Shoot sodium exclusion in salt stressed barley (Hordeum vulgare L.) is determined by allele specific increased expression of HKT1;5.

High affinity potassium transporters (HKT) are recognized as important genes for crop salt tolerance improvement. In this study, we investigated HvHKT1;5 as a candidate gene for a previously discovered quantitative trait locus that controls shoot Na+ and Na+/K+ ratio in salt-stressed barley lines on a hydroponic system. Two major haplotype groups could be distinguished for this gene in a barley collection of 95 genotypes based on the presence of three intronic insertions; a designated haplotype group A (HGA, same as reference sequence) and haplotype group B (HGB, with insertions). HGB was associated with a much stronger root expression of HKT1;5 compared to HGA, and consequently higher K+ and lower Na+ and Cl- concentrations and a lower Na+/K+ ratio in the shoots three weeks after exposure to 200 mM NaCl. Our experimental results suggest that allelic variation in the promoter region of the HGB gene is linked to the three insertions may be responsible for the observed increase in expression of HvHKT1;5 alleles after one week of salt stress induction. This study shows that in barley - similar to wheat and rice - HKT1;5 is an important contributor to natural variation in shoot Na+ exclusion.

Efficiency of transcriptional gene silencing of GBSSI in potato depends on the promoter region that is used in an inverted repeat.

Transcriptional gene silencing (TGS) of the endogenous GBSSI promoter in potato was induced by inverted repeat constructs containing different regions of the GBSSI promoter. Clear differences in silencing efficiency were observed. The 35SGBP-IR construct, containing sequences from -766 to -168 bp relative to the transcription initiation site (TIS), induced weak silencing effects in 57-60% of the transformants. Weak silencing effects were also induced by the ASP-IR construct harbouring allele-specific sequences covering the region from -531 to -330 bp relative to the TIS, but only in a low percentage (4-5.5%) of the transformants. These percentages are too low to distinguish effects between the two potato cultivars. Therefore, this approach cannot be used to induce allele-specific TGS. Strong silencing effects were obtained in 49% of the transformants harbouring the full promoter inverted repeat construct. This construct contained sequences from -766 to +194 bp relative to the TIS. In the strongly silenced transformants no GBSSI mRNA could be detected by Northern blot analysis. This was accompanied by the accumulation of GBSSI promoter-specific small interfering RNAs. Methylation studies revealed that, in the weakly silenced 35SGBP-IR transformants, the HpaII site at -213 bp relative to the TIS was methylated. Apparently, methylation of this sequence does not result in strong silencing effects. In the full promoter transformants, both CG methylation and CNN methylation were detected. We show that, to obtain strong TGS, it is important to include sequences in the vicinity of the TIS.