Foreword

This book reflects two decades of collaborative research on plant nematode interactions with a core of European teams that were brought together in the early nineties. When asked to write the foreword for this book, I wanted to document the origin of this group as it demonstrates that chance happenings can shape the future.

In the pioneering years of genetic engineering, I was working as a plant physiologist at the first plant biotech company in the Netherlands, Mogen International. In 1988 the Dutch potato processing industry approached us to solve their number one headache; potato cyst nematodes. It was a 4 year program with a team of 3-4 scientists to engineer resistance into their main cultivars. For a small biotech company that had no income except from investors, this was a big contract and we were keen to sign it in 1989.

To put this into historic perspective, these were the days where consumers had no clue yet about genetically modified food and we could routinely express a viral coat protein in tobacco and show systemic virus resistance... but anything else was far from routine or could even be classified as science fiction. We had just barely demonstrated transgenic potato plantlets and the first frail GM plants were cultivated in our high containment space age growth chambers, dazzling every visitor. We thought we were the new masters of the universe, carrying an unbelievable new toolbox that was growing every month with breathtaking inventions like PCR, reporter genes, genome sequencing, DNA synthesizers, etc. So, the fact that no one in the company had ever seen a live nematode before (my main background was in root physiology, but all the others were top-ranking molecular biologists) was considered a minor issue by management and by our contractors. They had faith in the scientists and the new toolbox. In retrospect, the naivety with which we entered this project was laughable, but without dreams there is no progress. It turned out to become the most fascinating period of my scientific career.

Parasitic nematodes are a problem in every country but are notoriously difficult to study. As a research subject they present many obstacles, delaying scientific progress to a pace that is no longer acceptable in the competitive world of grants and careers. On the other hand, the economic damage is significant everywhere and each country had at least one group of specialists to study their local threats and maintain a level of expertise, with relatively secure national funding. Therefore the European landscape was scattered with small but dedicated research groups, looking for ways to reduce chemical treatments, running breeding programs or just fascinated by the complex interactions between multicellular organisms. With one or two exceptions, molecular techniques were not widely used by these groups.

In the beginning of the project at Mogen, I visited many of these groups to get a better feeling for the state-of-the-art. Mogen in those days was just about 25 scientists, but being an "industry representative", I was met with scepticism and also with curiosity, as we were definitely a new kid-on-the-block. There was no budget for collaborations so I had very little to offer and my hosts kept their cards close to their chests. Seeing the work done by these groups, it gradually began to dawn on me that studying the life cycle of cyst nematodes on potato plants in vitro would already be a challenge, let alone interfering with its life cycle. A turning point in those visits was a trip to Kiel in Germany to meet Professor Urs Wyss. His enthusiasm was inspiring to say the least. After a crash course on nematode behaviour with his magnificent videos, I noticed that the Kiel group was able to grow cyst nematodes routinely on rapeseed in vitro, and from a root physiologist's point of view, these roots looked excellent, a far cry from the stunted potato roots we were growing at Mogen. The Kiel group offered me a few of their Petri dishes to bring back to Holland to use as a starter culture and to allow me get hands-on experience growing nematodes. No paperwork, no lawyers, no signatures, this was mutual trust only.

For other projects, I was growing Arabidopsis plants. At this time Arabidopsis was rapidly becoming the gold standard for plant molecular biologists, attracting the best and the brightest in plant science across the globe. One of the few areas where this model was not considered seriously was Phytopathology; Arabidopsis appeared to be resistant to most pathogens. Deviating way off from my project (I wouldn't dare tell my industrial partners that I was working on anything other than potato and I didn't dare tell my colleagues I was infecting Arabidopsis with a pathogen, which would have been considered a rather stupid venture in those days), I set up several experiments with Arabidopsis to see if I could get juveniles harvested from rapeseed cultures to infect the roots. I checked progress outside lab hours or during coffee breaks when I had the lab for myself. On several occasions I noticed behaviour similar to that which I had seen on the videos from Urs Wyss and realized that the worms recognized the presence of roots. Over the next few days I could even see movement within the translucent roots, indicating that the nematodes had managed to penetrate. I did not dare tell anyone yet. I remember vividly the first day I saw syncytia developing, the most prominent syncytia I had ever seen as they were developing in these really tiny roots. It was obvious that the nematodes were changing root growth in a way I had only seen with nitrogen fixing Rhizobia. But more astonishing, this was a pathogen infecting the model-plant Arabidopsis! Even before informing my colleagues, I phoned Urs to tell him what I had done. The message didn't really sink in and I took the next plane to Kiel, a bunch of Arabidopsis reviews in my bag, preparing a lecture on the model plant during the flight, it was my turn to inspire Urs and his group. The message did get through this time. We could jump on the fast train of Arabidopsis research. I left them with seeds and detailed protocols to repeat this in their lab.

A few more groups got involved and they all got the protocols to grow nematodes on Arabidopsis. Preparations were made to get European funding through a Concerted Action, bringing together 16 groups from all over Europe. All groups had basic funding already and we only applied for money to increase collaborations. Arabidopsis would be the common theme, a worthless weed so there were no issues about valuable crop species, exclusive fields and other potential roadblocks for such a large project. I could convince our industry partners that this was definitely a faster track to reach useful results that, at a later stage, we could transfer to potatoes. So fortunately, they stayed on board and I was allowed to continue. Brussels approved the program in 1992. For such a large group, it was a modest amount of money but just for travelling expenses, it was a staggering figure. With all expenses paid, any scientist from any of those groups could travel to any other group for the next 4 years and we organized large annual meetings where even the most junior members were able to attend. Obviously, collaborations flourished and gathered momentum with hundreds of exchange visits across the continent. The group had reached a critical mass that was unheard of in this field, resulting in excellent scientific publications in high ranking journals, patent applications, newspaper coverage, professorships, and last but not least, it attracted new scientists and students.

There were times where we thought that breakthroughs were close, as we were able to target gene expression directly in the syncytia and could beat the parasite using its own tricks; triggering plant promoters that were now coupled to toxic genes. But nature proved to be far more complex and within the time span of the Concerted Action, nobody came close to showing resistance even though the first field tests were done in 1995. The final annual meeting was staged in Toledo, Spain, and although we did not reach our ambitious milestones, it was clear that research on plant-nematode interactions had made a great leap forward. It was no longer a completely black box. The irony now is that to date, not one Arabidopsis ecotype could be identified with natural resistance against nematodes and this line of work still solely relies on crop species.

Even though I moved on to another job at that time, the momentum of this group remained and follow-up EU projects were prepared, submitted and granted throughout the following 15 years. People come and go and move on with their lives, but this book demonstrates that the backbone of our first Concerted Action is still prominently visible. No less then 21 of the 24 chapters include labs or scientists from the original group and the critical mass has been kept together for all these years. This is a vital ingredient for a niche in science that involves so many disciplines and focuses on such a complex biological interaction.

The book reviews progress that is impressive. Whole genome sequences of important plant-parasitic nematodes, application of new molecular tools for Arabidopsis, microanalysis of feeding cells, unravelling (suppression of) the host immune reaction, hormone regulation, cell cycle- and cell wall interference, cytoskeleton design, new breeding strategies and a series of field trials with GM-crops are all milestones within their specialized areas. Of course, Arabidopsis can not claim all the credit for this progress, but to have a non-commercial common interest was essential to start the initial collaboration and became the basis for the long term collaborations of which this book is the concrete proof.

It has been a privilege and a pleasure to work with this group of dedicated and enthusiastic scientists.

Peter C. Sijmons Szienz

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