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Acanthoscelides obtectus and Sitophilus zeamais are among the most damaging stored-product insect pests. This study was carried out to identify Beauveria bassiana and Metarhizium anisopliae and to evaluate their insecticidal potential against A. obtectus and S. zeamais adults. Fungal isolates were characterized molecularly based on ITS1, 5.8S and ITS2 regions of rDNA. B. bassiana YK23 and YK26, and M. anisopliae YK41 and YK45 were sprayed onto adults of A. obtectus and S. zeamais at 1×105, 1×106 and 1×107 conidia mL−1 doses. At the end of the 10th day of application, the numbers of dead adults were counted. Mortality rates (%) and lethal times (LT50 and LT90) were calculated for each isolate. Fungal isolates displayed promising insecticidal activity against A. obtectus and S. zeamais. B. bassiana and M. anisopliae caused 100% mortality against A. obtectus, even at the lowest dose. However, B. bassiana and M. anisopliae caused 100% mortality on S. zeamais at higher doses. Depending on the increasing doses, significant decreases were observed at the lethal times. Median lethal time (LT50) decreased up to 64.24 h (2.67 days) and 53.69 h (2.24 days) in A. obtectus exposed to B. bassiana and M. anisopliae, respectively. LT50 decreased to 61.04 h (2.54 days) and 53.89 h (2.25 days) in S. zeamais exposed to the same fungi. B. bassiana and M. anisopliae can be taken into account in the control of those pests.

Xanthomonas oryzae pv. oryzae (Xoo) is a destructive pathogen that causes bacterial blight disease of rice worldwide. Xoo uses T3SS (type III secretion system) effectors to subvert rice innate immunity. However, the comprehensive knowledge of rice genes involved in T3SS effectors-mediated interaction remains unclear. In this study, the transcriptome profiles of rice infected with a virulent Xoo strain from North-eastern region of India relatives to its avirulent strain (that lacks functional T3SS) were analyzed at early (2–6 hpi) and late (16–24 hpi) hours of infection. Out of total 255 differentially expressed genes (DEGs), during early infection, 62 and 70 genes were upregulated and downregulated, respectively. At late infection, 70 and 53 genes were upregulated and downregulated, respectively. The transcriptomic data identified many differentially expressed resistant genes, transposons, transcription factors, serine/threonine protein kinase, cytochrome P450 and peroxidase genes that are involved in plant defense. Pathway analysis revealed that these DEGs are involved in hormone signaling, plant defense, cellular metabolism, growth and development processes. DEGs associated with plant defense were also validated through quantitative real-time PCR. Our study brings a comprehensive picture of the rice genes that are being differentially expressed during bacterial blight infection. Nevertheless, the DEG-associated pathways would provide sensible targets for developing resistance to bacterial blight. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-022-03193-4.

Introduction: The endemic fishes of the ancient lakes of Sulawesi are under increasing threat. Objective: To evaluate the data and information available from a holistic management perspective and to formulate measures to conserve the endemic ricefish Oryzias nigrimas in Poso Lake, Indonesia. Methods: Collection of primary data from three stations around Lake Poso and literature study. Results: Threats to O. nigrimas include habitat degradation and loss, introduced alien species, and exploitation as a locally important food fish. Options to promote sustainable fisheries management include spatial and temporal limitations to minimise catch of gravid or brooding fish. Habitat protection should include measures to minimise impacts from activities which can reduce water quality and disturb or kill aquatic vegetation. Conclusion: Measures to prevent further O. nigrimas population decline are considered urgent and further research is recommended to fill identified knowledge gaps. Ex-situ conservation, including the development of captive breeding, could also contribute to a holistic O. nigrimas conservation strategy. Resumen Introducción: Los peces endémicos de los antiguos lagos de Sulawesi están cada vez más amenazados. Objetivo: Evaluar los datos y la información disponible desde una perspectiva de gestión holística y formular medidas para conservar el pez endémico Oryzias nigrimas en el lago Poso, Indonesia. Métodos: Recopilación de datos primarios de tres estaciones alrededor del lago Poso y revisión de la literatura. Resultados: Las amenazas para O. nigrimas incluyen la degradación y pérdida del hábitat, la introducción de especies exóticas y la explotación como producto pesquero de importancia local. Las opciones para promover la ordenación pesquera sostenible incluyen limitaciones espaciales y temporales para minimizar la captura de peces grávidos o reproductores. La protección del hábitat debe incluir medidas para minimizar los impactos de las actividades que pueden reducir la calidad del agua y perturbar o matar la vegetación acuática. Conclusión: Las medidas para prevenir una disminución de la población de O. nigrimas se consideran urgentes y se recomienda realizar más investigaciones para llenar los vacíos de conocimiento identificados. La conservación ex situ, incluido el desarrollo de la cría en cautividad, también podría contribuir a una estrategia de conservación holística de O. nigrimas.

