Journal Article
ARM1: application for engineered resistance to plant-parasitic nematodes [1999]
Karimi, M.; Porceddu, A.; Mironov, V.; Montagu, M. van; et al.
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To some extent plant-parasitic nematodes can be controlled by using conventional methods. However, these methods are either not efficient or cause environmental hazard. An alternative and promising way is making nematode-resistant plants by introducing engineered genes that are expressed specifically in the nematode feeding site. To identify plant promoters that are induced upon infected plant-parasitic nematodes a promoter trapping method has been used. An Agrobacterium tumefaciens strain harboring a beta-glucuronidase (gus) fusion vector has been used to transform Arabidopsis thaliana. We have regenerated a thousand transgenic Arabidopsis plants and part of them has been screened for gus expression after root-knot nematode infection. Several tagged nematode-inducible lines were identified and from them the ARM1 line was characterized in more detail. Microscopic analysis showed GUS activity inside the giant cells or syncytia induced by root knot and cyst nematodes; respectively. The ARM1 promoter was isolated by inverse PCR and after fusion to the gus-coding region, this chimeric gene was introduced into the A. thaliana genome. The same gus expression pattern as in the original ARM1 was observed in the transformed A. thaliana lines. We have also used the ARM1 promoter to test a resistance strategy in which the aim is to inhibit the formation or development of the feeding cells. More specifically, a strategy was designed to block cell cycle progression in
the nematode feeding site. Four dominant negative mutations of cdc2aAt referred to as DN, DL41, DL36 and DL50 were fused to the ARM1 promoter and constructs were introduced into A. thaliana.
