Intracellular Hyphae Research Articles

Three-Dimensional Ultrastructure of Arabidopsis Cotyledons Infected with Colletotrichum higginsianum.

We used serial block-face scanning electron microscopy (SBF-SEM) to study the host-pathogen interface between Arabidopsis cotyledons and the hemibiotrophic fungus Colletotrichum higginsianum. By combining high-pressure freezing and freeze-substitution with SBF-SEM, followed by segmentation and reconstruction of the imaging volume using the freely accessible software IMOD, we created 3D models of the series of cytological events that occur during the Colletotrichum-Arabidopsis susceptible interaction. We found that the host cell membranes underwent massive expansion to accommodate the rapidly growing intracellular hypha. As the fungal infection proceeded from the biotrophic to the necrotrophic stage, the host cell membranes went through increasing levels of disintegration culminating in host cell death. Intriguingly, we documented autophagosomes in proximity to biotrophic hyphae using transmission electron microscopy (TEM) and a concurrent increase in autophagic flux between early to mid/late biotrophic phase of the infection process. Occasionally, we observed osmiophilic bodies in the vicinity of biotrophic hyphae using TEM only and near necrotrophic hyphae under both TEM and SBF-SEM. Overall, we established a method for obtaining serial SBF-SEM images, each with a lateral (x-y) pixel resolution of 10 nm and an axial (z) resolution of 40 nm, that can be reconstructed into interactive 3D models using the IMOD. Application of this method to the Colletotrichum-Arabidopsis pathosystem allowed us to more fully understand the spatial arrangement and morphological architecture of the fungal hyphae after they penetrate epidermal cells of Arabidopsis cotyledons and the cytological changes the host cell undergoes as the infection progresses toward necrotrophy. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Synergism study of Bacopa monnieri and Piriformospora indica and its impact on Biomass and metabolite

BackgroundA symbiotic connection between Piriformospora indica and Bacopa monnieri (L.) Wettest, obtained through co-cultivation synergism, was found to improve growth, biomass production, and bacoside content in the plants. Brahmi (B. monnieri L.), a well-known Indian plant prized for its memory-boosting properties, has a lengthy history and a premium price tag. Because of its remarkable ability to colonize a wide variety of plant species, the axenically cultivable mycorrhiza-like endophytic fungus P. indica has gained a lot of interest recently.MethodsIn the current study, fungal spores from recently revived cultures were added to jam bottles next to rooted Brahmi plants for in vitro co-cultivation. The control plants were left without fungal discs. Pre-rooted micro-propagated Brahmi plants were treated with agar discs containing actively growing hyphae. For a period of 3 months, both trials were conducted with a fully randomized setup. Microscopy of the treated and control plant roots verified co-cultivation.ResultsMicroscopic examination of the roots of co-cultivated plants reveals a high degree of colonization with host plants. These endophytic fungal structures include intracellular chlamydospores, and arbuscules, an intercellular and intracellular hyphae network, and a mycelial network on the root surface. In both in vitro and in vivo co-cultivation studies, the plant extended the host plant’s lifespan in 3 months by displaying continuous regeneration; in contrast, the control plant displayed signs of senescence. With biomass exceeding the control by 1.18 times in vivo and 1.28 times in vitro. In vitro, co-cultivation circumstances also led to an increase in the rate of utilization of nutritional medium. In comparison to the control, the amount of bacoside increases to 100% in vivo after a month of co-cultivation and 33% in vitro after 3 months.ConclusionsIn the present investigation, in vivo co-cultivation showed a favorable interaction effect on biomass production as well as bacoside content, which can satisfy the raw material demands of Brahmi plants in pharmaceutical industries.

