Molecular cloning of the mRNA coding for the G protein of the viral haemorrhagic septicaemia (VHS) of salmonids

Viral haemorrhagic septicaemia virus (VHSV), a rhabdovirus, is a major threat for continental European trout fish farming. The development of a recombinant subunit vaccine could solve that problem. The neutralizing epitopes are located on the glycoprotein or G protein, the surface antigen. The G protein has a molecular weight of 65 kDa, reduced to 55 kDa by deglycosylation. cDNA was synthetized from mRNA of VHS virus infected cells, and cloned in E. coli. The viral cDNA was recognized by positive hybridization with a labelled probe made from infected cell RNA, and negative hybridization with labelled cDNA made from cellular RNA. The Northern blot hybridization with different clones on VHS infected cell RNA revealed two VHS mRNA whose lengths, 2.0 and 1.5 kb, were compatible with the mRNA length for G and N proteins respectively. This mRNA must contain about 400 bp of untranslated sequence.     

A pmp genes-based PCR as a valuable tool for the diagnosis of avian chlamydiosis

In a previous study we described the use of a new set of PCR primers (CpsiA/CpsiB) specific of the conserved pmp-family genes of Chlamydophila abortus as an efficient tool for the detection of these bacteria in ruminants including also preliminary results on avian strains. In this work, the use of this set of primers was extended to representative strains of the six major avian serovars (serovars A-F) and to field isolates of C. psittaci. For all the studied representative strains, using purified genomic DNA as a template, CpsiA/CpsiB primers allowed, as observed for C. abortus, a minimal 10-fold PCR signal increase compared to the one observed with ompA specific primers. In comparison to primers targeting the 16S-23S rRNA intergenic spacer, similar or increased sensitivity was observed depending on the strain. All the field isolates were amplified with CpsiA/CpsiB primers. On clinical samples, our primers are the best among those tested for detection of C. psittaci by simple conventional PCR. RFLP experiments performed using PCR fragments amplified with the CpsiA/CpsiB primers gave promising results demonstrating that these primers may provide an interesting tool for molecular typing when the bacterium cannot be grown from pathological samples.     

Nucleotide sequence and expression of an AIDS-associated retrovirus (ARV-2)

The nucleotide sequence of molecular clones of DNA from a retrovirus, ARV-2, associated with the acquired immune deficiency syndrome (AIDS) was determined. Proviral DNA of ARV-2 (9737 base pairs) has long terminal repeat structures (636 base pairs) and long open reading frames encoding gag (506 codons), pol (1003 codons), and env (863 codons) genes. Two additional open reading frames were identified. Significant amino acid homology with several other retroviruses was noted in the predicted product of gag and pol, but ARV-2 was as closely related to murine and avian retroviruses as it was to human T-cell leukemia viruses (HTLV-I and HTLV-II). By means of an SV-40 vector in transfected simian cells, the cloned gag and env genes of ARV-2 were shown to express viral proteins.     

Breeding for increased nitrogen‐use efficiency: a review for wheat (T. aestivum L.)

Nitrogen fertilizer is the most used nutrient source in modern agriculture and represents significant environmental and production costs. In the meantime, the demand for grain increases and production per area has to increase as new cultivated areas are scarce. In this context, breeding for an efficient use of nitrogen became a major objective. In wheat, nitrogen is required to maintain a photosynthetically active canopy ensuring grain yield and to produce grain storage proteins that are generally needed to maintain a high end‐use quality. This review presents current knowledge of physiological, metabolic and genetic factors influencing nitrogen uptake and utilization in the context of different nitrogen management systems. This includes the role of root system and its interactions with microorganisms, nitrate assimilation and its relationship with photosynthesis as postanthesis remobilization and nitrogen partitioning. Regarding nitrogen‐use efficiency complexity, several physiological avenues for increasing it were discussed and their phenotyping methods were reviewed. Phenotypic and molecular breeding strategies were also reviewed and discussed regarding nitrogen regimes and genetic diversity.     

