QTL associated with Fusarium head blight resistance in the soft red winter wheat Ernie

Fusarium head blight (FHB), mainly caused by Fusarium graminearum Schwabe [telomorph: Gibberella zeae Schw. (Petch)], is an increasingly important disease of wheat (Triticum aestivum L.). Host-plant resistance provides the best hope for reducing economic losses associated with FHB, but new sources of resistance are limited. The moderately resistant winter wheat cultivar, Ernie, may provide a source of resistance that differs from Sumai 3 but these genes have not been mapped. Also hindering resistance breeding may be associations of resistance with agronomic traits such as late maturity that may be undesirable in some production environments. This research was conducted to identify QTL associated with type II FHB resistance (FHB severity, FHBS), and to determine if they are associated with days to anthesis (DTA), number of spikelets (NOS), and the presence/absence of awns. Two hundred and forty-three F₈ recombinant inbred lines from a cross between the resistant cultivar, Ernie and susceptible parent, MO 94-317 were phenotyped for type II FHB resistance using point inoculation in the greenhouse during 2002 and 2003. Genetic linkage maps were constructed using 94 simple sequence repeat (SSR) and 146 amplified fragment length polymorphic (AFLP) markers. Over years four QTL regions on chromosomes 2B, 3B, 4BL and 5A were consistently associated with FHB resistance. These QTL explained 43.3% of the phenotypic variation in FHBS. Major QTL conditioning DTA and NOS were identified on chromosome 2D. Neither the QTL associated with DTA and NOS nor the presence/absence of awns were associated with FHB resistance in Ernie. Our results suggest that the FHB resistance in Ernie appears to differ from that in Sumai 3, thus pyramiding the QTL in Ernie with those from Sumai 3 could result in enhanced levels of FHB resistance in wheat.     

Production of ethanol by a flocculentSaccharomyces sp. in a continuous upflow reactor using sucrose,sugar-cane juice,and molasses as the carbon source

Summary A flocculentSaccharomyces sp., isolated from spontaneously fermenting sugar-cane juice, was used to produce ethanol in a continuous upflow reactor. Media of different composition were tested using the same fermentation conditions. The best results were obtained fermenting either sugar-cane juice or molasses. The differences observed could be due to the influence of the medium composition on the growth rate and on the expression of the flocculent capacity of the yeast, both factors which modified the cell biomass concentration within the reactor.

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Resumen Una cepa floculante deSaccharomyces, aislada de jugo de caña de azúcar, se utilizó para producir etanol fermentando medios de distinta composición en un reactor continuo de flujo ascendente. Los mejores resultados se obtuvieron con jugo de caña de azúcar y con melaza. Las diferencias observadas según el mosto utilizado pueden deberse a la influencia de la composición del medio sobre la velocidad de multiplicación y capacidad foculante de la levadura, modificando de esta manera la concentración de biomasa dentro del reactor.

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Résumé Une souche floculante deSaccharomyces isolée du jus de canne à sucre a été utilisée pour produire l'éthanol, dans un réacteur continu à flux ascendant, par fermentation de milieux de culture de différentes compositions. Les meilleurs résultats ont été obtenus avec le jus de canne à sucre et les molasses. Les différences qu'on a vues selon le moût utilisé peuvent être dues à l'influence de la composition du milieu sur la vitesse de multiplication et la capacité floculante de la levure, donnant une modification de la concentration de biomasse dans le réacteur.