On the Occurrence of Arbuscular Mycorrhizal Fungi in a Bryophyte Community of Punta Lara Natural Reserve, Buenos Aires, Argentina

The evolutionary history of the symbiotic association between arbuscular mycorrhizal fungi (AMF) and embryophytes dates back to the Devonian period. Previous ecological and physiological studies have described the presence of arbuscules, inter- and intracellular hyphae, vesicles, coils and spores, in liverworts and hornworts, which are considered absent in mosses. This study aimed to report the presence of AMF in a community of bryophytes (mosses and liverworts) from Punta Lara Natural Reserve, Argentina. Senescent and green sections of gametophytes were stained and, following microscopic observation, revealed AMF structures. We found intracellular hyphae, vesicles, spores and sporocarps associated with thallus and rhizoids of mosses and liverworts and senescent moss caulidia. The morphological characterization of spores resulted in the determination of Rhizophagus intraradices and Dominikia aurea. The species D. aurea is reported for the first time for Argentina. Sequencing of the D1 variable domain of the LSUrDNA from AMF spores mixes plus hyphae resulted in high similitude to the Dominikia sequences available from NCBI. This study reported the presence of AMF associated with declining and senescent gametophytes of bryophytes (mosses and liverworts) in a Natural Reserve in Argentina. These findings open up new lines of study, which should further investigate these associations and their diversity, physiology and significance.

Diversity of arbuscular mycorrhizal fungi and root colonization in Polygonatum verticillatum

Abstract. Kumar A, Tapwal A. 2022. Diversity of arbuscular mycorrhizal fungi and root colonization in Polygonatum verticillatum. Nusantara Bioscience 14: 53-63. Diversity of arbuscular mycorrhizal fungi (AMF) associated with Polygonatum verticillatum (L.) All. was investigated in two sites of Himachal Himalaya, India. A total of 15 AMF species were isolated and identified from the rhizosphere soil of P. verticillatum. The spore density was 1.48 ± 1.91 and 3.99 ± 3.78 per 20 gram of rhizosphere soil at site-I and site-II, respectively. Mycorrhizal colonization in the roots of P. verticillatum was recorded 46.12 and 52.23 percent at site-I and site-II, respectively. The mycorrhizal structures like darkly stained endophytic hyphae, coiled intracellular hyphae, Y-shaped hyphae and ‘H’ connection (Arum type) were recorded.

RNA-Seq and Gene Regulatory Network Analyses Uncover Candidate Genes in the Early Defense to Two Hemibiotrophic Colletorichum spp. in Strawberry.

Two hemibiotrophic pathogens, Colletotrichum acutatum (Ca) and C. gloeosporioides (Cg), cause anthracnose fruit rot and anthracnose crown rot in strawberry (Fragaria × ananassa Duchesne), respectively. Both Ca and Cg can initially infect through a brief biotrophic phase, which is associated with the production of intracellular primary hyphae that can infect host cells without causing cell death and establishing hemibiotrophic infection (HBI) or quiescent (latent infections) in leaf tissues. The Ca and Cg HBI in nurseries and subsequent distribution of asymptomatic infected transplants to fruit production fields is the major source of anthracnose epidemics in North Carolina. In the absence of complete resistance, strawberry varieties with good fruit quality showing rate-reducing resistance have frequently been used as a source of resistance to Ca and Cg. However, the molecular mechanisms underlying the rate-reducing resistance or susceptibility to Ca and Cg are still unknown. We performed comparative transcriptome analyses to examine how rate-reducing resistant genotype NCS 10-147 and susceptible genotype ‘Chandler’ respond to Ca and Cg and identify molecular events between 0 and 48 h after the pathogen-inoculated and mock-inoculated leaf tissues. Although plant response to both Ca and Cg at the same timepoint was not similar, more genes in the resistant interaction were upregulated at 24 hpi with Ca compared with those at 48 hpi. In contrast, a few genes were upregulated in the resistant interaction at 48 hpi with Cg. Resistance response to both Ca and Cg was associated with upregulation of MLP-like protein 44, LRR receptor-like serine/threonine-protein kinase, and auxin signaling pathway, whereas susceptibility was linked to modulation of the phenylpropanoid pathway. Gene regulatory network inference analysis revealed candidate transcription factors (TFs) such as GATA5 and MYB-10, and their downstream targets were upregulated in resistant interactions. Our results provide valuable insights into transcriptional changes during resistant and susceptible interactions, which can further facilitate assessing candidate genes necessary for resistance to two hemibiotrophic Colletotrichum spp. in strawberry.