Made available in DSpace on 2020-12-11T12:42:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-16 National Natural Science Foundation of China CAS President's International Fellowship Initiative (PIFI) China Postdoctoral Science Foundation SERB-Start-Up Research Grant (Young Scientist) Scheme, Department of Science and Technology, Government of India, New Delhi Antimicrobial activity of silver nanoparticles (AgNPs) has been well documented in earlier studies. As their efficient role in combating phytopathogens has begun recently, there is a huge scope to explore their effectiveness in agriculture. Considering the strong antifungal activity of biosynthesized AgNPs (as reported in our previous study), our main aim is to elucidate their antibacterial activity against bacterial plant pathogens to authenticate their wide range of agricultural applications. The present manuscript highlights the potential role of biosynthesized AgNPs against Xanthomonas oryzae pv. oryzae (Xoo) causing disastrous sheath blight disease of rice worldwide. We observed strong antibacterial activity of biosynthesized AgNPs (size similar to 12 nm) against Xoo at 20, 30 and 50 mu g/mL concentrations. The significant inhibitory impact of AgNPs on biofilm formation by Xoo was noted even at the lower dose of 5 mu g/mL (p = 0.001). Maximum biofilm inhibition (p = 0.000) was caused at 50 mu g/mL concentration of AgNPs in comparison to control. Furthermore, disease suppression by biosynthesized AgNPs was authenticated under greenhouse conditions. Foliar spray of AgNPs significantly reduced the blight symptoms in rice sheaths as shown by 9.25% DLA (% Diseased leaf area) as compared to 33.91% DLA in Xoo inoculated rice plants. Altogether, our data suggest that biosynthesized AgNPs based nanoformulation can be applied for successful management of blight disease of rice. In addition, the antibiofilm strategies instigated by AgNPs can be exploited against a wide range of bacterial phytopathogens. In light of rapidly emerging antibiotic-resistant microbial strains, the current work provides an alternate effective platform for the application of nanoformulation for augmenting sustainability in the agriculture. Chinese Acad Sci, Xishuangbanna Trop Bot Garden, CAS Key Lab Trop Forest Ecol, Menglun 666303, Yunnan, Peoples R China Banaras Hindu Univ, Dept Mycol & Plant Pathol, Inst Agr Sci, Varanasi 221005, Uttar Pradesh, India Sao Paulo State Univ, Lab Environm Nanotechnol, Inst Sci & Technol Sorocaba, Sao Paulo, Brazil Somvanshi Res Fdn, 13-21 Vikas Nagar, Lucknow 226022, Uttar Pradesh, India Sao Paulo State Univ, Lab Environm Nanotechnol, Inst Sci & Technol Sorocaba, Sao Paulo, Brazil National Natural Science Foundation of China: 31700457 CAS President's International Fellowship Initiative (PIFI): 2019PC0095 China Postdoctoral Science Foundation: 2018M631112 SERB-Start-Up Research Grant (Young Scientist) Scheme, Department of Science and Technology, Government of India, New Delhi: YSS/2015/000082

The rice XA21 and XA3 pattern receptor kinases, derived from Oryza longistaminata and an Oryza. sativa japonica cultivar Wase Aikoku 3, respectively, confer resistance to strains of the Gram-negative bacterium Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of rice bacterial blight disease. Previously, we showed that transfer of Xa21 to the model rice cultivar Kitaake enhances resistance to Xoo. In this manuscript we demonstrate that Kitaake expressing Xa3 confers resistance to Xoo strain PXO79 and that the stress-related marker genes PR10b and KO5 are upregulated in Xoo-infected Xa3 rice leaves. We also show that rice somatic embryogenesis receptor kinase 2 (OsSERK2) positively regulates XA3-mediated immunity in Kitaake. We found that overexpression of XA21 binding protein 15 (XB15) and XB24, two negative regulators of XA21-mediated immunity, do not affect XA3-mediated immunity in the Kitaake genetic background. Our results indicate that the rice immune receptors XA21 and XA3 employ both shared and distinct signaling components in their response to Xoo. The results are important to further understand pathogen-associated molecular pattern (PAMP)-triggered immunity in rice. Furthermore, the presence of Kitaake rice carrying Xa3 will facilitate genetic research to study the XA3-mediated immunity.

