Rice-Infecting Pseudomonas Genomes Are Highly Accessorized and Harbor Multiple Putative Virulence Mechanisms to Cause Sheath Brown Rot

Sheath rot complex and seed discoloration in rice involve a number of pathogenic bacteria that cannot be associated with distinctive symptoms. These pathogens can easily travel on asymptomatic seeds and therefore represent a threat to rice cropping systems. Among the rice-infecting Pseudomonas, P. fuscovaginae has been associated with sheath brown rot disease in several rice growing areas around the world. The appearance of a similar Pseudomonas population, which here we named P. fuscovaginae-like, represents a perfect opportunity to understand common genomic features that can explain the infection mechanism in rice. We showed that the novel population is indeed closely related to P. fuscovaginae. A comparative genomics approach on eight rice-infecting Pseudomonas revealed heterogeneous genomes and a high number of strain-specific genes. The genomes of P. fuscovaginae-like harbor four secretion systems (Type I, II, III, and VI) and other important pathogenicity machinery that could probably facilitate rice colonization. We identified 123 core secreted proteins, most of which have strong signatures of positive selection suggesting functional adaptation. Transcript accumulation of putative pathogenicity-related genes during rice colonization revealed a concerted virulence mechanism. The study suggests that rice-infecting Pseudomonas causing sheath brown rot are intrinsically diverse and maintain a variable set of metabolic capabilities as a potential strategy to occupy a range of environments.

Estimating production costs and profitability of strawberry genotypes in southeast Queensland using plant traits

In Queensland the subtropical strawberry (Fragaria ×ananassa) breeding program aims to combine traits into new genotypes that increase production efficiency. The contribution of individual plant traits to cost and income under subtropical Queensland conditions has been investigated. The study adapted knowledge of traits and the production and marketing system to assess the economic impact (gross margin) of new cultivars on the system, with the overall goal of improving the profitability of the industry through the release of new strawberry cultivars. Genotypes varied widely in their effect on gross margin, from 48% above to 10% below the base value. The advantage of a new genotype was also affected by the proportion of total area allocated to the new genotype. The largest difference in gross margin between that at optimum allocation (8% increase in gross margin) and an all of industry allocation (20% decrease in gross margin) of area to the genotype was 28%. While in other cases the all of industry allocation was also the optimum allocation, with one genotype giving a 48% benefit in gross margin.

Variation in susceptibility among macadamia genotypes and species to Phytophthora root decay caused by Phytophthora cinnamomi

Phytophthora cinnamomi is a major pathogen of cultivated macadamia (Macadamia integrifolia, Macadamia tetraphylla and their hybrids) worldwide. The susceptibility of the two non-edible Macadamia species (Macadamia ternifolia and Macadamia jansenii) to P. cinnamomi is not well-understood. Commercial macadamia trees are established on grafted seedling (seed propagation) or own-rooted cutting (vegetative propagation) rootstocks of hybrids of the cultivated species. There is little information to support the preferential use of rootstock propagated by either seedling or own-rooted cutting methods in macadamia. In this study we assessed roots of macadamia plants of the four species and their hybrids, derived from the two methods of propagation, for their susceptibility to P. cinnamomi infection. The roots of inoculated plant from which P. cinnamomi was recovered showed blackening symptoms. The non-cultivated species, M. ternifolia and M. jansenii and their hybrids were the most susceptible germplasm compared with M. tetraphylla and M. integrifolia. Of these two species, M. tetraphylla was less susceptible than M. integrifolia. Significant differences were observed among the accessions of their hybrids. A strong association (R2 > 0.75) was recorded between symptomatic roots and disease severity. Root density reduced with increasing disease severity rating in both own-rooted cuttings (R2 = 0.65) and germinated seedlings (R2 = 0.55). P. cinnamomi severity data were not significantly (P > 0.05) different between the two methods of plant propagation. The significance of this study to macadamia breeding and selection of disease resistant rootstocks is discussed.

Detection of virulence-associated genes in pathogenic and commensal avian Escherichia coli isolates

Poultry colibacillosis due to Avian Pathogenic Escherichia coli (APEC) is responsible for several extra-intestinal pathological conditions, leading to serious economic damage in poultry production. The most commonly associated pathologies are airsacculitis, colisepticemia, and cellulitis in broiler chickens, and salpingitis and peritonitis in broiler breeders. In this work a total of 66 strains isolated from dead broiler breeders affected with colibacillosis and 61 strains from healthy broilers were studied. Strains from broiler breeders were typified with serogroups O2, O18, and O78, which are mainly associated with disease. The serogroup O78 was the most prevalent (58%). All the strains were checked for the presence of 11 virulence genes: 1) arginine succinyltransferase A (astA); ii) E. coli hemeutilization protein A (chuA); iii) colicin V A/B (cvaA/B); iv) fimbriae mannose-binding type 1 (fimC); v) ferric yersiniabactin uptake A (fyuA); vi) iron-repressible high-molecular-weight proteins 2 (irp2); vii) increased serum survival (iss); viii) iron-uptake systems of E. coli D (iucD); ix) pielonefritis associated to pili C (papC); x) temperature sensitive haemaglutinin (tsh), and xi) vacuolating autotransporter toxin (vat), by Multiplex-PCR. The results showed that all genes are present in both commensal and pathogenic E. coli strains. The iron uptake-related genes and the serum survival gene were more prevalent among APEC. The adhesin genes, except tsh, and the toxin genes, except astA, were also more prevalent among APEC isolates. Except for astA and tsh, APEC strains harbored the majority of the virulence-associated genes studied and fimC was the most prevalent gene, detected in 96.97 and 88.52% of APEC and AFEC strains, respectively. Possession of more than one iron transport system seems to play an important role on APEC survival.

