Philippe C. Nicot

Identification of two powdery mildew fungi, Oidium neolycopersici sp. nov. and O. lycopersici , infecting tomato in different parts of the world

A world-wide study of the Oidium species causing economic damage on tomato has identified two taxa using classical morphological, scanning electron microscope (SEM) and molecular phylogenetic analyses. The material consisted of a total of 25 tomato powdery mildew isolates and 29 herbarium specimens coming from all continents where tomatoes are grown. A taxon with non-catenate conidia widespread in Europe, Africa, North and South America and Asia was identified as an O. subgen. Pseudoidium species (teleomorph: Erysiphe sect. Erysiphe). Formerly mistaken for O. lycopersicum (or O. lycopersici), it is now recognised as a distinct species, O. neolycopersici sp. nov. A phylogenetic analysis of the internal transcribed spacer (ITS) sequences of the ribosomal DNA (rDNA) indicated that O. neolycopersici is closely related to Erysiphe macleayae, E. aquilegiae and other Pseudoidium species. Only a taxon with catenate conidia was found on Australian specimens. This was identified as a species of O. subgen. Reticuloidium (teleomorph: Golovinomyces sp.). Phylogenetic analysis of the rDNA ITS sequences showed that this species is closely related to O. longipes infecting eggplant. Because it is most likely to be the same species as the original O. lycopersicum, which was actually first described in Australia, this is here neotypified as O. lycopersici.

Expanding the Paradigms of Plant Pathogen Life History and Evolution of Parasitic Fitness beyond Agricultural Boundaries

How do pathogens, whether they parasitize plants or animals, acquire virulence to new hosts and resistance to the arms we deploy to control disease? The significance of these questions for microbiology and for society at large can be illustrated by the recent worldwide efforts to track and limit the emergence of human transmissible strains of swine and avian influenza virus and of multidrug-resistant lines of human pathogenic bacteria, and to restrain the spread of Ug99, a strain of stem rust of wheat. Recent research in medical epidemiology has elucidated the impact of pathogen ecology in environmental reservoirs on the evolution of novel or enhanced pathogen virulence. In contrast, the evolution of virulence in plant pathogens has been investigated from a predominantly agro-centric perspective, and has focused overwhelmingly on evolutionary forces related to interactions with the primary plant host. Here, we argue that current concepts from the field of medical epidemiology regarding mechanisms that lead to acquisition of novel virulence, biocide resistance, and enhanced pathogenic fitness can serve as an important foundation for novel hypotheses about the evolution of plant pathogens. We present numerous examples of virulence traits in plant pathogenic microorganisms that also have a function in their survival and growth in nonagricultural and nonplant habitats. Based on this evidence, we make an appeal to expand concepts of the life history of plant pathogens and the drivers of pathogen evolution beyond the current agro-centric perspective.

Aerial plant surface microbiology

The Physical and Chemical Environment of Aerial Plant Surfaces: Cuticle Permeability Studies: A Model for Estimating Leaching of Plant Metabolites to Leaf Surfaces (J. Schoenherr, P. Baur). Nutrients on the Leaf Surface (S. Derridj). Interactions between Plant Surface Microorganisms and Their Hosts: Microbial Attachment to Plant Aerial Surfaces (M. Romantschuk et al.). Bacterial hrp and Avirulence Genes Are Key Determinants in Plant Pathogen Interactions (U. Bonas, G. Van den Ackerveken). Interactions among Microorganisms in Communities on Aerial Plant Surfaces: Gene Transfer between Microorganisms in the Phyllosphere (M.J. Bailey et al.). Microbial Interactions Preventing Fungal Growth on Senescent and Necrotic Aerial Plant Surfaces (W.F. Pfender). Plant Surface Microorganisms, Agricultural Practices and Food Quality: Role of Immigration of Other Processes in Determining Epiphytic Bacterial Populations: Implications for Disease Management (S.E. Lindow). Integrated Control of Botrytis cinerea on Greenhouse Tomatoes (P.C. Nicot, A. Baille). Quantification and Modeling of Population Dynamics of Microorganisms Associated with Aerial Plant Surfaces: The Effect of Leaf Age and Position on the Dynamics of Microbial Populations on Aerial Plant Surfaces (M.A. Jacques). Research Outlook: Phyllosphere Ecology: Past, Present and Future (J.H. Andrews). 9 additional articles. Index.

