Enterobacter asburiae Reduces Cadmium Toxicity in Maize Plants by Repressing Iron Uptake-Associated Pathways.

Soil microbes have recently been utilized to improve cadmium (Cd) tolerance and lower its accumulation in plants. Nevertheless, whether rhizobacteria can prevent Cd uptake by graminaceous plants and the underlying mechanisms remain elusive. In this study, inoculation with Enterobacter asburiae NC16 reduced transpiration rates and the expression of some iron (Fe) uptake-related genes including ZmFer, ZmYS1, ZmZIP, and ZmNAS2 in maize (Zea mays) plants, which contributed to mitigation of Cd toxicity. However, the inoculation with NC16 failed to suppress the transpiration rates and transcription of these Fe uptake-related genes in plants treated with fluridone, an abscisic acid (ABA) biosynthetic inhibitor, indicating that the impacts of NC16-inoculation observed were dependent on the actions of ABA. We found that NC16 increased the host ABA levels by mediating the metabolism of ABA rather than its synthesis. Moreover, the capacity of NC16 to inhibit plant uptake of Cd was greatly weakened in plants overexpressing ZmZIP, encoding a zinc/iron transporter. Collectively, our findings indicated that E. asburiae NC16 reduced Cd toxicity in maize plants at least partially by hampering the Fe uptake-associated pathways.

Bacillus subtilis STU6 Ameliorates Iron Deficiency in Tomato by Enhancement of Polyamine-Mediated Iron Remobilization.

Iron (Fe) deficiency often triggers arginine overproduction in plants. However, it remains elusive whether Fe deficiency-induced increases of arginine levels are involved in beneficial rhizobacteria recruitment and that the mechanism underlying rhizobacteria induced plant Fe deficiency tolerance. Here, Bacillus subtilis STU6 increased soluble Fe content in tomato, thereby alleviating Fe deficiency-induced chlorosis. In a split-root system, STU6 significantly induced arginine exudation by Fe-deficient roots, and increased arginine levels promoted spermidine (Spd) production by STU6 and bacterial colonization. Deletion of the STU6 speB gene inhibited Spd synthesis and abrogated STU6-induced increments of soluble Fe content in the Fe-deficient plants. Increased host Spd levels by STU6 greatly stimulated the NO accumulation in the Fe-deficient roots. Furthermore, disruption of NO signaling markedly repressed STU6-mediated cell wall Fe remobilization. Collectively, our data provide important evidence that chemical dialogues between tomato and STU6 contribute to enhancement of microbe-mediated plant adaptation to Fe deficiency.

Arabidopsis Transcription Factor MYB102 Increases Plant Susceptibility to Aphids by Substantial Activation of Ethylene Biosynthesis.

Induction of ethylene biosynthesis by aphids increases the susceptibility of several plant species to aphids. Recent studies have indicated that some MYB transcription factors regulate the phloem-based defense against aphid infestation by modulating ethylene (ET) signaling. Arabidopsis MYB102 has previously been shown to be induced by wound signaling and regulate defense response against chewing insects. However, it remains unclear whether ArabidopsisMYB102 takes part in the defense response of plants to aphids. Here, we investigated the function of MYB102 in the response of Arabidopsis to aphid infestation. ArabidopsisMYB102 was primarily expressed in vascular tissues, and its transcription was remarkably induced by green peach aphids (GPA; Myzus persicae). The results of RNA-Sequencing revealed that overexpression of MYB102 in Arabidopsis promoted ET biosynthesis by upregulation of some 1-aminocyclopropane-1-carboxylate synthase (ACS) genes, which are rate-limiting enzymes of the ET-synthetic pathway. Enhanced ET levels led to reduced Arabidopsis resistance to GPA. Furthermore, dominant suppression of MYB102 inhibited aphid-induced increase of ET levels in Arabidopsis. In agreement with a negative regulatory role for ET in aphid defense responses, the MYB102-overexpressing lines were more susceptible to GPA than wild-type (WT) plants. Overexpression of MYB102 in Arabidopsis obviously repressed aphid-induced callose deposition. Conversely, overexpression of MYB102 failed to increase aphid susceptibility in both the ET-insensitive mutants and plants treated with inhibitors of ET signaling pathways, demonstrating that the ET was critical for promoting aphid performance conferred by overexpression of MYB102. Collectively, our findings indicate that the Arabidopsis MYB102 increases host susceptibility to GPA through the ET-dependent signaling pathways.

Involvement of abscisic acid in microbe-induced saline-alkaline resistance in plants.

