The fungus Acremonium alternatum enhances salt stress tolerance by regulating host redox homeostasis and phytohormone signaling.

While endophytic fungi offer promising avenues for bolstering plant resilience against abiotic stressors, the molecular mechanisms behind this biofortification remain largely unknown. This study employed a multifaceted approach, combining plant physiology, proteomic, metabolomic, and targeted hormonal analyses to illuminate the early response of Brassica napus to Acremonium alternatum during the nascent stages of their interaction. Notably, under optimal growth conditions, the initial reaction to fungus was relatively subtle, with no visible alterations in plant phenotype and only minor impacts on the proteome and metabolome. Interestingly, the identified proteins associated with the Acremonium response included TUDOR 1, Annexin D4, and a plastidic K+ efflux antiporter, hinting at potential processes that could counter abiotic stressors, particularly salt stress. Subsequent experiments validated this hypothesis, showcasing significantly enhanced growth in Acremonium-inoculated plants under salt stress. Molecular analyses revealed a profound impact on the plant's proteome, with over 50% of salt stress response proteins remaining unaffected in inoculated plants. Acremonium modulated ribosomal proteins, increased abundance of photosynthetic proteins, enhanced ROS metabolism, accumulation of V-ATPase, altered abundances of various metabolic enzymes, and possibly promoted abscisic acid signaling. Subsequent analyses validated the accumulation of this hormone and its enhanced signaling. Collectively, these findings indicate that Acremonium promotes salt tolerance by orchestrating abscisic acid signaling, priming the plant's antioxidant system, as evidenced by the accumulation of ROS-scavenging metabolites and alterations in ROS metabolism, leading to lowered ROS levels and enhanced photosynthesis. Additionally, it modulates ion sequestration through V-ATPase accumulation, potentially contributing to the observed decrease in chloride content.

Characterization of Endophytic Fungi, Acremonium sp., from Lilium davidii and Analysis of Its Antifungal and Plant Growth-Promoting Effects.

The present study was aimed at isolating endophytic fungi from the Asian culinary and medicinal plant Lilium davidii and analyzing its antifungal and plant growth-promoting effects. In this study, the fungal endophyte Acremonium sp. Ld-03 was isolated from the bulbs of L. davidii and identified through morphological and molecular analysis. The molecular and morphological analysis confirmed the endophytic fungal strain as Acremonium sp. Ld-03. Antifungal effects of Ld-03 were observed against Fusarium oxysporum, Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi. The highest growth inhibition, i.e., 78.39 ± 4.21%, was observed for B. dothidea followed by 56.68 ± 4.38%, 43.62 ± 3.81%, and 20.12 ± 2.45% for B. cinerea, F. fujikuroi, and F. oxysporum, respectively. Analysis of the ethyl acetate fraction through UHPLC-LTQ-IT-MS/MS revealed putative secondary metabolites which included xanthurenic acid, valyl aspartic acid, gancidin W, peptides, and cyclic dipeptides such as valylarginine, cyclo-[L-(4-hydroxy-Pro)-L-leu], cyclo(Pro-Phe), and (3S,6S)-3-benzyl-6-(4-hydroxybenzyl)piperazine-2,5-dione. Other metabolites included (S)-3-(4-hydroxyphenyl)-2-((S)-pyrrolidine-2-carboxamido)propanoic acid, dibutyl phthalate (DBP), 9-octadecenamide, D-erythro-C18-Sphingosine, N-palmitoyl sphinganine, and hydroxypalmitoyl sphinganine. The strain Ld-03 showed indole acetic acid (IAA) production with or without the application of exogenous tryptophan. The IAA ranged from 53.12 ± 3.20 µg ml-1 to 167.71 ± 7.12 µg ml-1 under different tryptophan concentrations. The strain was able to produce siderophore, and its production was significantly decreased with increasing Fe(III) citrate concentrations in the medium. The endophytic fungal strain also showed production of organic acids and phosphate solubilization activity. Plant growth-promoting effects of the strain were evaluated on in vitro seedling growth of Allium tuberosum. Application of 40% culture dilution resulted in a significant increase in root and shoot length, i.e., 24.03 ± 2.71 mm and 37.27 ± 1.86 mm, respectively, compared to nontreated control plants. The fungal endophyte Ld-03 demonstrated the potential of conferring disease resistance and plant growth promotion. Therefore, we conclude that the isolated Acremonium sp. Ld-03 should be further investigated before utilization as a biocontrol agent and plant growth stimulator.

