Figure 1.
An unrooted phylogenetic tree of the barley Csl super family.
Phylogenetic analysis was done using MrBayes (codon position model) in TOPALi v2. The posterior probabilities have been multiplied by 100. The scale bar shows expected number of nucleotide substitutions per site.
Figure 2.
The genetic location of barley HvCslF genes.
Genetic map of barley chromosomes 1H, 2H, 5H, and 7H showing the positions of barley HvCslF genes as mapped in a ‘Morex’ × ‘Barke’ population [12]. cM = centimorgan.
Figure 3.
The protein structure of the HvCslF family members.
The shaded boxes indicate the positions of sequences encoding trans-membrane helices, which can be found in similar positions in all genes. The triangle marks the intron position with the size given in base pairs. The lines indicate the glycosyltransferase GT2-motif D,D,D,QxxRW.
Figure 4.
Part of the glycosyltransferase GT2-motif with surrounding amino acid residues.
The C-terminal part of the glycosyltransferase GT2-motif, encompassing ED and QxxRW, are marked by the red border. The colour coding is as follows: Aliphatic/hydrophobic: ILVAM (blue), Aromatic: FWY (orange), Positive: KRH (purple), Negative: DE (red), Hydrophilic: STNQ (green), conformationally special: PG (pink), Cysteine: C (yellow).
Figure 5.
Expression data of the HvCslF genes based on RNA-sequence data.
The RNA-sequence data for the HvCslF genes includes three biological replicates per tissue. The results are given in FPKM expression values (fragments per kilobase of exon per million fragments mapped). Values were obtained from the International Barley Genome Sequencing Consortium [12].
Figure 6.
Transient expression of different HvCslF-constructs in Nicotiana benthamiana leaves results in necrosis.
HvCslF6 (a), HvCslF11 (b), HvCslF12 (c) and empty vector (d) constructs were transiently expressed in 4-weeks old N. benthamiana leaves using Agrobacterium tumefaciens as a vector. Photographs were taken six days after infiltration. Necrosis symptoms were observed for all three constructs.
Figure 7.
Phylogenetic tree of the CslF gene family including five cereals.
Phylogenetic tree of 69 members of the CslF gene family including wheat, barley, Brachypodium, rice and sorghum. The tree was constructed with TOPALi v2 on a subset of the genes using MrBayes (codon position model). The posterior probabilities have been multiplied by 100. An outgroup (not shown) comprised 10 CslH/CslJ sequences. HvCslF13 is marked as a potential pseudogene ψ. The two potential mosaic sequences Sb02g022011 and Sb02g036030 are marked by #. The scale bar shows expected number of nucleotide substitutions per site.