Identificatie en functionele studie van effectormoleculen in Meloidogyne graminicola
Nematoden zijn er in veel maten en gewichten. M. graminicola, die bestudeerd werd in deze masterthesis, ziet er onder de microscoop uit als een doorzichtig wormpje, maar is met het blote oog amper waar te nemen. Deze nematode is een parasiet: hij vestigt zich in planten en leeft van de voedingsstoffen die hij in zijn gastheer vindt. Is de gastheer een landbouwgewas, dan is dit synoniem voor minder opbrengst. Daarbij komt nog dat de nematode zijn gastheer verwondt. Een plant waarin nematoden huizen is een zieke plant. Het spreekt voor zich dat men nematoden op de velden liever kwijt dan rijk is. Ze worden dan ook fel bestreden met pesticiden. Het ontwikkelen van nieuwe en betere pesticiden is één methode om de nematoden in de akkerbouw de baas te kunnen. Een andere manier is het ontwikkelen van resistente planten. Welke methode men ook verkiest, voor men kan beginnen met het ontwikkelen van pesticiden of resistente planten moet men eerst weten hoe de nematode een plant binnendringt en hoe ze erin slaagt te overleven in de plant. Specifieker: men moet te weten komen welke eiwitten de nematode nodig heeft om de plant te infecteren en erin te overleven. Eens één of meerdere van deze cruciale eiwitten – ook effectors genoemd – bekend zijn, kan men hierop inspelen om de nematode te saboteren.
Het vinden van deze effectors is een proces van lange adem. De resultaten van veel verschillende soorten experimenten moeten in elkaar gepuzzeld worden voor men voldoende aanwijzingen verzameld heeft. Deze masterscriptie had als doel bij te dragen tot het vinden van effectoreiwitten in de nematode M. graminicola, die voornamelijk in de rijstteelt voor grote opbrengstverliezen zorgt.
In een nematode zitten tienduizenden verschillende eiwitten; men kan ze onmogelijk één voor één onderzoeken. Daarom werd in de eerste plaats een selectie gemaakt van eiwitten die over de passende eigenschappen beschikken. Bijvoorbeeld weet men dat veel effectors door de nematode in de plant worden gespoten; eigenschappen die passen bij dit gegeven dienden als criteria voor de selectie. M.b.v. software werden eiwitten uit de nematode geselecteerd die aan de meeste criteria voldeden.
Verschillende eiwitten uit de selectie werden vervolgens aan allerlei experimenten onderworpen. Van één eiwit kwam ik te weten dat ze wordt gemaakt in de klieren van de nematode. In die klieren worden bijna alle eiwitten gemaakt die de nematode in de plantencellen spuit. Dat is dus een sterke aanwijzing dat het eiwit in kwestie inderdaad een effector is. Voor een aantal andere eiwitten heb ik in kaart gebracht wanneer de nematode ze het meest aanmaakt. Eiwitten die worden aangemaakt op het moment van de infectie van de plant, zullen waarschijnlijk enkel op dat moment nodig zijn. Eiwitten die pas worden aangemaakt op het moment dat de nematode zich voortplant, zullen hoogstwaarschijnlijk daarbij een rol spelen. Zo werden een aantal vermoedens geformuleerd over de functie van enkele eiwitten uit de selectie. Daarnaast heb ik in een ander experiment ook nog vastgesteld dat nematoden die meer van een bepaald eiwit maakten dan gewoonlijk, daardoor beter de rijstplantjes konden infecteren. Statistisch kon dit helaas niet worden gestaafd.
Er zal nog veel tijd en onderzoek geïnvesteerd moeten worden om beetje bij beetje meer informatie bloot te leggen. Als alle verzamelde aanwijzingen later in elkaar gepast kunnen worden en de effectors bekend zijn, kan het echte werk beginnen: de nematoden verjagen uit de rijstvelden!
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