Genera Acremonium and Sarocladium Cause Brown Spot on Bagged Apple Fruit in China.

Apple fruit spot disease has caused serious economic losses for years in China since the widespread application of fruit bagging in production. Although the three genera Trichothecium, Alternaria, and Acremonium have been reported to be the causal agents, studies on the disease etiology and pathogen biology are still sparse. Here, we report characterization of eight fungal isolates from lesions on 126 symptomatic fruit samples collected in Shaanxi Province, China. Pathogenicity of the isolates was assessed. DNA sequences were obtained at four loci, including D1/D2 domains of the large-subunit nrRNA gene, internal transcribed spacer regions 1 and 2, 5.8S nrDAN gene, a fragment of the actin gene, and a fragment of the ß-tubulin. Based on phylogenetic analysis and morphological features, three new species were found: Acremonium mali, Sarocladium liquanensis, and Sarocladium mali. In addition, we made the first report of Sarocladium terricola as a plant pathogen. Temperature and moisture significantly affected in vitro conidial germination of five Acremonium-like species, and their impact on infection of apple fruit was tested using Acremonium sclerotigenum. Conidia of five species germinated from 15 to 35°C in free water; four of the species had optimum temperature around 25°C, whereas conidia of S. terricola had an optimum temperature of 30°C. Conidial germination rate increased as relative humidity (RH) increased. The five isolates had relatively high conidial germination rates at RH > 97%, with a significant decline at 95% RH. Incidence of infection also increased in proportion to RH. In free water, conidial germination was relatively unaffected by temperature.

Insecticidal Activities of Chloramphenicol Derivatives Isolated from a Marine Alga-Derived Endophytic Fungus, Acremonium vitellinum, against the Cotton Bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

A great deal of attention has been focused on the secondary metabolites produced by marine endophytic fungi, which can be better alternatives to chemicals, such as biopesticides, for control of polyphagous pests. On the basis of its novel biocontrol attributes, chemical investigation of a marine alga-derived endophytic fungus, Acremonium vitellinum, resulted in the isolation of three chloramphenicol derivatives (compounds 1⁻3). Their chemical structures were elucidated by detailed analysis of their nuclear magnetic resonance spectra, high-resolution electrospray ionization mass spectrometry, and by comparison with the data available in the literature. In this paper, compound 2 was firstly reported as the natural origin of these fungal secondary metabolites. The insecticidal activities of compounds 1⁻3 against the cotton bollworm, Helicoverpa armigera, were evaluated. The natural compound 2 presented considerable activity against H. armigera, with an LC50 value of 0.56 ± 0.03 mg/mL (compared to matrine with an LC50 value of 0.24 ± 0.01 mg/mL). Transcriptome sequencing was used to evaluate the molecular mechanism of the insecticidal activities. The results presented in this study should be useful for developing compound 2 as a novel, ecofriendly and safe biopesticide.

The Elicitor Protein AsES Induces a Systemic Acquired Resistance Response Accompanied by Systemic Microbursts and Micro-Hypersensitive Responses in Fragaria ananassa.

