Mode of resistance to respiration inhibitors at the cytochrome bc1 enzyme complex of Mycosphaerella fijiensis field isolates

Field isolates of Mycosphaerella fijiensis, causing black Sigatoka of banana, were characterised for their sensitivity to different inhibitors of the cytochrome bc₁ enzyme complex (Qo respiration inhibitors, strobilurin fungicides), using physiological, biochemical and molecular genetic methods. Strobilurin‐resistant isolates exhibited very high resistance factors both in mycelial growth inhibition and NADH consumption assays. Cross‐resistance was observed among all Qo inhibitors, including trifloxystrobin, azoxystrobin, famoxadone, strobilurin B and myxothiazol. However, the Qi and the cytochrome aa₃ inhibitors, antimycin A and potassium cyanide, respectively, were not cross‐resistant to Qo inhibitors. In sensitive but not in resistant isolates, mixtures of Qo inhibitors and SHAM, an inhibitor of the alternative oxidase (AOX), were more active than the components alone, indicating that the alternative pathway is essential in metabolism, but not causal for resistance. In the cell‐free NADH‐consumption assay, the Qo inhibitors affected the sensitive but not the resistant isolates, suggesting that AOX was not active in sub‐mitochondrial particles. In whole cells, however, the AOX has a basic expression level and is probably not inducible by trifloxystrobin. Sequencing of the cytochrome b gene of sensitive and resistant M fijiensis isolates revealed a difference in the nucleotide sequence leading to a single amino acid change from glycine to alanine at position 143 in the resistant isolate. This change is known to occur also in the naturally tolerant basidiomycete Mycena galopoda. It is suggested that the field isolates of M fijiensis can acquire resistance to Qo inhibitors due to a target site alteration with a single base pair change. Resistant isolates do not seem to contain a mixture of mutated and non‐mutated DNA, indicating a complete selection of resistant mitochondria and a maternally donated mode of resistance. © 2000 Society of Chemical Industry     

Assessment of latent infection frequency in progeny tubers of advanced potatoclones resistant to bacterial wilt: A new selection criterion

Summary This paper reports results of a 3-year evaluation of CIP advanced potato clones in a bacterial wilt-infested field (race 3) in Peru. Clones resistant or moderately resistant to wilt were selected and all tubers harvested from each clone were tested for latent infection byRalstonia solanacearum using a sensitive serological technique developed at CIP. A sampling strategy to estimate accurately the frequency of infected tubers in the clones has been evaluated. This method will allow consideration of tuber latent infection as a new selection criterion in breeding for resistance to bacterial wilt. Thirteen clones were found resistant to wilt in all three evaluations (i.e.≤6% wilt), from which five had no wilt in all trials. However, all clones harboured latent infection in tubers averaging 30%. Analysing 30 tubers/clone provides an accurate estimation of the proportion of infected tubers with a high precision level.     

Early degradation of plant alkanes in soils: A litterbag experiment using C-labelled leaves

We monitored the carbon isotope composition of bulk leaves and specific long-chain alkanes during a four-year litterbag experiment using ¹³C-labelled leaves and unlabelled reference leaves of the European beech tree (Fagus sylvatica L.). Whereas the isotope composition of alkanes from ¹³C-enriched leaves exhibited a marked decrease in ¹³C-content, the isotope composition of unlabelled reference leaves remained nearly constant. We interpreted this difference as evidence for a microbial contribution to the long-chain alkane pool of the decomposing leaves and related it to the progressive invasion of leaves by soil organisms which was revealed upon microscopic examination. These results suggest that long-chain alkanes may not provide an unaltered record of organic carbon isotope composition in soils and sediments.     

Detection of linkage between RFLP markers and genes affecting anthocyanin pigmentation in maize (Zea mays L.)