Soil salinity-alkalinity is one of abiotic stresses that lead to plant growth inhibition and yield loss. It has recently been indicated that plant growth promoting rhizobacteria (PGPR) can enhance the capacity of plants to counteract negative effects caused by adverse environments. However, whether PGPR confers increased saline-alkaline resistance of plants and the underlying mechanisms remain unclear. We thus investigated the effects of Bacillus licheniformis (strain SA03) on Chrysanthemum plants grown under saline-alkaline conditions. Soil inoculation with SA03 significantly mitigated saline-alkaline stress in plants with augmented photosynthesis, biomass and survival rates. Moreover, the inoculated plants accumulated more Fe and less Na+ content than the non-inoculated plants under the stress. However, the inoculation with SA03 failed to trigger a series of saline-alkaline stress responses in abscisic acid (ABA)- and nitric oxide (NO)-deficient plants. Furthermore, NO acted as a secondary messenger of ABA to regulate the stress responses and tolerance in Chrysanthemum plants. Therefore, these findings indicated that B. licheniformis SA03 could be employed to improve saline-alkaline tolerance of plants by mediating cellular ABA levels.

Bacillus licheniformis SA03 Confers Increased Saline-Alkaline Tolerance in Chrysanthemum Plants by Induction of Abscisic Acid Accumulation.

Soil saline-alkalization is a major abiotic stress that leads to low iron (Fe) availability and high toxicity of sodium ions (Na+) for plants. It has recently been shown that plant growth promoting rhizobacteria (PGPR) can enhance the ability of plants to tolerate multiple abiotic stresses such as drought, salinity, and nutrient deficiency. However, the possible involvement of PGPR in improving saline-alkaline tolerance of plants and the underlying mechanisms remain largely unknown. In this study, we investigated the effects of Bacillus licheniformis (strain SA03) on the growth of Chrysanthemum plants under saline-alkaline conditions. Our results revealed that inoculation with SA03 alleviated saline-alkaline stress in plants with increased survival rates, photosynthesis and biomass. The inoculated plants accumulated more Fe and lower Na+ concentrations under saline-alkaline stress compared with the non-inoculated plants. RNA-Sequencing analyses further revealed that SA03 significantly activated abiotic stress- and Fe acquisition-related pathways in the stress-treated plants. However, SA03 failed to increase saline-alkaline tolerance in plants when cellular abscisic acid (ABA) and nitric oxide (NO) synthesis were inhibited by treatment with fluridone (FLU) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), respectively. Importantly, we also found that NO acted downstream of SA03-induced ABA to activate a series of adaptive responses in host plants under saline-alkaline stress. These findings demonstrated the potential roles of B. licheniformis SA03 in enhancing saline-alkaline tolerance of plants and highlighted the intricate integration of microbial signaling in regulating cellular Fe and Na+ accumulation.

Bacillus amyloliquefaciens SAY09 Increases Cadmium Resistance in Plants by Activation of Auxin-Mediated Signaling Pathways.

Without physical contact with plants, certain plant growth-promoting rhizobacteria (PGPR) can release volatile organic compounds (VOCs) to regulate nutrient acquisition and induce systemic immunity in plants. However, whether the PGPR-emitted VOCs can induce cadmium (Cd) tolerance of plants and the underlying mechanisms remain elusive. In this study, we probed the effects of Bacillus amyloliquefaciens (strain SAY09)-emitted VOCs on the growth of Arabidopsis plants under Cd stress. SAY09 exposure alleviates Cd toxicity in plants with increased auxin biosynthesis. RNA-Seq analyses revealed that SAY09 exposure provoked iron (Fe) uptake- and cell wall-associated pathways in the Cd-treated plants. However, SAY09 exposure failed to increase Cd resistance of plants after treatment with 1-naphthylphthalamic acid (NPA) or 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO). Under Cd stress, SAY09 exposure markedly promoted Fe absorption in plants with the increased hemicellulose 1 (HC1) content and Cd deposition in root cell wall, whereas these effects were almost abrogated by treatment with NPA or c-PTIO. Moreover, exogenous NPA remarkably repressed the accumulation of nitric oxide (NO) in the SAY09-exposed roots under Cd stress. Taken together, the findings indicated that NO acted as downstream signals of SAY09-induced auxin to regulate Fe acquisition and augment Cd fixation in roots, thereby ameliorating Cd toxicity.

Exogenous Melatonin Improves Plant Iron Deficiency Tolerance via Increased Accumulation of Polyamine-Mediated Nitric Oxide.