The elicitor AsES (Acremonium strictum elicitor subtilisin) is a 34-kDa subtilisin-like protein secreted by the opportunistic fungus Acremonium strictum. AsES activates innate immunity and confers resistance against anthracnose and gray mold diseases in strawberry plants (Fragaria × ananassa Duch.) and the last disease also in Arabidopsis. In the present work, we show that, upon AsES recognition, a cascade of defense responses is activated, including: calcium influx, biphasic oxidative burst (O2⋅- and H2O2), hypersensitive cell-death response (HR), accumulation of autofluorescent compounds, cell-wall reinforcement with callose and lignin deposition, salicylic acid accumulation, and expression of defense-related genes, such as FaPR1, FaPG1, FaMYB30, FaRBOH-D, FaRBOH-F, FaCHI23, and FaFLS. All these responses occurred following a spatial and temporal program, first induced in infiltrated leaflets (local acquired resistance), spreading out to untreated lateral leaflets, and later, to distal leaves (systemic acquired resistance). After AsES treatment, macro-HR and macro-oxidative bursts were localized in infiltrated leaflets, while micro-HRs and microbursts occurred later in untreated leaves, being confined to a single cell or a cluster of a few epidermal cells that differentiated from the surrounding ones. The differentiated cells initiated a time-dependent series of physiological and anatomical changes, evolving to idioblasts accumulating H2O2 and autofluorescent compounds that blast, delivering its content into surrounding cells. This kind of systemic cell-death process in plants is described for the first time in response to a single elicitor. All data presented in this study suggest that AsES has the potential to activate a wide spectrum of biochemical and molecular defense responses in F. ananassa that may explain the induced protection toward pathogens of opposite lifestyle, like hemibiotrophic and necrotrophic fungi.

Is the root-colonizing endophyte Acremonium strictum an ericoid mycorrhizal fungus?

In previous investigations, we found that Acremonium strictum (strain DSM 100709) developed intracellular structures with similarity to mycelia of ericoid mycorrhizal fungi in the rhizodermal cells of flax plants and in hair roots of Rhododendron plantlets. A. strictum had also been isolated from roots of ericaceous salal plants and was described as an unusual ericoid mycorrhizal fungus (ERMF). As its mycorrhizal traits were doubted, we revised the hypothesis of a mycorrhizal nature of A. strictum. A successful synthesis of mycorrhiza in hair roots of inoculated ericaceous plants was a first step of evidence, followed by fluorescence microscopy with FUN(®)1 cell stain to observe the vitality of the host cells at the early infection stage. In inoculation trials with in vitro-raised mycorrhiza-free Rhododendron plants in axenic liquid culture and in greenhouse substrate culture, A. strictum was never observed in living hair root cells. As compared to the ERMF Oidiodendron maius and Rhizoscyphus ericae that invaded metabolically active host cells and established a symbiotic unit, A. strictum was only found in cells that were dead or in the process of dying and in the apoplast. In conclusion, A. strictum does not behave like a common ERMF-if it is one at all. A comparison of A. strictum isolates from ericaceous and non-ericaceous hosts could reveal further identity details to generalize or specify our findings on the symbiotic nature of A. strictum. At least, the staining method enables to discern between true mycorrhizal and other root endophytes-a tool for further studies.

Transformation of the endophytic fungus Acremonium implicatum with GFP and evaluation of its biocontrol effect against Meloidogyne incognita.

Acremonium implicatum is an endophytic fungus with biocontrol potential against Meloidogyne incognita based on its opportunistic egg-parasitic, hatching inhibition, and toxic properties. To understand its mode of plant endophytism and opportunistic egg parasitism, GFP-tagged A. implicatum was constructed by PEG-mediated protoplast transformation. By laser scanning confocal microscopy (LSCM), we evaluated the endophytism and opportunistic egg parasitism of a stable gfp transformant (Acr-1). Acr-1 could colonize epidermal tissue, cortical tissue, and xylem of roots and form a mutualistic symbiosis with tomato host plants. LSCM of Acr-1 infecting M. incognita eggs revealed that hyphae penetrated the shell and grew inside eggs to form trophic hyphae. A large number of hyphae enveloped parasitized eggs. In addition, the egg shell integrity was destroyed by fungal penetration. The percentage of egg parasitism was 33.8 %. There were no marked differences between the wild type and mutant in nematode second-stage juvenile mortality and egg hatching and in fungal control efficiency in a pot experiment. In conclusion, gfp-transformation did not change the nematicidal activity of A. implicatum and is a tool to examine the mode of plant endophytism and opportunistic egg parasitism of A. implicatum.

