Transcriptome response of maize (Zea mays L.) seedlings to heat stress.

Among stresses, heat stress (HS) is a prime factor restricting plant growth and productivity. However, the molecular mechanisms of plants' response to HS need to be further uncovered. Here, the transcriptome response of maize seedlings to HS was dissected using transcriptome data analysis. The data exhibited that a total of 43,221 genes in maize seedlings had been found, 37,534 of which were referred, while 5686 were not. Under HS, comparison with the control without HS, there were 13,607 genes that were differentially expressed (DEGs, 6195 upregulated and 7412 downregulated). In addition, Gene Ontology (GO) enrichment analysis indicated that there were 220, 478, and 1300 terms that were enriched in cellular component, molecular function, and biological process, respectively. Significantly enriched GO terms were involved in 23 cellular components, 27 molecular functions, and 124 biological processes. Also, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis suggested that there were 2613 DEGs that were assigned to 131 pathways, 14 of which (enriched 1068 DEGs in total) were significantly upregulated. These pathways were mainly related to protein renaturation, biomembrane repair, osmotic adjustment, and redox balance. Among them, protein processing in endoplasmic reticulum was the most significantly upregulated. The transcriptome data decoded that protein renaturation, biomembrane repair, osmotic adjustment, and redox balance played a key role in the response of maize seedlings to HS.

Interplay between hydrogen sulfide and methylglyoxal initiates thermotolerance in maize seedlings by modulating reactive oxidative species and osmolyte metabolism.

Hydrogen sulfide (H2S) and methylglyoxal (MG) were supposed to be novel signaling molecules in plants. However, whether interplay between H2S and MG can initiate thermotolerance in maize seedlings and in relation to metabolism of reactive oxygen species (ROS) and osmolytes is little known. In this study, watering with MG and NaHS (H2S donor) alone or in combination elevated survival and tissue vigor of maize seedlings under heat stress and coped with an increase in the biomembrane injury (as indicated in membrane lipid peroxidation and electrolyte leakage). The above-mentioned effects were separately weakened by MG scavengers (N-acetyl cysteine: NAC; aminoguanidine: AG) and H2S inhibitor (DL-propargylglycine, PAG) and scavenger (hypotaurine, HT). These suggested that the interplay between H2S and MG initiated the thermotolerance in maize seedlings. The further data indicated that, under non-heat stress and heat stress conditions, MG and NaHS alone or in combination modulated ROS metabolism by regulating the activities of antioxidant enzymes (catalase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase, monodehydroascorbate reductase, and dehydroascorbate reductase) and the contents of non-enzymatic antioxidants (ascorbic acid, glutathione, flavonoids, and carotenoids) in maize seedlings. In addition, MG and NaHS alone or in combination also separately modulated the metabolism of osmolytes (proline, trehalose, glycine betaine, and total soluble sugar), H2S (L-cysteine desulfhydrase and O-acetylserine (thione) lyase), and MG (glyoxalase I, glyoxalase II, and MG reductase). These physiological effects also were separately impaired by NAC, AG, PAG, and HT. The current data illustrated that the interplay between H2S and MG initiated the thermotolerance in maize seedlings by modulating ROS, osmolyte, H2S, and MG metabolism.

Methylglyoxal triggers the heat tolerance in maize seedlings by driving AsA-GSH cycle and reactive oxygen species-/methylglyoxal-scavenging system.

Traditionally, methylglyoxal (MG) was looked upon as a toxic byproduct of cellular metabolism. Nowadays, MG has been found to be a novel signaling molecule. However, whether MG can trigger the heat tolerance in maize seedlings and the underlying mechanisms is still elusive. In this study, the maize seedlings irrigated with MG increased the survival percentage of seedlings under heat stress (HS), remitted a decrease in tissue vitality and an increase in electrolyte leakage, and reduced membrane lipid peroxidation, implying MG could trigger the heat tolerance of maize seedlings. The further experiments showed that MG drove the ascorbic acid (AsA)-glutathione (GSH) cycle by activating enzymes (glutathione reductase, monodehydroascorbate reductase, dehydroascorbate reductase, and ascorbate peroxidase) and increasing the contents of antioxidants (AsA and GSH) and the ratio of GSH/(GSH + oxidized glutathione) and AsA/(AsA + dehydroascorbate) under both non-HS and HS. Also, the reactive oxygen species (ROS)-scavenger system (catalase, guaiacol peroxidase, carotenoid, total phenols, and flavonoids) and MG-scavenger system (glyoxalase I and glyoxalas II) also were up-regulated in maize seedlings pretreated with MG under non-HS and HS. This work for the first time reported that MG could trigger the heat tolerance of maize seedlings by driving the AsA-GSH cycle and ROS-/MG-scavenging system.