Summary Linkages between molecular markers and genes involved in the expression of agronomical traits have already been described in all of the major crops. In most cases, the genetic model underlying the Quantitative Traits Loci (QTL) is discussed. Here, Restriction Fragment Length Polymorphisms (RFLPs) and Mapmaker-QTL have been used to pinpoint seven regions of the genome significantly correlated with four pigmentation qualitative traits of maize (Zea mays L.). Two of these, located on chromosomes 2 and 10, explain most of the variation of these traits. The R and B gene loci known to be involved in the regulation of the anthocyanin pathway map to the same regions and we suggest that these loci could be the candidate genes involved in the correlations detected with RFLPs. This type of result is in accordance with the hypothesis of the candidate gene which supposes that, if we have a very high density map of randomly-selected cDNA clones, it should theoretically be possible to associate a cloned genic sequence with a phenotypic trait where correlations are found.     

Enzyme-induced gelation of extensively hydrolyzed whey proteins by Alcalase: peptide identification and determination of enzyme specificity

Extensive hydrolysis of whey protein isolate by Alcalase was shown to induce gelation mainly via hydrophobic interactions. The aim of this work was to characterize the peptides released in order to better understand this phenomenon. The apparent molecular mass distribution indicated that aggregates were formed by small molecular mass peptides (<2000 Da). One hundred and thirty peptides with various lengths were identified by reversed-phase high-performance liquid chromatography coupled with electrospray ionization mass spectrometry. Alcalase was observed to have a high specificity for aromatic (Phe, Trp, and Tyr), acidic (Glu), sulfur-containing (Met), aliphatic (Leu and Ala), hydroxyl (Ser), and basic (Lys) residues. Most peptides had an average hydrophobicity of 1-1.5 kcal/residue and a net charge of 0 at the pH at which gelation occurred (6.0). Therefore, an intermolecular attractive force such as hydrophobic interaction suggests the formation of aggregates that further leads to the formation of a gel.     

Synthetic clonal reproduction through seeds

Cloning through seeds has potential revolutionary applications in agriculture, because it would allow vigorous hybrids to be propagated indefinitely. However, asexual seed formation or apomixis, avoiding meiosis and fertilization, is not found in the major food crops. To develop de novo synthesis of apomixis, we crossed Arabidopsis MiMe and dyad mutants that produce diploid clonal gametes to a strain whose chromosomes are engineered to be eliminated after fertilization. Up to 34% of the progeny were clones of their parent, demonstrating the conversion of clonal female or male gametes into seeds. We also show that first-generation cloned plants can be cloned again. Clonal reproduction through seeds can therefore be achieved in a sexual plant by manipulating two to four conserved genes.     

Consequences of adult and juvenile movement for marine protected areas

Adult and juvenile mobility has a considerable influence on the functioning of marine protected areas. It is recognized that adult and juvenile movement reduces the core benefits of protected areas, namely protecting the full age–structure of marine populations, while at the same time perhaps improving fisheries yield over the no-reserve situation through export of individuals from protected areas. Nevertheless, the study of the consequences of movement on protected area functioning is unbalanced. Significant attention has been paid to the influence of certain movement patterns, such as diffusive movement and home ranges, while the impacts of others, such as density-dependent movements and ontogenetic migrations, have been relatively ignored. Here we review the diversity of density-independent and density-dependent movement patterns, as well as what is currently known about their consequences for the conservation and fisheries effects of marine protected areas. We highlight a number of ‘partially addressed’ issues in marine protected area research, such as the effects of reserves targeting specific life phases, and a number of essentially unstudied issues, such as density-dependent movements, nomadism, ontogenetic migrations, behavioral polymorphism and ‘dynamic’ reserves that adjust location as a realtime response to habitat changes. Assessing these issues will be essential to creating effective marine protected area networks for mobile species and accurately assessing reserve impacts on these species.     