Virulence variation and DNA polymorphism in Sphaerotheca fuliginea, causal agent of powdery mildew of cucurbits

Strains of Sphaerotheca fuliginea, one of the causal agents of powdery mildew of cucurbits, were examined for differences in virulence, mating type and DNA polymorphism. The 28 strains were chosen to be diverse according to host and geographic origin. Characterization of virulence phenotypes was based on the expression of symptoms on 4 species of cucurbits and 6 cultivars of melon. Two pathotypes, capable of attacking either cucumber cv. ‘Marketer’ and melon cv. ‘IranH’ and squash cv. ‘Diamant’ or cucumber cv. ‘Marketer’ and melon cv. ‘IranH’ were observed. Tests on melon cultivars revealed 3 races. In tests of sexual compatibility with reference strains, heterothallism was observed for all isolates. Frequency of the two mating types differed significantly in the population. DNA polymorphism was determined both by restriction fragment length polymorphism (RFLP) of the ribosomal internal transcribed spacers (ITS) and 5.8S DNA amplified by the polymerase chain reaction and by random amplified polymorphic DNA (RAPD). For any one of the 11 restriction enzymes tested all strains presented an identical pattern of ITS RFLP. RAPD analysis, using 22 primers which provided reproducible patterns, revealed a relatively low degree of polymorphism. Furthermore, cluster analysis based on RAPD data (152 markers) did not separate groups within the species S. fuliginea. No association could be found between virulence, mating type, geographical and host origin and RAPD patterns. The lack of association between phenotypic and molecular markers and the close fit to linkage equilibrium for the characters examined suggest that recombination may play a role in populations of S. fuliginea.

Differential spore production by Botrytis cinerea on agar medium and plant tissue under near-ultraviolet light-absorbing polyethylene film

Plastic films containing additives that alter their transmission of the light spectrum may be useful tools for the control of aerial plant pathogens of greenhouse crops. Several samples of polyethylene films containing additives that absorb near ultraviolet (nUV) light in the range 280 to 380 nm were compared for their ability to affect spore germination, mycelial growth, and sporulation of Botrytis cinerea on agar medium. One film was selected and further evaluated. The kinetics of spore production by the pathogen was similar on agar medium and on tomato stem tissue, and whether incubation took place under the nUV-absorbing film or under a control film. However, spore production on both types of substrates under the nUV film remained at less than 0.05% that of the control for several weeks after inoculation, demonstrating that the nUV film inhibited rather than delayed sporulation. A sharp reduction of spore production was also observed on other plant tissues. However, the efficiency of the nUV film appeared different for different plants, and it was lower on flowers and cotyledons than on stem tissue. Two of the five strains of B. cinerea tested on tomato stem tissue were less sensitive to sporulation inhibition by the nUV film. To clarify the potential of nUV films for the control of gray mold on greenhouse crops, the epidemiological significance of these results needs to be further examined in light of the abundance of such strains in the environment.

Rapid Change in the Genetic Diversity of Botrytis cinerea Populations After the Introduction of Strains in a Tomato Glasshouse

In tomato glasshouses, the population structure of airborne inoculum of Botrytis cinerea depends on the production of endogenous inoculum on diseased plants as well as on incoming exogenous inoculum. Both types of inocula may contribute differently to the development of epidemics. Two strains of B. cinerea were introduced in each of four separate compartments of an experimental tomato glasshouse. We monitored their impact on disease development and on the genetic diversity of B. cinerea populations using microsatellite markers. The naturally occurring airborne inoculum of B. cinerea displayed a high level of genetic diversity and was rapidly displaced in the glasshouse, as isolates with microsatellite profiles identical to the introduced strains amounted to 66% of the inoculum sampled from the air 14 days after inoculation and 91% of those collected from stem lesions 60 days after inoculation. This suggested an important role of secondary inoculum in disease development, which is compatible with the hypothesis of a polycyclic development of gray mold epidemics in tomato glasshouses. In controlled-environment tests on tomatoes, a wide range of aggressiveness levels was observed, both for isolates sampled from the air and from lesions on plants. Hypotheses are proposed to explain the negligible impact of naturally incoming isolates on the epidemics observed inside the four glasshouse compartments.

Functional and structural comparison of pyrrolnitrin- and iprodione-induced modifications in the class III histidine-kinase Bos1 of Botrytis cinerea.

