Calcium-hydrogen sulfide crosstalk during K+-deficient NaCl stress operates through regulation of Na+/H+ antiport and antioxidative defense system in mung bean roots

Abstract

Present investigation reports the role of calcium (Ca²⁺) and hydrogen sulfide (H₂S) crosstalk associated with Vigna radiata seedlings subjected to K⁺ deficient conditions under short-term (24 h) and long-term (72 h) NaCl stress. Perusal of the data reveals that under short-term NaCl stress an initial decline in K⁺ level led to the elevation in Ca²⁺ and H₂S levels along with improvement in antioxidant system and reduction in reactive oxygen species (ROS) production. Under long-term NaCl stress a further decline in K⁺ content was deleterious that led to a lower K⁺/Na⁺ ratio. This was followed by reduction in antioxidant system along with excessive accumulation of ROS and methylglyoxal content, and increased membrane damage. However, supplementation of the seedling roots with Ca²⁺ enhanced biosynthesis of H₂S through enhancing cysteine pool. The present findings suggest that synergistic action of Ca²⁺ and H₂S induced the activity of H⁺-ATPase that created H⁺ gradient which in turn induced Na⁺/H⁺ antiport system that accelerated K⁺ influx and Na⁺ efflux. All of these together contributed to a higher K⁺/Na⁺ ratio, activation of antioxidative defense system, and maintenance of redox homeostasis and membrane integrity in Ca²⁺-supplemented stressed seedlings. Role of Ca²⁺ and H₂S in the regulation of Na⁺/H⁺ antiport system was validated by the use of sodium orthovanadate (plasma membrane H⁺-ATPase inhibitor), tetraethylammonium chloride (K⁺ channel blocker), and amiloride (Na⁺/H⁺ antiporter inhibitor). Application of Ca²⁺-chelator EGTA (ethylene glycol-bis(b-aminoethylether)-N,N,N',N'-tetraacetic acid) and H₂S scavenger hypotaurine abolished the effect of Ca²⁺, suggesting the involvement of Ca²⁺ and H₂S in the alleviation of NaCl stress. Moreover, use of EGTA and HT also substantiates the downstream functioning of H₂S during Ca²⁺-mediated regulation of plant adaptive responses to NaCl stress. To sum up, present findings reveal the association of Ca²⁺ and H₂S signaling in the regulation of ion homeostasis and antioxidant defense during K⁺-deficient NaCl stress.

Keywords: Ascorbate-glutathione cycle; Calcium signaling; Hydrogen sulfide; Na(+)/H(+) antiport; Proline; Redox homeostasis.