Populus tomentosa (Chinese white poplar) is well adapted to various extreme environments, and is considered an important species to study the effects of salinity stress on poplar trees. To decipher the mechanism of poplar’s rapid response to short-term salinity stress, we firstly detected the changes in H2O2 and hormone, and then profiled the gene expression pattern of ten-week-old seedling roots treated with 200 mM NaCl for 0, 6, 12 and 24 hours (h) by RNA-seq on the Illumina-Solexa platform. Physiological determination showed that the significant increase in H2O2 began at 6 h, while that in hormone ABA was at 24 h, under salt stress. Compared with controls (0 h), 3991, 4603 and 4903 genes were up regulated, and 1408, 2206 and 3461 genes were down regulated (adjusted P-value ≤ 0.05 and |log2Ratio|≥1) at 6, 12, and 24 h time points, respectively. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation revealed that the differentially expressed genes (DEGs) were highly enriched in hormone- and reactive oxygen species-related biological processes, including ‘response to oxidative stress or abiotic stimulus’, ‘peroxidase activity’, ‘regulation of transcription’, ‘hormone synthetic and metabolic process’, ‘hormone signal transduction’, ‘antioxidant activity’ and ‘transcription factor activity’. Moreover, K-means clustering demonstrated that DEGs (total RPKM value>12 from four time points) could be categorized into four kinds of expression trends: quick up/down over 6 h or 12 h, and slow up/down over 24 h. Of these, DEGs involved in H2O2- and hormone- producing and signal-related genes were further enriched in this analysis, which indicated that the two kinds of small molecules, hormones and H2O2, play pivotal roles in the short-term salt stress response in poplar. This study provides a basis for future studies of the molecular adaptation of poplar and other tree species to salinity stress.

γ-Aminobutyric acid (GABA) influences plant growth, but little is known about how this metabolite regulates adventitious root (AR) development. Here, we investigate the effects of GABA on ARs using poplar lines overexpressing glutamate decarboxilase 2 (GAD2) and by treating poplar stem cuttings with exogenous GABA or vigabatrin (VGB; a specific GABA transaminase inhibitor). Endogenous GABA accumulation not only inhibited AR growth, but it also suppressed or delayed AR formation. Anatomical observations revealed that the GABA and VGB treatments resulted in a 1 d delay in the formation of AR primordia and the appearance of ARs. This delay coincided with changes in primary metabolism, including transient increases in hexose and amino acid levels. GABA-dependent changes in the expression of genes related to hormone synthesis and signalling, as well as analysis of hormone levels revealed that ethylene-dependent pathways were decreased at the earliest stage of AR formation. In contrast, auxin and abscisic acid were increased at 1–5 d as well as GA4 over a 5 d period of AR formation. These results demonstrate that GABA plays a crucial role in AR development. Evidence is presented demonstrating that GABA can interact with hormone-related pathways as well as carbon/nitrogen metabolism. These findings also elucidate the functions of GABA in plant development.

Exogenous citric acid (CA), which acts as an important intermediate product of the tricarboxylic acid (TCA) cycle, can enhance the TCA cycle activity and activate the branched operation of the TCA cycle, thus providing energy required for resistance to adverse conditions. However, the effects of CA application on TCA cycle-related metabolism under cadmium (Cd) were less reported. To investigate the effects of CA on the Cd tolerance of Dahurian wildrye grass (Elymus dahuricus), the growth, Cd accumulation, antioxidant systems and metabolic pathways of leaves and roots were investigated by a potted soil experiment with Cd (50 mg/kg) and CA (4 mmol/L) treatments. The results showed that Cd stress seriously affected growth and induced the production of reactive oxygen in clover leaves and roots, leading to membrane peroxidation and activation of the antioxidant defense system. Exogenous CA could not only effectively relieve the inhibition of Cd stress on growth and reduce the amount of reactive oxygen by increasing the antioxidant capacities but could also promote an increase in root Cd content. Metabolomic results showed that the application of CA increased the contents of sugars, sugar alcohols, and resistant substances, and promoted the metabolism of amino acids including γ-aminobutyric acid (GABA). These alterations contributed the significant enhancement of the Cd resistance, which may be related to the changes in the TCA cycle activity and the metabolism of the shikimic acid pathway in leaves and roots as well as GABA shunt in roots.