Development of the infection strategy of the hemibiotrophic plant pathogen,Colletotrichum orbiculare,and plant immunity

The hemibiotrophic fungus Colletotrichum orbiculare forms appressoria as infection structures and primarily establishes biotrophic infection in cucumber epidermal cells. Subsequently, it develops necrotrophic infection. In the pre-invasion stage, morphogenesis of appressoria of C. orbiculare is triggered by signals from the plant surface. We found that C. orbiculare PAG1 (Perish-in-the-Absence-of-GYP1), a component of MOR [morphogenesis-related NDR (nuclear Dbf2-related) kinase network] plays an essential role as a key component of the plant-specific signaling pathway and that hydrolysis of cutin by a spore surface esterase creates a cutin monomer that constitutes a key plant-derived signal. Development of the infection structure of C. orbiculare is strictly regulated by the cell cycle and we found that proper regulation of G1/S progression via two-component GAP genes, consisting of BUB2 (Budding-Uninhibited-by-Benomyl-2) and BFA1 (Byr-Four-Alike-1) is essential for the establishment of successful infection. In the post-invasion stage, the establishment of the biotrophic phase of hemibiotrophic fungi is crucial for successful infection. We found that C. orbiculare WHI2 (WHIsky-2), an Saccharomyces cerevisiae stress regulator homolog, is involved in the phase transition from biotrophy to necrotrophy through TOR (Target of Rapamycin) signaling, and is thus essential for full pathogenesis.     

Proteomic studies of phytopathogenic fungi,oomycetes and their interactions with hosts

Proteomics, the systematic analysis of the proteome, is a powerful tool in the post-genomic era. Proteomics studies have examined global changes in proteomes of phytopathogenic fungi, oomycetes and their hosts during compatible or incompatible interactions. This article compiles proteomics reports in order to decipher the molecular mechanisms underlying fungal development (infection-related morphogenesis), fungal or oomycete—host plant interactions, and phytopathogenesis.     

Inhibitory potential of metabolites produced by Cercospora sp. strain ME202 isolated from Trifolium incarnatum against anthracnose caused by Colletotrichum orbicular

Chemical fungicides are mainly used to control Colletotrichum orbiculare, but this fungal pathogen usually develops resistance to the pesticides. Natural compounds are therefore desirable for controlling C. orbiculare. Here, a culture filtrate (CF) from ME202, a fungal strain isolated from Trifolium incarnatum in Shimane Prefecture in 2020, was tested in vitro and found to inhibit conidial germination of C. orbiculare. The inhibitory compound in ME202-CF was soluble in ethyl acetate. The ethyl acetate extract of ME202-CF (ME202-ECF) significantly inhibited in vitro conidial germination of C. orbiculare in a dose-dependent manner and lesion formation on cucumber leaves. The inhibitory compounds were heat stable at 40–121 °C and molecular mass?≥?500 and ≥?1000 Da. Thin layer chromatography–bioautography of ME202-ECF showed that the compounds inhibiting C. orbiculare growth had an Rf of 0.81 and 0.91. Furthermore, ME202 was found to be a member of the genus Cercospora through sequence analysis of the internal transcribed spacer and 5.8 S rDNA. These results suggest that secondary metabolites of Cercospora spp., such as ME202, can be used to develop new fungicides against anthracnose.

     

Molecular markers for genotyping anastomosis groups and understanding the population biology of Rhizoctonia species

