Reaction of sugarcane genotypes to Pratylenchus brachyurus and P. zeae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Reação de porta-enxertos de videira a Pratylenchus brachyurus e Pratylenchus zeae

Nesse trabalho, o objetivo foi avaliar a reação de oito porta-enxertos de videira aos nematoides das lesões radiculares. As estacas dos porta-enxertos Kober, SO4, 101-14, R99, 420-A, Rupestris du Lot, Riparia do Traviú e Telek 5C, cedidas pelo Centro APTA de Frutas/IAC, foram plantadas em vasos contendo mistura de solo:areia na proporção 2:1 (v:v) e mantidas em casa de vegetação. Após quatro meses, os porta-enxertos foram inoculados com 1.200 espécimes de Pratylenchus brachyurus ou P. zeae, e o milho foi usado para comprovar a viabilidade do inóculo. Aos 180 dias após a inoculação, avaliou-se o número de nematoides por sistema radicular e em 100 cm³ de solo. O milho foi avaliado aos 90 dias após a inoculação e cada vaso recebeu uma nova planta, que foi avaliada juntamente com os porta-enxertos. No milho, as populações finais de P. brachyurus e P. zeae foram respectivamente iguais a 8.040 e 6.940 indivíduos. Todos os porta-enxertos comportaram-se como imunes a P. brachyurus e P. zeae, isto é, a população final dos nematoides e o fator de reprodução foram iguais a zero. Recuperaram-se seis e 11 espécimes de P. zeae nas amostras de solo cultivadas com os porta-enxertos Kober e 420-A, respectivamente. Conclui-se que os porta-enxertos estudados apresentam potencial para serem usados em áreas infestadas com esses nematoides das lesões.

Potencial in vitro de rizobactérias para o controle de nematóides-chave da cana-de-açúcar, dinâmica populacional e manejo de Pratylenchus zeae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Agressividade comparada de Pratylenchus brachyurus com P. zeae e eficácia de métodos de controle de nematoides em cana-de-açúcar

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Aggressiveness of pratylenchus brachyurus to sugarcane, compared with key nematode p. zeae

Pratylenchus zeae, Meloidogyne javanica and M. incognita are considered key species of nematodes in sugarcane in Brazil, but P. brachyurus is also frequently found. This study was conducted to determine the aggressiveness of P. brachyurus compared with P. zeae to sugarcane. Plants were grown in pots (100 L) in an open area with initial inoculation of 10, 100, 1,000, 10,000 and 100,000/plant for P. brachyurus and P. zeae. The nematode inocula were from in vitro, carrot-cylinder cultures. Sampling was performed every 60 days until 300 days after inoculation. At harvest, we evaluated the population dynamics of the nematodes and plant growth characteristics. The population for the initial levels of 10 and 100,000 specimens/plant, for P. brachyurus and P. zeae at 300 days after inoculation were similar. This fact shows that, upon detection of nematodes in a certain place during the planting of sugarcane, the ratoon on this area should be treated so as to control populations of P. brachyurus and/or P. zeae. The damage caused by the initial population of 10 specimens of P. brachyurus was similar to those of 10.000 specimens of P. zeae. The variety CTC 2 was classified as susceptible to P. zeae and intolerant to P. brachyurus. Compared to the control, the losses as measured by the volume and fresh weight of shoots by the nematode species were 29.82% and 40.34%, respectively. Pratylenchus brachyurus was more aggressive than P. zeae to the CTC 2 sugarcane variety.

Comparison of lipase production on crambe oil and meal by Fusarium sp (Gibberella fujikuroi complex)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chicken fat and inorganic nitrogen source for lipase production by Fusarium sp. (Gibberella fujikuroi complex)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structure Variation And Dynamics of the Nuclear Ribosomal Intergenic Spacer Region in Key Gibberella Fujikuroi Complex Species

The intergenic spacer (IGS) region of the ribosomal DNA was cloned and sequenced in eight species within the Gibberella fujikuroi species complex with anamorphs in the genus Fusarium , a group that includes the most relevant toxigenic species. DNA sequence analyses revealed two categories of repeated elements: long repeats and short repeats of 125 and 8 bp, respectively. Long repeats were present in two copies and were conserved in all the species analyzed, whereas different numbers of short repeat elements were observed, leading to species-specific IGS sequences with different length. In Fusarium subglutinans and Fusarium nygamai , these differences seemed to be the result of duplication and deletion events. Here, we propose a model based on unequal crossing over that can explain these processes. The partial IGS sequence of 22 Fusarium proliferatum isolates was also obtained to study variation at the intraspecific level. The results revealed no differences in terms of number or pattern of repeated elements and detected frequent gene conversion events. These results suggest that the homogenization observed at the intraspecific level might not be achieved primarily by unequal crossing-over events but rather by processes associated with recombination such as gene conversion events.