Karakteristik Mikoriza Arbuskula Tanaman Serai Wangi (Cymbopogon nardus L.) di Lapangan Ternaungi dan Tidak Ternaungi

Arbuscular mycorrhiza (AM) characteristics of citronella grass in the field have not been reported. This research aimed to study the AM characteristics of citronella grass grown in unshaded and shaded fields. The roots of citronella grass were collected from citronella grass plantations in Cianjur, West Java. The root samples were analyzed for AM structures, namely entry points, intercellular hyphae, arbuscules, and vesicles. The results showed that the citronella grass form AM colonization. The quality of root colonization differed between the two cultivation systems. The unshaded citronella grass had higher root colonization compared to shaded citronella grass. In the unshaded citronella grass, the number of arbuscules was 7 per cm of root length, whereas in the shaded citronella grass was 4 per cm of root length. The types of arbuscules observed were arum and intermediate. There were no differences in the number of entry points in the two cultivated systems, which was 3,5 entry points per cm of root length. The numbers of vesicles and internal hyphae in unshaded citronella grass were lower than that of in the shaded citronella grass. In the unshaded citronella grass, the number of vesicles and intracellular hyphae were 1,5 and 8,5 per cm root length, whereas in the shaded citronella grass were 3,5 and 11 per cm root length, respectively. Shading plants grown in the field were bamboo, banana, coffee, tea, and sugar palm. All the shading plants formed AM symbiosis with a colonization value of 7 to 30%. This research indicates that arbuscular mycorrhiza is an important component in the citronella grass cultivation in unshaded and shaded fields.
 
 Keywords: Arbuscule, entry point, intercellular hyphae, root colonization, vesicle

Advances in the Role of Dark Septate Endophytes in the Plant Resistance to Abiotic and Biotic Stresses.

Endophytic fungi have been studied in recent decades to understand how they interact with their hosts, the types of relationships they establish, and the potential effects of this interaction. Dark septate endophytes (DSE) are isolated from healthy plants and form melanised structures in the roots, including inter- and intracellular hyphae and microsclerotia, causing low host specificity and covering a wide geographic range. Many studies have revealed beneficial relationships between DSE and their hosts, such as enhanced plant growth, nutrient uptake, and resistance to biotic and abiotic stress. Furthermore, in recent decades, studies have revealed the ability of DSE to mitigate the negative effects of crop diseases, thereby highlighting DSE as potential biocontrol agents of plant diseases (BCAs). Given the importance of these fungi in nature, this article is a review of the role of DSE as BCAs. The findings of increasing numbers of studies on these fungi and their relationships with their plant hosts are also discussed to enable their use as a tool for the integrated management of crop diseases and pests.

Epigenetic responses to Phytophthora citrophthora gummosis in citrus

Studies show that DNA methylation is associated with plant immunity but little is known as to how this epigenetic mechanism assists plants in adjusting their responses to biotic stress, especially when interacting with an hemibiotrophic pathogen such as citrus Phytophthora. The aim of the present study was to assess the effects of scion-rootstock interaction on plant resistance to P. citrophthora infection and DNA methylation patterns in ‘Pera’ sweet orange and ‘Tahiti’ acid lime grafted onto ‘Rangpur’ lime and ‘Tropical’ sunki rootstocks reinoculated with P. citrophthora. Results showed that reinoculated plants of the ‘Pera’ sweet orange/‘Rangpur’ lime and ‘Tahiti’ acid lime/‘Tropical’ sunki combinations with more and less sensitive varieties to Phytophthora, presented smaller stem lesions and increased frequency of full methylation and hemimethylation rates, compared to inoculated plants. In contrast, ‘Tahiti’ acid lime/‘Rangpur’ lime, two highly sensitive varieties, and ‘Pera’/‘Tropical’ sunki, two much less sensitive varieties, showed high increases in the frequency of hemimethylation and non-methylation levels. Results suggest that in citrus, both the scion-rootstock interaction and DNA methylation affect the response to P. citrophthora infection. Reinoculated plants, depending on the combination, showed changes in intracellular hyphae growth through the formation of sets of fibers and crystal accumulation in the periderm, cortex, and phloem. In addition, starch grain concentration was higher in reinoculated plants in comparison to inoculated plants. These findings support the assumption that DNA methylation is a plant defense mechanism and therefore may be exploited to improve the response of plants to the gummosis of P. citrophthora in citrus.

