Biocontrol of broomrape using Fusarium oxysporum f. sp. orthoceras in tomato crops under field conditions

The effects of Fusarium oxysporum f. sp. orthoceras as a mycoherbicide on broomrape, a weedy parasite of tomato crops, were examined during two growing seasons under field conditions. In contrast to emerged broomrapes in treated plots, none of the tomato plants inoculated with fungal pathogen showed Fusarium-wilt symptoms. In 2008, both suspension concentrations, 1?×?10⁶ and 1?×?10⁷ spore/ml, used via either root-dip or soil-inoculation three weeks after transplanting, and high-concentration-soil-inoculation one week after transplanting, reduced the number of broomrape plant and shoots per plant compared to the control. In 2009, single-point-low-dose and twice-high-dose applications of sulfosulfuron had significantly fewer broomrape plants than high-concentration-soil-inoculation three weeks after transplanting. High-concentration-root-dip inoculation decreased broomrape biomass weight significantly by 58% compared to the control in 2008. In 2009, mean biomass for single-point-application of sulfosulfuron at 50 and 75?g levels was lower than control. The highest yield was detected in high-concentration-soil-inoculation at transplanting in 2008 and in low-dose-twice-application of sulfosulfuron in 2009. Tomato yield was associated with the number of broomrape shoots per plant. There is promise that these field-scale findings will improve attempts to use mycoherbicides for biocontrol of broomrapes in tomato cropping systems.     

Suppressiveness to Fusarium oxysporum f. sp. radicis lycopersici in re-used perlite and perlite–peat substrates in soilless tomatoes

This work reports the occurrence of suppressiveness to Fusarium oxysporum f. sp. radicis lycopersici (FORL) on recycled perlite and perlite–peat mix from closed and open soilless systems. Nine soilless systems were sampled from three different sites in Northern and Southern Italy and different parameters, including sampling site, growing period before sampling, electric conductivity of the nutrient solution, tomato cultivar, and irrigation system were considered. The effects on seed germination and FORL incidence on tomato seedlings were finally assessed with or without additional artificial inoculation of the pathogen and with or without autoclaving the samples prior to inoculation. Suppression of FORL was experimentally evaluated with a technique already adopted for rockwool. Results collected on perlite and perlite–peat confirmed the possibility to reduce FORL severity on recycled substrates. Only the composition of the substrate (perlite, perlite–peat mix) and the disinfestation did affect the incidence of FORL. Suppression of FORL was observed in not disinfected recycled perlite–peat substrates while a reduction of FORL incidence was also been recorded in disinfected and recycled perlite. Disease incidence decreased from an average ranging from 44.4% to 61.9% in new perlite to 2.5–36.3% in recycled one. Similarly disease incidence decreased from an average ranging from 35.9% to 75.2% in new perlite–peat mix to 0.4–26.4% in recycled perlite–peat mix. In conclusion the recorded data suggest the possibility to consider FORL suppression as a predictable phenomenon when recycled substrates (perlite, perlite–peat mix) are adopted.     

Interaction of endophytic diazotrophic bacteria and Fusarium oxysporum f. sp. cubense on plantlets of banana ‘Maça’

The objective of this work was to evaluate the effect of endophytic diazotrophic bacteria Herbaspirillum and Burkholderia on the Fusarium oxysporum f. sp. cubense (Foc) establishment and on the plantlets growth of the ‘Maçã’ banana (Musa spp., group ABB). Two assays were carried out in a greenhouse at the National Center of Tropical Agroindustry in Fortaleza city, Ceará State (Brazil), using randomized block designs in the factorial arrangements 4?×?2 and 8?×?2. On the first trial plantlets were inoculated with the Burkholderia sp. AB202; Herbaspirillum sp., BA227, both of strains and controls bacteria, with and without Foc and cultivated in pots filled with an autoclaved mixture of washed sand and vermiculite (ratio 3:2 v v^?1), during 4 months. On a second assay, plants were subjected to following conditions: absence and presence of strains AB202, AB213 (Burkholderia spp.), BA227, BA234 (Herbaspirillum-like), AB202 plus BA227, AB213 plus BA234, the mixture of the four bacterial strain, absence and presence of the Foc; and cultivated in pots filled with autoclaved Haplic Arenosol, during 2 months. The plant association with diazotrophs and the Foc was confirmed, and factors interacted significantly on the most probable number of bacteria and the colony forming units of the pathogen on roots and plant rhizomes. The potential of the endophytes on the inhibition of Foc propagate units and on the plant growth promotion was demonstrated. The higher biomass was observed four and 2 months after plant inoculation with AB202 and BA234. Results showed that these endophytes may be used as potential biofertilizer and biocontrol agents.     

