Deoxyradicinin,a novel phytotoxin from Alternaria helianthi

A novel major metabolite which possesses phytotoxic and antifungal properties has been isolated from culture filtrates of Alternaria helianthi and its structure elucidated as deoxyradicinin.     

Drazepinone, a trisubstituted tetrahydronaphthofuroazepinone with herbicidal activity produced by Drechslera siccans

When grown in a minimal-defined medium, a strain of Drechslera siccans, a pathogenic fungus isolated from seeds of Lolium perenne, produced phytotoxic metabolites. This strain is one of the best toxin producers among several grass pathogenic fungal strains collected and tested to find phytotoxins to be used as natural herbicides of monocot weeds. From the culture filtrates of D. siccans, we isolated a new phytotoxic trisubstituted naphthofuroazepinone, named drazepinone, and characterised it as a 3,5,12a-trimethyl-2,5,5a,12a-tetrahydro-1H-naphtho[2',3':4,5]furo[2,3-b]azepin-2-one. Assayed at 2 microg microl(-1) solution the novel metabolite proved to have broad-spectrum herbicidal properties, without antibacterial and antifungal activities, and low zootoxic activity. Its original chemical structure and the interesting biological properties make drazepinone a potential natural herbicide.     

Tulasnein and podospirone from the coprophilous xylariaceous fungus Podosordaria tulasnei

Tulasnein (1), a new metabolite with strong antimicrobial and weaker cytotoxic and phytotoxic activity, was isolated from culture filtrates of three strains of the xylariaceous coprophilous fungus Podosordaria tulasnei. The producing strains were identified by their rhizomorphs and by ITS rDNA sequence analysis. A second new metabolite, podospirone (2), was also produced by all three strains whereas the weakly cytotoxic (+)-3,4-anhydroshikimic acid methyl ester (3) was detected in only one strain.     

Purification, characterization, and amino acid sequence of cerato-platanin, a new phytotoxic protein from Ceratocystis fimbriata f. sp. platani.

A new phytotoxic protein (cerato-platanin) of about 12.4 kDa has been identified in culture filtrates of the Ascomycete Ceratocystis fimbriata f. sp. platani, the causal agent of canker stain disease. The toxicity of the pure protein was bioassayed by detecting the inducing necrosis in tobacco leaves. The pure protein also elicited host synthesis of fluorescent substances in tobacco and plane (Platanus acerifolia) leaves. We purified the protein from culture medium to homogeneity. Its complete amino acid sequence was determined; this protein consists of 120 amino acid residues, contains 4 cysteines (S-S-bridged), and has a high percentage of hydrophobic residues. The molecular weight calculated from the amino acid sequence agrees with that determined by mass spectrometry, suggesting that no post-transnational modification occurs. Searches performed by the BLAST program in data banks (Swiss-Prot, EBI, and GenBank(TM)) revealed that this protein is highly homologous with two proteins produced by other Ascomycete fungi. One, produced during infection of wheat leaves, is codified by the snodprot1 gene of Phaeosphaeria nodorum (the causal agent of glume blotch of wheat), whereas the other is the rAsp f13 allergen from Aspergillus fumigatus. Furthermore, the N terminus of cerato-platanin is homologous with that of cerato-ulmin, a phytotoxic protein belonging to the hydrophobin family and produced by Ophiostoma (Ceratocystis) ulmi, a fungus responsible for Dutch elm disease.     

Bostrycin and 4-deoxybostrycin: two nonspecific phytotoxins produced by Alternaria eichhorniae.

