Ultrastructural analysis of responses of host and fungal cells during plant infection

Cellular responses in fungi and in susceptible or resistant hosts during fungus–plant interactions have been studied ultrastructurally to examine their role in pathogenicity. Pathogenicity is determined in some saprophytic fungi by various factors: the production of disease determinants such as the production of host-specific toxins (HSTs) or the extracellular matrix (ECM) by fungal infection structures and H₂O₂ generation from penetration pegs. Three different target sites for HSTs have been identified in host cells in many ultrastructural studies: plasma membranes, chloroplasts, and mitochondria. The mode of action of HSTs is characterized by the partial destruction of the target structures only in susceptible genotypes of host plants, with the result that the fungus can colonize the host. The infection structures of most fungal pathogen secrete ECM on plant surfaces during fungal differentiation, while the penetration pegs of some pathogens produce reactive oxygen species (ROS) in the cell walls and plasma membranes. The pathological roles of ECM and H₂O₂ generation are discussed here in light of ultrastructural evidence. Host and fungal characteristics in the incompatible interactions include the rapid formation of lignin in host epidermal cell walls, failure of penetration pegs to invade lignin-fortified pectin layers, the inhibition of subcuticular hyphal proliferation and the collapse of hyphae that have degraded cell walls within pectin layers of the host. Apoptosis-like host resistant mechanism is also discussed.     

Effect of temperature and duration of wetness during initial infection periods on disease development, fungal biomass and mycotoxin concentrations on wheat inoculated with single, or combinations of, Fusarium species

Experiments were conducted under controlled environment conditions to study the relationship between environmental conditions, development of fusarium head blight (FHB) and mycotoxin production. A single isolate from each of four Fusarium species ( F. avenaceum , F. culmorum , F. graminearum and F. poae ) was used to inoculate wheat ears separately. Combinations of two or three isolates were also used to inoculate ears simultaneously. Inoculated ears were subjected to various combinations of duration of wetness (6–48 h) and temperature (10–30°C). For all inoculations, both incidence of spikelets with FHB symptoms and concentration of mycotoxins generally increased with increasing length of wetness period and temperature. There were significant positive correlations among disease incidence, fungal biomass (quantified as total amount of fungal DNA) and mycotoxins. Mycotoxin production was also greatly enhanced by high temperatures (≥ 20°C) during initial infection periods. In single-isolate inoculations, F. poae was the least aggressive. There was no evidence to support synergetic interactions between fungal isolates in causing visual symptoms; rather the results suggest, in most cases, the presence of competitive interactions. Furthermore, the competition led to large reductions in fungal biomass compared to single-isolate inoculations, often > 90% reduction for the weaker isolate(s). In contrast, mycotoxin productivity increased dramatically in the co-inoculations, by as much as 1000 times, suggesting that competition resulted in greater production of trichothecene mycotoxins. The F. graminearum isolate was most competitive and isolates of the other three species were similar in their competitiveness.     

Molecular basis and regulatory mechanisms underlying fungal insecticides' resistance to solar ultraviolet irradiation

Resistance to solar ultraviolet (UV) irradiation is crucial for field-persistent control efficacies of fungal formulations against arthropod pests, because their active ingredients are formulated conidia very sensitive to solar UV wavelengths. This review seeks to summarize advances in studies aiming to quantify, understand and improve conidial UV resistance. One focus of studies has been on the many sets of genes that have been revealed in the postgenomic era to contribute to or mediate UV resistance in the insect pathogens serving as main sources of fungal insecticides. Such genetic studies have unveiled the broad basis of UV-resistant molecules including cytosolic solutes, cell wall components, various antioxidant enzymes, and numerous effectors and signaling proteins, that function in developmental, biosynthetic and stress-responsive pathways. Another focus has been on the molecular basis and regulatory mechanisms underlying photorepair of UV-induced DNA lesions and photoreactivation of UV-impaired conidia. Studies have shed light upon a photoprotective mechanism depending on not only one or two photorepair-required photolyases, but also two white collar proteins and other partners that play similar or more important roles in photorepair via interactions with photolyases. Research hotspots are suggested to explore a regulatory network of fungal photoprotection and to improve the development and application strategies of UV-resistant fungal insecticides. © 2021 Society of Chemical Industry.     

