Phenotypic evaluation of floral and flowering traits with relevance for hybrid breeding in wheat (Triticum aestivum L.)

Hybrid breeding is a promising approach to increase the yield potential in wheat (Triticum aestivum L.). The profitability of wheat hybrids highly depends on a cost‐efficient system for hybrid seed production for which an adequate outcrossing in the male pool is of utmost importance. Employing a set of 51 elite winter wheat lines, we developed and evaluated phenotyping methods for floral and flowering traits with relevance for improved cross‐pollination. We observed significant genotypic variances and high heritabilities for most traits, including important traits like pollen mass and anther extrusion. Our results suggest the utility of the developed phenotyping approaches for applied plant breeding and the potential of the traits to assist in the design of the male ideotype for increased cross‐fertilization.     

Evaluation of a semi‐controlled test as a selection tool for frost tolerance in durum wheat (Triticum durum)

Most durum wheat (Triticum durum) varieties possess only low winter hardiness due to their frost susceptibility. In North America and Central Europe, durum wheat is therefore typically sown in spring to circumvent the local winter conditions. However, the yield potential of durum in these regions could be much better exploited if durum varieties with increased frost tolerance were available, which could be sown in autumn. A factor limiting breeding for increased frost tolerance is the variation in the occurrence of frost stress across years. The ‘Weihenstephaner Auswinterungsanlage’ is a semi‐controlled test that exposes the plants to all weather conditions. Snow coverage of the plants, serving as frost protection, is prevented by the movable glass lid of the semi‐controlled test. In this study, different scorings for frost tolerance based on this semi‐controlled test were evaluated and compared with frost tolerance data in the field. Our results illustrate the potential of the ‘Weihenstephaner Auswinterungsanlage’ as an indirect selection tool for frost tolerance in durum breeding programmes, especially when regular frost tolerance data from the field are not available.     

Methicillin-resistant Staphylococcus aureus (MRSA) in veterinary medicine: a "new emerging pathogen"?

The problem of nosocomial infections is of increasing importance in veterinary medicine. As an example, this review summarizes current knowledge regarding methicillin-resistant Staphylococcus aureus (MRSA) as a typical example, as these pathogens are the most important agents of nosocomial infections in human medicine worldwide and are being increasingly reported in veterinary medicine. MRSA are classified by their ability to be resistant against oxacillin/methicillin, this feature being confered by mecA, a gene which was acquired by horizontal gene transfer of the staphylococcal gene cassette (SCCmec). It is this genetic information that enables MRSA to be resistant against all penicillins, cehalosporins and carbapenems. In addition, MRSA are often resistant against a variety of other antiinfectives, i.e. aminoglycosides, macrolides, lincosamide, streptomycins, tetracyclin, chloramphenicol, but also against fluorquinolones and rifampicin. Presumably, these highly adapted strains are particularly able to acquire resistance genes located on plasmids or transposons.They are also able to develop point mutations, further leading to resistant phenotypes. If these pathogens are leading to infectious diseases, veterinarians may be confronted with a worst-case scenario, being left without any antiinfective therapeutic. As Staphylococcus aureus is highly tenacid, professional hygiene management is of utmost importance. The increasing number of published sporadic MRSA infections, MRSA-infectious diseases as well as MRSA outbreaks in veterinary medicine justifies their recognition as a "New Emerging Pathogen". So far, horses and dogs are mostly affected by MRSA. Although transmission between humans and animals has been reported occasionally, the sources, routes of transmission or the epidemiological relevance of MRSA infections in animals are far from being understood. Therefore, epidemiological investigations utilizing molecular typing tools are mandatory. Typing tools like multilocus-sequence-typing (MLST), pulsefield-gelelectrophoresis (PFGE), sequence analysis of the gene encoding protein A (spa-typing) as well as SCCmec-typing are all at hand.     

Case control study to investigate risk factors for bovine neonatal pancytopenia (BNP) in young calves in southern Germany

