Fungus Lasiodiplodia Research Articles

Bioherbicidal effect of secondary metabolites from Lasiodiplodia theobromae for management of narrow-leaf and broad-leaf weeds

Herbicides are the most used form of weed management worldwide but their excessive use could lead to negative impacts on human health and the environment. Biological weed management utilizing microorganisms and their metabolites presents a promising alternative. The weed-associated fungus Lasiodiplodia theobromae TBRC 15112 produces a broad spectrum of biological molecules, and shows potential as a bioherbicide. In this study, L. theobromae crude extracts from cultures caused leaf lesions in crowfoot grass (Dactyloctenium aegyptium L.) and tridax daisy (Tridax procumbens L.). The treated tridax daisy plants wilted and collapsed in three days. Using HP-20SS column chromatography, the 20, 30 and 40% acetone fractions caused leaf lesions covering 54, 30 and 34% of whole leaf areas of D. aegyptium, respectively, whereas relatively lower severities were observed for the 50–70% fractions. Eight compounds, including one new metabolite, were isolated and characterized for their structures using NMR spectroscopic data. Of these, five exhibited bioherbicidal activity, namely uracil, tyrosol, N-(4-hydroxyphenyl) acetic acid, N-(4-hydroxyphenyl) acetamide (4HPA), and the new metabolite. This metabolite, which we named lasiodiplin, is similar to alboatrin but distinct at two positions. Intriguingly, the mixture of tyrosol and 4HPA had a greater effect on weed leaves than pure compounds alone, suggesting a synergistic effect. The crude extracts also caused leaf lesions on five of six tested crops. Our study revealed that L. theobromae crude extracts have bioherbicidal activity on the narrow-leaf and broad-leaf weeds. The data could pave a way for further development of bioherbicides based on microorganisms and microbial products.

Activity of a Recombinant Chitinase of the Atta sexdens Ant on Different Forms of Chitin and Its Fungicidal Effect against Lasiodiplodia theobromae.

This study evaluates the activity of a recombinant chitinase from the leaf-cutting ant Atta sexdens (AsChtII-C4B1) against colloidal and solid α- and β-chitin substrates. 1H NMR analyses of the reaction media showed the formation of N-acetylglucosamine (GlcNAc) as the hydrolysis product. Viscometry analyses revealed a reduction in the viscosity of chitin solutions, indicating that the enzyme decreases their molecular masses. Both solid state 13C NMR and XRD analyses showed minor differences in chitin crystallinity pre- and post-reaction, indicative of partial hydrolysis under the studied conditions, resulting in the formation of GlcNAc and a reduction in molecular mass. However, the enzyme was unable to completely degrade the chitin samples, as they retained most of their solid-state structure. It was also observed that the enzyme acts progressively and with a greater activity on α-chitin than on β-chitin. AsChtII-C4B1 significantly changed the hyphae of the phytopathogenic fungus Lasiodiplodia theobromae, hindering its growth in both solid and liquid media and reducing its dry biomass by approximately 61%. The results demonstrate that AsChtII-C4B1 could be applied as an agent for the bioproduction of chitin derivatives and as a potential antifungal agent.

New α-pyrone and unsaturated ketone analogues from the endophytic fungus Lasiodiplodia sp. and their cytotoxic effects

New α-pyrone and unsaturated ketone analogues from the endophytic fungus Lasiodiplodia sp. and their cytotoxic effects

The effectiveness of the doses of the biological agent Talaromyces pinophilus to control the dieback disease on nutmeg seedling

The nutmeg plant (Myristica fragrans houtt) is a native plant of Maluku Province, Indonesia, which has historical and economic value and is also an important export commodity. The decline in nutmeg productivity in Aceh Province was due to dieback disease caused by the pathogenic fungus Lasiodiplodia theobromae. Currently control with antagonistic agents has been widely developed, one of them is with Talaromyces pinophilus. The purpose of this study was to determine the effectiveness of the dose of Talaromyces pinophilus in controlling dieback disease on nutmeg seedlings. The design used was a non-factorial completely randomized design with six treatments and four replications. The variables observed in this study were incubation period, disease incidence, and length of xylem discoloration. The results showed that T. pinophilus 30 g could significantly prolong the incubation period of L. theobromae, which was 68.65 days, reduced the incidence of disease by 20% and the length of the lesion formed was 0.73 cm.