Genetic control of yield and yield components in winter oilseed rape (Brassica napus L.) grown under nitrogen limitation

Despite its high nitrogen absorption capacity, oilseed rape (OSR) has a low apparent nitrogen use efficiency (NUE), which makes its production highly dependent on nitrogen fertilization. Improving NUE in OSR is therefore a main target in breeding. The objectives of the present work were to determine the genomic regions (QTLs) associated with yield and to assess their stability under contrasted nitrogen nutrition regimes. One mapping population, AM, was tested in a French location for three growing seasons (2011, 2012 and 2013), under two nitrogen conditions (optimal and low). Eight yield-related traits were scored and nitrogen-responsive traits were calculated. A total of 104 QTLs were detected of which 28 controlled flowering time and 76 were related to yield and yield components. Very few genotype × nitrogen interactions were detected and the QTLs were highly stable between the nitrogen conditions. In contrast, only a few QTLs were stable across the years of the trial, suggesting a strong QTL × year interaction. Finally, eleven critical genomic regions that were stable across nitrogen conditions and/or trial years were identified. One particular region located on the A5 linkage group appears to be a promising candidate for marker assisted selection programs. The different strategies for OSR breeding using the QTLs found in the present study are discussed.     

An increase in the upper tree-limit of silver fir (Abiesalba Mill.) in the Alps since the mid-20th century: A land-use change phenomenon

Global environmental changes observed during recent decades are likely to have had an impact on the distribution of species. Currently, silver fir (Abies alba) is becoming established in the subalpine forests of the west central Alps at elevations higher than 2000 m a.s.l.; prior to the 1970s its upper altitudinal limit was 2000 m. Several hypotheses could explain this recent expansion of the upper tree-limit. Silver fir regeneration could be linked (1) to land-use changes or (2) to current climatic warming. Using dendrochronology, the age structure of 31 forest plots containing at least one silver fir was examined in order to elucidate the population dynamics of subalpine communities. This allowed us to decipher the timing of fir regeneration in relation to the mean age of the stands examined and of the other tree-canopy species present. The majority of the firs germinated sporadically since 1950, before the regional temperature increase. The pattern of fir recruitment did not appear to relate to altitude, but followed a pattern characteristic of secondary succession. The age structures identified showed an exponential increase in tree-density during the 20th century; the stands were first dominated by Larix decidua during the 18th and 19th centuries, and then by Pinus cembra during the 20th century. In most stands, fir regeneration occurred after Larix and before P. cembra dominated, following a similar pattern to Picea abies regeneration. The number of local inhabitants and temperature both exhibited a negative relationship with fir tree recruitment, thus supporting the land-use change hypothesis. There has been a significant upward shift of the altitudinal range of fir, amounting to an increase of about 300 m since 1950. This followed the abandonment of low-productivity land. This trend is likely to continue during the 21st century, because of new agricultural and forestry practices which involve limited intervention in low-productivity areas and may be because of the effects of global warming.     

Post‐capture survival of whale sharks encircled in tuna purse‐seine nets: tagging and safe release methods

1. Whale shark, the world's largest fish, is believed to be particularly vulnerable owing to its biological characteristics (slow growth, late maturation, great longevity) and is listed as Vulnerable by IUCN and included in Appendix II of CITES. 2. Whale sharks are occasionally encircled in tropical tuna purse‐seine nets, throughout this global fishery. Although apparent immediate survival rates following encirclement and release have recently been assessed through scientific onboard observer programmes, a more rigorous methodology is still required for studying post‐released survival. 3. This work provides a method for applying pop‐up satellite tags and reports an enhanced release procedure for whale sharks. The first assessment of survival after release from purse‐seine nets involved six whale sharks tagged between May and September 2014 in the eastern tropical Atlantic Ocean. Five tags transmitted data: three popped up as programmed (after 30 days), while two surfaced prematurely (one after 21 and the other after 71 days (programmed to pop off after 30 and 90 days, respectively)) but showed no sign of unusual behaviour. 4. Overall, whale sharks survived at least 21 days (one at least 71 days) after release from purse‐seine nets. These observations based on five large individuals (total length > 8 m), suggest that whale sharks have a good chance of survival when released with the proposed method. 5. Additional tagging in this and other oceans, especially of juveniles which may be more sensitive to encirclement and release operations, is essential to further assess whale shark post‐release survival rates in tuna purse‐seine fisheries. Copyright © 2016 John Wiley & Sons, Ltd.     