Genome sequence comparisons to infer likely gene functions require accurate ortholog assignments. In Pseudomonas spp., the sensor-regulator ColS-ColR two-component regulatory system responds to zinc and other metals to control certain membrane-related functions, including lipid A remodeling. In Xanthomonas spp., three different two-component regulatory systems, RaxH-RaxR, VgrS-VgrR, and DetS-DetR, have been denoted as ColS-ColR in several different genome annotations and publications. To clarify these assignments, we compared the sensor periplasmic domain sequences and found that those from Pseudomonas ColS and Xanthomonas RaxH share a similar size as well as the location of a Glu-X-X-Glu metal ion-binding motif. Furthermore, we determined that three genes adjacent to raxRH are predicted to encode enzymes that remodel the lipid A component of lipopolysaccharide. The modifications catalyzed by lipid A phosphoethanolamine transferase (EptA) and lipid A 1-phosphatase (LpxE) previously were detected in lipid A from multiple Xanthomonas spp. The third gene encodes a predicted lipid A glycosyl transferase (ArnT). Together, these results indicate that the Xanthomonas RaxH-RaxR system is orthologous to the Pseudomonas ColS-ColR system that regulates lipid A remodeling. To avoid future confusion, we recommend that the terms ColS and ColR no longer be applied to Xanthomonas spp., and that the Vgr, Rax, and Det designations be used instead.

Pseudomonas species are metabolically robust, with capacity to produce secondary metabolites including cyclic lipopeptides (CLPs). Herein we conducted a chemical analysis of a crude CLP extract from the cocoyam rhizosphere-derived biocontrol strain Pseudomonas sp. COW3. We performed in silico analyses on its whole genome, and conducted in vitro antagonistic assay using the strain and purified CLPs. Via LC-MS and NMR, we elucidated the structures of four novel members of the bananamide group, named bananamides D-G. Besides variability in fatty acid length, bananamides D-G differ from previously described bananamides A-C and MD-0066 by the presence of a serine and aspartic acid at position 6 and 2, respectively. In addition, bananamide G has valine instead of isoleucine at position 8. Kendrick mass defect (KMD) allowed the assignment of molecular formulae to bananamides D and E. We unraveled a non-ribosomal peptide synthetase cluster banA, banB and banC which encodes the novel bananamide derivatives. Furthermore, COW3 displayed antagonistic activity and mycophagy against Pythium myriotylum, while it mainly showed mycophagy on Pyricularia oryzae. Purified bananamides D-G inhibited the growth of P. myriotylum and P. oryzae and caused hyphal distortion. Our study shows the complementarity of chemical analyses and genome mining in the discovery and elucidation of novel CLPs. In addition, structurally diverse bananamides differ in their antimicrobial activity.

Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver an opinion on proline-specific oligopeptidase (Tolerase (R) G) as a novel food ingredient submitted pursuant to Regulation (EC) No 258/97 of the European Parliament and of the Council, taking into account the comments and objections of a scientific nature raised by Member States. The novel food is an enzyme preparation of prolyl-oligopeptidase produced with a genetically modified Aspergillus niger self clone strain. The target population is the general adult population. The results from a bacterial reverse mutation test and of an in vitro chromosome aberration test did not indicate genotoxicity. The Panel considers that the reported effects observed in a 90-day rat study are treatment-related effects and can be attributed to the higher energy consumption by these animals. Taking into account the intended maximum use level for Tolerase (R) G, its daily consumption would correspond to 2,746 mg TOS/person or to 39.2 mg TOS/kg body weight (bw) per day, when considering a default body weight of 70 kg for an adult person. The margin between this value and the dose in the rats, which caused effects attributable to the excess energy intake, is approximately 45. Noting this margin, the Panel considers that it is unlikely that such effects would occur in human at the intended use levels. The Panel concludes that the NF, Tolerase (R) G, is safe for the intended use at the intended use level. (C) 2017 European Food Safety Authority.