In vitro antifungal activity of lasiodiplodin, isolated from endophytic fungus Lasiodiplodia pseudotheobromae J-10 associated with Sarcandra glabra and optimization of culture conditions for lasiodiplodin production

A macrolide antibiotic, lasiodiplodin was isolated from the endophytic fungus (EF) Lasiodiplodia pseudotheobromae J-10 associated with the medicinal plant Sarcandra glabra. In vitro antifungal assay demonstrated the inhibitory activity of lasiodiplodin against the growth of six phytopathogenic fungi, with the IC50 values ranging between 15.50 and 52.30μg/mL. The highest antifungal activities were recorded against Exserohilum turcicum, Colletotrichum capsici, and Pestalotiopsis theae, with IC50 values of 15.50, 15.90, and 17.55μg/mL, respectively. The underlying mechanism of the antifungal activity of lasiodiplodin against E. turcicum included the alteration of its colony morphology and disturbance of its cell membrane integrity. In addition, the optimization of L. pseudotheobromae J-10 culture conditions increased lasiodiplodin yield to 52.33mg/L from 0.59mg/L at pre-optimization. This is the first report on the isolation and identification of antifungal compound from the EF L. pseudotheobromae J-10 associated with S. glabra, as well as on the optimization of L. pseudotheobromae J-10 culture conditions to increase lasiodiplodin yield. The results of this study support that lasiodiplodin is a natural compound with high potential bioactivity against phytopathogens, and provide a basis for further study of the EF associated with S. glabra.

Lasiodiplodiapyrones A and B, Pyrone-Preussomerin Adducts with Highly Strained Polycyclic Ring Systems from Lasiodiplodia pseudotheobromae.

Lasiodiplodiapyrones A and B (1 and 2), two new preussomerin derivatives, possessing an unexpected 6-methyl-4H-furo[3,2-c]pyran-4-one moiety and a highly functionalized conjoint and complicated polycyclic ring system, along with two known congeners (3 and 4), were isolated from the fungus Lasiodiplodia pseudotheobromae. Their structures including absolute configurations were determined by spectroscopic analyses, Mosher's method, and ECD calculations. A biosynthetic pathway was proposed to explain the origin of lasiodiplodiapyrones A and B as well as their relationship with preussomerins. Compounds 1-4 showed suppressive effects on the production of NO with IC50 values of 4.8 ± 0.3, 8.5 ± 1.1, 5.9 ± 0.8, and 12.8 ± 1.3 μM, respectively.

Isolation, purification and structural elucidation of Mellein from endophytic fungus Lasiodiplodia theobromae strain (SJF-1) and its broad-spectrum antimicrobial and pharmacological properties.

In an on-going investigation of bioactive metabolites producing potential endophytic fungi, the strain Lasiodiplodia theobromae (SJF-1) was isolated from a medicinal plant Syzygium cumini. The cultural, morphological and molecular identification was done with the SJF-1 strain. The obtained gene sequence was deposited in NCBI with accession number MG 938644. The methanolic extract of SJF-1 strain possessed one major bioactive fraction, and it was purified by column chromatography. Further, it was identified as Mellein by various spectroscopic studies (1 H, 13 C, DEPT-135°, FT-IR, ESI-HR-MS and 2D NMR). Biologically, Mellein showed potent anti-Xanthomonas activity with minimum inhibitory concentration (MIC) values ranging from 1·9 to 62·5 μg ml-1 against 11 Xanthomonas strains, a broad-spectrum antimicrobial activity with MIC 7·8-31·25 μg ml-1 and 1·9-31·25 μg ml-1 towards both bacterial and fungal strains, respectively. The scanning electron microscope analysis proved the antimicrobial efficacy of a Mellein by rupturing the cell walls of Xanthomonas sp. Molecular docking studies further supported that the Mellein showed good binding interactions with the proteins of Xanthomonas sp. to reduce pathogenicity. Further, in silico pharmacological studies showed that this metabolite exhibited high gastrointestinal absorption properties and promising oral drug bioavailability. We report, anti-Xanthomonas, in silico docking and pharmacological studies of Mellein from (SJF-1) strain for the first time.

Efficacy of the biocontrol agent Trichoderma hamatum against Lasiodiplodia theobromae on macadamia.

Macadamia (Macadamia integrifolia) trees are an important source of revenue in rainforest ecosystems. Their nuts are rich in vitamins, minerals, fiber, antioxidants, and monounsaturated oils. The fungus Lasiodiplodia theobromae, however, is a major disease problem, causing kernel rot and other disease symptoms. In the present study, a dual confrontation assay was used to evaluate the inhibitory effect of an endophytic strain of Trichoderma hamatum C9 from macadamia root against L. theobromae. Volatiles and cell-free culture filtrate of T. hamatum were also used to assess their antifungal activity against L. theobromae. Results suggested that T. hamatum exhibited a significant inhibitory effect against L. theobromae in vitro. Further results of a biocontrol assay indicated that a spray treatment of T. hamatum conidial suspension significantly decreased the size of lesions caused by artificially inoculated L. theobromae on macadamia leaves, as well as the disease index in young trees inoculated with L. theobromae, relative to sterile water controls. Collectively, our findings indicate that T. hamatum C9 represents a potential biocontrol agent that can be used to manage L. theobromae on macadamia.

