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Investigation of genes encoding calcineurin B-like protein family in legumes and their expression analyses in chickpea (Cicer arietinum L.).

Meena MK, Ghawana S, Sardar A, Dwivedi V, Khandal H, Roy R, Chattopadhyay D - PLoS ONE (2015)

Bottom Line: Most of the CaCBL genes exhibited high expression in flowers.Expression profile of CaCBL genes in response to different abiotic stresses and hormones related to development and stresses (ABA, auxin, cytokinin, SA and JA) at different time intervals suggests their diverse roles in development and plant defence in addition to abiotic stress tolerance.These data not only contribute to a better understanding of the complex regulation of chickpea CBL gene family, but also provide valuable information for further research in chickpea functional genomics.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, 110067, India.

ABSTRACT
Calcium ion (Ca2+) is a ubiquitous second messenger that transmits various internal and external signals including stresses and, therefore, is important for plants' response process. Calcineurin B-like proteins (CBLs) are one of the plant calcium sensors, which sense and convey the changes in cytosolic Ca2+-concentration for response process. A search in four leguminous plant (soybean, Medicago truncatula, common bean and chickpea) genomes identified 9 to 15 genes in each species that encode CBL proteins. Sequence analyses of CBL peptides and coding sequences (CDS) suggested that there are nine original CBL genes in these legumes and some of them were multiplied during whole genome or local gene duplication. Coding sequences of chickpea CBL genes (CaCBL) were cloned from their cDNAs and sequenced, and their annotations in the genome assemblies were corrected accordingly. Analyses of protein sequences and gene structures of CBL family in plant kingdom indicated its diverse origin but showed a remarkable conservation in overall protein structure with appearance of complex gene structure in the course of evolution. Expression of CaCBL genes in different tissues and in response to different stress and hormone treatment were studied. Most of the CaCBL genes exhibited high expression in flowers. Expression profile of CaCBL genes in response to different abiotic stresses and hormones related to development and stresses (ABA, auxin, cytokinin, SA and JA) at different time intervals suggests their diverse roles in development and plant defence in addition to abiotic stress tolerance. These data not only contribute to a better understanding of the complex regulation of chickpea CBL gene family, but also provide valuable information for further research in chickpea functional genomics.

No MeSH data available.


Tissue specific gene expression analysis of chickpea CBL genes.For tissue specific expression studies, total RNA was isolated from different tissues of chickpea plants. cDNA were prepared and expression pattern of all chickpea CBL genes were analysed by qRT-PCR and presented as heatmap. Chickpea elongation factor 1-α (EF-1α) mRNA was used as internal control for normalization. The scale bar represents relative expression values. Hierarchical clustering has been shown at left. Relative fold expression values are presented as bar diagram in S4 Fig.
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pone.0123640.g004: Tissue specific gene expression analysis of chickpea CBL genes.For tissue specific expression studies, total RNA was isolated from different tissues of chickpea plants. cDNA were prepared and expression pattern of all chickpea CBL genes were analysed by qRT-PCR and presented as heatmap. Chickpea elongation factor 1-α (EF-1α) mRNA was used as internal control for normalization. The scale bar represents relative expression values. Hierarchical clustering has been shown at left. Relative fold expression values are presented as bar diagram in S4 Fig.

Mentions: The expression pattern of the genes can provide important information for understanding their function. Expression pattern of CaCBL genes in different tissues were studied by qRT-PCR. The mRNA accumulation of each gene in root, stem, leaf and flower tissues was assessed. Except CaCBL8, other CaCBL genes are expressed at a higher level in flower in comparison to the other tissues, indicating important role of CBL-CIPK pathway in reproductive development (Fig 4). Specifically, expression ratio of CaCBL5 in flower to other tissues is exceptionally high. AtCBL1 and AtCBL9 were reported to function in pollen germination and pollen tube growth [49]. CaCBL4 is expressed more in root and flower. CaCBL8 was mostly expressed in roots. Expressions of CaCBL1, -2, -3, -5 and -6 in root were lowest among all the tissues. Although, gene and peptide structures of CBL family in higher plants are highly conserved, their expression patterns vary from plant to plant. Arabidopsis and B. napus are phylogenetically very close. Inspite of that their CBL genes showed entirely different patterns of expression. Likewise, tissue-specific expression pattern of CaCBL genes do not follow the pattern of those from Arabidopsis. However, higher expression of AtCBL1, -2 and -3 in flower corresponded with similar expression of their orthologs in chickpea [18, 50].