Consequences of antagonistic interactions between endophytic fungus and bacterium on plant growth and defense responses in Atractylodes lancea.

Many studies have examined pair-wise interactions between plants and endophytes, while overlooking the interplays among multiple endosymbionts and their combined impacts on hosts. In this study, Atractylodes lancea plantlets were inoculated with endophytic fungus Acremonium strictum AL16, or endophytic bacterium Acinetobacter sp., or both, to investigate the impacts of the three-way symbiosis on the host. Our results showed that defense-related responses of the co-inoculated plantlets were delayed and weakened relative to plantlets with single inoculants, but no detrimental effects on phyto-physiology (growth, photosynthesis) were observed after combined inoculations. Quantitative PCR analysis verified a decrease in AL16 colonization density within plants after co-inoculation with the endobacteria. An in vitro assay was then performed to elucidate the suppressed plant defense responses and reduced fungal colonization by dual inoculation. The data showed that the presence of Acinetobacter sp. reduced AL16 colony diameter and spore germination rate without negatively affecting fungal morphology. Additionally, direct hyphal attachment of the bacterium to AL16 in vitro was visualized by scanning electronic microscopy. Therefore, we propose that a balanced and compatible symbiosis might require constraints conferred by the antagonistic endophyte Acinetobacter sp. on the fungus AL16 in the tripartite endophytic bacterium-fungus-plant system.

Drought degree constrains the beneficial effects of a fungal endophyte on Atractylodes lancea.

AIMS: Plants, fungal endophytes (FEs) and the changing environment interact with each other forming an interlaced network. This study evaluates nonadditive and interactive effects of the FE Acremonium strictum and drought treatment on Atractylodes lancea plantlets. METHODS AND RESULTS: By applying FEs (meristem cultures of At. lancea, fungal inoculation of Ac. strictum and plantlet acclimatization) and drought treatment (regular watering, mild drought, severe drought), a research system of At. lancea ramets under different treatments was established. During 12 days of drought treatment, the plantlets' physiological responses and basic growth traits were measured and analysed. Although drought and FE presence affected plantlet traits to differing degrees, the interactive effects of the two were more pronounced. In particular under mild drought treatment, the FE conferred drought tolerance to plantlets by enhancing leaf soluble sugars, proteins, proline and antioxidant enzyme activity; decreasing the degree of plasmalemma oxidation; and increasing the host's abscisic acid level and root:shoot ratio. When exposed to regular watering or severe drought, these effects were not significant. CONCLUSIONS: Plant traits plasticity was conferred by dual effects of drought stress and FEs, and these factors are interactive. Although FEs can help plants cope with drought stress, the beneficial effects are strictly constrained by drought degree. SIGNIFICANCE AND IMPACT OF THE STUDY: During finite environmental stress, FEs can benefit plants, and for this reason, they may alleviate the effects of climate change on plants. However, because the benefits of FEs are highly context dependent, the role of FEs in a changing background should be re-assessed.

The autophagy-related gene Acatg1 is involved in conidiation and cephalosporin production in Acremonium chrysogenum.

Autophagy is a highly conserved eukaryotic mechanism for degradation of cellular components and nutrient recycling process. A serine/threonine kinase Atg1 is essential for autophagosome formation under starvation. Acatg1, the homologous gene of atg1 was cloned from the cephalosporin producing fungus Acremonium chrysogenum. Disruption of Acatg1 inhibited the autophagosome formation under starvation and significantly reduced conidia formation. However, exogenous supply of glucose, sucrose, mannitol or inositol restored the conidia formation of the Acatg1 disruption mutant to the wild-type level, suggesting that autophagy is involved in the carbon utilization which is required for cell growth and morphological differentiation. Unexpectedly, the Acatg1 disruption mutant showed strong resistance to exogenous hydrogen peroxide comparing with the wild-type strain. Disruption of Acatg1 also enhanced cephalosporin production at the late stage of fermentation. Consistent with cephalosporin production, the transcription of cephalosporin biosynthetic genes was increased in the Acatg1 disruption mutant and Western blotting demonstrated that the isopenicillin N synthase PcbC involved in cephalosporin biosynthesis was retained at the late stage of fermentation in the Acatg1 disruption mutant while it was sharply reduced in the wild-type strain. These results indicated that Acatg1 plays an important role in both autophagosome formation and cephalosporin production of A. chrysogenum.