Fungal-based bioherbicides for weed control: a myth or a reality?

The use of bioherbicides containing fungal active ingredients or natural fungal molecules is one of the possible solutions to reduce the use of chemical products. This paper focuses on studies of bioherbicides, including both living fungi and natural fungal molecules, published in the last 45 years, and their associated weed targets; current problems in the development of bioherbicides are also discussed. Bibliometric methods based on the Web of Science database were used to analyse relevant articles published between 1973 and 2018. Overall analysis suggested that interest in bioherbicides extends over the preceding thirty years, when many potential microorganisms and natural fungal molecules were proposed. Furthermore, analysis of about 229 articles indicated an encouraging exploitable potential, although there is a real gap between the number of experimental studies and the small number of products currently on the market. A dozen fungal-based bioherbicides are on the market in the United States and Canada, while countries, such as China and South Africa, have one, and none is available in Europe. The active ingredients in these bioherbicides are living fungi, but no fungal molecule-based product is thus far on the market. Reasons for this gap include production hurdles, formulation process, ecological fitness, duration of herbicidal effects, and costly and time-consuming registration procedures. However, it is clear that analysis of fungus–plant interactions provides a promising source of bioherbicides that may be applied to appropriate cropping systems for environment-friendly, sustainable weed control.     

Dynamics of infection-related morphogenesis and pathogenesis in Colletotrichum orbiculare

Colletotrichum orbiculare (syn. C. lagenarium) is the causal agent of anthracnose disease on cucurbit plants. This fungus forms dome-shaped, melanized appressoria as a host invasion structure. Strain 104-T (MAFF240422) of C. orbiculare, which was originally isolated from a cucumber plant in 1951 by Dr. Yasumori, Kyoto University, has proven to be an excellent experimental model for the study of fungal pathogenesis and morphogenesis because of its stable pathogenicity and synchronous infection-related morphogenesis. This review considers the discoveries made during 60 years of study on C. orbiculare. In particular, we focus on advances made in the last two decades, which have provided a basis for the molecular analysis not only of fungal morphogenesis, but also of plant–microbe interactions, including plant immunity to adapted and nonadapted Colletotrichum fungi. This substantial body of innovative research was originated by the Phytopathological Society of Japan and represents a major contribution to the international research communities working on plant pathology, plant–microbe interactions, and fungal molecular genetics. This review deals with the past achievements and future prospects in the study of Colletotrichum biology, focusing on the molecular genetics of C. orbiculare with regard to four aspects: (1) metabolic and functional development of infection structures, (2) signaling pathways required for fungal pathogenesis, (3) pathogen-associated molecular patterns (PAMPs) and host basal resistance, and (4) establishment of host specificity.     

Fungal communities in organically grown winter wheat affected by plant organ and development stage

The fungal community on the roots, stem bases, stems and grains of organically grown winter wheat was analysed using terminal restriction fragment length polymorphism (T-RFLP) combined with cloning and sequencing of the ITS region. The changes in the composition of fungi in different plant parts and over time as well as interactions between fungi were also investigated. Among 58 fungal taxa found the most common were Davidiella macrospora, Cladosporium spp., Tetracladium maxilliforme, Didymella exitialis, Microdochium nivale and an unidentified species within Ascomycetes. Several potential wheat pathogens were found: Fusarium spp. including F. poae and G. avenacea (F. avenaceum), Microdochium nivale, Oculimacula yallundae, Parastagonospora nodorum and Zymoseptoria tritici and most of them were present on all plant parts. Plant part affected the most the fungal colonization of wheat as was shown both by multivariate analysis of the whole fungal community as well as the analysis based on the identified species. The composition of fungal communities in different parts changed during the growing season but no pattern common for the whole crop could be observed. The most dynamic and significant changes were found among yeasts. Both positive and negative significant interactions between pairwise combinations of pathogens were observed. Positive significant associations were also found between pathogens and other fungi.     