Investigation of genes encoding calcineurin B-like protein family in legumes and their expression analyses in chickpea (Cicer arietinum L.).

Meena MK, Ghawana S, Sardar A, Dwivedi V, Khandal H, Roy R, Chattopadhyay D - PLoS ONE (2015)

Tissue specific gene expression analysis of chickpea CBL genes.For tissue specific expression studies, total RNA was isolated from different tissues of chickpea plants. cDNA were prepared and expression pattern of all chickpea CBL genes were analysed by qRT-PCR and presented as heatmap. Chickpea elongation factor 1-α (EF-1α) mRNA was used as internal control for normalization. The scale bar represents relative expression values. Hierarchical clustering has been shown at left. Relative fold expression values are presented as bar diagram in S4 Fig.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390317&req=5

pone.0123640.g004: Tissue specific gene expression analysis of chickpea CBL genes.For tissue specific expression studies, total RNA was isolated from different tissues of chickpea plants. cDNA were prepared and expression pattern of all chickpea CBL genes were analysed by qRT-PCR and presented as heatmap. Chickpea elongation factor 1-α (EF-1α) mRNA was used as internal control for normalization. The scale bar represents relative expression values. Hierarchical clustering has been shown at left. Relative fold expression values are presented as bar diagram in S4 Fig.
Mentions: The expression pattern of the genes can provide important information for understanding their function. Expression pattern of CaCBL genes in different tissues were studied by qRT-PCR. The mRNA accumulation of each gene in root, stem, leaf and flower tissues was assessed. Except CaCBL8, other CaCBL genes are expressed at a higher level in flower in comparison to the other tissues, indicating important role of CBL-CIPK pathway in reproductive development (Fig 4). Specifically, expression ratio of CaCBL5 in flower to other tissues is exceptionally high. AtCBL1 and AtCBL9 were reported to function in pollen germination and pollen tube growth [49]. CaCBL4 is expressed more in root and flower. CaCBL8 was mostly expressed in roots. Expressions of CaCBL1, -2, -3, -5 and -6 in root were lowest among all the tissues. Although, gene and peptide structures of CBL family in higher plants are highly conserved, their expression patterns vary from plant to plant. Arabidopsis and B. napus are phylogenetically very close. Inspite of that their CBL genes showed entirely different patterns of expression. Likewise, tissue-specific expression pattern of CaCBL genes do not follow the pattern of those from Arabidopsis. However, higher expression of AtCBL1, -2 and -3 in flower corresponded with similar expression of their orthologs in chickpea [18, 50].

Bottom Line: Most of the CaCBL genes exhibited high expression in flowers.Expression profile of CaCBL genes in response to different abiotic stresses and hormones related to development and stresses (ABA, auxin, cytokinin, SA and JA) at different time intervals suggests their diverse roles in development and plant defence in addition to abiotic stress tolerance.These data not only contribute to a better understanding of the complex regulation of chickpea CBL gene family, but also provide valuable information for further research in chickpea functional genomics.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, 110067, India.

ABSTRACT
Calcium ion (Ca2+) is a ubiquitous second messenger that transmits various internal and external signals including stresses and, therefore, is important for plants' response process. Calcineurin B-like proteins (CBLs) are one of the plant calcium sensors, which sense and convey the changes in cytosolic Ca2+-concentration for response process. A search in four leguminous plant (soybean, Medicago truncatula, common bean and chickpea) genomes identified 9 to 15 genes in each species that encode CBL proteins. Sequence analyses of CBL peptides and coding sequences (CDS) suggested that there are nine original CBL genes in these legumes and some of them were multiplied during whole genome or local gene duplication. Coding sequences of chickpea CBL genes (CaCBL) were cloned from their cDNAs and sequenced, and their annotations in the genome assemblies were corrected accordingly. Analyses of protein sequences and gene structures of CBL family in plant kingdom indicated its diverse origin but showed a remarkable conservation in overall protein structure with appearance of complex gene structure in the course of evolution. Expression of CaCBL genes in different tissues and in response to different stress and hormone treatment were studied. Most of the CaCBL genes exhibited high expression in flowers. Expression profile of CaCBL genes in response to different abiotic stresses and hormones related to development and stresses (ABA, auxin, cytokinin, SA and JA) at different time intervals suggests their diverse roles in development and plant defence in addition to abiotic stress tolerance. These data not only contribute to a better understanding of the complex regulation of chickpea CBL gene family, but also provide valuable information for further research in chickpea functional genomics.

No MeSH data available.