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Fig. 3 | BMC Plant Biology

Fig. 3

From: Diversity in boron toxicity tolerance of Australian barley (Hordeum vulgare L.) genotypes

Fig. 3

Genetic variation in barley at the 6H B tolerance locus (HvNIP2;1). a Schematic of HvNIP2;1 coding sequences from Clipper and Sahara showing positions of two identified SNPs. No other sequence variation was found among 68 barley genotypes. A KASPTM marker assay was designed against the Sahara-specific 5’UTR SNP. b Expression levels of HvNIP2;1 determined by q-RT-PCR, in roots of seedlings of barley genotypes classified into four haplotypes on the basis of Southern and cDNA sequence analyses: Class 1, Clipper RFLP, ORF and uORF; 2, California Mariout RFLP, Clipper ORF and uORF; 3, Sahara RFLP and ORF/ Clipper uORF; 4, Sahara RFLP, ORF and uORF. Refer to Additional file 8: Table S5 for a list of barley genotypes included in each class. Transcript levels were normalised using the RNA housekeeping genes for tubulin, GAPDH and HSP70. c Relative translation efficiencies of a luciferase gene driven by an SP6 RNA polymerase promoter, when 5’UTR sequences from Clipper and Sahara HvNIP2;1 are inserted immediately upstream of luciferase. Translation efficiency was measured as luciferase activity by luminescence, and is expressed relative to the activity of SP6 control luciferase DNA. Six transcription/translation reactions were set up for each construct, and four luciferase assays recorded for each reaction. Bar represents the 95 % confidence interval for the difference between Clipper and Sahara 5’UTR sequences

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