Glutamate signaling enhances the heat tolerance of maize seedlings by plant glutamate receptor-like channels-mediated calcium signaling.

Glutamate (Glu), a neurotransmitter in animal, is a novel signaling molecule in plants, which takes part in cellular metabolism, seed germination, plant growth, development, and long-distance information transfer. However, whether Glu can enhance the heat tolerance in maize seedlings and its relation to calcium signaling is still elusive. In this study, maize seedlings were pretreated with Glu and then exposed to heat stress. The results showed that Glu pretreatment enhanced the survival percentage of maize seedlings under heat tolerance, indicating that Glu could increase the heat tolerance of maize seedlings. The Glu-induced heat tolerance was weakened by exogenous calcium chloride, plasma membrane Ca2+ channel blocker (LaCl3), Ca2+ chelator (ethylene glycol-bis(b-aminoethylether)-N,N, N΄,N΄-tetraacetic acid), calmodulin antagonists (trifluoperazine and chlopromazine), and plant glutamate receptor-like antagonists (MgCl2 and 6,7-dinitroquinoxaline- 2,3-(1H,4H)- dione). These findings for the first time reported that Glu could increase the heat tolerance of maize seedlings by plant glutamate receptor-like channels-mediated calcium signaling.

Signaling Role of Glutamate in Plants.

It is well known that glutamate (Glu), a neurotransmitter in human body, is a protein amino acid. It plays a very important role in plant growth and development. Nowadays, Glu has been found to emerge as signaling role. Under normal conditions, Glu takes part in seed germination, root architecture, pollen germination, and pollen tube growth. Under stress conditions, Glu participates in wound response, pathogen resistance, response and adaptation to abiotic stress (such as salt, cold, heat, and drought), and local stimulation (abiotic or biotic stress)-triggered long distance signaling transduction. In this review, in the light of the current opinion on Glu signaling in plants, the following knowledge was updated and discussed. 1) Glu metabolism; 2) signaling role of Glu in plant growth, development, and response and adaptation to environmental stress; as well as 3) the underlying research directions in the future. The purpose of this review was to look forward to inspiring the rapid development of Glu signaling research in plant biology, particularly in the field of stress biology of plants.

Signaling Molecule Hydrogen Sulfide Improves Seed Germination and Seedling Growth of Maize (Zea mays L.) Under High Temperature by Inducing Antioxidant System and Osmolyte Biosynthesis.

Hydrogen sulfide (H2S) is a novel type signaling molecule in plants. Seed germination is a key stage of life cycle of plants, which is vulnerable to environmental stress including high temperature. However, under high temperature stress, whether pre-soaking of maize seeds with NaHS (a H2S donor) could improve seed germination and seedling growth and the possible mechanisms are not completely clear. In this study, maize seeds pre-soaked with NaHS enhanced germination percentage, sprout length, root length, and fresh weight compared with the control without NaHS treatment, illustrating that H2S could improve maize seed germination and seedling growth under high temperature. In addition, in comparison to the control, NaHS pre-soaking stimulated antioxidant enzymes [ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT)] activities and the contents of water soluble non-enzymatic antioxidants [ascorbic acid (AsA) and glutathione (GSH)], as well as the ratio of reduced antioxidant to oxidized antioxidant. Moreover, pre-soaking with NaHS activated Δ1-pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase [OAT; both are rate-limiting enzymes in proline (Pro) synthesis], betaine aldehyde dehydrogenase [BADH; a key enzyme in glycine betaine (GB)], and trehalose (Tre)-6-phosphate phosphatase (a key step in Tre synthesis), which in turn accumulated Pro, GB, and Tre in germinating seeds compared with the control. Also, an improved germination by NaHS under high temperature was reinforced by the above osmotic adjustment substances (osmolytes) alone, while deteriorated by the inhibitors of osmolyte biosynthesis [gabaculine (GAB), disulfiram (DSF), and sodium citrate (SC)]. These results imply that H2S could improve maize seed germination and seedling growth under high temperature by inducing antioxidant system and osmolyte biosynthesis.

Pharmacokinetics and pharmacodynamics of multiple-dose intravenous nemonoxacin in healthy Chinese volunteers.