Distinct phosphoinositides define the biotrophic interface of plant–microbe interactions

Distinct phosphoinositides define the biotrophic interface of plant–microbe interactions

A Histopathological Study Reveals New Insights Into Responses of Chestnut (Castanea spp.) to Root Infection by Phytophthora cinnamomi.

The European chestnut (Castanea sativa) is threatened by the hemibiotrophic oomycete Phytophthora cinnamomi, the causal agent of ink disease. Chestnut species have different susceptibility levels to P. cinnamomi, with the Asian species (C. crenata; C. mollissima) exhibiting the highest level of resistance. A histological approach was used to study the responses exhibited by susceptible and resistant chestnut genotypes by characterizing the early stages of P. cinnamomi infection and the cellular responses it induces in roots. C. sativa (susceptible) and C. crenata (resistant) plantlets were inoculated with a P. cinnamomi virulent isolate with a zoospore suspension or by direct contact with mycelia agar pieces. Root samples were collected at 0.5, 3.5, 24, 48, and 72 h after inoculation (hai) for microscopic observations. Penetration was observed in both species at 0.5 and 3.5 hai with mycelium and zoospore inoculations, respectively. In both inoculation methods, following penetration into the rhizodermis, P. cinnamomi hyphae grew inter- and intracellularly through the cortex and into the vascular cylinder. C. crenata cells displayed a delay in the pattern of infection by having fewer cell layers colonized compared with C. sativa. At 72 hai, the collapse of the first layers of C. sativa cortical cells was observed, indicating the beginning of necrotrophy. C. crenata was able to respond more efficiently to P. cinnamomi than C. sativa by restricting the pathogen's growth area through the early activation of resistance responses such as callose deposition around some intracellular hyphae, hypersensitive response-like cell death, cell wall thickening, and accumulation of phenolic-like compounds.

Ceratocystis fimbriata Employs a Unique Infection Strategy Targeting Peltate Glandular Trichomes of Sweetpotato (Ipomoea batatas) Plants.

The infection processes of Ceratocystis fimbriata BMPZ13 (BMPZ13) was elucidated on vegetative tissues of sweetpotato plants employing light and scanning electron microscopy. Vegetative tissues infected with C. fimbriata BMPZ13 by either wounding or nonwounding inoculation methods developed typical disease symptoms, establishing black rot in stems and necrosis on buds, young leaves, and stems of sprouts, in addition to wilt on leaves and shoot cuttings, typical of vascular associated diseases. The runner hyphae of C. fimbriata BMPZ13 formed from germinated conidia were able to directly penetrate the epidermal cuticle for initial infection and invade sweetpotato peltate glandular trichomes, specialized secretory structures to store and secrete metabolites. A two-step biotrophic phase was observed with nonwounding inoculation on leaves and stems, featuring both intercellular and intracellular invasive hyphae, with the latter found within living cells of the leaf epidermis. Subsequent to the biotrophic phase was a necrotrophic phase displaying cell death in infected leaves and veins. Additionally, this cell death was an iron-associated ferroptosis, supporting the notion that iron is involved in the necrotrophic phase of C. fimbriata BMPZ13 infection. Significantly, we establish that C. fimbriata employs a unique infection strategy: the targeting of peltate glandular trichomes. Collectively, our findings show that C. fimbriata is a plant fungal pathogen with a hemibiotrophic infection style in sweetpotato vegetative tissues.

Complexity of Brassica oleracea-Alternaria brassicicola Susceptible Interaction Reveals Downregulation of Photosynthesis at Ultrastructural, Transcriptional, and Physiological Levels.