A case control study on farm level was conducted at the Clinic for Ruminants, LMU Munich, to identify possible risk factors associated with the observed increase in numbers of calves showing clinical signs of Bovine Neonatal Pancytopenia (BNP) since 2006 in southern Germany. Interviews were conducted between August 2008 and June 2010. The characteristics of 56 dairy farms with at least one confirmed case of BNP (thrombocytopenia and leukocytopenia and/or typical findings in post-mortem examination and bone marrow histology) were compared with those of two sets of 50 control dairy farms each, with no history of BNP. The first set of 50 control farms was selected randomly from veterinary practices which had never observed a BNP case on the farms they serviced. The second set of 50 control farms was matched by the veterinary practices which had provided case farms. Two separate analyses were conducted: (1) case farms (n=56) vs. randomly selected control farms (n=50) and (2) case farms (n=56) vs. a matched set of control farms (n=50). All variables with p<0.2 in the univariable analysis were included in stepwise logistic regression models. In the first analysis, only the use of PregSure(?) BVD vaccine was positively associated with BNP with an odds ratio of 1292 (95% CI: 114-14707). In the second analysis, conditional logistic regression models did not converge, therefore non-conditional logistic regression models were conducted. In the non-conditional analysis five variables remained in the model, three of which were negatively associated with BNP: the use of vitamin E and selenium, the frequent use of mastitis tubes, and the use of stem growth regulators in grain production. The use of prophylactic measures (such as control of parasites or vaccination of calves against respiratory disease) was positively associated with BNP with an odds ratio of 14.3 as well as the use of PregSure(?) BVD vaccine with an odds ratio of 426 (95% CI: 20-9095).     

Biomass and nutrients allocation in pot cultured beech seedlings： influence of nitrogen fertilizer

Allocation of biomass and nutrient elements including Nitrogen to above and belowground compartments of beech seedlings (Fagus sylvatica L.) treated by labeled nitrogen fertilizer in the form of ¹⁵NH₄ and ¹⁵NO₃ were investigated at the end of two successive growing seasons. Pot cultured beech seedlings were grown at a green house on intact soil cores sampled from three adjacent stands including beech, Norway spruce and mixed beech-spruce cultures of Solling forest, Germany. Comparing biomass allocation and nutrients concentrations of the seedlings between the control and ¹⁵N-fertilized treatments revealed no significant effect of N fertilization on nutrients uptake by seedlings over the experiment. The form of N input influenced its movement into plant pools. It was demonstrated that beech seedlings take up nitrogen mainly in the form of nitrate, which is then reduced in the leaves, although the differences between the retention of NO₃ ^?-N and NH₄ ⁺-N in plants were not statistically significant. Percent recoveries of ¹⁵N in trees were typically greater after ¹⁵NO₃ than after ¹⁵NH₄ additions. It was indicated that immobilization of 15N tracer in fine roots was a slower process comparing other plant compartments such as stem and coarse roots, but a powerful sink for N during the course of study.     

Validation of a specific PCR screening test for Pantoea stewartii subsp. stewartii in maize (Zea mays) samples

Validation data is presented for a conventional PCR test that specifically detects the quarantine pathogen Pantoea stewartii subsp. stewartii in maize leaf and seed samples and does not cross‐react with the non‐pathogenic P. stewartii subsp. indologenes. The PCR tests currently recommended by the EPPO Diagnostic Protocol PM 7/60 (2) for initial screening bear the risk of false positives as they detect both subspecies of P. stewartii. The test presented here has high analytical sensitivity (10² cells mL^–1 in leaf extracts, at least 10³ cells mL^?1 in seed extracts), high repeatability and high reproducibility and performed well on the two matrices tested: maize seed and leaf extract. Its improved analytical specificity and analytical sensitivity compared to the currently recommended tests lead the authors to suggest that this test should be included as a first screening test in the diagnostic protocol for P. stewartii subsp. stewartii when it is next revised.     

Hybrid seed set in wheat is a complex trait but can be improved indirectly by selection for male floral traits

Efficient hybrid wheat breeding requires the redesign of the wheat floral architecure to enhance cross-pollination. Several studies evaluated the phenotypic variation and the genetic architecture of male floral traits, but their contribution to the most important trait, hybrid seed set on the female parent, has not yet been considered. To bridge this gap, we employed 31 male lines and evaluated the hybrid seed set on two female tester lines in crossing blocks. Hybrid seed set showed large genetic variance and high heritability, which demonstrates the potential for the improvement of this trait. However, the assessment of hybrid seed set is difficult as secondary traits like plant height and especially flowering time, as well as the environment largely influence the hybrid seed set. Nevertheless, a moderately high correlation between visual anther extrusion and hybrid seed set opens up the possibility to use visual anther extrusion as an indirect trait for preliminary male screenings. Further research evaluating traits influencing female receptivity coupled with genomics-assisted approaches are highly recommended to develop an improved selection portfolio for maximizing hybrid seed set.     