Two new depsidones from the fungus Lasiodiplodia pseudotheobromae

Lasiodiplodiaones A (1) and B (2), two new depsidones, along with five known depsidone compounds (3–7), were isolated from the fungus Lasiodiplodia pseudotheobromae. Their structures and absolute configurations were demonstrated based on NMR, HRESIMS, X-ray crystallography, and modified Mosher’s method. Especially, compounds 1 and 2 are the first examples of depsidones decorating with a 2,3-butanediol group.

Indole-3-Carboxylic Acid From the Endophytic Fungus Lasiodiplodia pseudotheobromae LPS-1 as a Synergist Enhancing the Antagonism of Jasmonic Acid Against Blumeria graminis on Wheat.

The discovery of natural bioactive compounds from endophytes or medicinal plants against plant diseases is an attractive option for reducing the use of chemical fungicides. In this study, three compounds, indole-3-carbaldehyde, indole-3-carboxylic acid (3-ICA), and jasmonic acid (JA), were isolated from the EtOAc extract of the culture filtrate of the endophytic fungus Lasiodiplodia pseudotheobromae LPS-1, which was previously isolated from the medicinal plant, Ilex cornuta. Some experiments were conducted to further determine the antifungal activity of these compounds on wheat powdery mildew. The results showed that JA was much more bioactive than indole-3-carbaldehyde and 3-ICA against Blumeria graminis, and the disease severity caused by B. graminis decreased significantly with the concentration increase of JA treatment. The assay of the interaction of 3-ICA and JA indicated that there was a significant synergistic effect between the two compounds on B. graminis in each of the ratios of 3-ICA to JA (3-ICA:JA) ranging from 1:9 to 9:1. When the compound ratio of 3-ICA to JA was 2:8, the synergistic coefficient was the highest as 22.95. Meanwhile, a histological investigation indicated that, under the treatment of JA at 500 μg/ml or 3-ICA:JA (2:8) at 40 μg/ml, the appressorium development and haustorium formation of B. graminis were significantly inhibited. Taken together, we concluded that JA plays an important role in the infection process of B. graminis and that 3-ICA as a synergist of JA enhances the antagonism against wheat powdery mildew.

Four undescribed ergostane-type steroids from Lasiodiplodia pseudotheobromae and their neuroprotective activity

Four undescribed ergostane-type steroids, (22E,24R)-4α,5α-epoxyergosta-9α,14β-dihydroxy-7,22-diene-3,6-dione, (22E,24R)-4α,5α-epoxyergosta-9α,14α-dihydroxy-7,22-diene-3,6-dione, 12α-hydroxyergosta-7,22,24(28)-triene-3-one, and 3β,12α-dihydroxyergosta-7,24(28)-diene, along with a known congener (22E,24R)-9α,14β-dihydroxyergosta-4,7,22-triene-3,6-dione, were isolated from the fungus Lasiodiplodia pseudotheobromae. Their structures were elucidated using NMR, HRESIMS, ECD calculation, and X-ray diffraction analyses. (22E,24R)-4α,5α-epoxyergosta-9α,14β-dihydroxy-7,22-diene-3,6-dione and (22E,24R)-4α,5α-epoxyergosta-9α,14α-dihydroxy-7,22-diene-3,6-dione are a pair of C-14 epimers possessing an unusual epoxy group between C-4 and C-5, which was demonstrated using single-crystal X-ray diffraction analyses. The absolute configurations of 12α-hydroxyergosta-7,22,24(28)-triene-3-one and 3β,12α-dihydroxyergosta-7,24(28)-diene were determined by ECD calculations. Moreover, 3β,12α-dihydroxyergosta-7,24(28)-diene exhibited neuroprotective activity in vitro in glutamate-treated SH-SY5Y cell lines.

Insights into Asparaginase from Endophytic Fungus Lasiodiplodia theobromae: Purification, Characterization and Antileukemic Activity.