Two crystalline red pigments with phytotoxic activity were isolated from culture filtrates of Alternaria eichhorniae, a pathogen of the water hyacinth Eichhornia crassipes. The pigments were present in the ratio of 4:1 and were identified as bostrycin and 4-deoxybostrycin, respectively. This is the first isolation of 4-deoxybostrycin from a natural source. Bostrycin, 4-deoxybostrycin, and their isopropylidene derivatives induced necrosis on tested plant leaves comparable to the A. eichhorniae-induced necrosis on water hyacinth. The lowest phytotoxic concentrations of crystalline bostrycin and 4-deoxybostrycin on water hyacinth leaves were about 7 and 30 microgram/ml, respectively. Both substances were inhibitory to Bacillus subtilis but were inactive against the fungus Geotrichum candidum.     

Production of Phytotoxic Sirodesmins by Aggressive Strains of Leptosphaeria maculans Differing in Interactions with Oil Seed Rape Genotypes

Sirodesmins were extracted from culture filtrates of 3 aggressive strains of Leptosphaeria maculans which showed different specific interactions with the oilseed rape cultivars Quinta and Jet Neuf. Up to 8 compounds were detectable showing reactions on thin layer plates which are characteristic for sirodesmins. Sirodesmin PL, sirodesmin C, deacetylsirodesmin PL and deacetyl-sirodesmin C were identified on the basis of their UV-spectra, by various conversion reactions and by cochromatography with known standards. Two further compounds were tentatively identified as sirodesmin B and deacetylsirodesmin B on the basis of conversion reactions only. In the cultures of all strains sirodesmin PL was the main component, followed by deacetylsirodesmin PL, sirodesmin C, and deacetylsirodesmin C. These 4 metabolities were mainly responsible for the phytotoxic activity of all culture filtrates. They were systemically transported to leaves of root treated rape seedlings. No correlation was found between the production of the 4 main sirodesmins and the differential host parasite interactions of the 3 aggressive strains of Leptosphaeria maculans.     

Deoxyradicinin and 3-epideoxyradicinol production by the sunflower pathogen Alternaria helianthi

The phytotoxic compound deoxyradicinin, a metabolite of Alternaria helianthi, was isolated from naturally infected leaves of sunflower (Helianthus annuus). The production of this toxin in liquid culture together with 3-epideoxyradicinol is discussed. Production of deoxyradicinin by liquid cultures was observed to decrease after successive sub-culture of the fungus on solid medium. All 10 accessions of Helianthus tested were sensitive to deoxyradicinin although some quantitative differences between lines were noted. Evidence is presented for the metabolism of deoxyradicinin by host leaf tissue.     

Identification and Phytotoxicity of 3-methylthiopropanoic and Trans-3-methylthiopropenoic Acids Produced in Culture by Xanthomonas campestris pv. vitians

Two phytotoxic metabolites were isolated from culture filtrates of Xanthomonas campestris pv, vitians , the causal agent of lettuce leaf spots and headrot. The two compounds were identified as 3-methylthiopropanoic (1) and trans-3-methylthiopropenoic (2) acids by chemical and spectroscopic methods. Toxic effects of the two compounds on leaf tissues and protoplasts of lettuce and cabbage were investigated. Solutions of 1 and 2 induced chlorosis and necrosis on lettuce leaves at minimum concentrations of 300 and 50 μg/ml, respectively. Infiltration in cabbage leaves did not produce any symptoms. The LD₅₀ values for 1 and 2 against lettuce protoplasts were 15 and 16 μg/ml, respectively. Activity of the two metabolites against cabbage protoplasts was very low (LD₅₀ > 500 μg/ml).     

Studies on Alternaria sesami pathogenic to sesame

Influence of pH on the growth of Alternaria sesami, its nutritional requirements and its ability to produce phytotoxic and antibacterial metabolites were tested. The isolate was cultured on Czapek-Dox broth and the culture filtrates were screened for phytotoxicity against seeds and seedlings of sesame. Chloroform extracts of the fungus exhibited antibacterial activity. Analysis of the culture filtrates for identifying toxins using chromatographic techniques revealed the presence of tenuazonic acid.     

