Plants are challenged by a variety of biotic or abiotic stresses, which can affect their growth and development, productivity and geographic distribution. In order to survive adverse environmental conditions, plants have evolved various adaptive strategies, among which is the accumulation of metabolites that play protective roles. A well-established example of the metabolites that are involved in stress responses, or stress tolerance, is the low-molecular-weight aliphatic polyamines, including putrescine,spermidine and spermine. The critical role of polyamines in stress tolerance is suggested by several lines of evidence: firstly, the transcript levels of polyamine biosynthetic genes, as well as the activities of the corresponding enzymes, are induced by stresses; secondly, elevation of endogenous polyamine levels by exogenous supply of polyamines, or overexpression of polyamine biosynthetic genes, results in enhanced stress tolerance; and thirdly, a reduction of endogenous polyamines is accompanied by compromised stress tolerance. A number of studies have demonstrated that polyamines function in stress tolerance largely by modulating the homeostasis of reactive oxygen species (ROS) due to their direct, or indirect, roles in regulating antioxidant systems or suppressing ROS production. The transcriptional regulation of polyamine synthesis by transcription factors is also reviewed here. Meanwhile, future perspectives on polyamine research are also suggested.

The basic helix-loop-helix (bHLH) transcription factors are involved in a variety of physiological processes. However, plant bHLHs functioning in cold tolerance and the underlying mechanisms remain poorly understood. Here, we report the identification and functional characterization of PtrbHLH isolated from trifoliate orange (Poncirus trifoliata). The transcript levels of PtrbHLH were up-regulated under various abiotic stresses, particularly cold. PtrbHLH was localized in the nucleus with transactivation activity. Overexpression of PtrbHLH in tobacco (Nicotiana tabacum) or lemon (Citrus limon) conferred enhanced tolerance to cold under chilling or freezing temperatures, whereas down-regulation of PtrbHLH in trifoliate orange by RNA interference (RNAi) resulted in elevated cold sensitivity. A range of stress-responsive genes was up-regulated or down-regulated in the transgenic lemon. Of special note, several peroxidase (POD) genes were induced after cold treatment. Compared with the wild type, POD activity was increased in the overexpression plants but decreased in the RNAi plants, which was inversely correlated with the hydrogen peroxide (H₂O₂) levels in the tested lines. Treatment of the transgenic tobacco plants with POD inhibitors elevated the H₂O₂ levels and greatly compromised their cold tolerance, while exogenous replenishment of POD enhanced cold tolerance of the RNAi line. In addition, transgenic tobacco and lemon plants were more tolerant to oxidative stresses. Yeast one-hybrid assay and transient expression analysis demonstrated that PtrbHLH could bind to the E-box elements in the promoter region of a POD gene. Taken together, these results demonstrate that PtrbHLH plays an important role in cold tolerance, at least in part, by positively regulating POD-mediated reactive oxygen species removal.

Background: Drought is one of the major abiotic stresses affecting plant growth, development and crop productivity. ABA responsive element binding factor (ABF) plays an important role in stress responses via regulating the expression of stress-responsive genes.Results: In this study, a gene coding for ABF (PtrABF) was isolated fromPoncirus trifoliata(L.) Raf.PtrABFhad a complete open reading frame of 1347 bp, encoding a 448 amino acid peptide, and shared high sequence identities with ABFs from other plants. PtrABF was subcellularly targeted to the nucleus, exhibited transactivation activity in yeast cell and could bind to ABRE, supporting its role as a transcription factor. Expression levels ofPtrABFwere induced by treatments with dehydration, low temperature and ABA. Ectopic expression ofPtrABFunder the control of a CaMV 35S promoter in transgenic tobacco plants enhanced tolerance to both dehydration and drought. Under dehydration and drought conditions, the transgenic plants accumulated lower levels of reactive oxygen species compared with wild type, accompanied by higher activities and expression levels of three antioxidant enzymes. In addition, steady-state mRNA levels of nine stress-responsive genes coding for either functional or regulatory proteins were induced to higher levels in the transgenic lines with or without drought stress.Conclusions: PtrABFis a bZIP transcription factor and functions in positive modulation of drought stress tolerance. It may be an important candidate gene for molecular breeding of drought- tolerant plants.

Polyamines, small aliphatic polycations, have been suggested to play key roles in a number of biological processes. In this paper, attempts were made to investigate the possibility of improving dehydration tolerance of citrus in vitro plants by exogenous application of spermine (Spm). ‘Red Tangerine’ (Citrus reticulata Blanco) in vitro plants pretreated with 1 mM Spm exhibited less wilted phenotype and lower water loss and electrolyte leakage than the control under dehydration. Spm-pretreated plants contained higher endogenous polyamine content during the course of the experiment relative to the control, particularly at the end of dehydration, coupled with higher expression levels of ADC and SPMS. Histochemical staining showed that the Spm-pretreated leaves were stained to a lower extent than those without Spm pretreatment, implying generation of less reactive oxygen species (ROS). On the contrary, activities of peroxidase (POD) and superoxide dismutase (SOD) in the Spm-pretreated samples were higher than the control at a given time point or during the whole experiment, suggesting that Spm exerted a positive effect on antioxidant systems. In addition, significantly smaller stomatal aperture size was observed in Spm-pretreated epidermal peels, which showed that stomatal closure was promoted by polyamines. All of these data suggest that Spm pretreatment causes accumulation of higher endogenous polyamines and accordingly leads to more effective ROS scavenging (less tissue damage) and stimulated stomatal closure (lower water loss) upon dehydration, which may function collectively to enhance dehydration tolerance.