     

Amino acids and protein profile in floral nectar: Much more than a simple reward

Nectar is considered a primary alimentary reward for a large variety of pollinators. Recent studies demonstrated that nectar may have other functions in addition to attracting pollinators. Mainly other two functions have been recognized: defense against microbial invasion and post-secretion modification of sugar profile. The floral nectar of Cucurbita pepo L. has been analyzed to confirm the presence of defense proteins, namely xylosidases that were identified by means of a proteomic approach in a previous study. An enzyme assay was also performed that revealed low invertase activity for which the optimal pH was determined. This invertase activity is not able to modify the sugar profile significantly during the short period of anthesis (6 h). The amino acids complement of the nectar of both sexes was also determined. Sixteen of the 20 protein amino acids have been detected. Proline comprises more than 30% of total amino acid content in male and female flowers. Three non-protein amino acids (taurine, β-alanine, and GABA) represent almost one third of the total amino acid content, and of these, GABA is the most abundant (16%). Several biological activities were attributed to these amino acids and further studies are needed to assess their presumed involvement in managing the foraging behavior of insects. More recent studies, including our own, demonstrate that the various functions of nectar are much more complex than previously thought.     

PCR-Activated Cell Sorting for Cultivation-Free Enrichment and Sequencing of Rare Microbes

Microbial systems often exhibit staggering diversity, making the study of rare, interesting species challenging. For example, metagenomic analyses of mixed-cell populations are often dominated by the sequences of the most abundant organisms, while those of rare microbes are detected only at low levels, if at all. To overcome this, selective cultivation or fluorescence-activated cell sorting (FACS) can be used to enrich for the target species prior to sequence analysis; however, since most microbes cannot be grown in the lab, cultivation strategies often fail, while cell sorting requires techniques to uniquely label the cell type of interest, which is often not possible with uncultivable microbes. Here, we introduce a culture-independent strategy for sorting microbial cells based on genomic content, which we term PCR-activated cell sorting (PACS). This technology, which utilizes the power of droplet-based microfluidics, is similar to FACS in that it uses a fluorescent signal to uniquely identify and sort target species. However, PACS differs importantly from FACS in that the signal is generated by performing PCR assays on the cells in microfluidic droplets, allowing target cells to be identified with high specificity with suitable design of PCR primers and TaqMan probes. The PACS assay is general, requires minimal optimization and, unlike antibody methods, can be developed without access to microbial antigens. Compared to non-specific methods in which cells are sorted based on size, granularity, or the ability to take up dye, PACS enables genetic sequence-specific sorting and recovery of the cell genomes. In addition to sorting microbes, PACS can be applied to eukaryotic cells, viruses, and naked nucleic acids.     

The Role of Grain Boundaries on Ionic Defect Migration in Metal Halide Perovskites

Halide perovskites are emerging as revolutionary materials for optoelectronics. Their ionic nature and the presence of mobile ionic defects within the crystal structure have a dramatic influence on the operation of thin‐film devices such as solar cells, light‐emitting diodes, and transistors. Thin films are often polycrystalline and it is still under debate how grain boundaries affect the migration of ions and corresponding ionic defects. Laser excitation during photoluminescence (PL) microscopy experiments leads to formation and subsequent migration of ionic defects, which affects the dynamics of charge carrier recombination. From the microscopic observation of lateral PL distribution, the change in the distribution of ionic defects over time can be inferred. Resolving the PL dynamics in time and space of single crystals and thin films with different grain sizes thus, provides crucial information about the influence of grain boundaries on the ionic defect movement. In conjunction with experimental observations, atomistic simulations show that defects are trapped at the grain boundaries, thus inhibiting their diffusion. Hence, with this study, a comprehensive picture highlighting a fundamental property of the material is provided while also setting a theoretical framework in which the interaction between grain boundaries and ionic defect migration can be understood.     

Xylan-degrading enzymes in male and female flower nectar of Cucurbita pepo

Background and Aims

Nectar is a very complex mixture of substances. Some components (sugars and amino acids) are considered primary alimentary rewards for animals and have been investigated and characterized in numerous species for many years. In contrast, nectar proteins have been the subject of few studies and little is known of their function. Only very recently have detailed studies and characterization of nectar proteins been undertaken, and then for only a very few species. This current work represents a first step in the identification of a protein profile for the floral nectar of Cucurbita pepo. In this regard, the species studied is of particular interest in that it is monoecious with unisexual flowers and, consequently, it is possible that nectar proteins derived from male and female flowers may differ.

