3-hydroxypropionic acid as a nematicidal principle in endophytic fungi

3- Hydroxypropionic acid was isolated by bioactivity-guided fractionation of extracts obtained from submerged cultures of several endophytic fungi isolated from above-ground plant organs. This compound showed selective nematicidal activity against the plant-parasitic nematode Meloidogyne incognita with LD50 values of 12.5-15 microg/ml. Activity against the saprophytic Caenorhabditis elegans was fivefold lower. No antimicrobial, cytotoxic or phytotoxic effects were observed. Propionic acid and D- and L-lactic acids were not active against either nematode species. Based on morphological features and ITS, 18S and 28S rDNA analyses, the producing strains were identified as Phomopsis phaseoli isolated from the leaf of a tropical tree, and four strains of Melanconium betulinum isolated from twigs of Betula pendula and B. pubescens in Germany. This is the first report of 3-hydroxypropionic acid in fungi, and of the nematicidal activity of this metabolite.     

Biocidal Compounds from Mentha sp. Essential Oils and Their Structure–Activity Relationships

Essential oils from Greek Mentha species showed different chemical compositions for two populations of M. pulegium, characterized by piperitone and pulegone. Mentha spicata essential oil was characterized by endocyclic piperitenone epoxide, piperitone epoxide, and carvone. The bioactivities of these essential oils and their components have been tested against insect pests (Leptinotarsa decemlineata, Spodoptera littoralis and Myzus persicae), root‐knot nematodes (Meloydogine javanica) and plants (Lactuca sativa, Lolium perenne, Solanum lycopersicum). The structure–activity relationships of these compounds have been studied including semi‐synthetic endocyclic trans‐carvone epoxide, exocyclic carvone epoxide, a new exocyclic piperitenone epoxide and trans‐pulegone epoxide. Leptinotarsa decemlineata feeding was affected by piperitenone and piperitone epoxide. Spodoptera littoralis was affected by piperitone epoxide and pulegone. The strongest nematicidal agent was piperitenone epoxide, followed by piperitone epoxide, piperitenone and carvone. Germination of S. lycopersicum and L. perenne was significantly affected by piperitenone epoxide. This compound and carvone epoxide inhibited L. perenne root and leaf growth. Piperitenone epoxide also inhibited the root growth of S. lycopersicum. The presence of a C(1) epoxide resulted in strong antifeedant, nematicidal and phytotoxic compounds regardless of the C(4) substituent. New natural crop protectants could be developed through appropriate structural modifications in the p‐menthane skeleton.     

Effect of urea and certain NPK fertilizers on the cereal cyst nematode (Heterodera avenae) on wheat

Two outdoor pot experiments were conducted in two consecutive years under outdoor conditions during the wheat growing season in Saudi Arabia to determine the effects of urea and certain compound fertilizers (NPK), compared to the effects of the nematicide fenamiphos on the cereal cyst nematode (CCN), Heterodera avenae, and wheat growth. The results showed that all of the treatments, except the fertilizer diammonium phosphate (DAP), reduced the number of nematode cysts/root system and increased (P ⩽ 0.05) the dry weight of nematode-infected wheat plants. Fenamiphos and urea resulted in the best control, followed by the NPK fertilizers. The combined application of urea and fenamiphos resulted in the most significant effect in decreasing (P ⩽ 0.05) the number of cysts/root system and increasing (P ⩽ 0.05) the growth of nematode-infected wheat plants.     

Effects of Microbial and Other Antagonistic Organism and Competition on Entomopathogenic Nematodes

