Improved short-term drought response of transgenic rice over-expressing maize C4 phosphoenolpyruvate carboxylase via calcium signal cascade

Abstract

To understand the link between long-term drought tolerance and short-term drought responses in plants, transgenic rice (Oryza sativa L.) plants over-expressing the maize C_4-pepc gene encoding phosphoenolpyruvate carboxylase (PC) and wild-type (WT) rice plants were subjected to PEG 6000 treatments to simulate drought stress. Compared with WT, PC had the higher survival rate and net photosynthetic rate after 16days of drought treatment, and had higher relative water content in leaves after 2h of drought treatment as well, conferring drought tolerance. WT accumulated higher amounts of malondialdehyde, superoxide radicals, and H₂O₂ than PC under the 2-h PEG 6000 treatment, indicating greater damages in WT. Results from pretreatments with a Ca²⁺ chelator and/or antagonist showed that the regulation of the early drought response in PC was Ca²⁺-dependent. The NO and H₂O₂ levels in PC lines were also up-regulated via Ca²⁺ signals, indicating that Ca²⁺ in PC lines also reacted upstream of NO and H₂O₂. 2-h drought treatment increased the transcripts of CPK9 and CPK4 in PC via positive up-regulation of Ca²⁺. The transcripts of NAC6 [NACs (NAM, ATAF1, ATAF2, and CUC2)] and bZIP60 (basic leucine zipper, bZIP) were up-regulated, but those of DREB2B (dehydration-responsive element-binding protein, DREB) were down-regulated, both via Ca²⁺ signals in PC. PEPC activity, expressions of C₄-pepc, and the antioxidant enzyme activities in PC lines were up-regulated via Ca²⁺. These results indicated that Ca²⁺ signals in PC lines can up-regulate the NAC6 and bZIP60 and the downstream targets for early drought responses, conferring drought tolerance for the long term.

Keywords: Drought; Endogenous Ca(2+); Phosphoenolpyruvate carboxylase; Rice (Oryza sativa L.); Signal transduction.