Variation in Metabolite Production by Septoria nodorum Isolates Adapted to Wheat or to Barley

The production of secondary metabolites by S. nodorum strains was examined in relation to their adaptation to wheat or to barley. Eleven strains of each type previously characterized were tested for the production in culture of (-)-(3R)-mellein (ochracin), (3R)-O-metbylmellein; (-)-(3R,4R)-4-hydroxyniellein, (-)-(3R,4S)-4-hydro-xymellein, mycophenolic acid, septorine, N-methoxy-septorine, N-methoxyseptorinol, and a new compound (=UN).
Mellein was produced by every strain. Both 4-hydro-xymellein isomers were yielded by all wheat-adapted strains and six barley-adapted strains. Most of the strains produced mycophenolic acid. On the other hand the pigmentation of culture filtrates of wheat-adapted strains was pale-yellow whereas the pigmentation of barley-adapted strains was grey-blue. The productions of O-methylmellein, septorine, N-metboxyseptorine and N-methoxyseptorinol were typical for wheat-adapted strains. The detection of UN was restricted to the filtrates of barley-adapted strains. If the pigmentation of filtrates and the production of septorines, UN and O-methyl-mellein are considered the strains fell into two groups. This classification is related to the host adaptation except for one barley-adapted strain that showed characteristics of wheat-adapted strains and one wheat-adapted strain which did not produce septorines.     

A New Insecticidal Pyrethroid Ester

Infected leaves of the noxious weed johnsongrass were collected in a field with a view to the isolation of toxigenic phytopathogens. Exserohilum turcicum (Pass.) Leonard et Suggs [≡ Drechslera turcica (Pass.) Subramanian et Jain] was isolated from leaf lesions and grown in a liquid culture to yield the known fungal metabolite monocerin (1). This compound was shown to possess (non-specific) phytotoxic activity. Monocerin inhibited seedling growth both of johnsongrass and, to a lesser extent, cucumber. The phytotoxic nature of this compound has not been described previously. Its activity against leaf tissue of two other species, from different plant families, was also demonstrated.     

Inhibition of Rust Diseases of Cereals by Metabolic Products of Verticillium chlamydosporium

Verticillium chlamydosporium produced in submers culture several antifungal and/or phytotoxic compounds which were detected in a bioassay by using the pathogen-host system Puccinia coronata and oat seedlings. The antifungal compounds were also tested against P. recondita on wheat and P. sorghi on corn seedlings. The production of the active metabolic compounds highly depended on the nutrient solution (peptone-Czapek [PC] and malt extract [ME]) and on the fermentation times. Cell-free filtrates of PC-cultures of the fungus were highly phytotoxic; the fungitoxic and phytotoxic compounds were heat-labile and dialyzable. The ethyl acetate extracts of the PC-culture filtrates contained only the antifungal active substances. The antifungal compounds in ME-culture filtrates proved to be heat-stable, could be dialyzed and extracted with ethyl acetate. Ethyl acetate extracts of PC- and ME-culture filtrates at concentrations of 500 μg/ml reduced rust disease incidence by up to 80 % compared to the control treatment. Further studies with extracts of ME-culture filtrates displayed a distinct protective but no systemic activity. The extract interfered with the development of several infection structures of the rust fungi, mostly with the growth of germ tubes as well as with the formation of the aappressoria and haustorial mother cells. Three rust-active fractions were obtained by preparative layer chromatography on silica gel. One of these fractions exhibited phytotoxic activity. The most active antifungal fraction is identical with the macrolid antibiotic monorden which caused a desorientated spiral growth in P. coronata germlings on oat leaves.     

