Bacterio-opsin gene overe×pression fails to elevate fungal disease resistance in transgenic poplar (Populus)

Abstract
Overexpression of the bacterio-opsin (b0) gene in tobacco had previously been shown to induce hypersensitiveresponse-like lesions, increase viral and bacterial disease resistance, and stimulate pathogenesis-related gene expression. To see if this gene enhanced resistance to fungal pathogens of poplar, we generated a total of 35 transgenic lines in two clones of Populus trichocarpa Torr. & A. Gray x Populus deltoides Bartr. ex Marsh. and one clone of P. trichocarpa x Populus nigra L. and challenged them with the fungal pathogens Melampsora occidentalis H. Jack (leaf rust), Uenturia populina (Vuill.) Fabric. (leaf and shoot blight), Septoria musiva Peck, and Septoria populicola Peck (leaf spot and stem canker) in greenhouse, field, or laboratory inoculations. Northern analysis showed that the b0 gene was expressed in the transgenic poplars; however, no increase in expression of phenylalanine ammonia-lyase (PALL) or two wound-inducible poplar chitinase genes (WIN6 and WIN8) were observed, even in one line that showed very high b0 expression, intensive lesion development, and retarded growth. Poplars required a high threshold of b0 expression for lesion development, and susceptibility to all of the pathogens tested was unaffected by b0 overexpression. Resume : Des etudes anterieures ont dfimontre que la surexpression du g6ne de la bactdrio-opsine (b0) chez le tabac induisait des lesions semblables a celles associ6es a la rdponse hypersensible, augmentait la resistance aux maladies virales et bactdriennes et stimulait 1'expression de genes lids a la pathogenese. Afin de verifier si ce gene augmente la resistance a 1'endroit d'agents pathogenes du peuplier, nous avons produit un total de 35 lignees transgeniques chez deux clones de Populus trichocarpa Torr. & A. Gray x Populus deltoides Bartr. ex Marsh. et chez un clone de P. tricho carpa x Populus nigra L. que nous avons inoculees avec les champignons pathogenes Melampsora occidentalis H. Jack (rouille foliaire), Ueuturia populiua (Vuill.) Fabric. (brnlure des feuilles et des pousses) ainsi que Septoria musiva Peck et Septoria populicola Peck (tache foliaire et chancre caulicole) en serre, au champ et au laboratoire. L'analyse Northern a confirme 1'expression du g6ne b0 chez les peupliers transgeniques; on n'a cependant pas observd d'augmentation de 1'expression de la phenylalanine ammonia-lyase (PALI) ni de deux genes de chitinase inductibles par blessure (WIN6 et WIN8) et ce, meme chez la lignee chez laquelle on observe une expression de b0 trbs elev6e, le developpement intense de lesions et une croissance ralentie. Les peupliers requierent un seuil d'expression de b0 eleve pour que les lesions se ddveloppent, tandis que la surexpression de b0 n'a changfi la susceptibilite a aucun des agents pathogenes etudids.