黄龙病对柑橘叶际微生物组的影响

为明确柑橘叶际微生物组对黄龙病发生的响应规律，采用荧光定量PCR技术检测采集柑橘叶片感染黄龙病菌的情况，通过扩增子测序分析法对PCR检测结果为阴性和阳性叶片的微生物群落进行比较，并通过相关性分析和网络分析对黄龙病菌和叶际微生物的相关性进行解析。结果显示，夏季采集的柑橘叶片中黄龙病菌的检出率为7.5%，而到秋季黄龙病菌的检出率上升到32.3%。黄龙病的发生显著改变了柑橘叶际细菌群落的组成和结构，对于细菌类群的丰度也有一定影响；而对叶际真菌群落的组成和结构无显著影响。黄龙病菌与叶际细菌存在普遍的负相关关系，正相关关系相对较少但相关性更强。黄龙病菌可能通过与厌氧棍状菌Anaerotruncus sp.、梭菌Clostridiales sp.、假单胞菌Pseudomonas sp.、鞘脂单胞菌Sphingomonas sp.、毛螺菌Lachnospiraceae sp.和链球菌Streptococcus sp.的负向互作对柑橘叶际整个细菌群落产生负影响。表明黄龙病发生对柑橘叶际真菌和细菌群落的影响规律不同，同时黄龙病菌可能通过与几种主要细菌的负向互作来实现对叶际微生物组的影响。     

Grapevine trunk diseases: complex and still poorly understood

This review presents an overview of eutypa dieback, esca and botryosphaeria dieback, the predominant grapevine trunk diseases worldwide. It covers their symptomatologies in the trunk, leaves and berries; the characteristics of the different fungal species associated with them; and host–pathogen interactions. Here, the host–pathogen relationship is defined at the cytological, physiological and molecular levels. Currently available experimental tools for studying these diseases, both in vitro and in the field, are discussed. Finally, a progress report on their control, which, since the ban of sodium arsenite, comprises chemical, biological and/or sanitation methods, is presented.     

Pathogen behaviour and plant cell reactions in interactions between Alternaria species and leaves of host and nonhost plants

The growth behaviour of several Alternaria species on leaf discs of host and nonhost plants was examined by u.v. microscopy. Specific patterns of development were found for the pathogens. Differences in development which correlated to host specificity, or which differentiated specialized pathogens from weak or opportunistic pathogens, were obvious only after attempted penetration. Thus, little variation was observed amongst different plants for individual Alternaria species in the rate of germination, in the extent of germ-tube growth or in the frequency of appressorium production. However, plant responses to attempted penetration, which included the formation of callose-containing papillae, callose deposition in the walls of attacked cells and their neighbours and cell necrosis, varied with specific pathogen–plant interactions. Callose deposition occurred at sites of both successful and unsuccessful penetrations and may, therefore, not be a determining factor in the plant–pathogen interactions examined. A biplot technique is used to illustrate the different degrees of host specificity apparent at pre- and post-penetration stages of fungal development.     

Morphological characterization of the interaction between Diplocarpon rosae and various rose species

Blackspot, caused by Diplocarpon rosae , is the most severe and ubiquitous disease of garden roses, but information is lacking about genotype-specific forms of resistance and susceptibility of the host. Macro- and microscopic analyses of 34 rose genotypes with a defined monoconidial culture black spot inoculum identified susceptible and resistant rose genotypes and further genotype-specific subdivisions, indicating the presence of partial forms of resistance and different resistance mechanisms. In total, eight interaction types were characterized, five representing compatible (types 1–5) and three representing incompatible interactions (types 6–8). The incompatible interactions were characterized by the lack of any visible fungal structures beneath the cuticle (type 8), single-cell necroses (type 7) or necroses of larger cell clusters (type 6), the latter two types with penetration hyphae and haustoria in epidermal cells.     