Does vacuum-packaging or co-delivered amendments enhance shelf-life of Striga-mycoherbicidal products containing Fusarium oxysporum f. sp. strigae during storage?

The effect of vacuum packaging on the shelf-life and handling of Pesta granules and seed treatment made with chlamydospores of Fusarium oxysporum strains Foxy2, PSM197 or their mixture was studied at 4°C and 22±3°C over 1 year. In addition, the effects of co-incorporated amendments [urea in Pesta or co-delivered fungicides (Ridomil Gold®, Apron XL®) on coated sorghum seeds], and coating material (Arabic gum ‘AG’, SUET Binder ‘SB’) on the viability of Striga-mycoherbicides were evaluated. Storage under vacuum packaging did not enhance shelf-life of the formulated Striga-mycoherbicidal products after 12 months of storage regardless of the treatment used. The co-incorporated urea into Pesta granules significantly reduced the viability of mycoherbicides, but less so at 4°C (58% strain-stability after 12 months). No significant differences between the coating materials in maintaining the viability of mycoherbicides were observed. The shelf-life of isolates on coated seeds significantly decreased when adding Ridomil Gold®. However, at 4°C, the fungicide Apron XL® allowed better survival of Foxy2 and PSM197 by maintaining their averaged half-lives (t _0.5) by an additional 6 months compared to Ridomil Gold®. In general, Striga-mycoherbicidal product combinations exhibited a significantly higher shelf-life when stored at 4°C than at 22±3°C. The absence of a positive effect of vacuum packaging on shelf-life of Striga-mycoherbicidal products reflects the tolerance of the formulated fungal propagules (chlamydospores) to withstand an oxygen enriched environment and allows their handling and distribution through ordinary packaging systems in Africa. The high compatibility between Striga-mycoherbicides and the co-delivered fungicide Apron XL®, and the fungal storage stability allows simultaneous control of Striga and fungal cereal diseases within an integrated pest management (IPM).     

Strain-specific and recessive QTLs involved in the control of partial resistance to Fusarium oxysporum f. sp. melonis race 1.2 in a recombinant inbred line population of melon

Fusarium oxysporum f. sp. melonis (FOM) causes serious economic losses in melon (Cucumis melo L.). Two dominant resistance genes have been identified, Fom-1 and Fom-2, which provide resistance to races 0 and 2 and races 0 and 1, respectively, however FOM race 1.2 overcomes these resistance genes. A partial resistance to FOM race 1.2 that has been found in some Far East accessions is under polygenic control. A genetic map of melon was constructed to tag FOM race 1.2 resistance with DNA markers on a recombinant inbred line population derived from a cross between resistant (Isabelle) and susceptible (cv. Védrantais) lines. Artificial root inoculations on plantlets of this population using two strains, one that causes wilting (FOM 1.2w) and one that causes yellowing (FOM 1.2y), resulted in phenotypic and genotypic data that enabled the identification of nine quantitative trait loci (QTLs). These QTLs were detected on five linkage groups by composite interval mapping and explained between 41.9% and 66.4% of the total variation. Four digenic epistatic interactions involving seven loci were detected and increased the total phenotypic variation that was explained. Co-localizations between QTLs and resistance gene homologs or resistance genes, such as Fom-2 and Vat, were observed. A strain-specific QTL was detected, and some QTLs appeared to be recessive.     

Structures of Conversion Products Formed from 18β-Glycyrrhetinic Acid by Streptomyces sp. G-20

Dynamic profiles of the rate of O₂ generation from press-injured and inoculated rice leaf slices, versus the time after inoculation, discriminated between the incompatible and compatible combination of blast fungus races with a cultivar. The application of sodium saccharin to rice seedlings via the root system for 6 days changed the compatible to incompatible profile. Even after press- injury and inoculation with the compatible conidia, the leaf application of sodium saccharin enhanced superoxide generation. The application of N-methylsaccharin in a similar manner, however, did not enhance the superoxide generation. Inoculation of press-injured leaves with incompatible conidia in the presence of an aqueous diffusate of the germinating compatible conidia changed the incompatible to compatible profile. The application to press-injured of concanavalin A or a lyophylized preparation from 5 m ammonia extracts of rice leaf homogenate prior to stimulating with a resistance-inducing factor (RIF) from the fungus also enhanced the superoxide generation. The RIF, either from the incompatible or compatible race, gave a quite similar profile of activation upon the generation of the superoxide anion.     

