Maria Beihaghi; Ahmad Reza Bahrami; Abdolreza Bagheri; Mohammad Zare Mehrjerdi
Abstract
Studies on the genes contributing to the seed filling in chickpea and its protein content might be valuable in engineering plants with seeds of a higher nutritional value. A gene of interest is phosphoenolpyruvate carboxykinase (pepck), encoding a protein with a substantial role in the gluconeogenesis ...
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Studies on the genes contributing to the seed filling in chickpea and its protein content might be valuable in engineering plants with seeds of a higher nutritional value. A gene of interest is phosphoenolpyruvate carboxykinase (pepck), encoding a protein with a substantial role in the gluconeogenesis pathway. In the present study, the protein content (percentage) was measured in a number of cultivated chickpea genotypes, followed by comparison of the expression levels of pepck gene at different stages of seed filling in some of the genotypes. This study aims at revealing the relation between pepck transcript level with protein content of chickpea seeds which might, in the longer term, end at protein quality improvement of the crop through the gene manipulation procedures. The results which were verified by Real Time-PCR and Western blot techniques in four genotypes of plant, showed that, the amount of pepck expression was significantly higher at the stage of seed fillingthan at other stages in all of genotypes and the lowest levels of expression belonged to flowering and seed formation and the PEPCK protein level was higher in the high protein genotypes compared to the low protein genotypes.
Ahmad Reza Bahrami; Maria Beihaghi; Abdolreza Bagheri; Richard Leegood; Mehdi Ghabooli; Jafar Zolala; Farajollah Shahriari
Abstract
Abstract
Phosphoenolpyruvate Carboxykinase, encoded by the pepck gene, plays an important role in
gluconeogenesis. It also seems to be important in metabolism of nitrogenous compounds in developing seeds of
legumes, including amides and ureides which are then transformed into amino acids, necessary ...
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Abstract
Phosphoenolpyruvate Carboxykinase, encoded by the pepck gene, plays an important role in
gluconeogenesis. It also seems to be important in metabolism of nitrogenous compounds in developing seeds of
legumes, including amides and ureides which are then transformed into amino acids, necessary for the
synthesis of storage proteins. In this research, pepck gene expression in mRNA level, in different genotypes of
chickpea (Cicer arietinum L.), was determined. Two low protein genotypes (MCC291 and MCC373) and two
high protein genotypes (MCC458 and MCC053) out of 20 chickpea genotypes were selected. Total RNA were
extracted through different stages of seed development, and the expression of the pepck gene was estimated by
semi-quantitative RT-PCR. The results of the RT-PCR showed that two isoforms of this gene are expressed in
high protein genotypes, whereas in the low protein genotypes, the expression of these isoforms was not
obvious. Also this method showed a differential expression of pepck gene in different stages of flowering and
seed development. pepck gene is expressed in higher levels during the sheet formation and developing seeds
compared to the flowering and seed formation stages. Probably, the differential expression of pepck gene is
related to its possible role in metabolism of seed components, particularly in determination of the protein
content of chickpea seeds.