The P450–1 gene of Gibberella fujikuroi encodes a multifunctional enzyme in gibberellin biosynthesis

Recent studies have shown that the genes of the gibberellin (GA) biosynthesis pathway in the fungus Gibberella fujikuroi are organized in a cluster of at least seven genes. P450–1 is one of four cytochrome P450 monooxygenase genes in this cluster. Disruption of the P450–1 gene in the GA-producing wild-type strain IMI 58289 led to total loss of GA production. Analysis of the P450–1-disrupted mutants indicated that GA biosynthesis was blocked immediately after ent-kaurenoic acid. The function of the P450–1 gene product was investigated further by inserting the gene into mutants of G. fujikuroi that lack the entire GA gene cluster; the gene was highly expressed under GA production conditions in the absence of the other GA-biosynthesis genes. Cultures of transformants containing P450–1 converted ent-[14C]kaurenoic acid efficiently into [14C]GA14, indicating that P450–1 catalyzes four sequential steps in the GA-biosynthetic pathway: 7β-hydroxylation, contraction of ring B by oxidation at C-6, 3β-hydroxylation, and oxidation at C-7. The GA precursors ent-7α-hydroxy[14C]kaurenoic acid, [14C]GA12-aldehyde, and [14C]GA12 were also converted to [14C]GA14. In addition, there is an indication that P450–1 may also be involved in the formation of the kaurenolides and fujenoic acids, which are by-products of GA biosynthesis in G. fujikuroi. Thus, P450–1 displays remarkable multifunctionality and may be responsible for the formation of 12 products.

SIG e distribuição espacial da ferrugem tropical do milho (Physopella zeae) no cenário de mudanças climáticas no Brasil.

A mudança climática é um processo decorrente do efeito acumulativo contínuo das emissões excessivas de gases de efeito estufa e de aerossóis provenientes da intensificação de algumas atividades antrópicas, produzindo mudanças no clima do planeta. Tais mudanças podem alterar a incidência de doenças nas plantas. O conhecimento de técnicas de geoprocessamento pode ser de grande importância para aplicações no setor agrícola, auxiliando na elaboração de estratégias para minimizar prejuízos futuros causados pelos impactos climáticos globais. A ferrugem tropical do milho, causada por Physopella zeae, é responsável por grandes danos na produção nacional. Este trabalho teve como objetivo avaliar a distribuição geográfica dessa doença com o uso de ferramentas SIG (Sistema de Informações Geográficas). Foram obtidos mapas de favorabilidade climática para o desenvolvimento da doença na cultura do milho, utilizando condições climáticas propícias definidas por Casela et al. (2006), entre os meses de janeiro a junho, para a normal climatológica (1961-1990) e para o clima futuro de 2080, do IPCC (Painel Intergovernamental de Mudanças Climáticas). Os resultados indicam que haverá um deslocamento da ncidência da doença das regiões Norte, Nordeste para as regiões Sudeste e Sul no futuro, permanecendo também na região Centro-Oeste.

Impact of tillage and residues from rotation crops on the nematode community in soil and surface mulch during the following sugarcane crop.