Response of some chickpea ( Cicer arietinum L.) genotypes to Orobanche foetida Poir. parasitism

In Tunisia, broomrape ( Orobanche foetida Poir.) causes major drawbacks especially in faba bean ( Vicia faba L.) Chickpea ( Cicer arietinum L.) suffers little damage compared to faba bean, but with the winter sowing chickpea cultivars, broomrape might become a serious problem for chickpea cultivation. The development of resistant cultivars remains the most efficient way to solve this problem. The behavior of six chickpea genotypes to O. foetida was studied under field natural infestation and artificial inoculation in pots and petri dishes in greenhouse conditions. During the cropping seasons 2010-2011 and 2012-2013 the level of infection was very low. The number of emerged parasites per host plant varied from 0.18 to 0.43 and the incidence from 6.5% to 23%. Among the six tested genotypes, G1, G2, and G4 showed partial resistance to O. foetida with low number and dry weight of emerged parasite and high grain yield compared to the other genotypes, although no significant differences were recorded. In pot experiments, the number and total dry weight of broomrape per plant were lower for G1 and G2 genotypes than the other genotypes. Parasitism does not affect significantly the shoot dry weight and number of pods of these genotypes. The total chlorophyll content was significantly reduced under infestation in all genotypes. In Petri dishes experiments, results showed that percent germination of O. foetida seeds varied from 49% to 65% and does not play a role in the resistance of chickpea genotypes. In contrast, broomrape attachment was lower and slower for the genotypes G1, G2, and G4 than the other genotypes. Resistance in chickpea genotypes was characterized by few parasite attachments on roots and a limited growth of established tubercles. No necrosis of attached tubercles was observed in the different experiments.

Heterosis in locally adapted sorghum genotypes and potential of hybrids for increased productivity in contrasting environments in Ethiopia

Increased productivity in sorghum has been achieved in the developed world using hybrids. Despite their yield advantage, introduced hybrids have not been adopted in Ethiopia due to the lack of adaptive traits, their short plant stature and small grain size. This study was conducted to investigate hybrid performance and the magnitude of heterosis of locally adapted genotypes in addition to introduced hybrids in three contrasting environments in Ethiopia. In total, 139 hybrids, derived from introduced seed parents crossed with locally adapted genotypes and introduced R lines, were evaluated. Overall, the hybrids matured earlier than the adapted parents, but had higher grain yield, plant height, grain number and grain weight in all environments. The lowland adapted hybrids displayed a mean better parent heterosis (BPH) of 19%, equating to 1160 kg ha− 1 and a 29% mean increase in grain yield, in addition to increased plant height and grain weight, in comparison to the hybrids derived from the introduced R lines. The mean BPH for grain yield for the highland adapted hybrids was 16% in the highland and 52% in the intermediate environment equating to 698 kg ha− 1 and 2031 kg ha− 1, respectively, in addition to increased grain weight. The magnitude of heterosis observed for each hybrid group was related to the genetic distance between the parental lines. The majority of hybrids also showed superiority over the standard check varieties. In general, hybrids from locally adapted genotypes were superior in grain yield, plant height and grain weight compared to the high parents and introduced hybrids indicating the potential for hybrids to increase productivity while addressing farmers' required traits.

ADAPTATION OF INTRODUCED MUNGBEAN GENOTYPES IN UGANDA

Mungbean ( Vigna radiata (L.) Wilczek) is an important source of nutrients and income for smallholder farmers in East Africa. Mungbean production in countries like Uganda largely depends on landraces, in the absence of improved varieties. In order to enhance productivity, efforts have been underway to develop and evaluate mungbean varieties that meet farmers’ needs in various parts of the country. This study was conducted at six locations in Uganda, to determine the adaptability of introduced mungbean genotypes, and identify mungbean production mega-environments in Uganda. Eleven genotypes (Filsan, Sunshine, Blackgram, Mauritius1, VC6148 (50-12), VC6173 (B-10),Yellowmungo, KPS1, VC6137(B-14),VC6372(45-60),VC6153(B-20P) and one local check were evaluated in six locations during 2013 and 2014. The locations were; National Semi Arid Resources Research Institute (NaSARRI), Abi Zonal Agricultural Research and Development Institute (AbiZARDI),Kaberamaido variety trial center, Kumi variety trial center, Nabuin Zonal Agricultural Research and Development Institute (NabuinZARDI), and Ngetta Zonal Agricultural Research and Development Institute (NgettaZARDI). G × E interactions were significant for grain yield. Through GGEBiplot analysis, three introduced genotypes (Filsan, Blackgram and Sunshine) were found to be stable and high yielding, and therefore, were recommended for release. The six test multi-locations were grouped into two candidate mega-environments for mungbean production (one comprising of AbiZARDI and Kaberamaido and the other comprising of NaSARRI, NabuinZARDI, Kumi, and NgettaZARDI). National Semi Arid Resources Research Institute (NaSARRI) was the most suitable environment in terms of both discriminative ability and representativeness and therefore can be used for selection of widely adaptable genotypes.

