Transgenic sterility in Populus: expression properties of the poplar PTLF, Agrobacterium NOS and two minimal 35S promoters in vegetative tissues.
Transgenic sterility is a desirable trait for containment of many kinds of transgenes and exotic species. Genetically engineered floral sterility can be imparted by expression of a cytotoxin under the control of a predominantly floral-tissue-specific promoter. However, many otherwise desirable floral promoters impart substantial non-floral expression, which can impair plant health or make it impossible to regenerate transgenic plants. We are therefore developing a floral sterility system that is capable of attenuating undesired background vegetative expression. As a first step towards this goal, we compared the vegetative expression properties of the promoter of the poplar (Populus trichocarpa Torr. & Gray) homolog of the floral homeotic gene LEAFY (PTLF), which could be used to impart male and female flower sterility, to that of three candidate attenuator-gene promoters: the cauliflower mosaic virus (CaMV) 35S basal promoter, the CaMV 35S basal promoter fused to the TMV omega element and the nopaline synthase (NOS) promoter. The promoters were evaluated via promoter::GUS gene fusions in a transgenic poplar hybrid (Populus tremula L. x P. alba L.) by both histochemical and fluorometric GUS assays. In leaves, the NOS promoter conveyed the highest activity and had a mean expression level 5-fold higher than PTLF, whereas the CaMV 35S basal promoter fused to the omega element and the CaMV 35S basal promoter alone directed mean expression levels that were 0.5x and 0.35x that of PTLF, respectively. Differential expression in shoots, leaves, stems and roots was observed only for the NOS and PTLF promoters. Strongest expression was observed in roots for the NOS promoter, whereas the PTLF promoter directed highest expression in shoots. The NOS promoter appears best suited to counteract vegetative expression of a cytotoxin driven by the PTLF promoter where 1:1 toxin:attenuator expression is required.
Wei
Hao
H
Department of Forest Science, Oregon State University, 321 Richardson Hall, Corvallis, OR 97331-5752, USA.
Meilan
Richard
R
Brunner
Amy M
AM
Skinner
Jeffrey S
JS
Ma
Caiping
C
Strauss
Steven H
SH
D000594:Amino Acid Oxidoreductases / Q000235:genetics*; D001483:Base Sequence; D017796:Caulimovirus / Q000235:genetics; D005298:Fertility / Q000235:genetics; D035264:Flowers / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics; D030821:Plants, Genetically Modified; D032107:Populus / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012231:Rhizobium / Q000235:genetics; D016415:Sequence Alignment
D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase; D005966:Glucuronidase