An ELISA method to identify the phytotoxic Pseudomonas syringae pv. actinidiae exopolysaccharides: A tool for rapid immunochemical detection of kiwifruit bacterial canker

The phytotoxic exopolysaccharides produced by Pseudomonas syringae pv. actinidiae, the causal agent of bacterial canker of kiwifruit, were isolated and partially identified. Their phytotoxic activity was evaluated on host and non-host plants and their role in the complex mechanisms of host-pathogen interaction was also discussed. The phytotoxic exopolysaccharides, which are natural antigens, were used to arise specific antibodies by rat immunization. The antibodies were used to develop a rapid and specific method to unambiguously detect P.s. pv. actinidiae exopolysaccharides isolated from bacterial culture and infected plant samples. Indeed, the antibodies recognized the exopolysaccharides produced by other two strains of P. s. pv. actinidiae but did not cross reacted with those isolated from P. s. pv. syringae and Pseudomonas viridiflava culture filtrates. Finally, the same antibodies significantly recognized the exopolysaccharides extracted from infected kiwi leaves.     

寄主植物对橄榄星室木虱自然种群的影响

通过野外调查和室内测试,结果表明:橄榄星室木虱Pseudophacopteron canarium在橄榄Canarium album和乌榄C.nigrum上均能完成生长发育,且其种群数量都有不同程度的增长,其中橄榄的2个品系间的星室木虱种群趋势指数(I)差异不大,分别为24.4165和20.0738,而乌榄的明显比橄榄的低,仅为9.7575,即乌榄的木虱数量增速较慢。在供试的寄主植物品种中,乌榄叶片中单宁含量是橄榄的3倍以上,而蛋白质含量相对较低。     

Secalonic acid a,a vivotoxin in pink root-infected onion

A highly potent phytotoxic compound isolated from liquid cultures of Pyrenochaeta terrestris was identified as secalonic acid A. This toxic metabolite was identified as a component of infected onion roots.     

An eco-metabolomic study of host plant resistance to Western flower thrips in cultivated,biofortified and wild carrots

Domestication of plants and selection for agronomic traits may reduce plant secondary defence metabolites relative to their ancestors. Carrot (Daucus carota L.) is an economically important vegetable. Recently, carrot was developed as a functional food with additional health-promoting functions. Biofortified carrots contain increased concentrations of chlorogenic acid as an antioxidant. Chlorogenic acid is involved in host plant resistance to Western Flower Thrips (Frankliniella occidentalis), one of the key agri- and horticultural pests worldwide. The objective of this study was to investigate quantitative host plant resistance to thrips in carrot and to identify candidate compounds for constitutive resistance. As such we explored whether cultivated carrot is more vulnerable to herbivore attack compared to wild carrot. We subjected a set of 14 biofortified, cultivated and wild carrot genotypes to thrips infestation. We compared morphological traits and leaf metabolic profiles of the three most resistant and susceptible carrots using nuclear magnetic resonance spectroscopy (NMR). In contrast to our expectation, wild carrots were not more resistant to thrips than cultivated ones. The most thrips resistant carrot was the cultivar Ingot which is known to be tolerant against carrot root fly (Psila rosae). Biofortified carrots were not resistant to thrips. Plant size, leaf area and number of leaf hairs did not differ between resistant and susceptible carrots. The metabolic profiles of the leaves of resistant carrots were significantly different from those of susceptible carrots. The leaves of resistant carrots contained higher amounts of the flavanoid luteolin, the phenylpropanoid sinapic acid and the amino acid β-alanine. The negative effect of these compounds on thrips was confirmed using in-vitro bioassays. Our results have potential implications for carrot breeders. The natural variation of metabolites present in cultivated carrots can be used for improvement of thrips resistance. This is especially promising in view of the candidate compounds we identified since they do not only confer a negative effect on thrips but as antioxidants also play an important role in the improvement of human health.     

