Weibiao Liao

Nitric oxide and hydrogen peroxide are involved in indole-3-butyric acid-induced adventitious root development in marigold

Summary Previous results have shown that both nitric oxide (NO) and hydrogen peroxide (H2O2) play crucial roles in the promotion of adventitious root development in explants of marigold (Tagetes erecta L.). In this study, the involvement of NO and H2O2 in indole-3-butyric acid (IBA)-induced adventitious root development in marigold explants, and the order of action of these molecules in the signal transduction pathway were investigated. The results indicated that, like NO and H2O2, IBA-induced adventitious rooting occurred in a dose-dependent manner. Two parameters of root growth in explants treated simultaneously with IBA and H2O2 were significantly higher than those in explants treated with IBA or H2O2 alone, suggesting that IBA and H2O2 may act synergistically to mediate adventitious rooting. NO and H2O2 were pre-requisites for the adventitious rooting induced by IBA. Moreover, treatment with IBA enhanced the endogenous levels of NO and H2O2 in explants, indicating that the induction of adventitious roots by IBA occurred through enhancing the levels of NO and H2O2. NO and H2O2 may be downstream signal molecules in the auxin signalling cascade, and NO may be involved as an upstream signalling molecule for H2O2 production.

Nitric oxide is required for hydrogen gas-induced adventitious root formation in cucumber

Hydrogen gas (H2) is involved in plant development and stress responses. Cucumber explants were used to study whether nitric oxide (NO) is involved in H2-induced adventitious root development. The results revealed that 50% and 100% hydrogen-rich water (HRW) apparently promoted the development of adventitious root in cucumber. While, the responses of HRW-induced adventitious rooting were blocked by a specific NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO), NO synthase (NOS) enzyme inhibitor N(G)-nitro-l-arginine methylester hydrochloride (l-NAME) and nitrate reductase (NR) inhibitor NaN3. HRW also increased NO content and NOS and NR activity both in a dose- and time-dependent fashion. Moreover, molecular evidence showed that HRW up-regulated NR genes expression in explants. The results indicate the importance of NOS and NR enzymes, which might be responsible for NO production in explants during H2-induced root organogenesis. Additionally, peroxidase (POD) and indoleacetic acid oxidase (IAAO) activity was significantly decreased in the explants treated with HRW, while HRW treatment significantly increased polyphenol oxidase (PPO) activity. In addition, cPTIO, l-NAME and NaN3 inhibited the actions of HRW on the activity of these enzymes. Together, NO may be involved in H2-induced adventitious rooting, and NO may be acting downstream in plant H2 signaling cascade.

Carbon Monoxide Is Involved in Hydrogen Gas-Induced Adventitious Root Development in Cucumber under Simulated Drought Stress

Hydrogen gas (H2) and carbon monoxide (CO) are involved in plant growth and developmental processes and may induce plant tolerance to several stresses. However, the independent roles and interaction effect of H2 and CO in adventitious root development under drought conditions have still not received the needed research attention. We hypothesize that there exists crosstalk between H2 and CO during adventitious root development under drought stress. The results of our current study revealed that 50% (v/v) hydrogen-rich water (HRW), 500 μM Hemin (the CO donor) and 30% (w/v) CO aqueous solution apparently promoted the development of adventitious roots in cucumber explants (Cucumis Sativus L.) under drought stress. H2 and CO increased relative water content (RWC), leaf chlorophyll content (chlorophyll a, b, and a+b), and chlorophyll fluorescence parameters [photochemical efficiency of photosystem II (PSII), PSII actual photochemical efficiency and photochemical quench coefficient] under drought condition. When the CO scavenger hemoglobin (Hb) or zinc protoporphyrin IX (ZnPPIX) was added to HRW/CO aqueous solution, the positive effect of HRW/CO aqueous solution on RWC, leaf chlorophyll content, and chlorophyll fluorescence parameters were reversed. Additionally, superoxide dismutases, peroxidase, catalase, and ascorbate peroxidase was significantly increased in the explants treated with HRW and CO aqueous solution under drought stress, thus alleviating oxidative damage, as indicated by decreases in thiobarbituric acid reactive substances (TBARS), hydrogen peroxide (H2O2), and superoxide radical (O2-) levels. H2 and CO also improved the levels of water soluble carbohydrate, total soluble protein, and proline content. However, the above CO/H2-mediated effects were reversed by CO scavenger Hb or CO specific synthetic inhibitor ZnPPIX. Therefore, CO may be involved in H2-induced adventitious rooting under drought stress and alleviate oxidative damage by enhancing RWC, leaf chlorophyll content, chlorophyll fluorescence parameters, metabolic constituent content, activating anti-oxidant enzymes and reducing TBARS, O2-, and H2O2 levels.