Management of Fusarium oxysporum f. sp. lycopersici and root-knot nematode disease complex in tomato by use of antagonistic fungi,plant resistance and neem

Simultaneous infestation with root-knot nematodes (RKN) and Fusarium oxysporum f. sp. lycopersici (FOL) leads to formation of a disease complex that increases crop losses than effect of either RKN or FOL. In this study a management programme involving plant resistance, biological control agents, and neem was carried out to manage RKN and fusarium wilt disease complex. The biological control agents were Purpureocillium lilacinum (PL) and Trichoderma harzianum (TH) while the RKN was Meloidogyne javanica. In vitro dual culture plates were set up to test the interaction of biological control agents and FOL. Greenhouse experiments were conducted using two tomato cultivars Rambo F1 and Prostar F1. The treatments were; PL, TH, PL–TH, neem, PL neem, TH neem, and PL–TH neem. Each treatment was replicated four times and the treatments set up in a randomised complete block design in the greenhouse. Inhibition of FOL mycelial growth by TH and PL was 51.9%, and 44% respectively by the ninth day in vitro culture plates. In the cultivar, Prostar F1, the treatments PL–TH, PL, and TH in the presence or absence of neem had a FOL disease severity score significantly lower than the untreated control. Host resistance sufficed to prevent infection of Rambo F1 with FOL. The treatments PL–TH, PL and TH reduced FOL propagules and M. javanica juveniles in the roots and performed even better when combined with neem in both tomato cultivars. Therefore, a host that is resistant combined with biological control agents and organic amendments can be used in the management of RKN and FOL in tomato production.     

Wiskott-Aldrich Syndrome Protein (WASp) Controls the Delivery of Platelet Transforming Growth Factor-β1

Platelets are immunologically competent cells containing cytokines such as TGF-β1 that regulate cell-mediated immunity. However, the mechanisms underlying cytokine secretion from platelets are undefined. The Wiskott-Aldrich syndrome protein (WASp) regulates actin polymerization in nucleated hematopoietic cells but has other role(s) in platelets. WASp-null (WASp^−/−) platelets stimulated with a PAR-4 receptor agonist had increased TGF-β1 release compared with WT platelets; inhibiting WASp function with wiskostatin augmented TRAP-induced TGF-β1 release in human platelets. TGF-β1 release is dissociated from α-granule secretion (P-selectin up-regulation) and occurs more gradually, with ∼10–15% released after 30–60 min. Blockade of Src family kinase-mediated WASp Tyr-291/Tyr-293 phosphorylation increased TGF-β1 release, with no additive effect in WASp^−/− platelets, signifying that phosphorylation is critical for WASp-limited TGF-β1 secretion. Inhibiting F-actin assembly with cytochalasin D enhanced secretion in WT platelets and further increased TGF-β1 release in WASp^−/− platelets, indicating that WASp and actin assembly independently regulate TGF-β1 release. A permeabilized platelet model was used to test the role of upstream small GTPases in TGF-β1 release. N17Cdc42, but not Rac1 mutants, increased TGF-β1 secretion and abrogated WASp phosphorylation. We conclude that WASp function restricts TGF-β1 secretion in a Cdc42- and Src family kinase-dependent manner and independently of actin assembly.     

Some Physicochemical Properties and Substrate Specificity of Acid Protease Isolated from Cladosporium sp. No. 45–2

Some physicochemical properties and substrate specificity of crystalline acid protease obtained from Cladosporium sp. No. 45–2 were investigated.

The molecular weight determined by the sedimentation equilibrium method and Sephadex G–75 gel filtration was 32,000 and 30,000, respectively. The isoelectric point was determined as 4.6 by using the Tiselius electrophoresis apparatus and the amino terminal amino acid was found to be alanine. The enzyme did not contain histidine and was composed of 294 amino acid residues. Substrate specificity against synthetic peptides was similar to that of pepsin. The enzyme was remarkably inactivated by a synthetic pepsin inhibitor such as α-Diazo-p-bromo acetophenone, Diazo acetyl glycine ethylester and N-Diazo acetyl norleucine methylester.     

Phylogenetic relationship between different race representative populations of Fusarium oxysporum f. sp. ciceris in respect of translation elongation factor-1α, β-tubulin,and internal transcribed spacer region genes

Genetic diversity of 70 isolates of Fusarium oxysporum f. sp. ciceris originated from various states of India representing eight races causing wilt in chickpea (Cicer arietinum) was analyzed using translation elongation factor-1α (TEF-1α), β-tubulin, and internal transcribed spacer (ITS) gene regions. TEF-1α, β-tubulin, and ITS gene-specific markers produced ~720-, ~500-, and ~550-bp amplicons, respectively, in all the isolates of the pathogen. A phylogenetic tree constructed from the sequences generated in the present study along with the sequences of foreign isolates of Fusarium species available in NCBI database sharing more than 90 % nucleotide sequence similarity grouped the isolates into two major clusters. Most of the isolates of the present study showed more or less similar grouping pattern in case of the three gene sequences. Each group had the isolates representing different races as well as place of origin indicating low level of diversity among the isolates in respect of these gene sequences. Except TEF-1α, the groups generated by β-tubulin and ITS gene sequences did not correspond to the state of origin and races of the pathogen. However, the groups of TEF-1α partially corresponded to the place of origin as well as races of the pathogen. The isolates did not show any race-specific grouping patterns; however, most of the isolates representing race 1 clustered separately.     