Black spot disease, caused by Alternaria brassicicola in Brassica species, is one of the most devastating diseases all over the world, especially since there is no known fully resistant Brassica cultivar. In this study, the visualization of black spot disease development on Brassica oleracea var. capitata f. alba (white cabbage) leaves and subsequent ultrastructural, molecular and physiological investigations were conducted. Inter- and intracellular hyphae growth within leaf tissues led to the loss of host cell integrity and various levels of organelle disintegration. Severe symptoms of chloroplast damage included the degeneration of chloroplast envelope and grana, and the loss of electron denseness by stroma at the advanced stage of infection. Transcriptional profiling of infected leaves revealed that photosynthesis was the most negatively regulated biological process. However, in infected leaves, chlorophyll and carotenoid content did not decrease until 48 hpi, and several chlorophyll a fluorescence parameters, such as photosystem II quantum yield (Fv/Fm), non-photochemical quenching (NPQ), or plant vitality parameter (Rdf) decreased significantly at 24 and 48 hpi compared to control leaves. Our results indicate that the initial stages of interaction between B. oleracea and A. brassicicola are not uniform within an inoculation site and show a complexity of host responses and fungal attempts to overcome host cell defense mechanisms. The downregulation of photosynthesis at the early stage of this susceptible interaction suggests that it may be a part of a host defense strategy, or, alternatively, that chloroplasts are targets for the unknown virulence factor(s) of A. brassicicola. However, the observed decrease of photosynthetic efficiency at the later stages of infection is a result of the fungus-induced necrotic lesion expansion.

Synthesis and ultrastructural observation of arbutoid mycorrhizae of black truffles (Tuber melanosporum and T. aestivum)

Arbutus unedo (the strawberry tree) is a Mediterranean shrub which forms arbutoid mycorrhizae with a variety of Asco- and Basidiomycetes. After the discovery of the mycorrhizal symbiosis between A. unedo and Tuber borchii, in this study, arbutoid mycorrhizae were synthetized in greenhouse with Tuber aestivum and Tuber melanosporum. Six months after inoculation, both species colonized the roots of all inoculated A. unedo seedlings, but mature mycorrhizae were only observed after 12 months. Ultrastructure analysis of Tuber arbutoid mycorrhizae was described for the first time, showing, as observed in typical endosymbiosis, a rearrangement of host cells and the creation of an interface compartment with both truffle species. Immunolabelling experiments suggested that pectins are not present in the interface matrix surrounding the intracellular hyphae. Thus, the ability to establish symbiosis with A. unedo seems to be a common feature in the genus Tuber, opening up the possibility to use this plant for mycorrhization with valuable truffles. This could represent an important economic opportunity in Mediterranean areas by combining the production of truffles, edible fruits and valued honey.

Ultrastructural changes during the symbiotic seed germination of Gastrodia elata with fungi, with emphasis on the fungal colonization region

BackgroundGastrodia elata is a fully mycoheterotrophic orchid and has long been used in traditional Chinese medicine. The life cycle of G. elata requires an association with two different fungi-Mycena for seed germination and Armillaria for tuber growth. The association with Armillaria is representative of the phytophagous type of orchid mycorrhiza: the intracellular hyphae are lysed without forming condensed pelotons. However, whether the association with Mycena during seed germination belongs to the same type of orchid mycorrhiza is unknown.ResultsHistological and ultrastructural studies revealed several notable features in different developmental stages. First, a thickened cell wall with papillae-like structures appeared during fungal penetration in the suspensor end cell, epidermal cells and cortical cells of germinating embryos. In addition, the formation of two distinctive cell types in the colonized region of a protocorm (i.e., the passage canal cell filled with actively growing fungal hyphae) can be observed in the epidermal cell, and the distinctive digestion cell with a dense cytoplasm appears in the cortex. Finally, within the digestion cell, numerous electron-dense tubules form a radial system and attach to degrading fungal hyphae. The fungal hyphae appear to be digested through endocytosis.ConclusionsThe present study provides important structural evidence for the phytophagous type of orchid mycorrhiza in the symbiotic germination of G. elata with Mycena. This case demonstrates a particular nutrient transfer network between G. elata and its litter-decaying fungal partner.

TPLATE Recruitment Reveals Endocytic Dynamics at Sites of Symbiotic Interface Assembly in Arbuscular Mycorrhizal Interactions.