The potential role of arbuscular mycorrhizal (AM) fungi in the bioprotection of plants against soil-borne pathogens in organic and/or other sustainable farming systems

Sustainable farming systems strive to minimise the use of synthetic pesticides and to optimise the use of alternative management strategies to control soil-borne pathogens. Arbuscular mycorrhizal (AM) fungi are ubiquitous in nature and constitute an integral component of terrestrial ecosystems, forming symbiotic associations with plant root systems of over 80% of all terrestrial plant species, including many agronomically important species. AM fungi are particularly important in organic and/or sustainable farming systems that rely on biological processes rather than agrochemicals to control plant diseases. Of particular importance is the bioprotection conferred to plants against many soil-borne pathogens such as species of Aphanomyces, Cylindrocladium, Fusarium, Macrophomina, Phytophthora, Pythium, Rhizoctonia, Sclerotinium, Verticillium and Thielaviopsis and various nematodes by AM fungal colonisation of the plant root. However, the exact mechanisms by which AM fungal colonisation confers the protective effect are not completely understood, but a greater understanding of these beneficial interactions is necessary for the exploitation of AM fungi within organic and/or sustainable farming systems. In this review, we aim to discuss the potential mechanisms by which AM fungi may contribute to bioprotection against plant soil-borne pathogens. Bioprotection within AM fungal-colonised plants is the outcome of complex interactions between plants, pathogens and AM fungi. The use of molecular tools in the study of these multifaceted interactions may aid the optimisation of the bioprotective responses and their utility within sustainable farming systems.     

昆虫体内不同微生物间互作关系的研究进展

昆虫在某些发育阶段可感染不同种类的共生微生物,这些共生物在宿主生长、繁殖、适应环境及进化的过程中起重要作用。大量研究发现,昆虫体内的不同共生微生物并不是孤立存在和发生作用的,而是相互间不同程度地存在某些联系。本文以共生细菌为重点,综述了最近10多年来有关不同种共生细菌之间的关系,及共生细菌与其他昆虫相关微生物(病原细菌、真菌和病毒及昆虫共生真菌)之间相互关系的研究进展。其中,重点回顾了不同细菌在昆虫体内的共存和对宿主功能上的协作,昆虫中已经发生或可能正在发生的细菌取代,以及共生细菌对宿主的病原物敏感性的影响等方面的研究。最后对各种生物学技术大量应用于共生细菌研究的背景下此领域的研究前景进行了讨论。     

稻纹枯病菌菌株与水稻品种的致病互作分析

 用离体叶接种方法进行稻纹枯病菌群体致病力和水稻抗病力测定,将结果进行统计学分析及图解分析发现,病菌群体的致病力呈现连续变异的数量性状特征,服从正态分布;菌株与品种的致病互作中,菌株间的差异、品种间的差异、以及菌株-品种互作间的差异均达到极显著水平,但互作效应对致病反应的变异所起的作用很小,致病反应的变异主要由菌株效应的差异或者品种效应的差异造成;大多数菌株在品种“中浙优1号”上的致病力排序与在“特优524”上的排序趋势是一致的;大多数品种对菌株2008-126-2的抗病力排序与对菌株2008-88-1的排序趋势是一致的。依结果认为寄主水稻的抗病力也服从正态分布。基于这些分布特征和致病互作特征,可以认为,一般要体现病害差异的试验,水稻寄主应选用中等抗病力的品种,病菌应选用中等致病力的菌株。抗病育种实践中,进行品种资源抗病性评价时,可采用二步行策略,第一步先用一个中等致病力菌株对所有品种先行鉴定,第二步用一个强致病力（或弱致病力）菌株对高抗病力（或低抗病力）的品种再行鉴定。同理,病菌材料的致病性评价也可采用二步行策略,第一步先用一个中等抗病力品种对所有菌株先行鉴定,第二步用一个高抗病力（或低抗病力）品种对强致病力（或弱致病力）的菌株再行鉴定。品种抗病性（或菌株的致病性）的鉴定结果可以采用正态分布函数计算法进行分组归类。     

Spatiotemporal changes in fungal growth and host responses of six yellow rust resistant near-isogenic lines of wheat