Cyclopiazonic acid (α-cyclopiazonic acid, α-CPA) is an indole-hydrindane-tetramic acid neurotoxin produced by various fungal species, including the notorious food and feed contaminant Aspergillus flavus. Despite its discovery in A. flavus cultures approximately 40 years ago, its contribution to the A. flavus mycotoxin burden is consistently minimized by our focus on the more potent carcinogenic aflatoxins also produced by this fungus. Here, we report the screening and identification of several CPA-type alkaloids not previously found in A. flavus cultures. Our identifications of these CPA-type alkaloids are based on a dereplication strategy involving accurate mass high resolution mass spectrometry data and a careful study of the α-CPA fragmentation pattern. In total, 22 CPA-type alkaloids were identified in extracts from the A. flavus strains examined. Of these metabolites, 13 have been previously reported in other fungi, though this is the first report of their existence in A. flavus. Two of our metabolite discoveries, 11,12-dehydro α-CPA and 3-hydroxy-2-oxo CPA, have never been reported for any organism. The conspicuous presence of CPA and its numerous derivatives in A. flavus cultures raises concerns about the long-term and cumulative toxicological effects of these fungal secondary metabolites and their contributions to the entire A. flavus mycotoxin problem., Valdet Uka was financially supported by Project Basileus V (Erasmus Mundus Action 2) funding from the European Commission.

Xanthomonas arboricola pv. pruni is the causal agent of the bacterial spot disease of stone fruits, almond and some ornamental Prunus species. In Spain it was first detected in 2002 and since then, several outbreaks have occurred in different regions affecting mainly Japanese plum, peach and almond, both in commercial orchards and nurseries. As the origin of the introduction(s) was unknown, we have assessed the genetic diversity of 239 X. arboricola pv. pruni strains collected from 11 Spanish provinces from 2002 to 2013 and 25 reference strains from international collections. We have developed an optimized multilocus variable number of tandem repeat analysis (MLVA) scheme targeting 18 microsatellites and five minisatellites. A high discriminatory power was achieved since almost 50% of the Spanish strains were distinguishable, confirming the usefulness of this genotyping technique at small spatio-temporal scales. Spanish strains grouped in 18 genetic clusters (conservatively delineated so that each cluster contained haplotype networks linked by up to quadruple-locus variations). Furthermore, pairwise comparisons among populations from different provinces showed a strong genetic differentiation. Our results suggest multiple introductions of this pathogen in Spain and redistribution through contaminated nursery propagative plant material. This work was supported by Instituto Nacional de Tecnología Agraria y Alimentaria. Project RTA-2011-00140-C03-01 (http://www.inia. es) to PLS MCM JP MML, Euphresco project (266505-FP7-ERANET Euphresco II) to PLS MCM JP APB MML, European Social Fund to PLS KB MCM JP APB CV MML OP, European Regional Published

Background Xanthomonas oryzae pv. oryzae (Xoo) is the causal agent of Bacterial Leaf Blight (BB), an emerging disease in rice in West-Africa which can induce up to 50 % of yield losses. So far, no specific resistance gene or QTL to African Xoo were mapped. The objectives of this study were to identify and map novels and specific resistance QTLs to African Xoo strains. Results The reference recombinant inbred lines (RIL) mapping population derived from the cross between IR64 and Azucena was used to investigate Xoo resistance. Resistance to African and Philippine Xoo strains representing different races was assessed on the RIL population under greenhouse conditions. Five major quantitative trait loci (QTL) for resistance against African Xoo were located on different chromosomes. Loci on chromosomes 1, 7, 9, 10 and 11 explained as much as 13 %, 37 %, 13 %, 11 % and 15 % of resistance variation, respectively. A major novel QTL located on chromosome 7 explained 37 % of the phenotypic variance to the African Xoo corresponding to race A3 whereas that on chromosome 11 is effective to all African races tested. Together with genes and QTLs for resistance to bacterial blight previously described, the QTLs described here were mapped onto the reference O. sativa subs japonica (var. Nipponbare) physical map. Conclusion We characterized new resistance QTLs. While some co-localize with known resistance genes/QTLs to Asian strains, others are specific to African strains. We result with new information on genes and QTLs for resistance to bacterial blight that will be useful for controlling the disease.