Introduction: Arbuscular mycorrhizal (AM) symbiosis between soil fungi and the majority of plants is based on a mutualistic exchange of organic and inorganic nutrients. This takes place inside root cortical cells that harbor an arbuscule: a highly branched intracellular fungal hypha enveloped by an extension of the host cell membrane—the perifungal membrane—which outlines a specialized symbiotic interface compartment. The perifungal membrane develops around each intracellular hypha as the symbiotic fungus proceeds across the root tissues; its biogenesis is the result of an extensive exocytic process and shows a few similarities with cell plate insertion which occurs at the end of somatic cytokinesis.Materials and Methods: We here analyzed the subcellular localization of a GFP fusion with TPLATE, a subunit of the endocytic TPLATE complex (TPC), a central actor in plant clathrin-mediated endocytosis with a role in cell plate anchoring with the parental plasma membrane.Results: Our observations demonstrate that Daucus carota and Medicago truncatula root organ cultures expressing a 35S::AtTPLATE-GFP construct accumulate strong fluorescent green signal at sites of symbiotic interface construction, along recently formed perifungal membranes and at sites of cell-to-cell hyphal passage between adjacent cortical cells, where the perifungal membrane fuses with the plasmalemma.Discussion: Our results strongly suggest that TPC-mediated endocytic processes are active during perifungal membrane interface biogenesis—alongside exocytic transport. This novel conclusion, which might be correlated to the accumulation of late endosomes in the vicinity of the developing interface, hints at the involvement of TPC-dependent membrane remodeling during the intracellular accommodation of AM fungi.

Spatiotemporal nature of Fusarium graminearum-wheat coleoptile interactions

Fusarium head blight (FHB) caused by the ascomycete fungus Fusarium graminearum can result in significant crop losses and render the crops harmful to human health due to contamination with mycotoxin. Although the pathogenesis of F. graminearum is widely investigated by molecular genetics approaches, detailed studies about its cellular and developmental processes at the initial stages of infection are very limited. We applied live-cell imaging approach to characterize the spatial and temporal development of growing hyphae and plant responses during F. graminearum and wheat coleoptile interactions. The present investigation demonstrates that F. graminearum uses two strategies to penetrate the host epidermal cells. The pathogen breaks through the host cell wall with appressoria-like structures derived from surficial hyphae, and also with the narrow pegs from thick and bulbous intracellular hyphae. Live cell imaging in the presence of the endocytic tracker FM4–64 showed that the plasma and intermembranes of the invaded wheat coleoptile cells were intact. Invasive hyphae exhibit branching, budding, pseudohyphae-like growth, cell-to-cell spreading ability, and were sealed within a plant membrane, indicating a biotrophic lifestyle of this fungus inside the invaded cells. Time-lapse imaging suggested that there were callose depositions at the plant cell walls in the form of continuous lines and also on the outer linings of the fungal invasive hyphae at colonization stage. In addition, our studies demonstrate that the activation of the toxisome-related gene (TRI4) requires external stimuli and is spatio-temporally modulated. Generally, this study provides new insights into the colonization strategies and host response features during F. graminearum-plant interactions. Further tracing of cellular details will significantly contribute to our understanding of molecular mechanisms of F. graminearum-host interactions.

Life at the edge – the cytology and physiology of the biotroph to necrotroph transition in Hymenoscyphus fraxineus during lesion formation in ash

The progress of colonization of ash stems from ascospore inocula of Hymenoscyphus fraxineus was examined by light and electron microscopy. The main aim of the study was to characterize the cytology of the biotroph to necrotroph transition during lesion formation. Following direct penetration into epidermal cells, the fungus produced intracellular hyphae that invaded up to five cells before plant cells died. A lack of close attachment between the hyphal cell wall and plant cell membrane was revealed by plasmolysis of epidermal cells. Plant cells died at the centre of the infection but hyphae at the edge were typically found in living plant cells even around large lesions. During biotrophic invasion, the cytoplasm of penetrated plant cells showed very little response despite the plant cell membrane being in direct contact with the fungal cell wall. Before plant cell death, dark staining of the cytoplasm and proliferation of small vesicles was noted, but organelles retained normal ultrastructure. Dead plant cells contained dark brown, osmiophilic droplets. Penetration between epidermal or collenchyma cells was usually targeted to shared pits and involved constriction of hyphae. The transition to necrotrophy was not associated with a clear change in hyphal morphology. Biotrophic intracellular hyphae contained dense cytoplasm but hyphae in dead plant cells were more vacuolated. Remarkably little plant cell wall degradation was observed despite the fungus penetrating up to 18 cells deep into stem tissue. Features of the development of the ash dieback fungus are compared with other hemibiotrophic pathogens.