Soil-borne Rhizoctonia fungi cause serious diseases in several plant species. For the classification of these fungi, the number of nuclei in a hyphal cell and the anastomosis reaction are important criteria. Although Rhizoctonia spp. has a wide host range, the causal agents have been reported to be selective for host plant families or species and lead to severe disease. Reports of new diseases, particularly in new host plants, and severe damage in agricultural fields incurred by subdivided or newly found groups of Ceratobasidium and Waitea circinata (a varied teleomorph of Rhizoctonia) have been increasing in recent years. The food production environment is altering because of climate change, introduction of potential new host plants, and heavy use of chemicals that reduce microbial diversity. These changes favor the occurrence of new diseases incurred by undefined anastomosis groups (AGs) or subgroups of Rhizoctonia spp. On the basis of the phylogenetic relationships of AGs and subgroups in Rhizoctonia spp., molecular markers for discriminating the groups of the Rhizoctonia species complex have been developed. The application of genetic markers, in the form of microsatellites or simple sequence repeats (SSR), has become increasingly important in fungal genetics. The analyses of population genetics for Rhizoctonia spp. using SSR markers elucidated the modes of sexual and asexual reproduction, phylogeographical distributions, and global migrations associated with adaptation to agroecosystems.     

Most Colletotrichum species associated with tree tomato (Solanum betaceum) and mango (Mangifera indica) crops are not host‐specific

An important constraint for crop production in Colombia is the high incidence of anthracnose caused by Colletotrichum species. Although several studies have focused on these fungi, the relationship between the different fungal species within the genus and their hosts and whether they display any host preference or host specificity has yet to be examined. In Colombia, diseases caused by Colletotrichum species are particularly severe in mango (Mangifera indica) and tree tomato (Solanum betaceum) crops. In a previous investigation, the Colletotrichum phylogenetic species attacking these crops were identified. The present study aimed to determine whether isolates collected from tree tomato and mango showed host preference or host specificity by assessing aggressiveness, spore density, latent period, and fitness of each strain on the two hosts. In the departments of Cundinamarca and Tolima, Colombia, isolates were collected from plants that presented typical anthracnose symptoms and were identified as C. acutatum, C. asianum, C. boninense, C. gloeosporioides, C. tamarilloi and C. theobromicola. Inoculation of conidia of each isolate onto both hosts showed isolates had no host preference and only the C. gloeosporioides isolate showed host specificity. However, in general, isolates produced a higher spore density when inoculated on the alternate host, which may indicate a difference in the degree of adaptation to each host. Statistical analyses of the assessed parameter values revealed that isolates use different infection strategies when infecting each host. In light of these results, the implications of using quantitative estimations of fitness when studying fungal pathogens are discussed.     

Chili anthracnose: Colletotrichum taxonomy and pathogenicity

Chili anthracnose is caused by Colletotrichum species mostly associated with the acutatum, truncatum and gloeosporioides complexes. Since 2009 the Colletotrichum taxonomy has been extensively revised based on multigene phylogenetics, which has had a large impact on the number of species known to cause anthracnose disease of chili. This review discusses (i) the taxonomy of Colletotrichum spp. infecting chili, and (ii) the impact of Colletotrichum pathotypes on breeding for resistance to anthracnose. To date, 24 Colletotrichum species have been identified as pathogens of chili anthracnose, with the three main pathogens being C. scovillei, C. truncatum and C. siamense. Identification of several pathotypes within these three Colletotrichum species, particularly pathotypes that can overcome resistance in the related Capsicum species, Ca. chinense and Ca. baccatum, will be of major concern to plant breeders as they develop resistant chili genotypes from the transfer of resistance genes from these Capsicum species into Ca. annuum. Accurate identification of the Colletotrichum species causing anthracnose and improved understanding of the biology of the Colletotrichum species and their interaction with the host will enable the application of improved integrated disease management techniques.     