Melatonin has recently been demonstrated to play important roles in the regulation of plant growth, development, and abiotic and biotic stress responses. However, the possible involvement of melatonin in Fe deficiency responses and the underlying mechanisms remained elusive in Arabidopsis thaliana. In this study, Fe deficiency quickly induced melatonin synthesis in Arabidopsis plants. Exogenous melatonin significantly increased the soluble Fe content of shoots and roots, and decreased the levels of root cell wall Fe bound to pectin and hemicellulose, thus alleviating Fe deficiency-induced chlorosis. Intriguingly, melatonin treatments induced a significant increase of nitric oxide (NO) accumulation in roots of Fe-deficient plants, but not in those of polyamine-deficient (adc2-1 and d-arginine-treated) plants. Moreover, the melatonin-alleviated leaf chlorosis was blocked in the polyamine- and NO-deficient (nia1nia2noa1 and c-PTIO-treated) plants, and the melatonin-induced Fe remobilization was largely inhibited. In addition, the expression of some Fe acquisition-related genes, including FIT1, FRO2, and IRT1 were significantly up-regulated by melatonin treatments, whereas the enhanced expression of these genes was obviously suppressed in the polyamine- and NO-deficient plants. Collectively, our results provide evidence to support the view that melatonin can increase the tolerance of plants to Fe deficiency in a process dependent on the polyamine-induced NO production under Fe-deficient conditions.

Rhizobacterial Strain Bacillus megaterium BOFC15 Induces Cellular Polyamine Changes that Improve Plant Growth and Drought Resistance.

Plant-growth-promoting rhizobacteria can improve plant growth, development, and stress adaptation. However, the underlying mechanisms are still largely unclear. We investigated the effects of Bacillus megaterium BOFC15 on Arabidopsis plants. BOFC15 produced and secreted spermidine (Spd), a type of polyamine (PA) that plays an important role in plant growth. Moreover, BOFC15 induced changes in the cellular PAs of plants that promoted an increase of free Spd and spermine levels. However, these effects were remarkably abolished by the addition of dicyclohexylamine (DCHA), a Spd biosynthetic inhibitor. Additionally, the inoculation with BOFC15 remarkably increased plant biomass, improved root system architecture, and augmented photosynthetic capacity. Inoculated plants also displayed stronger ability to tolerate drought stress than non-inoculated (control) plants. Abscisic acid (ABA) content was notably higher in the inoculated plants than in the control plants under drought stress and polyethylene glycol (PEG)-induced stress conditions. However, the BOFC15-induced ABA synthesis was markedly inhibited by DCHA. Thus, microbial Spd participated in the modulation of the ABA levels. The Spd-producing BOFC15 improved plant drought tolerance, which was associated with altered cellular ABA levels and activated adaptive responses.

Paenibacillus polymyxa BFKC01 enhances plant iron absorption via improved root systems and activated iron acquisition mechanisms.

Despite the high abundance of iron (Fe) in most earth's soils, Fe is the major limiting factor for plant growth and development due to its low bioavailability. With an increasing recognition that soil microbes play important roles in plant growth, several strains of beneficial rhizobactria have been applied to improve plant nutrient absorption, biomass, and abiotic or biotic stress tolerance. In this study, we report the mechanisms of microbe-induced plant Fe assimilation, in which the plant growth promoting rhizobacteria (PGPR) Paenibacillus polymyxa BFKC01 stimulates plant's Fe acquisition machinery to enhance Fe uptake in Arabidopsis plants. Mechanistic studies show that BFKC01 transcriptionally activates the Fe-deficiency-induced transcription factor 1 (FIT1), thereby up-regulating the expression of IRT1 and FRO2. Furthermore, BFKC01 has been found to induce plant systemic responses with the increased transcription of MYB72, and the biosynthetic pathways of phenolic compounds are also activated. Our data reveal that abundant phenolic compounds are detected in root exudation of the BFKC01-inoculated plants, which efficiently facilitate Fe mobility under alkaline conditions. In addition, BFKC01 can secret auxin and further improved root systems, which enhances the ability of plants to acquire Fe from soils. As a result, BFKC01-inoculated plants have more endogenous Fe and increased photosynthetic capacity under alkaline conditions as compared to control plants. Our results demonstrate the potential roles of BFKC01 in promoting Fe acquisition in plants and underline the intricate integration of microbial signaling in controlling plant Fe acquisition.