Fungal ovicidal activity on Toxocara canis eggs.

BACKGROUND: Visceral toxocariasis is a parasitic zoonosis caused by Toxocara canis. The prevalence of this parasite in dogs, soil contamination and the resistance of eggs increase human exposure to the disease. Moreover, the difficulties of the control measures justify the need for alternative ones. AIMS: The objective of this study was to evaluate the in vitro ovicidal activity of fungi isolated from soils from public places in the city of Pelotas, Rio Grande do Sul, Brazil, on Toxocara canis. METHODS: Samples of soil from ten localities were inoculated onto Petri dishes with 2% water-agar (WA) that contained antibiotics, and incubated at 25°C/21 days. Isolated fungi were tested in vitro for ovicidal activity, with five replicates. One mL of an embryonated Toxocara canis egg suspension (10(3) eggs) was poured over the fungal cultures after 10 days of growth. At intervals of 7, 14 and 21 days, 100 eggs were removed from each plaque and evaluated by optical microscopy. RESULTS: Acremonium, Aspergillus, Bipolaris, Fusarium, Gliocladium, Mucor and Trichoderma were isolated from the soil. A significant ovicidal type 3 effect was observed in Trichoderma, Fusarium solani complex and Acremonium. Those isolates from the genus Trichoderma showed their ovicidal effect on the 14th day of fungus-egg interaction. The other fungal genera tested showed a type 2 effect. CONCLUSIONS: These results suggest that the use of Trichoderma and Fusarium solani complex in biological control of T. canis is promising; however, further studies should be performed.

A septation related gene AcsepH in Acremonium chrysogenum is involved in the cellular differentiation and cephalosporin production.

T-DNA inserted mutants of Acremonium chrysogenum were constructed by Agrobacterium tumefaciens-mediated transformation (ATMT). One mutant 1223 which grew slowly was selected. TAIL-PCR and sequence analysis indicated that a putative septation protein encoding gene AcsepH was partially deleted in this mutant. AcsepH contains nine introns, and its deduced protein AcSEPH has a conserved serine/threonine protein kinase catalytic (S_TKc) domain at its N-terminal region. AcSEPH shows high similarity with septation H proteins from other filamentous fungi based on the phylogenetic analysis of S_TKc domains. In sporulation (LPE) medium, the conidia of AcsepH mutant was only about one-seventh of the wild-type, and more than 20% of conidia produced by the mutant contain multiple nuclei which were rare in the wild-type. During fermentation, the AcsepH disruption mutant grew slowly and its cephalosporin production was only about one quarter of the wild-type, and the transcription analysis showed that pcbC expression was delayed and the expressions of cefEF, cefD1 and cefD2 were significantly decreased. The vegetative hyphae of AcsepH mutant swelled abnormally and hardly formed the typical yeast-like cells. The amount of yeast-like cells was about one-tenth of the wild-type after fermentation for 5days. Comparison of hyphal viabilities revealed that the cells of AcsepH mutant died easily than the wild-type at the late stage of fermentation. Fluorescent stains revealed that the absence of AcsepH in A. chrysogenum led to reduction of septation and formation of multinucleate cells. These data indicates that AcsepH is required for the normal cellular septation and differentiation of A. chrysogenum, and its absence may change the cellular physiological status and causes the decline in cephalosporin production.

A novel impeller configuration to improve fungal physiology performance and energy conservation for cephalosporin C production.