Uncommonly thorough hydrolysis of peptides during ripening of Ragusano cheese revealed by tandem mass spectrometry

Ragusano is a pasta filata cheese produced from raw milk in Sicily. The proteolysis was extensively analyzed after stretching (day 0), at 4 and 7 months of ripening through soluble nitrogen, urea-PAGE, and peptide identification by tandem mass spectrometry. After stretching, 123 peptides were identified: 72 arising from β-casein, 34 from α(s1)-casein, and 17 from α(s2)-casein. The main protein splitting corresponded to the action of plasmin, chymosin, cathepsin D, cell envelope proteinase, and peptidase activities of lactic acid bacteria. Unlike other types of cheeses, <10% residual β- and α(s)-caseins remained intact at 7 months, indicating original network organization based on large casein fragments. The number of identified soluble peptides also dramatically decreased after 4 and 7 months of ripening, to 47 and 25, respectively. Among them, bioactive peptides were found, that is, mineral carrier, antihypertensive, and immunomodulating peptides and phosphopeptides.     

Assessing the effects of the clumping phenomenon on BRDF of a maize crop based on 3D numerical scenes using DART model

Inverting radiative transfer (R-T) models against remote sensing observations to retrieve key biogeophysical parameters such as leaf area index (LAI) is a common approach. Even if new inversion techniques allow the use of three-dimensional (3D) models for that purpose, one-dimensional (1D) models are still widely used because of their ease of implementation and computational efficiency. Nevertheless, they assume a random distribution of foliage elements whereas most canopies show a clumped organization. Due to that crude simplification in the representation of the canopy structure, sizeable discrepancies can occur between 1D simulations and real canopy reflectance, which may further lead to false LAI values. The present investigation aims to appraise to which extent the incorporation of a clumping index (noted λ) into 1D R-T model could improve the simulations of Bidirectional Reflectance Distribution Function (BRDF). Canopy BRDF is simulated here for three growth stages of a maize crop with the Discrete Anisotropic Radiative Transfer (DART) model in the visible and near infrared spectral bands, for two contrasted soil types (dark and bright) and different levels of heterogeneity to represent the canopy structure. 3D numerical scenes are based on in-situ structural measurements and associated BRDF simulations are thus considered as references. 1D scenarios assume either that leaves are randomly distributed (λ = 1) or clumped (λ < 1). If BRDF simulations seem globally reliable under the assumption of a random distribution in near infrared, it can also lead to relative errors on the total BRDF up to 30% in the red spectral band. It comes out that the use of a clumping index in a 1D reflectance model generally improves BRDF simulations in the red considering a bright soil, which seems relatively independent of LAI. In the near infrared, best results are usually obtained with homogeneous canopies, except with the dark soil. Clearly, influent factors are mainly the LAI and the spectral contrast between soil and leaves.     

Multivariate effects of plant canopy, soil physico-chemistry and microbiology on Sclerotinia stem rot of soybean in relation to crop rotation and urban compost amendment

The effects of canopy, soil physico-chemical and microbiological variables on Sclerotinia stem rot (SSR) on soybean were assessed in two soils (clay loam and sandy loam) using multiple regression and canonical redundancy analysis (RDA) and their partial form to control for the rotation (2 or 3-y-corn/soybean monoculture) and fertilization (mineral/urban compost) or spatial variables effects. The models revealed the minimal sets of variables that best explain the variance of the survival of Sclerotinia sclerotiorum’s sclerotia, carpogenic germination, disease severity and their associations. In clay loam, the 3-y-corn rotation reduced disease severity mainly through the reduction of weed biomass that favoured carpogenic germination. Urban compost has a conducive effect explained by a better soil surface drainage. Additionally, total N was found suppressive to sclerotial survival. In sandy loam, the carpogenic germination was negatively correlated with high C mineralization quotient and aggregate stability but correlated positively with Ca. Sclerotial survival was negatively correlated with pH and Ca, and positively correlated with biological fertility index. Aggregate stability, Ca and pH were associated with the urban compost. The regression and RDA analyses allowed to identify key variables that drived SSR development and explain their relationship with the cultural practices, soil health, as well as the spatial variation of disease variables.