Partial cloning, characterization, and analysis of expression and activity of plasma membrane H + -ATPase in Kallar gras
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ORIGINAL PAPER
Partial cloning, characterization, and analysis of expression and activity of plasma membrane H+‑ATPase in Kallar grass [Leptochloa fusca (L.) Kunth] under salt stress Hadi Hamidi Ravari1 · Hamid Reza Kavousi1 · Fereshteh Mohammadi1 · Shahram Pourseyedi1 Received: 28 October 2019 / Accepted: 15 May 2020 © Akadémiai Kiadó Zrt. 2020
Abstract Kallar grass (Leptochloa fusca) is a highly salt-tolerant C4 perennial halophytic forage. The regulation of ion movement across the plasma membrane (PM) to improve salinity tolerance of plant is thought to be accomplished with the aid of the proton electrochemical gradient generated by PM H+-ATPase. In this study, we cloned a partial gene sequence of the Lf PM H+-ATPase and investigated its expression and activity under salt stress. The amino acid sequence of the isolated region of Lf PM H+-ATPase possesses the maximum identity up to 96% to its ortholog in Aeluropus littoralis. The isolated fragment of Lf PM H+-ATPase gene is a member of the subfamily Π of plant PM H+-ATPase and is most closely related to the Oryza sativa gene OSA7. The transcript level and activity of the PM H+-ATPase were increased in roots and shoots in response to NaCl and were peaked at 450 mM NaCl in both tissues. The induction of activity and gene expression of PM H+-ATPase in roots and shoots of Kallar grass under salinity indicate the necessity for this pump in these organs during salinity adaptation to establish and maintain the electrochemical gradient across the PM of the cells for adjusting ion homeostasis. Keywords Salt tolerance · Leptochloa fusca · Plasma membrane H+-ATPase · Real-time qRT-PCR · Halophyte
Introduction Plants are continually exposed to a large variety of biotic and abiotic stresses during their whole life that severely reduces their productivity (Cao et al. 2017). Salinity is a significant abiotic stress affecting plant growth and development and is expected to increase because of global weather changes and as a consequence of many irrigation practices (Reddy et al. 2017). Usually, the high salt concentration can disturb vital physiological processes through inducing low osmotic potential of the soil solution (water deficits), hyperosmotic stress, ion imbalance, nutritional imbalance, metabolic disorders, and even death (Munns and Tester 2008). However, ionic imbalance caused by the accumulation of excessive amounts of Na+ and Cl− ions is a major contributory factor (AcostaMotos et al. 2017). Under conditions of elevated NaCl * Hamid Reza Kavousi [email protected]; [email protected] 1
Department of Biotechnology, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
levels outside the cell, N a + excessively accumulates in the cytoplasm, which results in the inhibition of plant growth and development (Janicka-Russak et al. 2013). The reactions of the plant to salinity stress are complex and involving responses to cellular osmotic and ionic stresses, subsequent secondary stresses (e.g., oxidative stress), and whole plant
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