H2O2 is known as a signal molecule in plant cells, but its role in the regulation of aqbscisic acid (ABA) and gibberellic acid (GA) metabolism and hormonal balance is not yet clear. In this study it was found that H2O2 affected the regulation of ABA catabolism and GA biosynthesis during seed imbibition and thus exerted control over seed dormancy and germination. As seen by quantitative RT-PCR (QRT-PCR), H2O2 up-regulated ABA catabolism genes (e.g. CYP707A genes), resulting in a decreased ABA content during imbibition. This action required the participation of nitric oxide (NO), another signal molecule. At the same time, H2O2 also up-regulated GA biosynthesis, as shown by QRT-PCR. When an ABA catabolism mutant, cyp707a2, and an overexpressing plant, CYP707A2-OE, were tested, ABA content was negatively correlated with GA biosynthesis. Exogenously applied GA was able to over-ride the inhibition of germination at low concentrations of ABA, but had no obvious effect when ABA concentrations were high. It is concluded that H2O2 mediates the up-regulation of ABA catabolism, probably through an NO signal, and also promotes GA biosynthesis. High concentrations of ABA inhibit GA biosynthesis but a balance of these two hormones can jointly control the dormancy and germination of Arabidopsis seeds.

In eukaryotes, mechanisms such as alternative splicing (AS) and alternative translation initiation (ATI) contribute to organismal protein diversity. Specifically, splicing factors play crucial roles in responses to environment and development cues; however, the underlying mechanisms are not well investigated in plants. Here, we report the parallel employment of short-read RNA sequencing, single molecule long-read sequencing and proteomic identification to unravel AS isoforms and previously unannotated proteins in response to abscisic acid (ABA) treatment. Combining the data from the two sequencing methods, approximately 83.4% of intron-containing genes were alternatively spliced. Two AS types, which are referred to as alternative first exon (AFE) and alternative last exon (ALE), were more abundant than intron retention (IR); however, by contrast to AS events detected under normal conditions, differentially expressed AS isoforms were more likely to be translated. ABA extensively affects the AS pattern, indicated by the increasing number of non-conventional splicing sites. This work also identified thousands of unannotated peptides and proteins by ATI based on mass spectrometry and a virtual peptide library deduced from both strands of coding regions within the Arabidopsis genome. The results enhance our understanding of AS and alternative translation mechanisms under normal conditions, and in response to ABA treatment.

Summary• Nitric oxide (NO) has been reported to be involved in breaking seed dormancy but its mechanism of action is unclear.• Here, we report that a rapid accumulation of NO induced an equally rapid decrease of abscisic acid (ABA) that is required for this action in Arabidopsis.• Results of quantitative real-time polymerase chain reaction (QRT-PCR) and Western blotting indicate that the NO-induced ABA decrease correlates with the regulation of CYP707A2 transcription and (+)-abscisic acid 8′-hydroxylase (encoded by CYP707A2) protein expression. By analysing cyp707a1, cyp707a2 and cyp707a3 mutants, we found that CYP707A2 plays a major role in ABA catabolism during the first stage of imbibition.• Fluorescent images demonstrate that NO is released rapidly in the early hours at the endosperm layer during imbibition. Evidently, such response precedes the enhancement of ABA catabolism which is required for subsequent seed germination.

The antagonism between abscisic acid (ABA) and gibberellin (GA) plays a key role in controlling seed germination, but the mechanism of antagonism during this process is not known. The possible links among ABA, reactive oxygen species (ROS), ascorbic acid (ASC), and GA during rice seed germination were investigated. Unlike in non-seed tissues where ROS production is increased by ABA, ABA reduced ROS production in imbibed rice seeds, especially in the embryo region. Such reduced ROS also led to an inhibition of ASC production. GA accumulation was also suppressed by a reduced ROS and ASC level, which was indicated by the inhibited expression of GA biosynthesis genes, amylase genes, and enzyme activity. Application of exogenous ASC can partially rescue seed germination from ABA treatment. Production of ASC, which acts as a substrate in GA biosynthesis, was significantly inhibited by lycorine which thus suppressed the accumulation of GA. Consequently, expression of GA biosynthesis genes was suppressed by the low levels of ROS and ASC in ABA-treated seeds. It can be concluded that ABA regulates seed germination in multiple dimensions. ROS and ASC are involved in its inhibition of GA biosynthesis.