Fusarium oxysporum (Schlecht.) Snyd. et Hans. — A dominant fungus species on the root-surface of woody plant seedlings

Summary and conclusions The imperfect fungusFusarium oxysporum dominated the root surface of first-year seedlings of woody plants grown in the forest nursery in eany near Pardubice, Czechoslovakia. The second most frequent species wasCylindrocarpon radicicola whose relative frequency increased towards the end of the growing season. The morphological variability of isolated strains ofFusarium oxysporum was studied; notwithstanding their great variability they all fall within the range of this complex species. The ecological conditions for the occurrence ofFusarium oxysporum on the root surface and the saprophytic or parasitic relations between this fungus and the higher plants are discussed on the basis of personal observations and reports in the literature.     

Maesaipsyche mekongensis sp. n. – the Third Species of the Genus from South-East Asia (Trichoptera,Arctopsychidae)

Maesaipsyche mekongensis sp. n. is described from Laos. The type series was collected near the Mekong River, which is suggestive of a potamobiontic habitat of the larvae. The male genitalic apparatus is illustrated.     

Tabanidae (Diptera) of Amaz?nia XXI. Descriptions of Elephantotus gen. n. and E. tracuateuensis sp. n. (Diachlorini) from the Brazilian coast

Elephantotus, a new genus of Tabanidae from the Amazon coast, Brazil, is described based on a new species E. tracuateuensis. Five females were collected in Pará State, and a male in Maranhão State. Arguments are presented for separating the new genus from Dasybasis, as well as the possibility of its occurrence being related to the nesting sites of coastal birds. The new species is characterized by its large size (x = 2.15 cm, n = 5 females), glabrous eyes, reddish-brown tegument, light brown frontal callus not touching the edges of the eyes, extending up to the vertex that has traces of ocelli, basal plate of the antennal flagellum with obtuse angle, without a tooth or spine, wings hyaline, with brown basal cells, without appendix in the fork of vein R₄₊₅, and genital furca wide with extended flaps.     

Morphology and life cycle of Ceratium rhomvoides nov. sp. (Dinophyceae) from the lake Plußsee (Federal Republic)

A new species of freshwater dinoflagellates, Ceratium rhomvoides nov. sp., is described from the Plußsee, northern West Germany, where it occurs regularly. C. rhomvoides nov. sp. is distinguished from closely related taxa by various aspects of the morphology of the vegetative cell and the cyst and by its sexual reproduction and seasonal distribution.     

The C-terminal Domain of the Long Form of Cellular FLICE-inhibitory Protein (c-FLIPL) Inhibits the Interaction of the Caspase 8 Prodomain with the Receptor-interacting Protein 1 (RIP1) Death Domain and Regulates Caspase 8-dependent Nuclear Factor κB (NF-κB) Activation

Caspase 8 plays an essential role in the regulation of apoptotic and non-apoptotic signaling pathways. The long form of cellular FLICE-inhibitory protein (c-FLIP_L) has been shown previously to regulate caspase 8-dependent nuclear factor κB (NF-κB) activation by receptor-interacting protein 1 (RIP1) and TNF receptor-associated factor 2 (TRAF2). In this study, the molecular mechanism by which c-FLIP_L regulates caspase 8-dependent NF-κB activation was further explored in the human embryonic kidney cell line HEK 293 and variant cells barely expressing caspase 8. The caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone greatly diminished caspase 8-dependent NF-κB activation induced by Fas ligand (FasL) when c-FLIP_L, but not its N-terminal fragment c-FLIP(p43), was expressed. The prodomain of caspase 8 was found to interact with the RIP1 death domain and to be sufficient to mediate NF-κB activation induced by FasL or c-FLIP(p43). The interaction of the RIP1 death domain with caspase 8 was inhibited by c-FLIP_L but not c-FLIP(p43). Thus, these results reveal that the C-terminal domain of c-FLIP_L specifically inhibits the interaction of the caspase 8 prodomain with the RIP1 death domain and, thereby, regulates caspase 8-dependent NF-κB activation.