Hydrogen peroxide is involved in abscisic acid-induced adventitious rooting in cucumber (Cucumis sativus L.) under drought stress

Abscisic acid (ABA) and hydrogen peroxide (H2O2) are important regulatory factors involved in plant development under adversity stress. Here, the involvement of H2O2 in ABA-induced adventitious root formation in cucumber (Cucumis sativus L.) under drought stress was determined. The results indicated that exogenous ABA or H2O2 promoted adventitious rooting under drought stress, with a maximal biological response at 0.5 μM ABA or 800 μM H2O2. The promotive effects of ABA-induced adventitious rooting under drought stress were suppressed by CAT or DPI, suggesting that endogenous H2O2 might be involved in ABA-induced adventitious rooting. ABA increased relative water content (RWC), leaf chlorophyll content, chlorophyll fluorescence parameters (Fv/Fm, ΦPS II and qP), water soluble carbohydrate (WSC) and soluble protein content, and peroxidase (POD), polyphenol oxidase (PPO) and indoleacetate oxidase (IAAO) activities, while decreasing transpiration rate. However, the effects of ABA were inhibited by H2O2 scavenger CAT. Therefore, H2O2 may be involved in ABA-induced adventitious root development under drought stress by stimulating water and chlorophyll content, chlorophyll fluorescence, carbohydrate and nitrogen content, as well as some enzyme activities.

Calcium and Calmodulin Are Involved in Nitric Oxide-Induced Adventitious Rooting of Cucumber under Simulated Osmotic Stress

Osmotic stress is a major form of abiotic stress that adversely affects growth and development of plants and subsequently reduces yield and quality of crops. In this study, the effect of nitric oxide (NO) and calcium (Ca2+) on the process of adventitious rooting in cucumber (Cucumis sativus L.) under simulated osmotic stress was investigated. The results revealed that the effect of exogenous NO and Ca2+ in promoting the development of adventitious roots in cucumber seedlings under simulated osmotic stress was dose-dependent, with a maximal biological response at 10 μM NO donor nitroprusside (SNP) or 200 μM Ca2+. The application of Ca2+ chelators or channel inhibitors and calmodulin (CaM) antagonists significantly reversed NO-induced adventitious rooting, implying that endogenous Ca2+/CaM might be involved in NO-induced adventitious rooting under osmotic stress. Moreover, intracellular Ca amount was also increased by NO in cucumber hypocotyls during the development of adventitious roots under osmotic stress. This increase of endogenous Ca2+ was inhibited by NO specific scavenger 2-(4-carboxyphenyl) -4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO), nitrate reductase inhibitors tungstate (Na2WO4) and sodium azide (NaN3). This gives an indication that Ca2+ might be a downstream signaling molecule in the adventitious root development by NO under osmotic condition. The results also show that NO or Ca2+ play a positive role in improving plant water status and photosynthetic system by increasing chlorophyll content and photochemical activity in leaves. Furthermore, NO and Ca2+ treatment might alleviate the negative effects of osmotic stress by decreasing membrane damage and reactive oxygen species (ROS) production by enhancing the activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX). Therefore, Ca2+/CaM may act as a downstream signaling molecule in NO-induced development of adventitious root under simulated osmotic stress through improving the photosynthetic performance of leaves and activating antioxidative system in plants.

Effect of nitric oxide on dormancy release in bulbs of Oriental lily (Lilium orientalis) ‘Siberia’

SUMMARY Nitric oxide (NO) is an essential endogenous plant signalling molecule involved in a wide range of plant developmental processes. To investigate the effect of NO on breaking dormancy in bulbs, bulbs of Oriental lily (Lilium orientalis) ‘Siberia’ were treated with various concentration of the NO donor, sodium nitroprusside (SNP; 0.0,1.0,3.0, or 5.0 mM). The results showed that the effect of NO was dose-dependent, with the maximum biological response at 1.0 mM SNP. When applied exogenously, the 1.0 mM SNP treatment reduced the time required to release dormancy in Oriental lily bulbs. Meanwhile, 1.0 mM SNP significantly increased the shoot length:bulb height ratio. In addition, 1.0 mM SNP significantly lowered starch concentrations and increased water soluble carbohydrate (WSC) and reducing sugar concentrations. These results indicate that NO treatment, at the correct dose, reduced the time required to release dormancy in bulbs by accelerating the degradation of starch and increasing the accumulation of WSC and reducing sugars in Oriental lily bulbs.

Effect of exogenous nitric oxide on vegetative and reproductive growth of oriental lily ‘Siberia’

Recent studies have shown that nitric oxide (NO) may be involved in diverse plant developmental processes as an important signal molecule. However, the effects of NO on vegetative and reproductive growth of intact plants are not fully understood. In our study, the NO donor sodium nitroprusside (SNP) at 3,000, 6,000 and 9,000 μM was used to characterize the roles of NO in vegetative and reproductive growth of “Siberia” lily. The results showed that the effects of NO on growth in “Siberia” lily were dose-dependent. Compared with the control (distilled water treatment), 3,000 μM SNP caused a significant increase in plant height and internode length. Significant differences were recorded in flowering period, lifetime of individual flowers, number of flowers per plant, flower diameter, and bud length with 6,000 μM SNP. However, 9,000 μM SNP resulted in negative effects on these parameters as compared to the control. Moreover, 3,000 μM SNP caused a significant increase in the contents of leaf chlorophyll, water soluble carbohydrate (WSC), starch, total carbohydrate, total soluble protein, and total nitrogen, all of which were decreased by high levels of SNP (9,000 μM). Together, these results indicated that NO treatments at the proper dosage can promote vegetative and reproductive growth, possibly by improving the levels of leaf chlorophyll, WSC, starch, total carbohydrate, total soluble protein and total nitrogen.