Genomic prediction and GWAS of Gibberella ear rot resistance traits in dent and flint lines of a public maize breeding program

Gibberella ear rot (GER) is a serious threat to maize cultivation, causing grain yield losses and contamination with mycotoxins. Genomic prediction (GP) has great potential to accelerate resistance breeding against GER. However, small training sets (TS) consisting of both phenotyped and genotyped individuals are a challenge for obtaining high prediction accuracy (ρ) in GP. A potential solution would be combining small-size populations across heterotic pools. However, genetic heterogeneity between populations in terms of segregating QTL, linkage disequilibrium (LD) pattern and genomic relationships can impair ρ of GP. In this study, we investigated the genetic architecture of GER severity, deoxynivalenol concentration (DON) and days to silking with genome-wide association studies within two elite panels of 130 dent and 114 flint lines from the maize breeding program of the University of Hohenheim tested in four environments. We also assessed the consistency of LD pattern and genomic relationships between the two heterotic pools. Furthermore, we compared four GP approaches differing in the composition of the TS with lines from a single or combined pool(s) and statistical models with marker effects identical or different but correlated between pools. We detected two and six QTL for DON within the dent and flint pool, respectively, but no common QTL. The LD pattern was consistent between pools for marker pairs ≤ 10 kb apart. GP across pools yielded low or even negative ρ. Combined-pool GP had no higher ρ than within-pool GP, regardless of the statistical model. Our findings underline the importance of investigating the genetic heterogeneity between populations prior to implementing GP using combined TS.     

Higher grain yield and higher grain protein deviation underline the potential of hybrid wheat for a sustainable agriculture

Grain yield and protein content are traits of major importance in wheat breeding, but their combination is challenging due to a tight negative correlation. Protein yield and grain protein deviation have been proposed as selection criteria to simultaneously improve both traits. Sedimentation volume is an indicator of protein quality, which plays an important role for bread‐making quality in wheat. All these traits have been investigated in our study with 135 parental inbred lines, their 1,604 hybrids and 10 commercial check varieties evaluated at five environments. The focus of our study was to investigate the usefulness of the grain protein deviation and to define a bivariate model for calculating the grain protein deviation. Further, we compared line and hybrid wheat for grain yield and quality‐related parameters such as protein content and sedimentation volume. The grain protein deviation determined with a bivariate model delivered robust estimates of variance components and enabled a balanced selection of genotypes with improved protein content and grain yield across different quality classes. Although heterosis for protein content and sedimentation volume was negative, hybrids had a higher grain protein deviation as well as higher grain yield at a given sedimentation volume or a given protein content than line varieties.     

Deconjugation and degradation of flavonol glycosides by pig cecal microbiota characterized by Fluorescence in situ hybridization (FISH)

As the bioavailability of flavonoids is influenced by intestinal metabolism, we have investigated the microbial deconjugation and degradation of several flavonols and flavonol glycosides using the pig cecum in vitro model system developed in our group. For this model system the microbiota was directly isolated from the cecal lumen of freshly slaughtered pigs. The characterization of the cecal microbiota by fluorescence in situ hybridization (FISH) with 16S rRNA-based oligonucleotide probes confirmed the suitability of the model system for studying intestinal metabolism by the human microbiota. We have investigated the microbial degradation of quercetin-3-O-beta-d-rutinoside 1, quercetin-3-O-beta-d-glucopyranoside 2, quercetin-4'-O-beta-d-glucopyranoside 3, quercetin-3-O-beta-d-galactopyranoside 4, quercetin-3- O-beta-d-rhamnopyranoside 5, quercetin-3- O-[alpha-l-dirhamnopyranosyl-(1-->2)-(1-->6)-beta-d-glucopyranoside 6, kaempferol-3-O-[alpha-l-dirhamnopyranosyl-(1-->2)-(1-->6)-beta-d-glucopyranoside 7, apigenin 8, apigenin-8- C-glucoside (vitexin) 9, and feruloyl-O-beta-d-glucopyranoside 10 (100 microM), representing flavonoids with different aglycones, sugar moieties, and types of glycosidic bonds. The degradation rate was monitored using HPLC-DAD. The flavonol O-glycosides under study were almost completely metabolized by the intestinal microbiota within 20 min and 4 h depending on the sugar moiety and the type of glycosidic bond. The degradation rates of the quercetin monoglycosides showed a clear dependency on the hydroxyl pattern of the sugar moiety. The degradation of 2 with all hydroxyl groups of the glucose in the equatorial position was the fastest. The intestinal metabolism of di- and trisaccharides was much slower compared to the monoglycosides. The structure of the aglycone has not much influence on the intestinal metabolism; however, the type of glycosidic bond ( C- or O-glycoside) has substantial influence on the degradation rate. The liberated aglycones were completely metabolized within 8 h. Phenolic compounds such as 3,4-dihydroxyphenylacetic acid 12, 4-hydroxyphenylacetic acid 13, and phloroglucinol 18 were detected by GC-MS as main degradation products.