Resveratrol Protects against Helicobacter pylori-Associated Gastritis by Combating Oxidative Stress.

Helicobacter pylori (H. pylori)-induced oxidative stress has been shown to play a very important role in the inflammation of the gastric mucosa and increases the risk of developing gastric cancer. Resveratrol has many biological functions and activities, including antioxidant and anti-inflammatory effect. The purpose of this study was to probe whether resveratrol inhibits H. pylori-induced gastric inflammation and to elucidate the underlying mechanisms of any effect in mice. A mouse model of H. pylori infection was established via oral inoculation with H. pylori. After one week, mice were administered resveratrol (100 mg/kg body weight/day) orally for six weeks. The mRNA and protein levels of iNOS and IL-8 were assessed using RT-PCR, Western blot and ELISA. The expression levels of IκBα and phosphorylated IκBα (which embodies the level and activation of NF-κB), Heme Oxygenase-1 (HO-1; a potent antioxidant enzyme) and nuclear factor-erythroid 2 related factor 2 (Nrf2) were determined using Western blot, and lipid peroxide (LPO) level and myeloperoxidase (MPO) activity were examined using an MPO colorimetric activity assay, thiobarbituric acid reaction, and histological-grade using HE staining of the gastric mucosa. The results showed that resveratrol improved the histological infiltration score and decreased LPO level and MPO activity in the gastric mucosa. Resveratrol down-regulated the H. pylori-induced mRNA transcription and protein expression levels of IL-8 and iNOS, suppressed H. pylori-induced phosphorylation of IκBα, and increased the levels of HO-1 and Nrf2. In conclusion, resveratrol treatment exerted significant effects against oxidative stress and inflammation in H. pylori-infected mucosa through the suppression of IL-8, iNOS, and NF-κB, and moreover through the activation of the Nrf2/HO-1 pathway.

Design of signal peptide bombyxin and its effect on secretory expression efficiency and levels of Helicobacter pylori urease subunit B in silkworm cells and larvae

This study employed a Bac-to-Bac/Bombyx mori bioreactor to mass-produce immunogenic urease subunit B (UreB) from Helicobacter pylori. The signal peptide bombyxin from B. mori was used to promote secretory expression to improve expression levels and was designed and integrated into the UreB gene to generate the Bacmid/BmNPV/(signal peptide)-UreB baculovirus expression system. To determine whether the bombyxin signal peptide resulted in secretory expression of recombinant UreB (rUreB) and to determine the secretory efficiency, we tested the secretory expression level of rUreB in Bm5 cells using ELISA. To further investigate whether secretory expression affected cell viability, cells were evaluated using 0.4% trypan blue staining, and Bacmid/BmNPV/UreB without the signal peptide served as a control. The above recombinant bacmid constructs were injected to silkworm larvae, and the secretory expression level of rUreB was detected using SDS-PAGE and semi-quantitative western blot analysis. The results indicated that the bombyxin signal peptide directed the secretory expression of rUreB and that this expression improved the viability of Bm5 cells. Moreover, the results showed that the expression level of rUreB was 1.5 times higher with the Bacmid/BmNPV constructs containing the bombyxin signal sequence than those without the signal sequence. These results demonstrate that secretory expression can enhance rUreB expression levels and is likely to aid in the large-scale expression and yield of rUreB in silkworm larvae.

The synthetic antimicrobial peptide pexiganan and its nanoparticles (PNPs) exhibit the anti-helicobacter pylori activity in vitro and in vivo.

The aim of this study was to probe the potential anti-H. pylori activity of the synthetic antimicrobial peptide pexiganan, which is an analog of the peptide magainin, and its nanoparticles (PNPs) that were prepared in our laboratory. To compare their antibacterial effects in vitro and in vivo, studies of H. pylori growth inhibition, kinetics and resistance assays were undertaken. The gastric mucoadhesive efficiency and H. pylori clearance efficiency of pexiganan and PNPs were evaluated in rats and mice infected with H. pylori. The eradication of H. pylori was determined using urease tests and a microbial culture method. We observed that PNPs adhered to gastric mucosa more effectively owing to a prolonged stay in the stomach, which resulted in a more effective H. pylori clearance. In addition, PNPs had greater anti-H. pylori effect than pexiganan in infected mice. The amount of pexiganan required to eradicate H. pylori was significantly less using PNPs than the corresponding pexiganan suspension. The results confirmed that PNPs improved peptide stability in the stomach and more effectively eradicated H. pylori from mice stomachs than pexiganan.