Responses of cucumber cultivars to induction of systemic resistance against anthracnose by plant growth promoting fungi

Initial experiment on the reactions of five Japanese cultivars of cucumber toColletotrichum orbiculare infection in the greenhouse revealed that cv Suyo and Gibai were susceptible and moderately susceptible, respectively, while cv Shogoin fushinari and Sagami hanjiro were resistant to infection byC. orbiculare; cv Ochiai fushinari was moderately resistant. The ability of 16 plant growth promoting fungi (some isolates belonged to species ofPhoma and some non-sporulating isolates) isolated from zoysiagrass rhizospheres to induce systemic resistance in the above five cucumber cultivars was tested by growing plants in potting medium infested with barley grain inocula of PGPF in the greenhouse. The second true leaves of 21-day-old plants were challenge inoculated withC. orbiculare and disease assessed. Nine, out of 16 isolates, caused significant reduction of disease caused byC. orbiculare in at least two cultivars.Phoma isolates (GS8-1 and GS8-2) and non-sporulating isolates (GU21-2, GU23-3, and GU24-3) significantly reduced the disease in all the five cultivars. The disease suppression in cucumber was due to the induction of systemic resistance, since the inducer(s) and the pathogen were separated spatially and that the inducer did not colonize aerial portions. The resistance induced by certain isolates in a susceptible cultivar was less than that in a resistant cultivar. Disease suppression caused by isolate GU21-2 was similar to theC. orbiculare induced control in certain cultivars. The average rate of expansion of lesion diameter on leaves due toC. orbiculare was slower due to induction with the selected plant growth promoting fungi compared to the uninduced control plants. Roots of four cultivars were colonized by only three isolates, however, roots of one cultivar (Suyo) was colonized by five isolates suggesting the cultivar-specific root colonization ability.Abbreviations cv cultivar(s) - PGPF plant growth promoting fungal isolates - PGPR plant growth promoting rhizobacteria     

Nonhost resistance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> against <Emphasis Type="Italic">Colletotrichum</Emphasis> species

Arabidopsis thaliana exhibits a durable resistance called nonhost resistance against nonadapted fungal pathogens. A. thaliana activates preinvasive resistance and terminates entry attempts by nonadapted fungi belonging to the genus Colletotrichum, which cause anthracnose disease in many plants. In the interaction between A. thaliana and nonadapted C. tropicale, the preinvasive resistance involves the PENETRATION 2-related antifungal secondary metabolite pathway and the ENHANCED DISEASE RESISTANCE 1-dependent antifungal peptide pathway. The development of invasive hyphae by C. tropicale owing to the reduction of preinvasive resistance then triggers the blockage of further hyphal expansion via the activation of the second layer of resistance, i.e., postinvasive resistance, which guarantees the robustness of the nonhost resistance of A. thaliana against Colletotrichum pathogens. Both the tryptophan-derived metabolic pathway and glutathione synthesis play critical roles in the postinvasive resistance against C. tropicale, although the molecular mechanism of postinvasive resistance remains to be elucidated. In this review, we describe the current understanding of the molecular background of the Arabidopsis nonhost resistance against Colletotrichum fungi and discuss perspectives for future research on this durable resistance.     

Cyanobacteria mediated plant growth promotion and bioprotection against Fusarium wilt in tomato

Cyanobacteria - phytopathogenic fungi - tomato plant interactions were evaluated for developing suitable biological options for combating biotic stress (Fusarium wilt) and enhancing plant vigour. Preliminary evaluation was undertaken on the fungicidal and hydrolytic enzyme activity of the cyanobacterial strains (Anabaena variabilis RPAN59, A. laxa RPAN8) under optimized environmental/nutritional conditions, followed by amendment in compost-vermiculite. Such formulations were tested against Fusarium wilt challenged tomato plants, and the Anabaena spp. (RPAN59/8) amended composts significantly reduced mortality in fungi challenged treatments, besides fungal load in soil. Cyanobacteria amended composts also led to an enhancement in soil organic C, nitrogen fixation, besides significant improvement in growth, yield, fruit quality parameters, N, P and Zn content. The tripartite interactions also enhanced the activity of defence and pathogenesis related enzymes in tomato plants. A positive correlation (r?=?0.729 to 0.828) between P content and pathogenesis/defense enzyme activity revealed their role in enhancing the resistance of the plant through improved nutrient uptake. Light and scanning electron microscopy (SEM) revealed cyanobacterial colonization, which positively correlated with reduced fungal populations. The reduced disease severity coupled with improved plant growth/ yields, elicited by cyanobacterial treatments, illustrated the utility of such novel formulations in integrated pest and nutrient management strategies for Fusarium wilt challenged tomato crop.     

