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Bacillus velezensis strain GB03 is a Gram-positive rhizosphere bacterium known for its ability to promote plant growth and immunity. This review provides a comprehensive overview of the research on GB03 from its initial discovery in Australian wheat fields in 1971 to its current applications. Recognized as a model plant growth-promoting rhizobacterium (PGPR), GB03 has exhibited outstanding performance in enhancing the growth and protection of many crop plants including cucumber, pepper, wheat, barley, soybean, and cotton. Notably, GB03 has been reported to elicit plant immune response, referred to as induced systemic resistance (ISR), against above-ground pathogens and insect pests. Moreover, a pivotal finding in GB03 was the first-ever identification of its bacterial volatile compounds, which are known to boost plant growth and activate ISR. Research conducted over the past five decades has clearly demonstrated the potential of GB03 as an eco-friendly substitute for conventional pesticides and fertilizers. Validating its safety, the U.S. Environmental Protection Agency endorsed GB03 for commercial use as Kodiak ® in 1998. Subsequently, other compounds, such as BioYield™, were released as a biological control agent against soil-borne pathogens and as a biofertilizer, utilizing a durable spore formulation. More recently, GB03 has been utilized as a keystone modulator for engineering the rhizosphere microbiome and for eliciting microbe-induced plant volatiles. These extensive studies on GB03 underscore its significant role in sustainable agriculture, positioning it as a safe and environmentally-friendly solution for crop protection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The authors declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Jang, Choi, Zhang, Zhang, Ryu and Kloepper.)

Antagonistic activity of strains from Bacillus species has made them among the preferred agricultural biological control agents against phytopathogenic fungi. These microorganisms' success is mostly based on the production of antagonistic secondary metabolites, mainly those of the non-ribosomal cyclic lipopeptides (CLPs) nature, which can affect phytopathogens directly (iturins and fengycins) or indirectly (surfactins and fengycins). However, abiotic factors in the target site can influence the behavior of the biocontrol traits, but to date, few studies attempting to decipher this kind of interaction have been conducted. This work aimed to evaluate the effect of temperature and culture medium on growth, antagonistic activity against Fusarium oxysporum f. sp. physali (Foph), and the profile of CLPs produced by Bacillus velezensis Bs006. The data showed that measured traits in Bs006 varied with temperature and medium interaction. The concentration of CLPs, as well as the antagonistic activity against Foph, was increased as the nutritional wealth, temperature, and time of incubation increased. The concentration of fengycins and iturins was higher than surfactins at high temperatures. However, a bacteriostatic effect was detected with a combination of Landy medium and 15 °C, which prevented both the biosynthesis of CLPs and the antagonistic activity. The results of this work highlight the importance of abiotic conditions of the target site where a biocontrol agent will be applied to stay active and develop its full antagonistic potential. This response by Bs006 could partly explain the variability of its biocontrol efficacy in the Foph-golden berry pathosystem., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Bacteria produce a huge diversity of metabolites, many of which mediate ecological relations. Among these, volatile compounds cause broad-range effects at low doses and, therefore, may be exploited for plant defence strategies and agricultural production, but such applications are still in their early development. Here, we review the latest technologies involving the use of bacterial volatile compounds for phytosanitary inspection, biological control, plant growth promotion, and crop quality. We highlight a variety of effects with a potential applicative interest, based on either live biocontrol and/or biostimulant agents, or the isolated metabolites responsible for the interaction with hosts or competitors. Future agricultural technologies may benefit from the development of new analytical tools to understand bacterial interactions with the environment., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

LC-MS analysis of plant growth-promoting rhizobacterium (PGPR) Bacillus velezensis AP203 supernatants indicated the presence of nematode-inhibiting compounds that increased in abundance when B. velezensis AP203 was grown on orange peel. Meloidogyne incognita J2 were incubated with B. velezensis AP203 spores and orange peel, spores alone, orange peel alone, or with a non-inoculated control, and the combination of B. velezensis AP203 with orange peel resulted in 94% mortality of M. incognita juveniles ( p  ≤ 0.05). The J2 mortality rate for B. velezensis alone was 53%, compared to 59% mortality with orange peel, and the non-inoculated control exhibited 7% mortality. When tested on soybeans raised in a greenhouse, it was observed that when grown in the presence of orange peel, B. velezensis AP203 culture broth, cell suspension or supernatant reduced the numbers of M. incognita eggs per g of root at 45 days after planting (DAP) compared to inoculated controls in soybean and cotton ( p  ≤ 0.05). Likewise, soybean root length and fresh root weight significantly increased after inoculation with B. velezensis AP203 amended with orange peel. In cotton, shoot and root length significantly increased after inoculation with cell pellets of B. velezensis AP203 amended with orange peel compared to the M. incognita inoculated control. These data indicate that B. velezensis AP203 responds to growth on pectin-rich orange peel by production of biologically active secondary metabolites that can promote plant growth and inhibit root-knot nematode viability., (© 2021 Authors.)