Antagonistic factors, broadly identified as antibiosis, competition and natural enemies, impact on entomopathogenic nematodes. Antibiosis can occur through the release of plant chemicals from the roots into the soil, which may adversely affect the host-finding behavior of the infective stage nematode, or the presence of these chemicals in the host insect may negatively affect nematode reproduction. In laboratory studies, intra-specific and inter-specific competition reduces nematode fitness, and inter-specific competition can cause local extinction of a nematode species. For example, after concomitant infection of a host, a steinernematid species usually excludes a heterorhabditid species. The mechanism for the steinernematid superiority has been postulated to be a bacteriocin(s) produced by Xenorhabdus, the symbiotic bacterium of the steinernematid, which prevents Photorhabdus, the symbiotic bacterium of the heterorhabditid, from multiplying. Inter-specific competition between two steinernematid species shows that both can co-exist in a host, but one species will eventually prevail in the environment. By having different foraging strategies, however, both steinermatid species may co-exist in the same habitat. An important issue is whether the introduction of an exotic entomopathogenic nematode species will competitively displace an indigenous nematode species. Although the environmental risks are small, the recommended policy is that the introduction of exotic nematodes be regulated. With other pathogens, entomopathogenic nematodes can out-compete entomopathogenic fungi, but not Bacillus thuringiensis, for the same host individual when both the nematode and entomopathogen are applied simultaneously. The best studied natural enemy is the nematophagous fungus, Hirsutella rhossiliensis, which causes higher mortality in Steinernema glaseri compared with Heterorhabditis bacteriorphora. Differential susceptibility to the fungus may be associated with the retention of the second-stage cuticle by H. bacteriophora. Invertebrate predators including mites and collembolans feed on entomopathogenic nematodes. Although a number of studies have been conducted with antagonists, there is a dearth of field data. We suggest that long-term research plots be established where natural populations of entomopathogenic nematodes occur and include antagonists as a component of such studies.     

Chemical constituents of Cordia latifolia and their nematicidal activity

Following nematicidal activity‐guided isolation studies on the fruits, bark, and leaves of Cordia latifolia, two new constituents, cordinoic acid (=11‐oxours‐12‐ene‐23,28‐dioic acid; 1 ) and cordicilin (=2‐{[(E)‐3‐(3,4‐dihydroxyphenyl)prop‐2‐enoyl]oxy}‐3‐[4‐hydroxy‐3‐(stearoyloxy)phenyl]propanoic acid; 2 ) were isolated from the stem and leaves, respectively, together with nine known compounds, namely cordioic and cordifolic acid from the stem bark, latifolicin A–D and rosmarinic acid from the fruits, and cordinol and cordicinol from the leaves. Their structures were determined by means of spectroscopic analyses including 1D‐ and 2D‐NMR techniques. The nematicidal activities of these constituents were determined against the root‐knot nematode Meloidogyne incognita. Hundred percent mortality was caused by all of these after 72 h at a 0.125% concentration. Compound 1 and cordioic acid were most active and caused 100% mortality after 24 h at a 0.50% concentration. Furthermore, compound 2 , the ester of rosemarinic acid, was found to be more active than the free acid.     

Pentacyclic triterpenoids from the aerial parts of Lantana camara and their nematicidal activity

Two new olean-12-ene triterpenoids, camarolic acid (1) and lantrigloylic acid (2), have been isolated from the aerial parts of Lantana camara, along with ten known triterpenes, namely, camaric acid, lantanolic acid, lantanilic acid, pomolic acid, camarinic acid, lantoic acid, camarin, lantacin, camarinin, and ursolic acid. The new compounds have been characterized as 3,25-epoxy-3alpha-hydroxy-22beta-{[(S)-3-hydroxy-2-methylidenebutanoyl]oxy}olean-12-en-28-oic acid (1) and 3,25-epoxy-3alpha-hydroxy-22beta-[(3-methylbut-2-enoyl)oxy]olea-9(11),12-dien-28-oic acid (2) through spectroscopic studies and a chemical transformation. Seven of the constituents, namely pomolic acid, lantanolic acid, lantoic acid, camarin, lantacin, camarinin, and ursolic acid, were tested for nematicidal activity against root-knot nematode Meloidogyne incognita. Pomolic acid, lantanolic acid, and lantoic acid showed 100% mortality at 1 mg/ml concentration after 24 h, while camarin, lantacin, camarinin, and ursolic acid exhibited 100% mortality at this concentration after 48 h. These results are comparable to those obtained with the conventional nematicide furadan (100% mortality at 1 mg/ml concentration after 24 h).     

Nematicidal metabolites produced by the endophytic fungus Geotrichum sp. AL4

From the endophytic fungal strain Geotrichum sp. AL4, cultivated from the leaves of the neem tree (Azadirachta indica), four compounds, 1-4, were isolated from the AcOEt extract, including two new, chlorinated, epimeric 1,3-oxazinane derivatives. All compounds were assessed for their nematicidal activities against the nematodes Bursaphelenchus xylophilus and Panagrellus redivivus, and three out of the four isolates showed noticeable bioactivities.     