The impact of three cropping histories (sugarcane, maize and soybean) and two tillage practices (conventional tillage and direct drill) on plant-parasitic and free-living nematodes in the following sugarcane crop was examined in a field trial at Bundaberg. Soybean reduced populations of lesion nematode (Pratylenchus zeae) and root-knot nematode (Meloidogyne javanica) in comparison to previous crops of sugarcane or maize but increased populations of spiral nematode (Helicotylenchus dihystera) and maintained populations of dagger nematode (Xiphinema elongatum). However the effect of soybean on P zeae and M. javanica was no longer apparent 15 weeks after planting sugarcane, while later in the season, populations of these nematodes following soybean were as high as or higher than maize or sugarcane. Populations of P zeae were initially reduced by cultivation but due to strong resurgence tended to be higher in conventionally tilled than direct drill plots at the end of the plant crop. Even greater tillage effects were observed with M. javanica and X. elongatum, as nematode populations were significantly higher in conventionally tilled than direct drill plots late in the season. Populations of free-living nematodes in the upper 10 cm of soil were initially highest following soybean, but after 15, 35 and 59 weeks were lower than after sugarcane and contained fewer omnivorous and predatory nematodes. Conventional tillage increased populations of free-living nematodes in soil in comparison to direct drill and was also detrimental to omnivorous and predatory nematodes. These results suggest that crop rotation and tillage not only affect plant-parasitic nematodes directly, but also have indirect effects by impacting on natural enemies that regulate nematode populations. More than 2 million nematodes/m(2) were often present in crop residues on the surface of direct drill plots. Bacterial-feeding nematodes were predominant in residues early in the decomposition process but fungal-feeding nematodes predominated after 15 weeks. This indicates that fungi become an increasingly important component of the detritus food web as decomposition proceeds, and that that the rate of nutrient cycling decreases with time. Correlations between total numbers of free-living nematodes and mineral N concentrations in crop residues and surface soil suggested that the free-living nematode community may provide an indication of the rate of mineralisation of N from organic matter.

Fungal community structure in disease suppressive soils assessed by 28S LSU gene sequencing

Natural biological suppression of soil-borne diseases is a function of the activity and composition of soil microbial communities. Soil microbe and phytopathogen interactions can occur prior to crop sowing and/or in the rhizosphere, subsequently influencing both plant growth and productivity. Research on suppressive microbial communities has concentrated on bacteria although fungi can also influence soil-borne disease. Fungi were analyzed in co-located soils 'suppressive' or 'non-suppressive' for disease caused by Rhizoctonia solani AG 8 at two sites in South Australia using 454 pyrosequencing targeting the fungal 28S LSU rRNA gene. DNA was extracted from a minimum of 125 g of soil per replicate to reduce the micro-scale community variability, and from soil samples taken at sowing and from the rhizosphere at 7 weeks to cover the peak Rhizoctonia infection period. A total of ∼994,000 reads were classified into 917 genera covering 54% of the RDP Fungal Classifier database, a high diversity for an alkaline, low organic matter soil. Statistical analyses and community ordinations revealed significant differences in fungal community composition between suppressive and non-suppressive soil and between soil type/location. The majority of differences associated with suppressive soils were attributed to less than 40 genera including a number of endophytic species with plant pathogen suppression potentials and mycoparasites such as Xylaria spp. Non-suppressive soils were dominated by Alternaria , Gibberella and Penicillum. Pyrosequencing generated a detailed description of fungal community structure and identified candidate taxa that may influence pathogen-plant interactions in stable disease suppression. © 2014 Penton et al.

Status, challenges and tools for identification and diagnosis of Peronosclerospora and Sclerophthora of regulatory concern for graminicolous crops

Graminicolous Downy Mildew (GDM) diseases caused by the genera Peronosclerospora (13 spp.) and Sclerophthora (6 spp. and 1 variety) are poorly studied but destructive diseases of major crops such as corn, sorghum, sugarcane and other graminoids. Eight of the 13 described Peronosclerospora spp. are able to infect corn. In particular, P. philippinensis (= P. sacchari), P. maydis, P. heteropogonis, and S. rayssiae var. zeae cause major losses in corn yields in tropical Asia. In 2012 a new species, P. australiensis, was described based on isolates previously identified as P. maydis in Australia; this species is now a pathogen of major concern. Despite the strong impact of GDM diseases, there are presently no reliable molecular methods available for their detection. GDM pathogens are among the most difficult Oomycetes to identify using molecular tools, as their taxonomy is very challenging, and little genetic sequence data are available for development of molecular tools to detect GDM pathogens to species level. For example, from over 15 genes used in identification, diagnostics or phylogeny of Phytophthora, only ITS1 and cox2 show promise for use with GDM pathogens. Multiplex/multigene conventional and qPCR assays are currently under evaluation for the detection of economically important GDM spp. Scientists from the USA, Germany, Canada, Australia, and the Philippines are collaborating on the development and testing of diagnostic tools for these pathogens of concern.