Endobiotic fungi are considered as a reservoir of numerous active metabolites. Asparaginase is used as an antileukemic drug specially to treat acute lymphoblastic leukaemia. The presented study aims to optimize the media conditions, purify, characterize, and test the antileukemic activity of the asparaginase induced from Lasiodiplodia theobromae. The culture medium was optimized using an experiment designed by The Taguchi model with an activity ranging from 10 to 175 IU/mL. Asparaginase was induced with an activity of 315 IU/mL. Asparaginase was purified with a specific activity of 468.03 U/mg and total activity of 84.4 IU/mL. The purified asparaginase showed an approximate size of 70 kDa. The purified asparaginase showed an optimum temperature of 37 °C and an optimum pH of 6. SDS reduced the activity of asparaginase to 0.65 U/mL while the used ionic surfactants enhanced the enzyme activity up to 151.92 IU/mL. The purified asparaginase showed a Km of 9.37 µM and Vmax of 127.00 µM/mL/min. The purified asparaginase showed an IC50 of 35.2 ± 0.7 IU/mL with leukemic M-NFS-60 cell lines and CC50 of 79.4 ± 1.9 IU/mL with the normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for metabolites such as asparaginase, provides an opportunity to develop a facile, eco-friendly, cost-effective, and rapid synthesis of antileukemic drugs, which have the potential to be used as alternative and reliable sources for potent anticancer agents.

Fermentative Production of Lasiodiplodan by Lasiodiplodia theobromae CCT3966 from Pretreated Sugarcane Straw

Lasiodiplodan is a β-glucan polymer with different interesting characteristics, including therapeutic properties. It is an extracellular product, which is produced by the filamentous fungus Lasiodiplodia theobromae, using glucose as a substrate. In the present work, the production of lasiodiplodan was studied by the utilization of sugarcane straw as a low-cost carbon source. Glucose-rich sugarcane straw hydrolysate was obtained by a sequential pretreatment with dilute nitric acid (1% v/v) and sodium hydroxide (1% w/v), followed by enzymatic hydrolysis. The fermentation process was conducted by the cultivation of the strain Lasiodiplodia theobromae CCT3966 in sugarcane straw hydrolysate in a shake flask at 28 °C for 114 h. It was found that hydrolysate obtained after enzymatic hydrolysis contained 47.10 gL−1 of glucose. Fermentation experiments of lasiodiplodan synthesis showed that the peak yield and productivity of 0.054 gg−1 glucose consumed and 0.016 gL−1 h−1, respectively, were obtained at 72 h fermentation time. Fungal growth, glucose consumption, and lasiodiplodan production from sugarcane straw hydrolysate presented a similar pattern to kinetic models. The study on the chemical structure of lasiodiplodan produced showed it had a β-glucan construction. The current study revealed that sugarcane straw is a promising substrate for the production of lasiodiplodan.

Potential biocontrol of endophytic fungi against Lasiodiplodia pseudotheobromae causal agent of cocoa dieback on cocoa seedling

Endophytic fungi associated with cocoa are one of the effective agents to control plant pathogens on cocoa, including the fungus Lasiodiplodia pseudotheobromae. L. pseudotheobromae cause a significant disease on cocoa, cocoa dieback. The disease is considered an emerged disease on cocoa in Sulawesi, Indonesia. The research aimed to determine the effect of the endophytic fungi on L. pseudotheobromae on cocoa seedlings. This study used a randomized block design consisting of 6 treatments and 3 replications. Five different fungi isolates were used that belong to Aspergillus genera. The treatment is three isolates of endophytes fungi (isolate T2S2LT, T245LT, T645LT), Aspergillus isolates combination, and control (with and without pathogen). The results showed that the combination of isolates and T645LT isolate was the best treatment for suppressing necrotic symptoms (leaf spot) incidence and severity. Also, T645LT isolate performed well to inhibit leaf blight symptoms incidence severity. Meanwhile, the chlorotic symptom was reduced by isolates combination, followed by T2S2LT, T645LT, and T245LT. Endophytic fungi can potentially be an important option to control cocoa dieback disease.

Isolation and antagonism of chitinolytic bacteria from Ipomea pes caprae against Lasiodiplodia pseudotheobromae

Chitin is the main component of fungal cell walls. The growth of fungi can be inhibited by chitinolytic bacteria because they can produce chitinase enzymes which play a role in degrading cell walls. The purpose of this study was to test the activity of bacteria isolated from the roots of Ipomea pes caprae in inhibiting the growth of the fungus Lasiodiplodia pseudothebromae. The research method was carried out by isolating as much as 5 g of I. pes caprae roots using specific media and testing for bacterial antagonism using a dual culture method. The results showed that there were four isolate codes obtained namely IPaR1, IPaR4, IPpA2, and IPpA3 and could inhibit the growth of Lasiodiplodia pseudtheobromae.