Effects of impeller configuration on fungal physiology and cephalosporin C production were investigated by an industrial strain Acremonium chrysogenum in a 12 m(3) bioreactor equipped with conventional and novel impeller configuration, respectively. The cell growth and oxygen uptake rate (OUR) profiles were little affected by the impeller configurations. However, differing impeller combinations significantly affected the morphology, which in turn influenced cephalosporin C production. Under the novel impeller configuration, the production of cephalosporin C was 10% higher and an excessive amount of dispersed arthrospores was also observed. Computational fluid dynamics (CFD) simulation further revealed that poor mass and energy exchange as well as inhomogeneous environment existed in the bioreactor equipped with conventional impeller configuration. For equivalent power dissipation, the volume oxygen transfer coefficient (K(L)a) could be enhanced by 15% compared with that of conventional impeller configuration. Power consumption was dramatically decreased by 25% by using novel impeller configuration.

Aggressiveness of Cephalosporium maydis causing late wilt of maize in Spain.

Late wilt of maize, caused by the vascular and soilborne pathogen Cephalosporium maydis, was identified in the Iberian Peninsula in 2008. During the last years the incidence and economical impact of the disease has importantly increased both in Portugal and Spain. Varieties of maize displaying tolerance to the pathogen are available, but the effectiveness can be dependent on the virulence of the fungus (i.e. ability to cause disease on a specific genotype). On the other hand, strains of crop pathogens from different geographic origins can differ with regard to the degree of disease caused on a specific genotype (i.e. aggressiveness). Our working hypothesis was that isolates of C. maydis from different maize growing areas may differ in aggressiveness towards maize plants. Seven fungal strains were isolated in 2009 from diseased plants collected in the most important maize growing regions of Spain and used to inoculate two susceptible maize varieties grown in shadehouse from March to July 2010. The experimental unit consisted of two 4-day-old seedlings planted in an 8-liter pot filled with sand/silt previously infested with 200 g of wheat grains colonized by the fungi. Non colonized wheat grains were used for the control treatments. Six replications (pots) were established for each variety/isolate combination according to a complete randomized 2 x 8 factorial design. The percentage of necrotic and dry aboveground tissues was recorded 14 weeks after inoculation and thereafter weekly until physiological senescence of the control plants. At the end of the experiment, weights of roots and aboveground parts of the plants were recorded. Initial occurrence of symptoms in the plants was significantly dependent on the isolate of C. maydis and on the maize variety. However, final severity of aboveground symptoms (leaf necroses and drying up) was only dependent on the fungal isolate. All the isolates significantly reduced the root weight of both varieties of maize. The highest root weight reductions were also associated to a significant low weight of above-ground parts. Considering all the symptoms analysed and their progression in the maize plants, our results reveal that a diversity of aggressiveness exists among isolates of C. maydis. The need for a characterization of maize genotypes by their reaction against highly aggressive isolates of the fungus in the Iberian Peninsula is suggested. This study is a first step towards a recommendation of crop varieties that are tolerant to C. maydis in different areas of the Iberian Peninsula. Future research aims at studying the relationship between aggressiveness levels, molecular characteristics and geographical origin whithin C. maydis.

A novel fungal pathogen under the spotlight--Acremonium spp. associated fungaemia in an immunocompetent host.

Acremonium spp. are filamentous, cosmopolitan fungi frequently isolated from plant debris and soil, they are known to result in invasive infections in the setting of severe immunosuppression. In this letter, we present a case of catheter-related fungaemia associated with Acremonium spp. in a patient with chronic renal failure. After removal of the subclavian catheter, the patient was treated successfully with voriconazole, with a loading dose of 400 mg followed by a maintenance dose of 200 mg bid. To the best of our knowledge, this is the first paper reporting Acremonium spp. associated fungaemia in a relatively immunocompetent host. We also discuss the diagnosis and treatment of Acremonium spp. associated infections in the context of current literature.

Engineering of Penicillium chrysogenum for fermentative production of a novel carbamoylated cephem antibiotic precursor.