Methods

Manually excised spots from two-dimensional (2-D) electrophoresis were subjected to in-gel protein digestion. The resulting peptides were sequenced using nanoscale LC–ESI/MS-MS (liquid chromatography–electrospray ionization/tandem mass spectrometry). An MS/MS ions search was carried out in Swiss-Prot and NCBInr databases using MASCOT software.

Key Results

Two-dimensional electrophoresis revealed a total of 24 spots and a different protein profile for male and female flower nectar. Four main proteins recognized by 2-D electrophoresis most closely resemble β-d-xylosidases from Arabidopsis thaliana and have some homology to a β-d-xylosidase from Medicago varia. They were present in similar quantities in male and female flowers and had the same molecular weight, but with slightly different isoelectric points.

Conclusions

A putative function for xylosidases in floral nectar of C. pepo is proposed, namely that they may be involved in degrading the oligosaccharides released by the nectary cell walls in response to hydrolytic enzymes produced by invading micro-organisms. Several types of oligosaccharides have been reported to increase the pathogenic potential of micro-organisms. Thus, it is possible that such a mechanism may reduce the virulence of pathogens present in nectar.     

Simul taneous production of alpha and beta amylases byBacillus subtilis MIR-5 in batch and continuous culture

Summary Simultaneous production of - and -amylase was studied in batch and continuous culture using starch as substrate inB. subtilis MIR-5. By mainipulating the cultural condition, both enzymes could then be produced by the same strain.Footnote: dedicated to 75^th aniverssary of Dr. Raul E. Trucco.     

Helminth species specific expansion and increased TNF-alpha production of non-classical monocytes during active tuberculosis

Both Mycobacterium tuberculosis infection and helminths may affect innate immune mechanisms such as differential effects on monocytes towards the non-classical and intermediate subsets that favor bacterial persistence. Our aim, was to investigate helminth species specific effects on the frequency and functional activity of monocyte subsets in patients with active tuberculosis and healthy subjects. HIV-negative patients with active pulmonary tuberculosis (PTB) and community controls (CCs) in Gondar, Ethiopia were screened for helminth infection by stool microscopy. Flow cytometric analysis of peripheral blood mononuclear cells (PBMCs) and ex vivo stimulation with purified protein derivative (PPD) and helminth antigens were used to characterize the distribution of monocyte subsets and their function. A total of 74 PTB patients and 57 CCs with and without helminth infection were included. Non-classical monocytes were increased in PTB patients with Ascaris and hookworm infection but not in Schistosoma-infected patients. Ascaris had the strongest effect in increasing the frequency of non-classical monocytes in both PTB patients and CCs, whereas PTB without helminth infection did not affect the frequency of monocyte subsets. There was a helminth specific increase in the frequency of TNF-α producing non-classical monocytes in hookworm infected PTB patients, both with and without PPD-stimulation. Low-to-intermediate TB disease severity associated with increased frequency of non-classical monocytes only for helminth-positive PTB patients, and the frequency of TNF-α producing monocytes were significantly higher in intermediate and non-classical monocytes of helminth positive PTB patients with an intermediate disease score. Helminth infection affected the frequency of monocyte subsets and function both in TB patients and controls which was helminth species dependent in TB patients. The clinical role of this potential immunomodulatory effect needs further study and may affect the response and protection to tuberculosis in areas where helminth infections are endemic.     

Cyclic AMP as an inducer of the cell differentiation of Trypanosoma cruzi

The addition to epimastigotes cultures of T. cruzi, of either cAMP, monobutyryl-cAMP, dibutyryl-cAMP, 8-Br-cAMP (at 2 mM each), or the cAMP-phosphodiesterase inhibitor, papaverine (0.2 mM), promoted the in vitro differentiation of these parasite forms into metacyclics. This effect of cAMP may also be exerted in vivo in the insect vector, since cAMP was detected in the urine and in the Malpighi secretion fluids of Rodnius prolixus.