Molecular cloning and characterization of CrNCED1, a gene encoding 9-cis-epoxycarotenoid dioxygenase in Citrus reshni, with functions in tolerance to multiple abiotic stresses

Abstract

Abscisic acid (ABA) is an important stress phytohormone that plays an essential role in mediating the signaling networks associated with plant responses to various abiotic stresses. In the present study, we isolated a gene CrNCED1 encoding the rate-limiting enzyme of ABA synthesis, 9-cis-epoxycarotenoid dioxygenase (NCED), from Citrus reshni. Expression patterns of CrNCED1 varied among different tissues, in which higher levels were measured in the leaves than in the roots. The steady-state mRNA levels of CrNCED1 were progressively elevated by dehydration and ABA, only transiently induced by cold, but not affected by salt. To examine its function in stress tolerance, transgenic tobacco (Nicotiana nudicaulis) plants constitutively overexpressing CrNCED1 were generated, which contained higher ABA levels than the wild type (WT) under both normal growth conditions and drought stress. The transgenic lines displayed enhanced tolerance to dehydration, drought, salt and oxidative stresses when compared with the WT. Lower levels of reactive oxygen species (H₂O₂ and O₂⁻) were detected in the transgenic plants than in the WT under dehydration and salt stress. On the contrary, transcript levels of several genes associated with ROS scavenging, osmoticum adjustment, and water maintenance, and activities of two antioxidant enzymes were higher in the transgenic plants relative to the WT under the dehydration stress. Taken together, CrNCED1 overexpression confers enhanced tolerance to multiple abiotic stresses, which may be, at least in part, ascribed to the positive activation of the stress-responsive genes.