Constitutive expression of human lactoferrin and its N-lobe in rice plants to confer disease resistance

Authors: Takase, Kenji; Hagiwara, Kiyoshi; Onodera, Haruko; Nishizawa, Yaeko; Ugaki, Masashi; Omura, Toshihiro; Numata, Shinichi; Akutsu, Katsuki; Kumura, Haruto; Shimazaki, Kei-ichi

Source: Biochemistry and Cell Biology, Volume 83, Number 2, April 2005 , pp. 239-249(11)

Publisher: NRC Research Press

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Abstract:

The milk protein, lactoferrin, is known to have antibacterial, antiviral, and antifungal activities. To explore the possibility of conferring disease resistance in plants by expressing this protein, the gene for the full-length human lactoferrin (HLF), as well as the N-lobe, the N-terminal half molecule (HLFN), was introduced into rice plants and expressed constitutively under the control of the cauliflower mosaic virus 35S promotor. Western blot analysis of leaves from HLF-transgenic rice plants showed an 80 kDa-band, which was about 1–2 kDa less than human milk lactoferrin. HLFN was expressed as a 45-kDa protein and retained its heparin-binding property. Deglycosylation experiments suggested that both proteins produced by the plants had plant-type oligosaccharide chains. The transgenic rice plants were assessed for resistance against disease-causing bacteria, virus, and fungi. Of the pathogens tested, significant resistance against Burkholderia (Pseudomonas) plantarii, the causative agent of bacterial seedling blight disease, was observed in the transgenic plants expressing HLF or HLFN.Key words: expression of domains; glycosylation; lactoferrin; plant disease resistance; transgenic rice.

La lactoferrine, une protéine du lait, est reconnue pour ses activités antibactériennes, antivirales et antifongiques. Afin d'explorer la possibilité de conférer à des plantes une résistance aux maladies par l'expression de cette protéine, le gène codant la lactoferrine humaine pleine longueur (HLF), ainsi que le lobe N, c'est-à-dire la moitié N-terminale de la molécule (HLFN), ont été introduits dans des plants de riz et exprimés de façon constitutive sous le contrôle du promoteur du virus de la mosaïque du tabac 35S. Des analyses en immunobuvardage de feuilles provenant de plants de riz transgéniques pour la HLF ont révélé la présence d'une bande de 80 kDa, soit environ 1–2 kDa de moins que le poids de la lactoferrine humaine du lait. La HLFN était exprimée sous forme d'une protéine de 45 kDa ayant retenu ses propriétés de liaison à l'héparine. Des expériences de déglycosylation ont suggéré que les deux protéines produites par les plantes possédent des chaînes d'oligosaccharides typiques à celles des plantes. Les plants de riz transgéniques ont été évalués relativement à leur résistance vis-à-vis des bactéries, virus et champignons pathogènes. Parmi les pathogènes testés, une résistance significative a été obtenue vis-à-vis Burkholderia (Pseudomonas) plantarii, l'agent responsable de la rouille des semis, chez les plants transgéniques exprimant la HLF ou la HLFN.Mots clés : expression de domaines, glycosylation, lactoferrine, résistance aux maladies chez les plantes, riz transgénique.[Traduit par la Rédaction]

Document Type: Research article

Publication date: 2005-04-01

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Constitutive expression of human lactoferrin and its N-lobe in rice plants to confer disease resistance

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