This study evaluated the safety and pharmacokinetic/pharmacodynamic profiles of nemonoxacin in healthy Chinese volunteers following multiple-dose intravenous infusion once daily for 10 consecutive days. The study was composed of two stages. In the open-label stage, 500 mg or 750 mg of nemonoxacin (n = 12 each) was administered at an infusion rate of 5.56 mg/min. In the second stage, with a randomized double-blind placebo-controlled design, 500, 650, or 750 mg of nemonoxacin (n = 16 in each cohort; 12 subjects received the drug and the other 4 subjects received the placebo) was given at an infusion rate of 4.17 mg/min. The results showed that, in the first stage, the maximal nemonoxacin concentrations (mean ± SD) at steady state (Cmax_ss) were 9.60 ± 1.84 and 11.04 ± 2.18 µg/ml in the 500-mg and 750-mg cohorts, respectively; the areas under the concentration-time curve at steady state (AUC0-24_ss) were 44.03 ± 8.62 and 65.82 ± 10.78 µg · h/ml in the 500-mg and 750-mg cohorts, respectively. In the second stage, the nemonoxacin Cmax_ss values were 7.13 ± 1.47, 8.17 ± 1.76, and 9.96 ± 2.23 µg/ml in the 500-mg, 650-mg, and 750-mg cohorts, respectively; the AUC0-24_ss values were 40.46 ± 9.52, 54.17 ± 12.10, and 71.34 ± 17.79 µg · h/ml in the 500-mg, 650-mg, and 750-mg cohorts, respectively. No accumulation was found after the 10-day infusion with any regimen. The drug was well tolerated. A Monte Carlo simulation indicated that the cumulative fraction of response of any dosing regimen was nearly 100% against Streptococcus pneumoniae. The probability of target attainment of nemonoxacin therapy was >98% when the MIC of nemonoxacin against S. pneumoniae was ≤1 mg/liter. It is suggested that all of the studied intravenous nemonoxacin dosing regimens should have favorable clinical and microbiological efficacies in future clinical studies. (This study has been registered at ClinicalTrials.gov under registration no. NCT01944774.).

[Comparative analysis of variable region of white spot syndrome virus genome in Penaeus vannamei in Guangxi, China].

Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank.

[Study of pharmacokinetics/pharmacodynamics of levofloxacin].

OBJECTIVE: To evaluate the dosing regimens of levofloxacin. METHODS: The drug concentrations in serum and urine were assayed by HPLC method and the pharmacokinetic parameters were calculated after intravenous infusion of a single dose of 200 mg, 300 mg and 500 mg levofloxacin to healthy volunteers. The in vitro activity MIC of levofloxacin against 823 clinical isolates were determined and compared with other antimicrobial agents. Based on the above results, the PK/PD parameters C(max)/MIC and AUC/MIC were calculated and the dosing regimens of levofloxacin were proposed for infections caused by different pathogens. RESULTS: The results of clinical pharmacokinetic study showed that the C(max) of levofloxacin was 3.4 mg/L +/- 0.8 mg/L, 4.8 mg/L +/- 1.4 mg/L and 7.6 mg/L +/- 1.1 mg/L respectively, AUC(0-infinity) was 14.4 mg.h/L +/- 2.5 mg.h/L, 21.9 mg.h/L +/- 4.5 mg.h/L and 38.3 mg.h/L +/- 4.9 mg.h/L respectively, T(1)/2 beta was 6.2 h +/- 0.4 h, 6.4 h +/- 0.9 h and 6.5 h +/- 0.6 h respectively, and 69% +/- 5%, 69% +/- 6%, and 65% +/- 4% of the doses were excreted in urine within 24 h after intravenous infusion of a single dose of levofloxacin 200 mg, 300 mg and 500 mg. The in vitro pharmacodynamic study showed that levofloxacin was highly active against Hemolytic streptococcus and Moraxella catarrhalis. It was active against Streptococcus pneumoniae (including penicillin nonsusceptible strains), Hemophilus influenzae, methicillin-sensitive Staphylococcus aureus (MSSA), Klebsiella pneumoniae and Stenotrophomonas maltophilia. Levofloxacin also had good activity against Pseudomonas aeruginosa and K.pneumoniae. However, most of isolates of enterococcal spp. were resistant to levofloxacin. Above 50% of Escherichia coli isolates were resistant to levofloxacin. Based on the results of PK/PD parameters, the adequate dosing regimens of levofloxacin should be once daily 200 mg once daily of levofloxacin was expected to be effective for the treatment of infections caused by M. catarrhalis. The regimen of 300 mg once daily could be effective for the treatment of infections caused by Hemolytic streptococcus. 500 mg once daily of levofloxacin was expected to be effective for the treatment of respiratory tract infections caused by S. pneumoniae or H. influenzae. For treatment of respiratory tract infections and urinary tract infections caused by levofloxacin-susceptible organisms including K. pneumoniae, E. coli, P. aeruginosa, and S.maltophilia, 500 mg once daily of levofloxacin was needed to obtain good clinical efficacy. But PK/PD parameters predicted that 500 mg daily of levofloxacin was not effective for infections caused by methicillin-resistant Staphylococcus aureus (MRSA) and Enterococci. CONCLUSION: The proposed dosing regimens of levofloxacin based on PK/PD concepts are expected to provide good efficacy in clinical practice.