Cytotoxic,larvicidal, nematicidal,and antifeedant activities of piperidin-connected 2-thioxoimidazolidin-4-one derivatives

The objective of this study was to investigate brine shrimp cytotoxicity, larvicidal, nematicidal, and antifeedant activities of novel piperidin-connected 2-thioxo-imidazolidin-4-one derivatives. The activities of target compounds were compared with some naturally occurring (?)-pinidinol, hydantocidin, and positive controls. Target compounds were synthesized via cyclocondensation method. The compounds were synthesized and then characterized by infrared spectroscopy, ¹H NMR, ¹³C NMR, mass spectral, and elemental analyses. Brine shrimp cytotoxicity assay was investigated using freshly hatched, free-swimming nauplii of Artemiasalina. Larvicidal screening was performed against urban mosquito larvae (Culex quinquefasciatus). Nematicidal activity was evaluated using juvenile nematodes of Meloidogyne javanica. Regarding antifeedant activity, marine-acclimated Oreochromis mossambicus fingerlings were used. Compounds 3a-c (piperidin-connected 2-thioxoimidazolidin-4-one) were found to be lethal to the second instar larvae of mosquito, which produced LD₅₀ values of 1.37, 6.66, 6.51 μg/mL, compared to compounds (?) pinidinol and hyantocidin LD₅₀ values of 18.28 and 22.11 μg/mL respectively. Compound 3a-c was found to kill 100% of fish fingerlings within 6 h at 20 µg/mL, with LD₅₀ values of 1.54, 1.79, 1.52 µg/mL, compared to compounds (?) pinidinol and hyantocidin with LD₅₀ values of 10.21 and 21.05 μg/mL respectively. Compound 3c with LD₅₀ value of 1.57 μg/mL demonstrated high nematicidal activity compared to compound 3a, 3b, (?) Pinidinol and Hyantocidin LD₅₀ values of 6.45, 2.42, 14.25, 26.30 μg/mL respectively. Therefore, the 2-thioxoimidazolidin-4-one with piperidin ring showed high potential cytotoxic, larvicidal, nematicidal, and antifeedent activities.     

Antimicrobial and antinematicidal metabolites from Streptomyces cuspidosporus strain SA4 against selected pathogenic bacteria,fungi and nematode

The exploration of novel therapeutic agents and other bioactive secondary metabolite from Streptomyces species, for possible agricultural farming, pharmaceutical and industrialized applications, has been, and still is, essential. The existing studies were aimed with biologically potential Streptomyces species and its antagonistic activity against dreadful microorganisms. Totally, morphological three different actinomycetes were selected from the fertile agricultural lands. Among the three, the isolate SA4 exhibited significant antimicrobial and anti-nematicidal activity towards selected microbial pathogens such as E Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, Salmonella typhi, Bacillus subtilis, Proteus vulgaris, Shigella flexineri, Candida albicans, and Fusarium sp. The prospective strain SA4 was identified as Streptomyces cuspidosporus. The isolate SA4 optimized for secondary metabolites production with International Streptomyces project 4 (ISP 4) medium, pH 7.0 at 37 °C for 14 days. Gas Chromatography-Mass spectrometry (GCMS) analysis of strain SA4 bioactive extract publicized the existence of 1,2-Benzenedicarboxylic acid, bis(2-Methylpropyl) ester compound and occupied by high peak area and its possessed significant biological properties.     

Changes in secondary metabolism during stromatal ontogeny of Hypoxylon fragiforme

Stromata of Hypoxylon fragiforme were studied during the vegetation period by hplc profiling, revealing changes in the composition during stromatal development. Cytochalasin H and two new cytochalasins named fragiformins A–B were identified as major constituents of the young, maturing stromata, whereas mature, ascogenous material yielded large amounts of mitorubrin-type azaphilones. The above compounds, further cytochalasins from Xylariaceae and other fungi, and additional azaphilones of the mitorubrin type were assayed for their nematicidal effects against Caenorhabditis elegans and their antimicrobial activities against Bacillus subtilis, Yarrowia lipolytica, and various filamentous fungi. The results confirmed data in the literature on broad-spectrum non-selective activities of azaphilones and cytochalasins in biological systems. Most interestingly, laboratory cultures of the above Hypoxylon spp. mainly produced dihydroisocoumarin derivatives and were found devoid of mitorubrins and cytochalasins. These rather drastic changes in the secondary metabolism of H. fragiforme and the above biological activities are discussed in relation to the possible biological functions of secondary metabolites (extrolites) in the Hypoxyloideae.