Characterization of the fungicidal activity of Calothrix elenkinii using chemical methods and microscopy

An investigation was directed towards biochemical characterization of cyanobacterium Calothrix elenkinii and analysis of the chemical nature and mode of action of its fungicidal metabolite(s) against oomycete Pythium debaryanum. Biochemical characterization of the culture in terms of carbohydrate utilization revealed the facultative nature of C. elenkinii. Unique antibiotic markers were also found for this strain. 16S rDNA sequencing of the strain revealed 98% similarity with Calothrix sp. PCC7101. The fungicidal activity was tested by disc diffusion assay of different fractions of the culture filtrate. A minimum inhibitory concentration of 10 microl was recorded for ethyl acetate fraction of the 7-weeks old culture filtrates. HPLC, followed by NMR spectral analysis demonstrated the presence of a substituted benzoic acid in the ethyl acetate fraction. Microscopic examination revealed distinct granulation, followed by disintegration of the hyphae of Pythium sp., indicating the presence of an active metabolite in the culture filtrates of Calothrix sp. The fungicidal activity of C. elenkinii can be attributed to the presence of 3-acetyl-2-hydroxy-6-methoxy-4-methyl benzoic acid. This is the first report of a benzoic acid derivative having fungicidal activity in cyanobacteria.     

The production of a phytotoxin,nectrolide, by some root-surface isolates ofCylindrocarpon radicicola,Wr.

Summary A phytotoxic compound, for which the name nectrolide has been proposed, was isolated fromCylindrocarpon radicicola. Analysis of nectrolide and its physical and chemical properties show that the compound is identical with brefeldin A, an antibiotic produced byPenicillium brefeldianum.Twenty different isolates ofC. radicicola from plant roots were examined for their ability to produce nectrolide in liquid culture. Twelve of these yielded nectrolide at concentrations varying from 31 to 82 µg per ml when grown for 14 days on Czapek-Dox liquid medium. Eight isolates did not yield the metabolite in culture.At a concentration of 6 µg per ml nectrolide arrested the growth of germinated blackbutt seedlings, whilst at lower concentrations (2–4 µg per ml) the compound caused severe stunting and blackening of the roots of seedling plants. Nectrolide watered onto blackbutt seedlings growing in forest soils can be detected in the roots after 24 hrs. The compound has been crystallised from ether extracts of tomato roots supplied with pure nectrolide and can be detected in stem tissues within 24 hours of watering it onto the roots.As a result of the widespread association ofC. radicicola with unthrifty plants and the phytotoxic nature of nectrolide, it is suggested that this compound may function in a number of plant diseases, the causes of which have not been elucidated.     

Phytotoxic Metabolites of Phyllosticta sp.

The additional new metabolites, named phyllostine (II) and 3-chloro-2,5-dihydroxybenzyl alcohol (III) from the cultural filtrates of Phyllosticta sp. have been isolated. The chemical structure of II have been established by spectral studies and chemical conversion from the known I, and the chlorine-containing metabolite (III) by spectral and synthetic studies. The metabolite (II) exhibits the similar phytotoxic effects as I, but the metabolite III does less phytotoxicity than I and II on the leaf test.     

Elicitin 172 from an isolate of Phytophthora nicotianae pathogenic to tomato

Elicitin 172, an acid protein with elicitor activity, has been isolated in true form from culture filtrates of Phytophthora nicotianae, the causal agent of crown and root rot of tomato (Lycopersicon esculentum). The M(r) (10,349 +/- 1) of the purified protein, determined by ES-MS, is identical to that calculated for parasiticein using the mean isotopic composition and assuming the occurrence of three disulfide bridges. The primary structure of elicitin 172, determined using also MALDI-MS experiments, shows complete identity with parasiticein, with elicitin 310 and a cloned elicitin gene from P. parasitica (= P. nicotianae), confirming conservation of the elicitin sequence within a single species. The protein induces necrosis (hypersensitive reaction) on tobacco, but no symptoms on tomato, when applied on the leaves. Tomato pretreated with elicitin 172 was affected by P. nicotianae, as well as by the phytotoxic aggregates, naturally occurring with the elicitin in the non permeated dialysis fraction of culture filtrates. Finally, the elicitin induce protection of capsicum (Capsicum annuum) and vegetable marrow (Cucurbita pepo) from P. capsici.