The BIOEXPLOIT Project

The EU Framework 6 Integrated Project BIOEXPLOIT concerns the exploitation of natural plant biodiversity for the pesticide-free production of food. It focuses on the pathogens Phytophthora infestans, Septoria tritici, Blumeria graminis, Puccinia spp. and Fusarium spp. and on the crops wheat, barley, tomato and potato. The project commenced in October 2005, comprises 45 laboratories in 12 countries, and is carried out by partners from research institutes, universities, private companies and small-medium enterprises. The project has four strategic objectives covered in eight sub-projects. These objectives relate to (i) understanding the molecular components involved in durable disease resistance, (ii) exploring and exploiting the natural biodiversity in disease resistance, (iii) accelerating the introduction of marker-assisted breeding and genetic engineering in the EU plant breeding industry, and (iv) coordinating and integrating resistance breeding research, providing training in new technologies, disseminating the results, and transferring knowledge and technologies to the industry.     

Potato root diffusate-induced secretion of soluble, basic proteins originating from the subventral esophageal glands of potato cyst nematodes

ABSTRACT In preparasitic second-stage juveniles (J(2)) of potato cyst nematode Globodera rostochiensis, six proteins with molecular masses of 30, 31a/b, 32, 39, and 49 kDa were recognized on Western blots by a monoclonal antibody (MGR48) specific for the subventral esophageal glands. All of these subventral gland proteins (svp's) focused in the basic range (pI 6.8 to 8.6) of an immobilized pH gradient. Western blotting showed that the svp's were present in preparasitic and parasitic J(2) and not in later juvenile stages and adult females. Minor svp quantities also were observed in adult males. Immunogold labeling of preparasitic J(2) showed that the svp's were localized in the rough endoplasmic reticulum and secretory granules of the subventral esophageal glands. Potato root diffusate triggered the secretion of svp's through the stylet, and 5-methoxy-N,N-dimethyltryptamine-hydrogen-oxalate had only a quantitative, additional effect. The forward flow of svp's through the metacorporal pump chamber was confirmed by the presence of svp's in the circular lumen of the esophagus (procorpus), as established by immunoelectron microscopy. Our data provide conclusive evidence that secretory proteins of the subventral glands of G. rostochiensis can be secreted through the stylet and support the hypothesis that the subventral esophageal glands play an important role in the early events of this nematode-plant interaction.     

Nuclear and Mitochondrial DNA Polymorphisms in Three Mitotic Parthenogenetic Meloidogyne Spp.

In order to expand our understanding of the genetics of root-knot nematodes, variation in nuclear DNA and mitochondrial DNA in Meloidogyne incognita, M. arenaria and M. javanica was investigated. Despite the obligate mitotic parthenogenetic mode of reproduction, a large number of AFLP polymorphisms were observed among all 16 populations studied. Both UPGMA and principle coordinate analyses revealed three distinct groups that corresponded with the respective species identities of the 16 populations. M. incognita was genetically most distinct. Amplification of 63-bp tandem repeats (TR) in mtDNA from single individuals enabled the calculation of diversity measures at three hierarchical levels: within individuals, among individuals of a single population and among populations. For all three species, the highest diversity was observed within individuals explaining 43–65% of the total diversity. Many individuals contained more than one mtDNA size variant. M. incognita harboured the most heteroplasmic individuals and was the most homogenous at the population level. Only 13% of the total diversity was observed among populations, while this figure was 35% for M. arenaria. Both TR and AFLP data showed that M. arenaria is the most heterogeneous species. The comparison of the genetic distances based on AFLPs and mtDNA size variants revealed a significant correlation for the six M. arenaria populations, whereas no consistent correlation was observed for the populations of the other two species.     

Rhizosphere bacteria from sites with higher fungal densities exhibit greater levels of potential antifungal properties

A field study was performed to examine whether an increased density of saprotrophic fungi in the rhizosphere selects for bacteria with traits advantageous to living in a fungal-rich environment. Fast-growing bacteria were isolated from the rhizosphere of Carex arenaria (sand sedge) plants growing in fungal-poor (sand drifts) and fungal-rich (forest) sites in the Netherlands and screened for several potential antifungal properties, namely in vitro antagonism and the production of siderophores, cyanide and lytic enzymes. A higher incidence of putatively antifungal traits was generally found in bacteria isolated from fungal-rich soils, thus supporting the hypothesis that high fungal densities can impose a selection pressure on rhizosphere bacteria.     

Rhizosphere bacterial community composition in natural stands of Carex arenaria (sand sedge) is determined by bulk soil community composition

The relative importance of specific plant properties versus soil characteristics in shaping the bacterial community structure of the rhizosphere is a topic of considerable debate. Here, we report the results of a study on the bacterial composition of the rhizosphere of the wild plant Carex arenaria (sand sedge) growing at 10 natural sites in The Netherlands. The soil properties of the sandy soils at these sites were highly disparate, most notably in pH, chloride and organic matter content. Rhizosphere and bulk soil bacterial communities were examined by culture-independent means, namely, 16S rDNA-directed PCR-DGGE profiling. Large differences were observed between the bacterial communities of the different sites for both bulk and rhizosphere soil. Cluster analysis of bacterial profiles revealed that the rhizosphere community of each site was generally more closely related to the bulk soil community of that site rather than to rhizosphere communities of other sites. Hence, bacterial community structure within the rhizosphere of C. arenaria appeared to be determined to a large extent by the bulk soil community composition. This conclusion was supported by a reciprocal planting experiment, where C. arenaria shoots of different sites yielded highly similar rhizosphere communities when planted in the same soil.