Farmers rely heavily on the use of strobilurin fungicides to manage sheath blight (ShB) caused by Rhizoctonia solani AG1-IA, the most important disease in rice in the southern United States. Greenhouse and field studies were conducted to evaluate the potential use of plant growth-promoting rhizobacteria (PGPRs) in combination with a reduced rate of azoxystrobin application as a strategy to improve the current fungicide-reliant management. Of the nine antagonistic PGPR strains screened in the greenhouse, Bacillus subtilis strain MBI600 provided the most significant and consistent suppression of ShB. Efficacy of strain MBI600 was further evaluated at the concentrations of 0, 10 3 , 10 6 , 10 9 , and 10 11 CFU/ml alone or in combinations with 0, 17, 33, 50, 67, 83, and 100% of the recommended application rate (0.16 kg a.i./ha) of azoxystrobin. Strain MBI600 applied at 10 6 ,10 9 , and 10 11 CFU/ml alone was effective in reducing ShB severity. Combinations of this strain at these rates with ≥33% of the recommended application rate of azoxystrobin further reduced ShB severity. A dose-response model defining the relationships between strain MBI600, azoxystrobin, and ShB severity was established. Estimates of the effective concentrations (EC 50 and EC 90 ) of strain MBI600 when applied in combination with 50% of the recommended application rate of azoxystrobin were 10 4 and 10 9 CFU/ml, respectively. A field trial was conducted over 4 years to verify the efficacy of their combinations. Strain MBI600 alone, when applied at 10 9 CFU/ml at the boot stage, reduced ShB severity but did not significantly increase grain yields each year. Combination of strain MBI600 with azoxystrobin at half of the recommended application rate improved efficacy of strain MBI600, reducing ShB severity to a level comparable to that of azoxystrobin applied at the full rate in all 4 years. The combined treatment also increased grain yield by 14 to 19%, comparable to the fungicide applied at the full rate in 3 of 4 years. Combined use of PGPR strain MBI600 with a reduced rate of azoxystrobin application can be a viable management option for control of ShB while allowing producers to use less fungicide on rice.