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB) in rice (Oryza sativa L.). In this study, we investigated the genome-wide transcription patterns of two Xoo strains (KACC10331 and HB1009), which showed different virulence patterns against eight rice cultivars, including IRBB21 (carrying Xa21). In total, 743 genes showed a significant change (p-value < 0.001 in t-tests) in their mRNA expression levels in the HB1009 (K3a race) strain compared with the Xoo KACC10331 strain (K1 race). Among them, four remarkably enriched GO terms, DNA binding, transposition, cellular nitrogen compound metabolic process, and cellular macromolecule metabolic process, were identified in the upregulated genes. In addition, the expression of 44 genes was considerably higher (log2 fold changes > 2) in the HB1009 (K3a race) strain than in the Xoo KACC10331 (K1 race) strain. Furthermore, 13 and 12 genes involved in hypersensitive response and pathogenicity (hrp) and two-component regulatory systems (TCSs), respectively, were upregulated in the HB1009 (K3a race) strain compared with the Xoo KACC10331 (K1 race) strain, which we determined using either quantitative real-time PCR analysis or next-generation RNA sequencing. These results will be helpful to improve our understanding of Xoo and to gain a better insight into the Xoo–rice interactions.

AmyR is commonly considered a regulator of starch degradation whose activity is induced by the presence of maltose, the disaccharide building block of starch. In this study, we demonstrate that the role of AmyR extends beyond starch degradation. Enzyme activity assays, genes expression analysis and growth profiling on d-glucose- and d-galactose-containing oligo- and polysaccharides showed that AmyR regulates the expression of some of the Aspergillus niger genes encoding α- and β-glucosidases, α- and β- galactosidases, as well as genes encoding α-amlyases and glucoamylases. In addition, we provide evidence that d-glucose or a metabolic product thereof may be the inducer of the AmyR system in A. niger and not maltose, as is commonly assumed. Electronic supplementary material The online version of this article (doi:10.1007/s00253-011-3550-6) contains supplementary material, which is available to authorized users.

Outer membrane vesicles (OMVs) are small nanoscale structures that are secreted by bacteria and that can carry nucleic acids, proteins and small metabolites. They can mediate intracellular communication and play a role in virulence. In this study, we show that treatment with the β-lactam antibiotic imipenem leads to a dramatic increase in the secretion of outer membrane vesicles in the nosocomial pathogen Stenotrophomonas maltophilia. Proteomic analysis of their protein content demonstrated that the OMVs contain the chromosomal encoded L1 metallo-β-lactamase and L2 serine-β-lactamase. Moreover, the secreted OMVs contain large amounts of two Ax21 homologues, i.e. outer membrane proteins known to be involved in virulence and biofilm formation. We show that OMV secretion and the levels of Ax21 in the OMVs are dependent on the quorum sensing diffusible signal system (DSF). More specific, we demonstrate that the S. maltophilia DSF cis-Δ2-11-methyl-dodecenoic acid and, to a lesser extent, the Burkholderia cenocepacia DSF cis-Δ2-dodecenoic acid, stimulate OMV secretion. By a targeted proteomic analysis, we confirmed that DSF-induced OMVs contain large amounts of the Ax21 homologues, but not the β-lactamases. This work illustrates that both quorum sensing and disturbance of the peptidoglycan biosynthesis provoke the release of OMVs and that OMV content is context dependent.

Diversiteten bland gener som ger resistens (R) mot infektionssjukdomar är mycket stor i växtriket, särskilt i den effektorutlösta klassen av försvaret. Denna artikel beskriver interaktioner mellan växters effektorutlösta fösvar och patogeners effektorproteiner och undersöker evolutionära och genetiska mekanismer för uppkomsten av ny resistens. R-genprodukter har en typisk domänstruktur och interagerar både direkt och indirekt med patogeners effektorproteiner.Vid kontakt med ett effektorprotein utlöses en försvarsrespons som kan förhindra fortsatt patogen tillväxt. Flera genetiska mekanismer verkar samtidigt på R-gener, något som resulterar i en hög hastighet för uppkomst av nya R-varianter. Den intima samevolution som existerar hos många patogen-växtsystem formar det evolutionära utvecklingsmönstret av R-gener. Diversifierande positiv selektion via den biologiska kapprustningsmodellen och bevarande negativ frekvensberoende selektion är båda viktiga samevolutionära processer. Denna artikel indikerar att växters immunförsvar är anpassningsbart och robust, men belyser också den kunskapsbrist som råder inom resistensforskningen. The diversity among genes conferring resistance (R) against infectious diseases is very high in the plant kingdom, especially in the effector triggered class of defense. This article describes the interactions between the plant effector triggered immunity and pathogen effectors and examines the evolutionary and genetic mechanisms for the emergence of new resistance. R gene products have a typical domain structure and interacts both directly and indirectly with pathogen effectors. Upon contact with an effector a defense response is triggered that may prevent the further pathogen growth. Multiple genetic mechanisms act simultaneously on the R-genes, which results in a rapid diversification of novel R variants. The intimate co-evolution of many existing plant-pathogen systems form the evolutionary pattern of R genes. Diversifying positive selection via the biological arms race model and conservative negative frequency-dependent selection are both important coevolutionary processes. This article indicates that the plant immune system is adaptive and robust, but also highlights the lack of knowledge in plant resistance research.

