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. 2017 Jan;24(1):80-89.
doi: 10.1016/j.sjbs.2016.05.011. Epub 2016 May 25.

Faba bean drought responsive gene identification and validation

Megahed H Ammar  1 , Altaf M Khan  2 , Hussein M Migdadi  2 , Samah M Abdelkhalek  3 , Salem S Alghamdi  2
Affiliations

Faba bean drought responsive gene identification and validation

Megahed H Ammar et al. Saudi J Biol Sci. 2017 Jan.

Abstract

This study was carried out to identify drought-responsive genes in a drought tolerant faba bean variety (Hassawi 2) using a suppressive subtraction hybridization approach (SSH). A total of 913 differentially expressed clones were sequenced from a differential cDNA library that resulted in a total of 225 differentially expressed ESTs. The genes of mitochondrial and chloroplast origin were removed, and the remaining 137 EST sequences were submitted to the gene bank EST database (LIBEST_028448). A sequence analysis identified 35 potentially drought stress-related ESTs that regulate ion channels, kinases, and energy production and utilization and transcription factors. Quantitative PCR on Hassawi 2 genotype confirmed that more than 65% of selected drought-responsive genes were drought-related. Among these induced genes, the expression levels of eight highly up-regulated unigenes were further analyzed across 38 selected faba bean genotypes that differ in their drought tolerance levels. These unigenes included ribulose 1,5-bisphosphate carboxylase (rbcL) gene, non-LTR retroelement reverse related, probable cyclic nucleotide-gated ion channel, polyubiquitin, potassium channel, calcium-dependent protein kinase and putative respiratory burst oxidase-like protein C and a novel unigene. The expression patterns of these unigenes were variable across 38 genotypes however, it was found to be very high in tolerant genotype. The up-regulation of these unigenes in majority of tolerant genotypes suggests their possible role in drought tolerance. The identification of possible drought responsive candidate genes in Vicia faba reported here is an important step toward the development of drought-tolerant genotypes that can cope with arid environments.

Keywords: Drought; Drought-responsive genes; Hassawi 2; Suppressive subtraction hybridization; Vicia faba.

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Figures

Figure 1
Figure 1
Blastx statistics of 137 Vicia faba sequences analyzed by Blast2go.
Figure 2
Figure 2
Functional classifications of the 137 unigenes that were assigned to GO terms (second level GO terms). The three GO categories, biological process (BP), molecular function (MF), and cellular component (CC) are presented.
Figure 3
Figure 3
Expression pattern of 35 selected unigenes against Hassawi 2 genotype of faba bean grown under stress conditions. The upper bars above 1 show the level of up-regulation in genotypes, while lower bars below 1 show the level of down-regulation. The bar at 1 indicates no differential expression.
See this image and copyright information in PMC

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