Expression profiles of <i>Ca-2LIM</i> genes across different developmental tissues.

<p>The expression data of <i>Ca-2LIM</i> genes in root, stem, leaf and flower were achieved through qRT-PCR. The expression values (Y-axis) were presented relative to root samples and EF-1α was taken as reference gene. Standard deviation of three replicates was indicated by error bars.</p

Genome Wide Identification of <i>LIM</i> Genes in <i>Cicer arietinum</i> and Response of <i>Ca-2LIMs</i> in Development, Hormone and Pathogenic Stress

<div><p>The eukaryotic lineage-specific LIM protein (<u><i>L</i></u><i>IN11</i>, <u><i>I</i></u><i>SL1</i>, and <u><i>M</i></u><i>EC3</i>) family play pivotal role in modulation of actin dynamics and transcriptional regulation. The systematic investigation of this family has not been carried in detail and rare in legumes. Current study involves the mining of <i>Cicer arietinum</i> genome for the genes coding for LIM domain proteins and displayed significant homology with LIM genes of other species. The analysis led to the identification of 15 members, which were positioned on chickpea chromosomes. The phylogenetic and motif analysis suggested their categorization into two sub-families <i>i</i>.<i>e</i>., Ca-2LIMs and Ca-DA1/DAR, which comprised of nine and six candidates, respectively. Further sub-categories of <i>Ca-2LIMs</i> were recognised as αLIM, βLIM, δLIM and γLIM. The LIM genes within their sub-families displayed conserved genomic and motif organization. The expression pattern of <i>Ca-2LIMs</i> across developmental and reproductive tissues demonstrated strong correlation with established consensus. The <i>Ca-2LIM</i> belongs to <i>PLIM</i> and <i>GLIM (XLIM)</i> was found highly expressed in floral tissue. Others showed ubiquitous expression pattern with their dominance in stem. Under hormonal and pathogenic conditions these LIMs were found to up-regulate during salicylic acid, abscisic acid and <i>Ascochyta rabiei</i> treatment or infection; and down-regulated in response to jasmonic acid treatment. The findings of this work, particularly in terms of modulation of LIM genes under biotic stress will open up the way to further explore and establish the role of chickpea LIMs in plant defense response.</p></div

Phylogenetic tree demonstrating the evolutionary relation of the deduced full-length amino acid sequences of 15 CaLIMs with LIM proteins of <i>A</i>. <i>thaliana</i>, <i>B</i>. <i>rapa</i> and <i>O</i>. <i>sativa</i>.

<p>The unrooted phylogenetic tree was constructed using MEGA 6.0 by Maximum likelihood method with 1000 bootstraps. Bootstrap values are presented next to branch node. Green closed circles were used to show CaLIM proteins. Two major divisions were presented in different colours.</p

Principal attributes of LIM gene members and their deduced proteins in <i>Cicer arietinum</i>.

<p>*Predicted as Pseudo-gene in Bioproject: PRJNA175619</p><p>Principal attributes of LIM gene members and their deduced proteins in <i>Cicer arietinum</i>.</p

Phylogenetic analysis, gene structure and motif analysis of LIM family in <i>Cicer arietinum</i>.

<p>(A) Phylogenetic tree of CaLIMs was constructed using Maximum-likelihood method and the bootstrap test was performed for 1,000 repetitions. (B) The exon/intron organization of LIM genes of <i>C</i>. <i>arietinum</i>. Blue boxes represent exons and black lines indicate introns. (C) The conserved protein motifs in the LIM proteins identified using MEME program. Each motif is indicated with a specific color.</p

Phylogenetic tree demonstrating the evolutionary relation of the deduced full-length amino acid sequences of Ca-2LIMs with members of <i>A</i>. <i>thaliana</i>, <i>B</i>. <i>rapa</i>, <i>O</i>. <i>sativa</i> and <i>P</i>. <i>trichocarpa</i>.

<p>The unrooted phylogenetic tree was constructed using MEGA 6.0 by Maximum likelihood method with 1000 bootstraps. Bootstrap values are presented next to branch node. Different sub-groups are enclosed in separate coloured boxes.</p

Functional relevance of LIM domain containing proteins in plants and structure of LIM domain in CaLIMs.

<p>Functional relevance of LIM domain containing proteins in plants and structure of LIM domain in CaLIMs.</p

Heatmap representation of <i>Ca-2LIMs</i> expression in response to (A) SA, JA and ABA treatment (B) after spore inoculation of <i>Ascochyta rabiei</i>.

<p>Data were obtained through qRT-PCR for indicated time periods and presented relative to 0 h treatment. β-tubulin was taken as reference gene. Green and red color gradients indicate lower or higher transcript abundance, respectively.</p

Phylogenetic tree of 16S rRNA gene in cyanobacteria generated through BLASTN search in NCBI cyanobacterial database using the neighbor joining algorithm provided by MEGA 5.

<p>Phylogenetic tree of 16S rRNA gene in cyanobacteria generated through BLASTN search in NCBI cyanobacterial database using the neighbor joining algorithm provided by MEGA 5.</p

Screening for anticancer activity on cancer cell line.

<p>Calcein assay using a renal cell carcinoma (A498) cell line. Significant difference (p<0.001) between pairs, control with <i>Geitlerinema</i> CCC728 and <i>Arthrospira</i> CCC729 <b>(a),</b> Control, C2 and C3 with C4 and C5 <b>(b)</b> and Control and D1 with D3 and D4, D2 with D4 <b>(c)</b>. Control was lacking with crude extracts or TLC fractions.</p