SUMMARY We show that OsSERK2 is a regulator of innate immune signaling mediated by multiple non-RD receptor kinases (RKs) including XA21, XA3, and OsFLS2. OsSerk2-silenced rice lines are impaired in XA21-mediated immunity to Xoo PXO99, XA3-mediated immunity to Xoo PXO86, and OsFLS2-mediated defense responses. Thus, OsSERK2 is broadly involved in PRR-mediated immunity in rice., The rice XA21 immune receptor kinase and the structurally related XA3 receptor confer immunity to Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial leaf blight. Here we report the isolation of OsSERK2 (rice somatic embryogenesis receptor kinase 2) and demonstrate that OsSERK2 positively regulates immunity mediated by XA21 and XA3 as well as the rice immune receptor FLS2 (OsFLS2). Rice plants silenced for OsSerk2 display altered morphology and reduced sensitivity to the hormone brassinolide. OsSERK2 interacts with the intracellular domains of each immune receptor in the yeast two-hybrid system in a kinase activity-dependent manner. OsSERK2 undergoes bidirectional transphosphorylation with XA21 in vitro and forms a constitutive complex with XA21 in vivo. These results demonstrate an essential role for OsSERK2 in the function of three rice immune receptors and suggest that direct interaction with the rice immune receptors is critical for their function. Taken together, our findings suggest that the mechanism of OsSERK2-meditated regulation of rice XA21, XA3, and FLS2 differs from that of AtSERK3/BAK1-mediated regulation of Arabidopsis FLS2 and EFR.

In filamentous fungi, most of the strategies used for the improvement of protein yields have been based on an increase in the transcript levels of a target gene. Strategies focusing at the translational level have been also described, but are far less explored. Here the 5' untranslated sequence of the glaA mRNA, a widely used expression system for the expression of recombinant proteins, was modified by the introduction of different nucleotide elements that have positive role in the translation process. Five Aspergillus niger laccase-like multicopper oxidases (MCOs) coding genes were fused to the native glaA 5'UTR and the three synthetic versions (sUTR1, sUTR2, and sUTR3) as well, and placed under the control of the glucoamylase gene promoter. Afterwards, a total of 20 fungal transformations were done using A. niger N593 as a recipient strain and 50 transformants per transformation were isolated and analyzed. The result of the incorporation of the synthetic 5'UTRs on the overall productivity of the transformants was assessed, on one hand by monitoring the laccase activity of all the isolated transformants, and on the other hand by quantifying and comparing the activity of those secreting the highest level of each MCO. For this purpose, a high-throughput method for the screening and selection of the best producers was developed. Once the best transformants producing the highest yield of McoA, McoB, McoC, McoD, and McoJ laccases were selected, their production level was quantified in supernatants of liquid cultures. The results obtained in this work indicate that modifications in the native glaA 5'UTR can lead to improvements in protein yields. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc

Pflanzen und Mikroorganismen haben im Lauf der Evolution eine Vielzahl an Interaktionen ausgebildet. Diese reichen von mutualistischen bis zu parasitären Beziehungen. Während der Ausbildung dieser Interaktionen werden von Mikroorganismen Effektorproteine eingesetzt. Dabei handelt es sich um sekretierte Proteine, die den Wirt beeinflussen sollen, um seine Kolonisierung zu erleichtern. In dieser Arbeit wurden mögliche Effektorproteine aus Magnaporthe oryzae deletiert und untersucht.