The objective of the present study was to investigate to what extent the macroscopic phenotype of incompatible host–pathogen interactions reflects differences in fungal development and host responses at the histological level. This was done by conventional and advanced microscopic analysis of six wheat near-isogenic lines differing by individual R genes and inoculated with an avirulent isolate of Puccinia striiformis. Wheat line AvocetYr15 had the lowest macroscopic infection type (IT) 0–1, in which fungal growth was stopped at early stages due to extensive expression of a hypersensitive response (HR) at all time points (4, 8 and 16 days post-inoculation, dpi) without any sign of haustorial bodies. AvocetYr5 and AvocetYr1 had IT 1, in which most fungal colonies developed secondary hyphae. Many colonies were encased by HR at 16 dpi, more extensively in AvocetYr1 than AvocetYr5. In AvocetYr6 (IT 2), HR was expressed after the formation of secondary hyphae at 4 dpi, after which fungal growth and HR increased simultaneously until most colonies became encased by HR. AvocetYr27 (IT 2–3) and AvocetYr17 (IT 4–5) showed similar fungal growth and HR at 4 dpi, where HR was only weakly expressed in a few host cells. Encasement of secondary and runner hyphae increased significantly in AvocetYr27, but at 16 dpi, HR was often circumvented by large, intermingled fungal colonies in both lines. No resistance responses were observed in Avocet S (susceptible control). The very different histological patterns conferred by the six R genes suggests differences in the timing of the host–pathogen recognition and onset of host defence pathways.     

The development of different pathotype groups of Mycosphaerella pinocles in susceptible and partially resistant pea leaves

The progress of infection by high- and low-virulence isolates of Mycosphaerella pinodes was examined in susceptible and partially resistant pea leaves. Conidia germinated with one or more germ tubes which frequently branched and formed appressorium-like structures on the leaf surface. Penetration occurred through the epidermal walls. A structure similar to an infection vesicle was formed, lying partly in the epidermal wail and partly in the cell lumen. From this structure, a penetration hypha was derived which initiated the development of the intra- and intercellularly-growing fungal colony. Infections led to rapid tissue collapse in both susceptible and resistant interactions. In resistant interactions, the formation of infection vesicles and penetration hyphae was reduced, and the development and spread of lesions was retarded.     

Interaction of a bioherbicide and glyphosate for controlling hemp sesbania in glyphosate-resistant soybean

The bioherbicidal fungus, Colletotrichum truncatum (Schwein.) Andrus & Moore, was tested at different inoculum concentrations alone and in combination with, prior to or following treatment with different rates of glyphosate ( N -[phosphonomethyl]glycine) (Roundup Ultra) for the control of hemp sesbania ( Sesbania exaltata [Raf.] Rydb. ex A.W. Hill) in Roundup Ready soybean field plots. Colletotrichum truncatum and glyphosate were applied in all pair-wise combinations of 0, 1.25, 2.5, 5.0, and 10.0 × 10⁶ spores mL⁻¹ (i.e. 3.125, 6.25, 12.5, and 25 × 10¹¹ spores ha⁻¹), and 0.15, 0.30, 0.60, and 1.2 kg ha⁻¹, respectively. Weed control and disease incidence were enhanced at the two lowest fungal and herbicidal rates when the fungal spores were applied after glyphosate treatment. The application of the fungus in combination with or prior to glyphosate application at 0.30 kg ha⁻¹ resulted in reduced disease incidence and weed control regardless of the inoculum's concentration. At the highest glyphosate rates, the weeds were controlled by the herbicide alone. These results suggest that it might be possible to utilize additive or synergistic herbicide and pathogen interactions to enhance hemp sesbania control.     

Improved attachment and parasitism of <Emphasis Type="Italic">Trichoderma</Emphasis> on <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> in vitro