Penicillium chrysogenum was successfully engineered to produce a novel carbamoylated cephalosporin that can be used as a synthon for semi-synthetic cephalosporins. To this end, genes for Acremonium chrysogenum expandase/hydroxylase and Streptomyces clavuligerus carbamoyltransferase were expressed in a penicillinG high-producing strain of P.chrysogenum. Growth of the engineered strain in the presence of adipic acid resulted in production of adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid (ad7-ACCCA) and of several adipoylated pathway intermediates. A combinatorial chemostat-based transcriptome study, in which the ad7-ACCCA-producing strain and a strain lacking key genes in beta-lactam synthesis were grown in the presence and absence of adipic acid, enabled the dissection of transcriptional responses to adipic acid per se and to ad7-ACCCA production. Transcriptome analysis revealed that adipate catabolism in P.chrysogenum occurs via beta-oxidation and enabled the identification of putative genes for enzymes involved in mitochondrial and peroxisomal beta-oxidation pathways. Several of the genes that showed a specifically altered transcript level in ad7-ACCCA-producing cultures were previously implicated in oxidative stress responses.

Asexual cephalosporin C producer Acremonium chrysogenum carries a functional mating type locus.

Acremonium chrysogenum, the fungal producer of the pharmaceutically relevant beta-lactam antibiotic cephalosporin C, is classified as asexual because no direct observation of mating or meiosis has yet been reported. To assess the potential of A. chrysogenum for sexual reproduction, we screened an expressed sequence tag library from A. chrysogenum for the expression of mating type (MAT) genes, which are the key regulators of sexual reproduction. We identified two putative mating type genes that are homologues of the alpha-box domain gene, MAT1-1-1 and MAT1-1-2, encoding an HPG domain protein defined by the presence of the three invariant amino acids histidine, proline, and glycine. In addition, cDNAs encoding a putative pheromone receptor and pheromone-processing enzymes, as well as components of a pheromone response pathway, were found. Moreover, the entire A. chrysogenum MAT1-1 (AcMAT1-1) gene and regions flanking the MAT region were obtained from a genomic cosmid library, and sequence analysis revealed that in addition to AcMAT1-1-1 and AcMAT1-1-2, the AcMAT1-1 locus comprises a third mating type gene, AcMAT1-1-3, encoding a high-mobility-group domain protein. The alpha-box domain sequence of AcMAT1-1-1 was used to determine the phylogenetic relationships of A. chrysogenum to other ascomycetes. To determine the functionality of the AcMAT1-1 locus, the entire MAT locus was transferred into a MAT deletion strain of the heterothallic ascomycete Podospora anserina (the PaDeltaMAT strain). After fertilization with a P. anserina MAT1-2 (MAT(+)) strain, the corresponding transformants developed fruiting bodies with mature ascospores. Thus, the results of our functional analysis of the AcMAT1-1 locus provide strong evidence to hypothesize a sexual cycle in A. chrysogenum.

Management of root knot nematode on tomato through application of fungal antagonists, Acremonium strictum and Trichoderma harzianum.

Management of root knot nematode disease infecting tomato, by the use of fungal bioagents Acremonium strictum and Trichoderma harzianum isolated from egg masses of M. incognita infecting tomato has been carried out. The rhizosphere and rhizoplane of root knot nematode infested tomato revealed consistent association of Acremonium strictum. In the present study A. strictum and other fungal bioagents viz. Aspergillus niger, Paecilomyces lilacinus, Rhizoctonia solani and Trichoderma harzianum isolated earlier from egg-masses of M. incognita, identified and maintained have been investigated through in-vitro and in-vivo trials for their potentiality against M. incognita. Out of the above, isolated mycoflora A. niger was identified to be toxic against M. incognita while A. strictum and T. harzianum was found to possess both egg parasitic or opportunistic and toxic properties. A field trial with all the above fungal bioagents both alone and together showed significant promising performance by the dual treatment of A. strictum and T. harzianum in improving the health of the tomato plant with a remarkable reduction in M. incognita population.