GBSS T-DNA inserts giving partial complementation of the amylose-free potato mutant can also cause co-suppression of the endogenous GBSS gene in a wild-type background.
The wild-type gene encoding granule-bound starch synthase (GBSS) is capable of both complementing the amylose-free (amf) potato mutant and inhibiting the endogenous GBSS gene expression in wild-type potato. Co-suppression of the endogenous GBSS gene, easily visualised by staining the starch with iodine, occurred when the full-size GBSS sequence (genomic), GBSS cDNA or even the mutant amf allele were introduced into the wild-type potato. Conversely, introduction of the GBSS promoter sequence alone, did not result in co-suppression in the 80 analysed transformants. Neither the orientation of the GBSS gene with respect to kanamycin resistance nor the presence of an enhancer influenced the frequency of plants showing a co-suppression phenotype. After crossing a partially complemented amf mutant with a homozygous wild-type plant, the F1 offspring segregated into plant phenotypes with normal and decreased expression of the GBSS gene. This decreased expression correlated with the presence of a linked block of five T-DNA inserts which was previously shown to be correlated with partial complementation of the amf mutant. This crossing experiment indicates that co-suppression can cause inhibition of gene expression of both inserted and endogenous wild-type GBSS genes. The frequency of partially complemented amf plants was equal to the frequency of co-suppressed wild types when a construct, with an enhancer in front of the GBSS promoter, was used (pWAM 101E). This might suggest that partial complementation of the amf genotype caused by unstable expression of the transgene can be overcome by inserting an enhancer in front of the GBSS promoter.
Flipse
E
E
Graduate School of Experimental Plant Sciences, Department of Plant breeding, Agricultural University, Wageningen, Netherlands.
Straatman-Engelen
I
I
Kuipers
A G
AG
Jacobsen
E
E
Visser
R G
RG
D000688:Amylose / Q000032:analysis; D004269:DNA, Bacterial / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics; D005816:Genetic Complementation Test; D018517:Plant Roots / Q000737:chemistry; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D011198:Solanum tuberosum / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D013213:Starch / Q000737:chemistry; D013214:Starch Synthase / Q000235:genetics* / Q000378:metabolism; D013489:Suppression, Genetic
D004269:DNA, Bacterial; D012333:RNA, Messenger; D018749:RNA, Plant; C036483:T-DNA; D013213:Starch; D000688:Amylose; D013214:Starch Synthase