Monoclonal and polyclonal antibodies that bind to eggs and/or second-stage juveniles of the nematode Meloidogyne javanica were tested for their effects on the parasitic interactions between this nematode and the fungus Trichoderma. Parasitism of Trichoderma asperellum-203 and Trichoderma atroviride on nematode egg masses, eggs and juveniles was enhanced when antibodies were incorporated into in vitro parasitism bioassays. Parasitism on separated eggs (without gelatinous matrix) and their hatched juveniles was also improved, compared to controls without antibodies that did not attach fungal conidia. Improved parasitism could be due to bilateral binding of the antibodies to the nematodes and conidia, enabling better conidial attachment to the nematodes. Enhanced germination of antibody-bound conidia further improved parasitism. Differences were observed among antibodies in their effects on fungal parasitism and their interaction with Trichoderma species. We focused mainly on the egg- and juvenile-binding monoclonal antibody MISC that exhibited a stronger reaction with T. asperellum-203 than with T. atroviride. Pretreatment of this antibody with fucose inhibited its binding to nematodes and conidial attachment to nematodes, as well as conidial agglutination in the presence of the antibody. Antibody binding to juveniles affected their movement and viability, especially gelatinous matrix-originated juveniles. The fucose-specific lectin Ulex europaeus-I enhanced conidial attachment to nematode life-stages, and conidial agglutination occurred in its presence. These phenomena were inhibited by preincubating lectin with fucose. Our results suggest that carbohydrate residues, such as fucose, on the surface of the nematode and fungal conidia are involved in the antibody- and lectin-mediated improved parasitism.     

Fungal endophyte protects Atractylodes lancea from root rot caused by Fusarium oxysporum

Endophytic fungi, which stimulate a variety of defence reactions in host plants without causing visible disease symptoms, have been isolated from almost every plant. However, beneficial interactions between fungal endophytes and pathogens from the same habitat remain largely unknown. An inoculation of Atractylodes lancea plantlets with Gilmaniella sp. AL12 (AL12) prior to infection with Fusarium oxysporum prevented the necrotization of root tissues and plant growth retardation commonly associated with fusarium root rot. Quantification of F. oxysporum infections using real‐time PCR revealed a correlation between root rot symptoms and the relative amount of fungal DNA. Pretreatment with AL12 reduced the accumulation of reactive oxygen species stimulated by F. oxysporum. An in vitro analysis of their interactions under axenic culture conditions showed AL12 could inhibit F. oxysporum growth. Additionally, F. oxysporum infections were shown to decrease salicylic acid (SA) production compared with control plantlets. SA biosynthesis inhibitors, 2‐aminoindan‐2‐phosphonic acid and paclobutrazol, abolished the inhibition of F. oxysporum growth in A. lancea even after inoculation with AL12. The results indicated that the fungal endophyte protected A. lancea not only by direct antibiosis, but also by reversing the F. oxysporum‐mediated suppression of SA production.     

Factors influencing growth,sporulation and virus transfer in <Emphasis Type="Italic">Cryphonectria parasitica</Emphasis> isolates from Castilla and León (Spain)

Biological control with Cryphonectria hypovirus CHV1 of the chestnut blight, caused by the fungus Cryphonectria parasitica, has reduced the impact of the disease in Europe. The virus reduces the virulence of the fungus so that it causes non-lethal cankers, thus enabling the chestnut trees to overcome the disease. The virus can be transmitted horizontally by hyphal anastomosis or vertically to the conidia. In this study, we investigated growth and sporulation of the fungus as well as rates of horizontal transmission of the virus at different temperatures. We used fungal isolates of the vegetative compatibility types (vc types) that are most prevalent in Castilla and León (central northern Spain) to evaluate the effects of fungal strain on the parameters tested. In addition, we infected four isolates of C. parasitica with hypovirus subtypes CHV1-F1 and CHV1-I, to determine the influence of virus subtype on growth, sporulation and virus transfer. We assessed growth of fungal colonies and horizontal transmission of the virus at 15 °C and 25 °C. Colony growth was affected by an interaction between fungal isolates included in vc type EU1 or EU11 and virus at both 15 °C and 25 °C. However, horizontal transmission of the virus was only influenced by the fungal genotype of isolates included in vc type EU1 or EU11, and spore production was only affected by the virus subtype. Vertical transmission was also influenced by the fungal isolate and virus subtype. Growth of the fungal isolates varied depending on the virus subtype with which they were infected. This supports the theory that fungal host and virus subtype influence transmission and dissemination of hypovirulence. The fungal genotype affects colony growth and horizontal transmission of the virus. It is common to expect a good dissemination of the hypovirus with a low vc type diversity but the selection of the best combination of hypovirus and fungal isolate is crucial for the success of biological control not only for small areas but in larger chestnut populations as well.