Here we report the cloning of the Pa_3_10940 gene from the coprophilic fungus Podospora anserina , which encodes a C-terminal family 1 carbohydrate binding module (CBM1) linked to a domain of unknown function. The function of the gene was investigated by expression of the full-length protein and a truncated derivative without the CBM1 domain in the yeast Pichia pastoris . Using a library of polysaccharides of different origins, we demonstrated that the full-length enzyme displays activity toward a broad range of β-glucan polysaccharides, including laminarin, curdlan, pachyman, lichenan, pustulan, and cellulosic derivatives. Analysis of the products released from polysaccharides revealed that this β-glucanase is an exo-acting enzyme on β-(1,3)- and β-(1,6)-linked glucan substrates and an endo-acting enzyme on β-(1,4)-linked glucan substrates. Hydrolysis of short β-(1,3), β-(1,4), and β-(1,3)/β-(1,4) gluco-oligosaccharides confirmed this striking feature and revealed that the enzyme performs in an exo-type mode on the nonreducing end of gluco-oligosaccharides. Excision of the CBM1 domain resulted in an inactive enzyme on all substrates tested. To our knowledge, this is the first report of an enzyme that displays bifunctional exo-β-(1,3)/(1,6) and endo-β-(1,4) activities toward beta-glucans and therefore cannot readily be assigned to existing Enzyme Commission groups. The amino acid sequence has high sequence identity to hypothetical proteins within the fungal taxa and thus defines a new family of glycoside hydrolases, the GH131 family.

Citrate synthase is a central player in the acidogenic metabolism of Aspergillus niger. The 5' upstream sequence (0.9 kb DNA) of citrate synthase gene (citA) from A. niger NCIM 565 was analyzed and its promoter function demonstrated through the heterologous expression of two proteins. The cloned citrate synthase promoter (PcitA) sequence was able to express bar coding sequence thereby conferring phosphinothricin resistance. This sequence was further analyzed by systematic deletions to define an effective but compact functional promoter. The PcitA driven egfp expression showed that PcitA was active in all differentiation cell-stages of A. niger. EGFP expression was highest on non-repressible carbon sources like acetate and glycerol. Mycelial EGFP levels increased during acidogenic growth suggesting that PcitA is functional throughout this cultivation. A. niger PcitA is the first Krebs cycle gene promoter used to express heterologous proteins in filamentous fungi. (C) 2011 Elsevier B.V. All rights reserved.

This article 'Hardinath 1, an early-maturing rice genotype released in Nepal' appeared in the International Rice Research Notes series, created by the International Rice Research Institute (IRRI) to expedite communication among scientists concerned with the development of improved technology for rice and rice-based systems. The series is a mechanism to help scientists keep each other informed of current rice research findings. The concise scientific notes are meant to encourage rice scientists to communicate with one another to obtain details on the research reported.

A set of aspergillus strains from culture collections and wild-type black aspergilli isolated on non-selective media were used to validate the use of media with 20 % tannic acid for exclusive and complete selection of the black aspergilli. The 20% tannic acid medium proved useful for both quantitative and qualitative selection of all different black aspergilli, including all recognized species: A. carbonarius, A. japonicus, A. aculeatus, A foetidus, A. heteromorphus, A. niger, A. tubingensis and A. brasiliensis haplotypes. Even higher concentrations of tannic acid can be utilized by the black aspergilli suggesting a very efficient tannic acid-degrading system. Colour mutants show that the characteristic ability to grow on high tannic acid concentrations is not causally linked to the other typical feature of these aspergilli, i.e. the formation of brown-black pigments. Sequence analysis of the A. niger genome using the A. orvzae tannase gene yielded eleven tannase-like genes, far more than in related species. Therefore, a unique ecological niche in the degradation of tannic acid and connected nitrogen release seems to be reserved for these black-spored cosmopolitans.

This article 'Polymorphism of DNA markers linked to bacterial blight resistance genes in useful rice germplasm' appeared in the International Rice Research Notes series, created by the International Rice Research Institute (IRRI) to expedite communication among scientists concerned with the development of improved technology for rice and rice-based systems. The series is a mechanism to help scientists keep each other informed of current rice research findings. The concise scientific notes are meant to encourage rice scientists to communicate with one another to obtain details on the research reported.

This article 'Improvement of conjugation methods for Xanthomonas oryzae pv. oryzae strain DY89031 to identify avrXa21 clones' appeared in the International Rice Research Notes series, created by the International Rice Research Institute (IRRI) to expedite communication among scientists concerned with the development of improved technology for rice and rice-based systems. The series is a mechanism to help scientists keep each other informed of current rice research findings. The concise scientific notes are meant to encourage rice scientists to communicate with one another to obtain details on the research reported.