Phytotoxic β-resorcylic acid derivatives from the endophytic fungus Lasiodiplodia theobromae in the mangrove plant

• New β -resorcylic acid derivatives were isolated from an endophyte L . theobromae . • Absolute configuration was established using Mosher's ester analysis and chemical ECD calculations. • (3 S, 7 R )-17- O -methyl- de - O -methyllasiodiplodin showed growth inhibitory effects against D. ciliaris. Three new β -resorcylic acid derivatives, 1 – 3 , were obtained from the endophytic strain Lasiodiplodia theobromae GI-1005, which was isolated from the mangrove plant Rhizopora mucronata. The structures of new compounds together with five known compounds were elucidated by NMR and MS analyses, and chemical means. The isolated compounds were evaluated for phytotoxicity against Digitaria ciliaris (southern crabgrass) and Lactuca sativa (lettuce). Compound 1 showed growth inhibitory effects against D. ciliaris in a dose-dependent manner.

Photodynamic inactivation of Lasiodiplodia theobromae: lighting the way towards an environmentally friendly phytosanitary treatment.

The fungus Lasiodiplodia theobromae is one of the main causal agents of trunk canker and dieback of grapevine. The objective of this work was to evaluate the efficiency of photodynamic inactivation (PDI) of L. theobromae with synthetic and natural photosensitizers and irradiation with either sunlight or artificial photosynthetically active radiation. Although the growth of the mycelium could not be completely prevented with natural sunlight irradiation, phenothiazine dyes (methylene blue, MB; toluidine blue O, TBO), riboflavin and a cationic porphyrin (Tetra-Py+-Me) caused complete inhibition under continuous irradiation with artificial light. Free radicals were the main cytotoxic agents in the PDI with MB, indicating the predominance of the type I mechanism. PDI with MB or Tetra-Py+-Me may represent a promising approach for the sanitation of vine material in greenhouse nurseries, in order to reduce the risk of infection upon grafting.

Aldsulfin, a novel unusual anti-mannheimiosis epithiodiketopiperazine antibiotic produced by Lasiodiplodia pseudotheobromae FKI-4499.

An anti-mannheimiosis agent, aldsulfin, was isolated from a culture broth of the fungus Lasiodiplodia pseudotheobromae FKI-4499, together with a known compound, lasiodipline C, using bioassay-guided fractionation. Spectroscopic analysis of aldsulfin, using NMR, mass spectrometry, and CD analyses revealed it to be an epithiodiketopiperazine with an unstable and unusual hemithioaminal moiety. Aldsulfin showed antibacterial activity against Mannheimia haemolytica and Pasteurella multocida.

Production of β-glucan exopolysaccharide lasiodiplodan by Lasiodiplodia theobromae CCT 3966 from corn bran acid hydrolysate.

The potential utilization of corn bran acid hydrolysate (CBAH) was evaluated as an inexpensive feedstock for the production of a rich carbohydrate and protein medium for lasiodiplodan (LAS) production using the filamentous fungus Lasiodiplodia theobromae CCT 3966. Experiments were performed according to a 22 CCRD experimental design aiming to evaluate the influence of agitation speed (rpm) and temperature (°C) over the production of total cell biomass (TCB) and LAS concentration released to the medium (LAS-M), adhered to biomass (LAS-C), and total (LAS-T). Under the selected conditions (temperature of 28°C and agitation of 200 rpm), 8.73 g·L-1 of LAS-T and 4.47 g·L-1 of TCB were obtained. Recovery of LAS-C with hot water was shown as an alternative to increase the production concentration, although it might require further purification steps. CBAH potential for substitution of synthetic media was demonstrated, indicating that it is an adequate raw material containing all necessary nutrients for LAS production.Key points• Corn bran acid hydrolysate is presented as a suitable substrate for β-glucan production.• Lasiodiplodia theobromae CCT 3966 have the potential for the industrial β-glucan production.• Simple recovering of biomass-adhered lasiodiplodan by hot water extraction.

Dimeric γ-lactone derivatives from the soil-derived fungus Lasiodiplodia theobromae NSTRU-PN1.4

Investigation of the soil-derived fungus Lasiodiplodia theobromae NSTRU-PN1.4 resulted in the isolation of five dimeric γ-lactones including two new botryosphaerilactones D and E (4 and 5) and three known structurally related analogoues (1–3) along with seven known compounds. Their structures were elucidated by extensive spectroscopic analysis. The absolute configuration of 1–5 was determined by comparison of the ECD data with those of the structurally related monomeric γ-lactones. For biological evaluation, this is the first report on antifungal activity of the known (3 R,4R)-4-acetyl-3-methyl-2(3H)-dihydrofuranone which displayed weak antifungal activity against Cryptococcus neoformans with an MIC value of 200 µg/mL.