Hydrogen Sulfide: A Gaseous Molecule in Postharvest Freshness

Hydrogen sulfide (H2S), as a signaling molecule, is involved in the regulation of growth and development in plants. Recent studies have indicated that H2S also plays important roles in regulating postharvest senescence of horticultural products. The focus of this review is to summarize the synthesis of H2S in plants and its potential roles in alleviating the senescence of cut flowers, fruits, and vegetables during postharvest storage. During postharvest of horticultural products, H2S could scavenge reactive oxygen species via promoting the activity of antioxidant enzymes, thereby, sustaining the integrity of the membrane. In fruits, H2S effectively enhanced the tolerance of chilling by increasing the content of proline and polyphenol compounds. During postharvest storage of perishable fruits and vegetables, H2S significantly alleviated decay, which was caused by fungi by inhibiting the growth of fungi spores. Moreover, H2S interacted with other molecules synergistically (NO) or antagonistically (ethylene) to alleviate senescence of horticultural products. At the transcriptional level, H2S regulated the expression of senescence-related genes, which were related to degradation of proteins and chlorophyll, to delay the senescence of horticultural products. Thus, H2S does not only possess positive antioxidant and antifungal properties, but also significantly regulates the senescence-related gene during postharvest of horticultural products. Future studies of H2S in postharvest storage should focus on its molecular mechanism in the posttranslational modifications of proteins as well as its safety attributes in treated fruits and vegetables.

5-Aminolevulinic Acid (ALA) Alleviated Salinity Stress in Cucumber Seedlings by Enhancing Chlorophyll Synthesis Pathway

5-Aminolevulinic acid (ALA) is a common precursor of tetrapyrroles as well as a crucial growth regulator in higher plants. ALA has been proven to be effective in improving photosynthesis and alleviating the adverse effects of various abiotic stresses in higher plants. However, little is known about the mechanism of ALA in ameliorating the photosynthesis of plant under abiotic stress. In this paper, we studied the effects of exogenous ALA on salinity-induced damages of photosynthesis in cucumber (Cucumis sativus L.) seedlings. We found that the morphology (plant height, leave area), light utilization capacity of PS II [qL, Y(II)] and gas exchange capacity (Pn, gs, Ci, and Tr) were significantly retarded under NaCl stress, but these parameters were all recovered by the foliar application of 25 mg L-1 ALA. Besides, salinity caused heme accumulation and up-regulation of gene expression of ferrochelatase (HEMH) with suppression of other genes involved in chlorophyll synthesis pathway. Exogenously application of ALA under salinity down-regulated the heme content and HEMH expression, but increased the gene expression levels of glutamyl-tRNA reductase (HEMA1), Mg-chelatase (CHLH), and protochlorophyllide oxidoreductase (POR). Moreover, the contents of intermediates involved in chlorophyll branch were increased by ALA, including protoporphyrin IX (Proto IX), Mg-protoporphyrin IX (Mg-Proto IX, protochlorophyllide (Pchlide), and chlorophyll (Chl a and Chl b) under salt stress. Ultrastructural observation of mesophyll cell showed that the damages of photosynthetic apparatus under salinity were fixed by ALA. Collectively, the chlorophyll biosynthesis pathway was enhanced by exogenous ALA to improve the tolerance of cucumber under salinity.

Effect of hydrogen-rich water on vase life and quality in cut lily and rose flowers

Hydrogen gas (H2) functions as an important signaling molecule in diverse plant developmental processes. H2 is thought to delay postharvest ripening and senescence in fruit. However, little is known about the influence of H2 on flower senescence. This study was conducted to determine whether H2 treatment could improve vase life and quality in cut lily (Lilium spp.) and rose (Rosa hybrid L.) flowers. Treatment with 0.5% and 1% hydrogen-rich water (HRW) increased vase life and maximum flower diameter in lily. In addition, 50% HRW treatment significantly increased vase life and maximum flower diameter in cut rose flowers. The fresh weight and leaf relative water content in cut lilies and roses were enhanced by proper doses of HRW. Compared with the control, HRW treatment decreased leaf stomata size in cut lily and rose flowers. HRW treatment significantly reduced leaf malondialdehyde contents and electrolyte leakage in cut lilies. Antioxidant enzyme activities were also improved by HRW treatment in cut lily and rose flowers. These results suggest that exogenously applied H2 might improve vase life and postharvest quality in cut flowers by maintaining proper water balance and membrane stability and by reducing stomata size and oxidative damage.