Study of Lomentospora prolificans pathogenicity: Host immune response and fungal virulence factors.

188 p.

Dual Quorum Quenching Capsules: Disrupting two bacterial communication pathways that lead to virulence

Healthcare Associated Infections (HAIs) in the United States, are estimated to cost nearly $10 billion annually. And, while device-related infections have decreased, the 60% attributed to pneumonia, gastrointestinal pathogens and surgical site infections (SSIs) remain prevalent. Furthermore, these are often complicated by antibacterial resistance that ultimately cause 2 million illnesses and 23,000 deaths in the US annually. Antibacterial resistance is an issue increasing in severity as existing antibiotics are losing effectiveness, and fewer new antibiotics are being developed. As a result, new methods of combating bacterial virulence are required. Modulating communications of bacteria can alter phenotype, such as biofilm formation and toxin production. Disrupting these communications provides a means of controlling virulence without directly interacting with the bacteria of interest, a strategy contrary to traditional antibiotics. Inter- and intra-species bacterial communication is commonly called quorum sensing because the communication molecules have been linked to phenotypic changes based on bacterial population dynamics. By disrupting the communication, a method called ‘quorum quenching’, bacterial phenotype can be altered. Virulence of bacteria is both population and species dependent; each species will secrete different toxic molecules, and total population will affect bacterial phenotype9. Here, the kinase LsrK and lactonase SsoPox were combined to simultaneously disrupt two different communication pathways with direct ties to virulence leading to SSIs, gastrointestinal infection and pneumonia. To deliver these enzymes for site-specific virulence prevention, two naturally occurring polymers were used, chitosan and alginate. Chitosan, from crustacean shells, and alginate, from seaweed, are frequently studied due to their biocompatibility, availability, self-assembly and biodegrading properties and have already been verified in vivo for wound-dressing. In this work, a novel functionalized capsule of quorum quenching enzymes and biocompatible polymers was constructed and demonstrated to have dual-quenching capability. This combination of immobilized enzymes has the potential for preventing biofilm formation and reducing bacterial toxicity in a wide variety of medical and non-medical applications.

First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes

This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS)1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs) to amphotericin B (AMB), caspofungin (CAS), and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS) and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilmgrowing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001). AMB promoted significant reductions (≈50%) in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management.

The prevalence of genotypes that determine resistance to macrolides, lincosamides, and streptogramins B compared with spiramycin susceptibility among erythromycin-resistant Staphylococcus epidermidis

Coagulase-negative staphylococci, particularly Staphylococcus epidermidis , can be regarded as potential reservoirs of resistance genes for pathogenic strains, e.g., Staphylococcus aureus . The aim of this study was to assess the prevalence of different resistance phenotypes to macrolide, lincosamide, and streptogramins B (MLSB) antibiotics among erythromycin-resistant S. epidermidis, together with the evaluation of genes promoting the following different types of MLSB resistance: ermA, ermB, ermC, msrA, mphC, and linA/A’. Susceptibility to spiramycin was also examined. Among 75 erythromycin-resistant S. epidermidis isolates, the most frequent phenotypes were macrolides and streptogramins B (MSB) and constitutive MLSB (cMLSB). Moreover, all strains with the cMLSB phenotype and the majority of inducible MLSB (iMLSB) isolates were resistant to spiramycin, whereas strains with the MSB phenotype were sensitive to this antibiotic. The D-shape zone of inhibition around the clindamycin disc near the spiramycin disc was found for some spiramycin-resistant strains with the iMLSB phenotype, suggesting an induction of resistance to clindamycin by this 16-membered macrolide. The most frequently isolated gene was ermC, irrespective of the MLSB resistance phenotype, whereas the most often noted gene combination was ermC, mphC, linA/A’. The results obtained showed that the genes responsible for different mechanisms of MLSB resistance in S. epidermidis generally coexist, often without the phenotypic expression of each of them.

The effect of chaperone compounds on glucocerebrosidase stability and activity in fibroblasts from Gaucher Disease patients with different genotypes

Dissertação de mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2014

Phenotypic and molecular characterization of common bean genotypes for resistance to anthracnose.

This study aimed to perform phenotypic and molecular characterization of cultivars and breeding lines of common bean for resistance to anthracnose.

Evidence for reductive genome evolution and lateral acquisition of virulence functions in two Corynebacterium pseudotuberculosis strains.

2011