Jihua Yu

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.

Transcriptome Analysis of Pepper (Capsicum annuum) Revealed a Role of 24-Epibrassinolide in Response to Chilling

Brassinosteroids (BRs) have positive effects on many processes during plant growth, development, and various abiotic stress responses. However, little information is available regarding the global gene expression of BRs in response to chilling stress in pepper. In this study, we used RNA sequencing to determine the molecular roles of 24-epibrassinolide (EBR) during a chilling stress response. There were 39,829 transcripts, and, among them, 656 were differently-expressed genes (DEGs) following EBR treatment (Chill+EBR) compared with the control (Chill only), including 335 up-regulated and 321 down-regulated DEGs. We selected 20 genes out of the 656 DEGs for RT-qPCR analysis to confirm the RNA-Seq. Based on GO enrich and KEGG pathway analysis, we found that photosynthesis was significantly up-enriched in biological processes, accompanied by significant increases in the net photosynthetic rate (Pn), Fv/Fm, and chlorophyll content. Furthermore, the results indicate that EBR enhanced endogenous levels of salicylic acid (SA) and jasmonic acid (JA) while suppressing the ethylene (ETH) biosynthesis pathway, suggesting that BRs function via a synergistic cross-talk with SA, JA, and ETH signaling pathways in response to chilling stress. In addition, EBR induced cellulose synthase-like protein and UDP-glycosyltransferase, suggesting a contribution to the formation of cell wall and hormone metabolism. EBR also triggered the calcium signaling transduction in cytoplasm, and activated the expression of cellular redox homeostasis related genes, such as GSTX1, PER72, and CAT2. This work, therefor, identified the specific genes showed different expression patterns in EBR-treated pepper and associated with the processes of hormone metabolism, redox, signaling, transcription, and defense. Our study provides the first evidence of the potent roles of BRs, at the transcription level, to induce the tolerance to chilling stress in pepper as a function of the combination of the transcriptional activities, signaling transduction, and metabolic homeostasis.

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.

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.

Autotoxicity in cucumber (Cucumis sativus L.) seedlings is alleviated by silicon through an increase in the activity of antioxidant enzymes and by mitigating lipid peroxidation

Autotoxicity in plants limits their growth and that of nearby plants of the same species, which has obvious implications in crop yield and quality. Silicon (Si) has been shown to increase plant tolerance to autotoxic stress. However, the physiological mechanisms underlying the effects of Si in alleviating autotoxicity during germination in cucumber (Cucumis sativus L.) are unknown. Cinnamic acid derivatives, such as 3-phenylpropionic acid (PA), are a class of autotoxins present in cucumber root exudates. Our objective was to investigate Si-induced autotoxic stress tolerance in cucumber seedlings by focusing on the effects of Si on the induction of antioxidant defense pathways. We found that PA treatment significantly reduced seed germination, radicle length, lateral root number, fresh weight, AsA and GSH contents, and the activities of SOD, CAT, and APX in cucumber seedlings, while it increased membrane permeability and levels of MDA, proline, O2-, and H2O2. Application of Si enhanced growth of PA-treated plants and significantly increased germination rate, radicle length, lateral root number, fresh weight, AsA and GSH levels, and SOD, CAT, POD, and APX activities. These results suggest that exogenous Si alleviates autotoxicity caused by PA during seed germination by increasing antioxidant enzyme activities and mitigating lipid peroxidation.

Reversed-Phase High-Performance Liquid Chromatography for the Quantification and Optimization for Extracting 10 Kinds of Carotenoids in Pepper (Capsicum annuum L.) Leaves

Carotenoids are considered to be crucial elements in many fields and, furthermore, the significant factor in pepper leaves under low light and chilling temperature. However, little literature focused on the method to determinate and extract the contents of carotenoid compositions in pepper leaves. Therefore, a time-saving and highly sensitive reversed-phase high-performance liquid chromatography method for separation and quantification of 10 carotenoids was developed, and an optimized technological process for carotenoid composition extraction in pepper leaves was established for the first time. Our final method concluded that six xanthophylls eluted after about 9-26 min. In contrast, four carotenes showed higher retention times after nearly 28-40 min, which significantly shortened time and improved efficiency. Meanwhile, we suggested that 8 mL of 20% KOH-methanol solution should be added to perform saponification at 60 °C for 30 min. The ratio of solid-liquid was 1:8, and the ultrasound-assisted extraction time was 40 min.

Nitric oxide is involved in ethylene-induced adventitious rooting in marigold (Tagetes erecta L.)

Abstract: The plant hormone ethylene and the gaseous signaling molecule nitric oxide (NO) are involved in numerous plant growth and development processes. However, the mechanisms by which their interaction affects adventitious root development in plants is still not adequately studied. In this experiment, the interaction of ethylene and NO in the adventitious rooting process of marigold (Tagetes erecta L. cv. Marvel) was investigated. Treatments with different dosages of ethylene-releasing ethephon (Ethrel®, Bayer Crop Science, Leverkusen, Germany) significantly affected the formation of adventitious roots. The greatest rooting ability was observed in 10 μM ethephon-treated explants. It was further shown that the effect of ethylene could be blocked by the specific NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO), nitric oxide synthase (NOS), enzyme inhibitor NG-nitro-l-Arg-methyl ester (l-NAME), and nitrate reductase inhibitor (NaN3). Moreover, ethephon treatments induced an increase in the endogenous NO levels and significantly improved activities of NOS and nitrate reductase (NR) during rooting. Thus, the induction of adventitious roots by ethylene may be through enhancing the levels of NO. Enzymatic pathway NOS and NR could be responsible for ethylene-induced NO production. Furthermore, ethylene and NO treatments at the appropriate dosage may increase the activities of indoleacetic acid oxidase, peroxidase, and polyphenol oxidase. Our work suggests that the stimulation of adventitious roots by ethylene relies on internal generation of NO.