Characterization of plum procyanidins by thiolytic depolymerization

The phenolic compounds of 'Green Gage' (GG) plums ( Prunus domestica L.), "Rainha Claudia Verde", from a 'protected designation of origin' (PDO), in Portugal, were quantified in both flesh and skin tissues of plums collected in two different orchards (GG-V and GG-C). Analyzes of phenolic compounds were also performed on another GG European plum obtained in France (GG-F) and two other French plums, 'Mirabelle' (M) and 'Golden Japan' (GJ). Thiolysis was used for the first time in the analysis of plum phenolic compounds. This methodology showed that the flesh and skin contain a large proportion of flavan-3-ols, which account, respectively, for 92 and 85% in GJ, 61 and 44% in GG-V, 62 and 48% in GG-C, 54 and 27% in M, and 45 and 37% in GG-F. Terminal units of procyanidins observed in plums are mainly (+)-catechin (54-77% of all terminal units in flesh and 57-81% in skin). The GJ plums showed a phenolic composition different from all of the others, with a lower content of chlorogenic acid isomers and the presence of A-type procyanidins as dimers and terminal residues of polymerized forms. The average degree of polymerization (DPn) of plum procyanidins was higher in the flesh (5-9 units) than in the skin (4-6 units). Procyanidin B7 was observed in the flesh of all GG plums and in the skin of the Portuguese ones. Principal component analysis of the phenolic composition of the flesh and skin of these plums obtained after thiolysis allowed their distinction according to the variety and origin, opening the possibility of the use of phenolic composition for variety/origin identification.     

Development of an oat-based biorefinery for the production of L(+)-lactic acid by Rhizopus oryzae and various value-added coproducts

A novel oat-based biorefinery producing L(+)-lactic acid and various value-added coproducts (e.g., beta-glucan, anti-irritant solution) is proposed. Pearling is employed for sequential separation of bran-rich fractions for the extraction of value-added coproducts. Lactic acid production is achieved via fungal fermentation of Rhizopus oryzae on pearled oat flour. Maximum lactic acid concentration (51.7 g/L) and starch conversion yield (0.68 g/g) were achieved when an oat flour concentration of 116.5 g/L was used. Oxygen transfer played a significant role with respect to lactic acid production and starch conversion yield. Rhizopus oryzae produced a range of enzymes (glucoamylase, protease, phosphatase) for the hydrolysis of cereal flour macromolecules. Enzyme production during fungal fermentation has been reported. The proposed biorefining strategy could lead to significant operating cost reduction as compared to current industrial practices for lactic acid production from pure glucose achieved by bacterial fermentations.     

Breakdown of the Scab Resistance Gene Vf in Apple Leads to a Founder Effect in Populations of the Fungal Pathogen Venturia inaequalis

ABSTRACT The recent breakdown of Vf, a major resistance gene to apple scab, provided an opportunity to analyze a population genetic process within the matching virulent subpopulation of the fungus Venturia inaequalis. We utilized the amplified fragment length polymorphism technique and allelic variation at four microsatellite loci to assess genetic structure of 133 isolates of V. inaequalis from a single commercial apple orchard sampled from one cultivar carrying the Vf gene (Judeline) and three cultivars devoid of the Vf gene. Both analyses indicated a strong decrease of the genetic diversity among isolates from the Vf cultivar compared with the high level of diversity among isolates from the three other cultivars. This leads to a high genetic differentiation between virVf and avrVf groups (F(ST) > 0.17). Analyses of the genetic distance between AFLP patterns based on the Jaccard index indicate that all virVf isolates could be assigned to a single clonal lineage. These results lead us to conclude that the clonal structure of the population isolated from the Vf cultivar is an example of a founder effect in response to a resistance gene breakdown and it is likely that this event occurred in the orchard during the sampling year.