Lactobacillus bacteremia, with special focus on the safety of probiotic Lactobacillus rhamnosus GG

Lactobacilli are gram positive rods, which belong to normal oropharyngeal, gastrointestinal and urogenital flora. They are widely used in food industry and as food additives. Although their virulence is presumed to be very low, opportunistic bacteremic infections, especially in immunocompromised hosts, have been detected. In the present study, the possible effects of increasing probiotic use of Lactobacillus rhamnosus GG (LGG) on the occurrence of bacteremia due to lactobacilli was evaluated on population level. In Finland, 90 Lactobacillus bacteremia cases were reported to the National Infectious Disease Register maintained by National Public Health Institute, during 1995-2000. Their proportion of all bacteremia cases was on average 0.24%, corresponding to 0.3 cases/100 000 inhabitants annually. In the Helsinki University Central Hospital district the corresponding proportion of all bacteremia cases was observed during 1990-2000. Despite LGG intake increased six folded no increasing trend in the occurrence of lactobacilli bacteremia was seen. A total of 85 Lactobacillus sp. blood isolates collected from different human bacteremic cases were characterised and compared with the commercial probiotic LGG strain. In species characterisation 46 L. rhamnosus strains, 12 L. fermentum and L. casei strains each, three each of L. gasseri, L. salivarius and L. jensenii species, two L. curvatus, and one each of L. plantarum, L. sakei, L. zeae and L. reuteri species were detected. Nearly half of the L. rhamnosus findings turned out to be indistinguishable from the probiotic LGG strain. Common predisposing factors to Lactobacillus bacteremia were immunosuppression, prior prolonged hospitalisation and prior surgical interventions. Severe or fatal comorbidities were found in 82% of the patients. Mortality at one month was 26% and severe underlying diseases were a significant predictor of death (OR 15.8). Antimicrobial susceptibility of Lactobacillus strains was species dependent. The Lactobacillus isolates were generally susceptible to imipenem, piperacillin-tazobactam, clindamycin and erythromycin, whereas all other than L. gasseri and L. jensenii species were not at all susceptible to vancomycin. The susceptibility to cephalosporin varied greatly even within species why they might not be recommended for treatment of Lactobacillus infections. The effect and safety of probiotic LGG preparation in amelioration of gastric symptoms and diarrhea in HIV-infected patients was evaluated. No significant differences in gastrointestinal symptoms or diarrhoea in LGG treated patients compared to placebo could be found. LGG was well tolerated with no adverse effects including bacteremic outbreaks could be observed. The use of probiotic LGG can be regarded safe in this immunocompromised patient group.

Indoleacetaldoxime hydro-lyase : II. Purification and properties

The purification and some properties of the enzyme indoleacetaldoxime hydrolyase (EC 4.2.1.29) from the fungus Gibberella fujikuroi, which dehydrates indoleacetaldoxime (IAOX) to indoleacetonitrile (IAN), are described. The enzyme activity in the fungus is present only under certain culture conditions. It is a soluble enzyme, has an optimum pH at 7, shows an energy of activation of —15,670 cal/mole, and has a Michaelis constant of 1.7 × 10−4 Image at 30 °. It appears to be specific for IAOX, and 1 mole of IAN is produced per mole of IAOX utilized. The enzyme is inhibited by a number of aldoximes of which phenylacetaldoxime (PAOX) is the most potent inhibitor. Inhibition by PAOX is competitive (Ki = 2.2 × 10−8 Image ). The enzyme is inhibited by SH reagents such as p-hydroxymercuribenzoate and N-ethylmaleimide, and by a number of SH compounds such as cysteine, β-mercaptoethanol, and 2,3-dimercaptopropanol (BAL). However, glutathione activates the enzyme. Metal chelating agents such as 8-OH-quinoline and diethyl dithiocarbamate inhibit the enzyme; the inhibition is partly reversed by ferric citrate. Ascorbic acid, and particularly dehydroascorbic acid (DHA), are good activators of the enzyme. Several other biological oxidants had either no action or had a slight effect. Potassium cyanide activates the enzyme at low concentration but inhibits at higher concentrations. Reduction of the enzyme with NaBH4 reduces activity, and the effect is partly reversed by pyridoxal phosphate and also by DHA. The above properties indicate that both an SH function and an oxidized function are required for activity.