Hydrogen gas (H2) was first identified as a unique molecular messenger in animals. Since H2 was reported as a novel antioxidant, it has been proven effective in treating many diseases. However, the studies concerning H2 in plants are just beginning to emerge. Here, two paths of H2 production in plants have been reported, namely, hydrogenase and nitrogenase. H2 has positive effects on seed germination, seedling growth, adventitious rooting, root elongation, harvest freshness, stomatal closure and anthocyanin synthesis. H2 also can enhance plant symbiotic stress resistance commonly through the enhancement of antioxidant defence system. Moreover, H2 shows cross talk with nitric oxide, carbon monoxide and other signalling molecules (for example, abscisic acid, ethylene and jasmonate acid). H2 can regulate the expression of responsive genes under abiotic stress and during adventitious roots formation and anthocyanin biosynthesis. Future work will need to focus on the molecular mechanism of H2 and its crosstalk with other signalling molecules in plants. With its promising application in agriculture, hydrogen agriculture will be welcomed in the near future.

Hydrogen gas (H 2 ) plays an important role in plant development and stress responses. Here, cucumber ( Cucumis sativus L.) explants were used to investigate the roles of H 2 in adventitious root development under cadmium (Cd) stress and its physiological mechanism. The results showed that hydrogen-rich water (HRW) promoted adventitious rooting under Cd stress and 50% HRW obtained the maximal biological response. Compared with Cd treatment, HRW + Cd treatment significantly reduced the content of malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ), superoxide radical (O 2 - ), thiobarbituric acid reactive substances (TBARS), ascorbic acid (AsA) and reduced glutathione (GSH), as well as relative electrical conductivity (REC), lipoxygenase (LOX) activity, AsA/docosahexaenoic acid (DHA) ratio, and GSH/oxidized glutathione (GSSG) ratio, while increasing DHA and GSSG content. HRW + Cd treatment also significantly increased in the activity and related gene expression of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR). Additionally, HRW + Cd treatment increased the contents of osmotic adjustment substances, as well as the activities of peroxidase (POD) and polyphenol oxidase (PPO), while significantly decreasing indoleacetic acid oxidase (IAAO) activity. In summary, H 2 could induce adventitious rooting under Cd stress by decreasing the oxidative damage, increasing osmotic adjustment substance content and regulating rooting-related enzyme activity.

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Previous studies have shown that both hydrogen gas (H2) and ethylene (ETH) play positive roles in plant adventitious rooting. However, the relationship between H2and ETH during this process has not been explored and remains insufficiently understood. In this study, cucumber (Cucumis sativus L.) was used to explore the proteomic changes in ETH-H2-induced rooting. Our results show that hydrogen-rich water (HRW) and ethylene-releasing compound (ethephon) at proper concentrations promote adventitious rooting, with maximal biological responses occurring at 50% HRW or 0.5 µM ethephon. ETH inhibitors aminoethoxyvinylglycine (AVG) and AgNO3 cause partial inhibition of adventitious rooting induced by H2, suggesting that ETH might be involved in H2-induced adventitious rooting. According to two-dimensional electrophoresis (2-DE) and mass spectrometric analyses, compared with the control, 9 proteins were up-regulated while 15 proteins were down-regulated in HRW treatment; four proteins were up-regulated while 10 proteins were down-regulated in ethephon treatment; and one protein was up-regulated while nine proteins were down-regulated in HRW+AVG treatment. Six of these differentially accumulated proteins were further analyzed, including photosynthesis -related proteins (ribulose-1,5-bisphosphate carall boxylase smsubunit (Rubisco), sedoheptulose-1,7-bisphosphatase (SBPase), oxygen-evolving enhancer protein (OEE1)), amino and metabolism-related protein (threonine dehydratase (TDH)), stress response-related protein (cytosolic ascorbate peroxidase (CAPX)), and folding, modification and degradation-related protein (protein disulfide-isomerase (PDI)). Moreover, the results of real-time PCR about the mRNA levels of these genes in various treatments were consistent with the 2-DE results. Therefore, ETH may be the downstream signaling molecule during H2- induced adventitious rooting and proteins Rubisco, SBPase, OEE1, TDH, CAPX and PDI may play important roles during the process.