Dicarboximides and phenylpyrroles are commonly used fungicides against plant pathogenic ascomycetes. Although their effect on fungal osmosensing systems has been shown in many studies, their modes-of-action still remain unclear. Laboratory- or field-mutants of fungi resistant to either or both fungicide categories generally harbour point mutations in the sensor histidine kinase of the osmotic signal transduction cascade. In the present study we compared the mechanisms of resistance to the dicarboximide iprodione and to pyrrolnitrin, a structural analogue of phenylpyrrole fungicides, in Botrytis cinerea. Pyrrolnitrin-induced mutants and iprodione-induced mutants of B. cinerea were produced in vitro. For the pyrrolnitrin-induced mutants, a high level of resistance to pyrrolnitrin was associated with a high level of resistance to iprodione. For the iprodione-induced mutants, the high level of resistance to iprodione generated variable levels of resistance to pyrrolnitrin and phenylpyrroles. All selected mutants showed hypersensitivity to high osmolarity and regardless of their resistance levels to phenylpyrroles, they showed strongly reduced fitness parameters (sporulation, mycelial growth, aggressiveness on plants) compared to the parental phenotypes. Most of the mutants presented modifications in the osmosensing class III histidine kinase affecting the HAMP domains. Site directed mutagenesis of the bos1 gene was applied to validate eight of the identified mutations. Structure modelling of the HAMP domains revealed that the replacements of hydrophobic residues within the HAMP domains generally affected their helical structure, probably abolishing signal transduction. Comparing mutant phenotypes to the HAMP structures, our study suggests that mutations perturbing helical structures of HAMP2-4 abolish signal-transduction leading to loss-of-function phenotype. The mutation of residues E529, M427, and T581, without consequences on HAMP structure, highlighted their involvement in signal transduction. E529 and M427 seem to be principally involved in osmotic signal transduction.

Resistance to bio-insecticides or how to enhance their sustainability: a review

After more than 70 years of chemical pesticide use, modern agriculture is increasingly using biological control products. Resistances to conventional insecticides are wide spread, while those to bio-insecticides have raised less attention, and resistance management is frequently neglected. However, a good knowledge of the limitations of a new technique often provides greater sustainability. In this review, we compile cases of resistance to widely used bio-insecticides and describe the associated resistance mechanisms. This overview shows that all widely used bio-insecticides ultimately select resistant individuals. For example, at least 27 species of insects have been described as resistant to Bacillus thuringiensis toxins. The resistance mechanisms are at least as diverse as those that are involved in resistance to chemical insecticides, some of them being common to bio-insecticides and chemical insecticides. This analysis highlights the specific properties of bio-insecticides that the scientific community should use to provide a better sustainability of these products.

Causal agents of powdery mildew of cucurbits in Sudan.

In a survey of cucurbit crops in Sudan, 234 samples of melon, cucumber, and squash showing symptoms of powdery mildew were infected with Sphaerotheca fuliginea, while Erysiphe cichoracearum was only observed on two samples of watermelon showing mild symptoms. Leveillula taurica was not observed on any sample. Field trials with differential lines of melon revealed the presence of race 1 of S. fuliginea in Gezira (Central Sudan) in the summer of 1993, and race 2 in the following winter. Virulence tests in controlled conditions on a limited number of isolates suggest that a shift in prevalence of those races may have occurred on that site over time. The geographic scale of this shift was probably limited as race 0 was observed in late autumn in northeast Sudan in the Gash Delta

Is the efficacy of biological control against plant diseases likely to be more durable than that of chemical pesticides

The durability of a control method for plant protection is defined as the persistence of its efficacy in space and time. It depends on (i) the selection pressure exerted by it on populations of plant pathogens and (ii) on the capacity of these pathogens to adapt to the control method. Erosion of effectiveness of conventional plant protection methods has been widely studied in the past. For example, apparition of resistance to chemical pesticides in plant pathogens or pests has been extensively documented. The durability of biological control has often been assumed to be higher than that of chemical control. Results concerning pest management in agricultural systems have shown that this assumption may not always be justified. Resistance of various pests to one or several toxins of Bacillus thuringiensis and apparition of resistance of the codling moth Cydia pomonella to the C. pomonella granulovirus have, for example, been described. In contrast with the situation for pests, the durability of biological control of plant diseases has hardly been studied and no scientific reports proving the loss of efficiency of biological control agents against plant pathogens in practice has been published so far. Knowledge concerning the possible erosion of effectiveness of biological control is essential to ensure a durable efficacy of biological control agents on target plant pathogens. This knowledge will result in identifying risk factors that can foster the selection of strains of plant pathogens resistant to biological control agents. It will also result in identifying types of biological control agents with lower risk of efficacy loss, i.e., modes of action of biological control agents that does not favor the selection of resistant isolates in natural populations of plant pathogens. An analysis of the scientific literature was then conducted to assess the potential for plant pathogens to become resistant to biological control agents.