Alternaria host-selective toxins: determinant factors of plant disease

Seven diseases caused by pathotypes of Alternaria alternata, which produced host-selective toxins (HSTs), a diverse group of low-molecular-weight secondary metabolites, are known, and each HST has an essential role as a determinant of pathogenicity in all interactions between the plant host and A. alternata. Although these HST-producing pathotypes are morphologically indistinguishable, each has a distinct host range and can be distinguished by its specificity on the respective host plant, hence their designation as pathotypes of A. alternata. In 1933, Tanaka made the first discovery of a HST; fungus-free culture filtrates of A. kikuchiana (now called A. alternata Japanese pear pathotype) were toxic to susceptible cultivar Nijisseiki, but not to resistant cultivars. Over the 80 years since then, the structure of HST molecules, target sites and mode of actions of HSTs, and the molecular genetics of HST production regulating by supernumerary chromosomes encoding HST gene clusters have been studied extensively. We focus this review on studies of low-molecular-weight HSTs produced by A. alternata and give an overview of various types of HST studies.     

Phenolic compounds as defence response of pepper fruits to Colletotrichum coccodes

Qualitative and quantitative changes of individual and total phenolics induced by Colletotrichum coccodes fungal infection have been studied in two susceptible sweet pepper cultivars ‘Soroksari’ and ‘Bagoly’, and the role of soluble phenolic compounds in plant's defence mechanism has been evaluated. Three distinct parts were analysed on pepper fruit: healthy tissue, anthracnose lesion, and bordering tissue, and individual phenolic compounds have been identified with the use of HPLC-MS system. In pepper fruit pericarp 21 phenolic compounds have been determined; the prevalent apigenin, quercetin and luteolin glycosides, chlorogenic acid and one chrysoeriol glucoside. C. coccodes infection increased the accumulation of chlorogenic acid, chrysoeriol glucoside, quercetin and luteolin glycosides in infected bordering tissue of both analysed pepper cultivars compared to healthy pepper tissue or symptomatic spot. Total apigenin derivatives did not show a significant increase in bordering tissue compared to the healthy pepper fruit in contrast to other groups of phenolics. This suggests a lesser role of apigenin glycosides in pepper plant defence against the Colletotrichum fungus. Intense phenolic synthesis was characteristic for the bordering zone between the healthy and infected plant tissue resulting in higher total phenolic content which might hinder the fungus to spread from the infected cells into the healthy tissue.     

Molecular and phenotypic characterization revealed six <Emphasis Type="Italic">Colletotrichum</Emphasis> species responsible for anthracnose disease of small cardamom in South India

Small cardamom (Elettaria cardamomum Maton) is extensively cultivated in the Western Ghats of South India either as a monocrop under the forest trees or as an intercrop along with arecanut and coffee plantations. Colletotrichum species responsible for severe outbreaks of anthracnose on small cardamom in South India are reported. Small cardamom anthracnose, popularly known as “Chenthal”, manifests itself on the foliage as yellowish lesions, which later coalesce to form large blighted areas. In advanced stages, the affected leaves dry up giving a burnt appearance to the plant. Twenty-five isolates of Colletotrichum were isolated from leaves of small cardamom in Karnataka, Kerala and Tamil Nadu states of India. The isolates were characterized through morphological studies and multilocus phylogenetic analysis (ITS, ACT, CHS-1, GAPDH, TUB2, CYLH3, GS and ApMat gene regions) to test whether different species are present and identified: C. karstii (2 isolates), C. gloeosporioides (1), C. siamense (7), C. syzygicola (6), Colletotrichum sp (5), and C. guajavae (4), as the cause of anthracnose on small cardamom for the first time. Pathogenicity of the six species was confirmed. To our knowledge, this is the first detailed study of Colletotrichum species which cause anthracnose diseases on small cardamom.     

