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Soil microbial community changes imposed by the cumulative effects of root-secreted phenolic acids (PAs) promote soil-borne pathogen establishment and invasion under monoculture systems, but the disease-suppressive soil often exhibits less soil-borne pathogens compared with the conducive soil. So far, it remains poorly understood whether soil disease suppressiveness is associated with the alleviated negative effects of PAs, involving microbial degradation. Here, the long-term monoculture particularly shaped the rhizosphere microbial community, for example by the enrichment of beneficial Pseudomonas species in the suppressive soil and thus enhanced disease-suppressive capacity, however this was not observed for the conducive soil. In vitro PA-degradation assays revealed that the antagonistic Pseudomonas species, together with the Xanthomonas and Rhizobium species, significantly increased the efficiency of PA degradation compared to single species, at least partially explaining how the suppressive soil accumulated lower PA levels than the conducive soil. Pot experiments further showed that this consortium harboring the antagonistic Pseudomonas species can not only lower PA accumulation in the 15-year conducive soils, but also confer stronger Fusarium wilt disease suppression compared with a single inoculum with the antagonistic bacteria. Our findings demonstrated that understanding microbial community functions, beyond the single direct antagonism, facilitated the construction of active consortia for preventing soil-borne pathogens under intensive monoculture.

Soil bacteria can assist plant growth and increase uptake of nutrient elements, the question arises as to whether beneficial soil microbes confer augmented iron (Fe) content of host plants under Fe limited conditions. Herein, a novel strain of Burkholderia cepacia (strain JFW16) was isolated from rhziospheric soils of Astragalus sinicus grown under alkaline conditions. Inoculation of plants with B. cepacia JFW16 displayed increased endogenous Fe content compared with non-inoculated plants. Growth promotion and enhanced photosynthetic capacity were also observed for the inoculated plants. The inoculation with JFW16 significantly increased the rhizospheric acidification, and up-regulated the transcription of some Fe acquisition-associated genes in Astragalus sinicus. Moreover, the metabolic pathways of flavins were remarkably enhanced in the inoculated plants, showing the increased biosynthesis and release of flavins in roots. Collectively, these findings demonstrated the potential of B. cepacia JFW16 to improve Fe assimilation in agricultural crops.

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.

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 Arabidopsis MYB102 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. Arabidopsis MYB102 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.

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.

Volatile organic compounds (VOCs) released by plant growth-promoting rhizobacteria (PGPR) are involved in promoting growth and triggering systemic resistance (ISR) in plants. Importantly, the release of VOCs by some PGPR strains confers improved plant uptake of nutrient elements from the soil. However, the underlying mechanisms of VOCs-regulated nutrient acquisition remain elusive. In this study, VOCs were extracted and identified from Bacillus amyloliquefaciens (strain BF06) using gas chromatography–mass spectrometry (GC–MS). BF06 VOCs exposure significantly promoted the growth and photosynthesis of Arabidopsis plants. To explore how microbial VOCs stimulate growth in plants, gene expression profiles of Arabidopsis seedlings exposed to BF06 VOCs were examined using transcriptomic analyses. In screening differentially expressed genes (DEGs), most upregulated DEGs were found to be related to amino acid transport, iron (Fe) uptake and homeostasis, and sulfate transport. Furthermore, BF06 VOCs significantly enhanced Fe absorption in plants under Fe-limited conditions. However, when nitric oxide (NO) synthesis was inhibited, BF06 VOCs exposure could not substantially augment Fe acquisition in plants under alkaline stress, indicating that VOCs-mediated plant uptake of Fe was required for induction of root NO accumulation. In addition, BF06 VOCs exposure led to a marked increase in some genes encoding for sulfate transporters, and further increased Se accumulation in plants. Intriguingly, BF06 VOCs exposure failed to increase Se uptake in sultr1;2 mutants, which may indicate that high-level transcription of these sulfate transporters induced by BF06 VOCs was essential for enhancing Se absorption by plants. Taken together, our results demonstrated the potential of VOCs released by this strain BF06 to increase Fe and Se uptake in plants.

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.

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.

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.

It is well documented that the homeodomain-leucine zipper (HD-Zip) transcription factors play diverse roles during plant growth and development. The Class I HD-Zip genes are shown to be involved in the regulation of abiotic stress responses and tolerance. Herein, a novel Class I HD-Zip gene, EsHD1, was isolated from Eutrema salsugineum, and an investigation was performed of its physiological functions in response to drought stress. The analyses of gene expression profiles revealed that the EsHD1 transcripts quickly accumulated upon exposure to various abiotic stress conditions or abscisic acid treatments. Furthermore, the overexpression of EsHD1 in tobacco plants conferred improved tolerance to drought stress. The EsHD1-overexpressing lines had lower levels of reactive oxygen species (ROS), ion leakage, and malondialdehyde, but they manifested higher activities of antioxidant enzymes and the transcription of pathogenic-related genes as compared to wild-type plants under drought stress conditions. Therefore, our findings demonstrated that EsHD1 positively regulated plant drought tolerance via alleviation of ROS-mediated membrane damage.

Previous studies have indicated that homeodomain-leucine zipper (HD-Zip) transcription factors play important roles during abiotic stress, but there is no information on the functions of HD-Zip genes in a new model plant Eutrema salsugineum for studying plant abiotic stress tolerance. Here, EsHdzip1 (GenBank No. XM_006390503) belonging to the Class IV subgroup of HD-Zip transcription factor family was isolated from E. salsugineum and characterized for its physiological roles under drought stress conditions. Transgenic tobacco plants overexpressing EsHdzip1 exhibited increased drought resistance with promoted root growth and reduction of water loss. Furthermore, these transgenic plants had lower ion leakage (IL), malondialdehyde (MDA), and reactive oxygen species (ROS) accumulation, but higher content of osmotic solutes (proline and total soluble sugars) and activities of antioxidant enzymes including superoxide dismutase (SOD) and ascorbate peroxidase (APX) relative to wild-type (WT) plants when subjected to drought stress treatments. The content of abscisic acid (ABA) was also observed to be remarkably higher in the transgenic lines than WT plants under drought stress conditions. In addition, the expression levels of three important stress-related genes (NtP5CS, NtERD10C, and NtLEA5) involved in the osmotic adjustment and water maintenance were significantly higher than WT plants under drought stress conditions. Therefore, we have revealed important roles of the EsHdzip1 gene in response to drought stress, suggesting that this gene has a great potential for improving plant drought tolerance by engineering manipulation.

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.

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.

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.

Travel time is a very important indicator which reflects the situation of road traffic. This article designed a method for computing the road travel time by the driving path which was obtained based on license plate recognition technology. With analysis of the on-site verification results, this paper showed that the algorithm can satisfy the accuracy and timeliness in the complex road conditions.