Table 1.
Characteristics of CarC3H proteins.
Fig 1.
Comparison of CCCH ZnF containing TFs in Arabidopsis, Medicago and chickpea.
(A) Total number of CCCH proteins and CCCH motifs present in the three plants. (B) Distribution of CCCH motifs into various consensus arrangements in the three plants
Table 2.
Prediction of conserved motifs in CarC3H proteins.
Fig 2.
Phylogenetic classification of CarC3H members.
The CarC3H members were divided into 12 groups based on their phylogenetic positions. The structural similarity of their corresponding exon-intron organisation as well as distribution of functional domains in their amino acid sequences validates this clustering
Fig 3.
Phylogenetic comparison of CCCH Znf proteins in chickpea, Arabidopsis and Medicago.
Fig 4.
Synteny analysis of CarC3H genes with the genomes of M.truncatula, G.max, P. vulgaris and A. thaliana.
Fig 5.
Location of CarC3H genes on the chromosomes of kabuli chickpea and duplication events in members of CarC3H gene family.
Tandemly duplicated genes are highlighted with same colour background and segmental duplications are represented by the type of blocks against duplicated members
Table 3.
Details of duplicated CarC3H genes and their Ka/Ks vaalues.
Fig 6.
(A) Digital expression analysis of CarC3H genes in different tissues of chickpea. Scale represents the log2 normalised RPKM values. (B) qRT PCR based analysis of CarC3H genes having preferential expression in chickpea seed tissue. Values on the Y-axis indicate relative expression values as calculated by 2 –ΔΔCt method
Fig 7.
(A) Digital expression analysis of CarC3H genes in control and stressed tissues. Scale represents the log2 normalised RPKM values. (B) qRT-PCR based expression analysis of CarC3H genes in control and stressed chickpea seedlings (3 weeks old) at different time points. Scale represents the relative expression values as calculated by 2 –ΔΔCt method.
Fig 8.
(A) Alignment of CarC3H45 protein sequence with known tandem CCCH zinc finger proteins from Arabidopsis. Sequence within green box indicates the CCCH motif and red box indicates conserved arginine rich region. (B) Southern blotting analysis showed that the CarC3H45 was present as a single copy in chickpea.
Fig 9.
Quantitative expression of CarC3H45 in (A) different tissues of chickpea, L = leaf, R = root, F = flower, DAA = Days after anthesis, 24 Hr = 24 hours old seedling, 48 Hr = 48 hours old seedling and 72 Hr = 72 hours old seedling and (B) in germinating seeds treated with GA and ABA at various time points.
Fig 10.
(A) Yeast one hybrid assay for determination of CarABI3-CaC3H45Prom interaction. p53-pGADT7/p53-pAbAi was used as positive control and CarABI3-pGADT7/p53-pAbAi was used as negative control. (B) Transactivation assay in yeast shows no significant transactivation activity for CarC3H45.