中国茶树炭疽菌属病害研究进展及展望

茶树是一种重要的经济作物,广泛种植于热带和亚热带地区。随着茶产品经济价值的提高,茶叶生产安全备受关注。炭疽菌是引起茶树病害的病原菌之一,严重影响茶叶产量和茶产品品质,威胁我国茶产业的良性发展。利用抗病品种是防治茶树病害最为经济、有效的措施。目前,茶树炭疽菌种名使用混乱,茶树与炭疽菌的互作研究也较为薄弱,而简单依靠药剂防治病害也对茶叶质量安全造成一定的影响。本文梳理并明确了危害茶树的炭疽菌种类;总结了现有的茶树和炭疽菌互作的研究成果,提出识别免疫(effector triggered immunity,ETI)模式是茶树防御炭疽菌的主要机制;对茶树病理学存在的问题进行了归纳并提出解决办法,并展望了深入开展茶树抗病机理研究的思路,以期为制定茶园绿色防控技术及选育茶树抗病良种提供科学参考。     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of Indian mulberry (Morinda citrifolia) in tropical islands of Andaman and Nicobar, India

This study identifies the Colletotrichum spp. which cause Morinda anthracnose in the Andaman and Nicobar Islands, India. A survey of Indian mulberry plantations was carried out to ascertain the identity of isolates associated with Morinda anthracnose. These were identified as C. gloeosporioides based on morphology and sequencing analysis of internal transcribed spacer (ITS) region, demonstrating that this species is currently the causal agent of Morinda anthracnose in a tropical ecosystem. The pathogenicity test of C. gloeosporioides isolates on Morinda seedlings for 1?month revealed that the five isolates MC1, MC8, MC2 (1), MC4 and MC12 were pathogenic to the respective host with different levels of anthracnose lesion.     

Functional characterization of CgPBS<Subscript>2</Subscript>, a MAP kinase kinase in <Emphasis Type="Italic">Colletotrichum gloeosporioides,</Emphasis> using osmotic stress sensitivity as a selection marker

Colletotrichum leaf disease of Hever brasiliensis (rubber tree) caused by C. gloeosporioides is one of the major causes of declining rubber tree yields. Little is known about the fungal molecular characters that are important for pathogenicity on rubber tree and fungicide resistance. In this study, we cloned the CgPBS₂ gene, the key component of the Hog1 pathway which controls various aspects of osmoregulation and fungicide resistance in various fungal pathogens, including the causal agent of Colletotrichum leaf disease of rubber tree. We characterized the function of the CgPBS₂ gene by reverse genetics. Because the Hog1 pathway plays an important role in stress responses, we obtained a CgPBS₂ gene deletion mutant by PEG-mediated transformation of protoplasts after reducing the concentration of sucrose in the screening medium from 1.0 M to 0.2 M. Then, the complemented transformants and GFP-labelled CgPBS₂ gene transformants were selected directly under highly hyperosmotic medium (PDA?+?1.5 M sorbitol) without using other selectable gene markers. Phenotypic observations showed that the CgPBS₂ protein was mainly localized in the conidial cytoplasm of the CgPBS₂-GFP transformants. In addition, disruption of CgPBS₂ led to sensitivity to hyperosmosis and high salt concentration as well as resistance to the fungicide fludioxonil. No obvious difference in virulence was observed between the null mutant and the wild-type strain. These results provide insights into the role of the CgPBS₂ gene in osmotic stress, salt stress and fludioxonil resistance and suggest that osmotic stress sensitivity can be used as a selection marker.     