Arbuscular Mycorrhizal and Dark Septate Endophyte Fungal Association in Cassava (Manihot esculenta Crantz) Varieties, Southern India

The study examined the root characteristics, type of arbuscular mycorrhizal (AM) and dark septate endophyte (DSE) fungal associations and their extent of colonization in 13 widely cultivated cassava (Manihot esculenta Crantz) varieties in southern India. Root and associated soil samples of each cassava variety were collected from Salem and Kolli Hills of the Eastern Ghats in south India. In addition, we also investigated the relationship between the extent of colonization of AM, DSE fungal and AM fungal spore communities and soil characteristics for each cassava variety. Roots of all the cassava varieties were colonized by AM fungi and 12 varieties had co-colonization of DSE fungi. The AM morphology in all the cassava varieties was of an intermediate type with intercellular and intracellular linear hyphae and arbusculate coils. In addition, we found high AM colonization rates (73-89%) but only moderate rates by DSE (22-68%) fungi. We also found 17 AM fungal spore morphotypes and their abundance varied significantly among cassava varieties. In addition, different physicochemical properties of soils may possibly increase AMF diversity, while different varieties of cassava are commonly associated with differing levels of AM and DSE fungi colonization with soil factors potentially influencing the symbiosis.

Histopathology of infection and colonisation of Elsinoë ampelina on grapevine leaves

Elsinoe ampelina, the causal agent of grapevine anthracnose, infects all aboveground succulent parts of the plant, causing serious yield losses. There are no histopathological reports on this pathogen interaction with grapevine tissues. This study aimed at understanding the infection and colonisation processes of E. ampelina on Vitis labrusca cv. Niagara Rosada leaves by light, transmission and scanning microscopy. From one to five germ tubes per conidia were formed during conidia germination, predominantly one or two. Cuticle degradation was observed on plant surface underneath germ tubes and appressorium-like structures were rare. Monilioid and branched hyphae were observed 48 h post inoculation. Direct penetration occurred only on the adaxial leaf surface due to high trichome density on the abaxial leaf surface and conidia trapped on trichomes were frequently observed. The colonisation process was similar in the leaf blade, veins and petiole with the formation of inter- and intracellular hyphae. Phenolic compounds accumulated in infected areas; however, they did not prevent the colonisation progress. Tissues of foliar blade were totally changed by cell collapse and conidiogenous cells were observed on the surface of necrotic lesions. Ascus-like structures were observed within deeply unstructured tissues of leaf veins and necrotic areas of the petiole.

Structural plasticity in root-fungal symbioses: diverse interactions lead to improved plant fitness.

Root-fungal symbioses such as mycorrhizas and endophytes are key components of terrestrial ecosystems. Diverse in trophy habits (obligate, facultative or hemi-biotrophs) and symbiotic relations (from mutualism to parasitism), these associations also show great variability in their root colonization and nutritional strategies. Specialized interface structures such as arbuscules and Hartig nets are formed by certain associations while others are restricted to non-specialized intercellular or intracellular hyphae in roots. In either case, there are documented examples of active nutrient exchange, reinforcing the fact that specialized structures used to define specific mycorrhizal associations are not essential for reciprocal exchange of nutrients and plant growth promotion. In feremycorrhiza (with Austroboletus occidentalis and eucalypts), the fungal partner markedly enhances plant growth and nutrient acquisition without colonizing roots, emphasizing that a conventional focus on structural form of associations may have resulted in important functional components of rhizospheres being overlooked. In support of this viewpoint, mycobiome studies using the state-of-the-art DNA sequencing technologies have unearthed much more complexity in root-fungal relationships than those discovered using the traditional morphology-based approaches. In this review, we explore the existing literature and most recent findings surrounding structure, functioning, and ecology of root-fungal symbiosis, which highlight the fact that plant fitness can be altered by taxonomically/ecologically diverse fungal symbionts regardless of root colonization and interface specialization. Furthermore, transition from saprotrophy to biotrophy seems to be a common event that occurs in diverse fungal lineages (consisting of root endophytes, soil saprotrophs, wood decayers etc.), and which may be accompanied by development of specialized interface structures and/or mycorrhiza-like effects on plant growth and nutrition.