The role of wheat germ agglutinin in the attachment of Pseudomonas sp. WS32 to wheat root.

Wheat germ agglutinin (WGA), which is secreted on the surface of wheat root, has been defined as a protein that reversibly and non-enzymatically binds to specific carbohydrates. However, little attention has been paid to the function of WGA in the attachment of bacteria to their host plants. The aim of this study was to investigate the role of WGA in the attachment of Pseudomonas sp. WS32 to wheat roots. Wheat roots were initially treated with double-distilled water, WGA-H (WGA solution that was heated at 100°C for 15 min) and WGA, independently. Subsequently, the roots were co-incubated with cell solutions (109 cells/ml). A dilution plate method using a solid nutrient medium was employed to determine the adsorption of WS32 to wheat roots. WGA was labeled with fluorescein isothiocyanate and detected using the fluorescent in situ hybridization (FISH) technique. The number of adsorptive WS32 cells on wheat roots was significantly increased when the wheat roots were pretreated with WGA, compared with the control treatment (p = 0.01). However, WGA-H failed to increase the amount of bacterial cells that attached to the wheat roots because of the loss of its physiological activity. The FISH assay also revealed that more cells adhered to WGA-treated wheat roots than to control or WGA-H-treated roots. The results indicated that WGA can mediate Pseudomonas strain WS32's adherence to wheat seedling roots. The findings of this study provide a better understanding of the processes involved in plant-microbe interactions.

Isolation and characterization of plant growth-promoting rhizobacteria from wheat roots by wheat germ agglutinin labeled with fluorescein isothiocyanate.

Thirty-two isolates were obtained from wheat rhizosphere by wheat germ agglutinin (WGA) labeled with fluorescein isothiocyanate (FITC). Most isolates were able to produce indole acetic acid (65.6%) and siderophores (59.3%), as well as exhibited phosphate solubilization (96.8%). Fourteen isolates displayed three plant growth-promoting traits. Among these strains, two phosphate-dissolving ones, WS29 and WS31, were evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum Wan33). Strain WS29 and WS31 significantly promoted the development of lateral roots by 34.9% and 27.6%, as well as increased the root dry weight by 25.0% and 25.6%, respectively, compared to those of the control. Based on 16S rRNA gene sequence comparisons and phylogenetic positions, both isolates were determined to belong to the genus Bacillus. The proportion of isolates showing the properties of plant growth-promoting rhizobacteria (PGPR) was higher than in previous reports. The efficiency of the isolation of PGPR strains was also greatly increased by WGA labeled with FITC. The present study indicated that WGA could be used as an effective tool for isolating PGPR strains with high affinity to host plants from wheat roots. The proposed approach could facilitate research on biofertilizers or biocontrol agents.

[Study on the extraction of mycelium polysaccharides from selenium-rich Morchella esculenta with neutral proteinase method].

OBJECTIVE: To study the extraction conditions of mycelium polysaccharides from selenium-rich Morchella esculenta with neutral proteinase for increasing of yield. METHODS: On the basis of single factor tests, orthogonal experiment design was applied to analyze the influence of factors including enzyme dosage, enzymolysis time, enzymolysis temperature and material/liquid ratio on the extraction rate of the polysaccharides. RESULTS: The optimal extraction conditions of mycelium polysaccharides from selenium-rich Morchella esculenta was as follows: enzyme dosage was 1.5%, enzymolysis time was 2 h, enzymolysis temperature was 40 degrees C, and material/liquid ratio was 1:15. The extraction rate of polysaccharide was 11.26% under the extraction conditions. CONCLUSION: The process is simple, stable and practicable, and can be used for the extraction of mycelium polysaccharides from selenium-rich Morchella esculenta.

[Comparison of genetic linkage maps based on F2F6 populations derived from rice subspecies cross].

A F2 population containing 180 lines, which was derived from the cross between the partially sequenced indica variety "Pei'ai 64S" and the completely sequenced japonica variety "Nipponbare" , was used to construct a genetic linkage framework map (referred to as F2 map), which included 138 microsatellite sites and covered 1737.81 cM of total genomic length, an average distance of 11.90 cM. Single seed descent F2:6 population with 330 lines was used to construct a genetic linkage map (known as F6 map) using 92 markers. The total genomic length and average distance were 2563.5 cM and 27.86 cM, respectively. The F2 and F6 maps differed in linkage groups, mapped markers, sequenced order of markers, ge-netic distance and average distance on the maps. Preliminary analysis about these difference was carried out.