Virulence factors of Botrytis cinerea

Botrytis cinerea is responsible for gray mold disease in more than 200 host plant species. The infection of host plants is mediated by numerous extracellular enzymes, proteins and metabolites. Each of these compounds may play a role in different stages of the infection process. Cell wall-degrading enzymes may facilitate the penetration into the host surface, while toxins, oxalic acid and reactive oxygen species may contribute to killing of the host cells. Cell wall-degrading enzymes contribute to the conversion of host tissue into fungal biomass. On the other hand, B. cinerea infection induces biosynthesis of phytoalexins. Therefore, the ability to overcome a wide spectrum of phytoalexins contributes to the pathogenicity of the fungus with a broad host range. The cloning of the corresponding genes has facilitated studies on gene expression and targeted mutagenesis. This review gives an overview of the research performed on virulence factors that play the roles in pathogenesis.     

Factors influencing biological traits and aggressiveness of Colletotrichum species associated with cashew anthracnose in Brazil

Anthracnose is the main fungal disease on cashew orchards in Brazil, occurring on both vegetative and reproductive organs of cultivated and noncultivated host plants. Understanding the effect of physical and chemical exogenous factors on the biological traits of Colletotrichum spp. and determining their host range are key to developing appropriate anthracnose control measures. The present study aimed to estimate the optimum temperatures for mycelial growth, sporulation, and conidial germination of seven Colletotrichum species (C. chrysophilum, C. fragariae, C. fructicola, C. gloeosporioides, C. queenslandicum, C. siamense, and C. tropicale) associated with cashew anthracnose in Brazil. Their aggressiveness on cashew leaves and six alternative host fruits, and their sensitivity to three fungicides were also investigated. The optimum temperatures for mycelial growth, sporulation, and conidial germination ranged from about 25 to about 33°C. All Colletotrichum species induced anthracnose symptoms on wounded cashew leaves, but none of them caused lesions on intact leaf surfaces. The Colletotrichum species, except for C. fragariae and C. fructicola, were pathogenic to wounded fruits of avocado, banana, guava, mango, and papaya, and some isolates also produced lesions on nonwounded fruit tissues. No symptoms were observed on passion fruits, regardless of the inoculation method. Mycelial growth, sporulation, conidial germination, and/or appressorial formation of the seven Colletotrichum species were inhibited by azoxystrobin, difenoconazole, and thiophanate-methyl to varying degrees. The present study will contribute to the development of forecasting models based on prevailing weather of cashew cropping zones and improve cashew anthracnose management in Brazil.     

Phylogenetic and morphological identification of Colletotrichum boninense: a novel causal agent of anthracnose in avocado

In recent years, anthracnose has become a significant disease affecting avocado fruit in the state of Michoacan, Mexico, where it significantly reduces fruit quality and commercial yield. Anthracnose has been assumed to involve Colletotrichum gloeosporioides and C. acutatum as causal agents. However, because of the increasing incidence of anthracnose, a more precise identification of the Colletotrichum spp. involved in this disease has become desirable. During the years 2004–2007, avocado fruits of different sizes exhibiting brown‐black and reddish spots on the pericarp and soft rot in the mesocarp, were gathered from orchards in nine counties. Fungal isolates were cultured on potato dextrose agar, and among these, 31 were selected for molecular, morphological and pathogenicity analyses. The molecular approaches used sequence typing of the internal transcribed spacer region and the partial nuclear large ribosomal subunit, allowing the unequivocal identification of C. gloeosporioides (71%), C. acutatum (16%) and C. boninense (13%). This last species has not been previously reported as being associated with anthracnose symptoms in avocado fruits anywhere in the world. Various morphological characteristics such as the size and shape of conidia were determined, as well as the conidial mass colour. Pathogenicity tests performed with all three species were conducted by inoculating healthy fruits. In each case, identical symptoms developed within 3 days of inoculation. Knowledge of the Colletotrichum populations in the Michoacan state, including the newly encountered avocado pathogen C. boninense, will facilitate further studies addressing the relationships between these Colletotrichum spp. and their avocado host.