiums are not reproductive barrier but serve as a bridge in the formation of polyploid plants. [ABSTRACT FROM AUTHOR]