Background: Inoculation of bermudagrass with rhizobacterial biostimulants can increase plant growth and influence relationships with grass-feeding herbivores. Tunneling and root-feeding behaviors of tawny mole crickets cause severe damage to grass in pastures, golf courses, and lawns. The goal of this study was to determine if inoculation of bermudagrass by a rhizobacteria blend (plant growth-promoting rhizobacteria, PGPR) increases the tolerance of hybrid bermudagrass to tawny mole crickets in captivity and under field conditions., Results: Bermudagrass in arenas treated with a rhizobacteria blend then infested with tawny mole crickets produced significantly greater root lengths compared to bermudagrass that was either fertilized and infested with mole crickets, or bermudagrass without mole crickets. Bermudagrass treated with either the rhizobacteria blend or fertilizer produce similar top growth, and both treatments in the presence of mole crickets produced greater top growth than bermudagrass without mole crickets. Bermudagrass field plots infested naturally with mole crickets and treated twice with the rhizobacteria blend, or the rhizobacteria blend mixed with bifenthrin produced similar damage ratings as plots treated twice with bifenthrin. The rhizobacteria blend mixed with bifenthrin provided more consistent reductions in the surface activity of mole crickets relative to non-treated plots., Conclusion: Enhanced growth of bermudagrass from fertilizer or rhizobacteria increased tolerance of bermudagrass to damage from tawny mole crickets. Application of PGPR or a PGPR-bifenthrin mixture to turfgrass damaged by mole crickets provided comparable reductions to a short residual, synthetic pyrethroid insecticide. Rhizobacterial products have potential utility for IPM of root herbivores. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Limited information is available on the performance of plant growth-promoting inoculants or bioproducts under different soil nutritional or fertility conditions. Consequently, the objective of this study was to evaluate the effects of a commercially available Bacillus-containing bioproduct, Microlife Abundance, at concentrations of 5.5 and 6.5 log cfu/ml on early growth, fertilizer use-efficiency, and fruit yield of bell pepper (Capsicum annuum L.) under two different soil fertility conditions (25% and 100% recommended N rates). Two pot experiments were conducted with bell pepper: (a) a 4-week-long early growth test with inoculant treatments applied once at transplanting; and (b) a 13-week-long yield test with inoculant treatments applied at transplanting and again at first blossom-set. Results from the early growth test indicated that at both N fertilization levels, applying Abundance once at transplanting at 6.5 log cfu/ml rather than 5.5 log cfu/ml significantly increased root dry weight, total root length, root volume, root surface area, and total length of very fine roots compared with the noninoculated control by 20%, 13%, 17%, 15%, and 12%, respectively. In contrast to the early growth, results from the yield test showed that only at the 100% recommended N rate, applying Abundance twice at both concentrations significantly enhanced N fertilizer use-efficiency and marketable yield of bell pepper over the noninoculated control by 34% (5.5 log cfu/ml) and 30% (6.5 log cfu/ml). Therefore, the efficacy of the Bacillus-containing bioproduct Abundance in enhancing fertilizer use-efficiency and marketable yield of bell pepper varied between soil nutritional conditions, but the early growth promotion effect of Abundance did not. Our results also demonstrate that selected microbial-based bioproducts, like Abundance, can be compatible with chemical fertilizers to enhance fertilizer use-efficiency and crop yields, but cannot be used as complete substitutes for chemical fertilizers., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Inoculation of hybrid bermudagrass with plant growth-promoting rhizobacteria (PGPR) can increase plant growth and influence relationships with above-ground herbivores like fall armyworms. However, few experiments have evaluated PGPR applications relative to root herbivory. Root-feeding white grubs cause severe damage to grasses, especially in tall fescue pastures, golf courses, and lawns. Since bacterial inoculants enhance root growth, the goal of this study was to determine if the inoculation of hybrid bermudagrass by rhizobacteria can increase the tolerance of tall fescue and hybrid bermudagrass to damage from white grub feeding, and if PGPR are compatible with neonicotinoid insecticides commonly used for white grub control., Results: In trials with tall fescue and hybrid bermudagrass, grasses were treated with the PGPR strain mixture Blend 20 or nitrogen or left non-treated and were then infested with Japanese beetle grubs. Grasses treated with PGPR and nitrogen fertilizer produced significantly more top growth than the non-treated grub-infested controls. Tall fescue and hybrid bermudagrass treated with Blend 20 produced root mass similar to or greater than nitrogen fertilized grasses. Both grasses treated with Blend 20 had greater root mass than non-treated infested grass. No treatment negatively impacted grub survival, and weight gains of grubs were similar for all treatments. Bacterial strains were typically compatible with insecticides used to control white grubs., Conclusion: PGPR and nitrogen fertilization stimulate root growth resulting in tolerance of tall fescue and hybrid bermudagrass to white grub infestation. PGPR, acting as biostimulants to increase root biomass on grasses, may have utility for IPM of root herbivores. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Plant growth-promoting rhizobacteria (PGPR) are increasingly used in crops worldwide. While selected PGPR strains can reproducibly promote plant growth under controlled greenhouse conditions, their efficacy in the field is often more variable. Our overall aim was to determine if pectin or orange peel (OP) amendments to Bacillus velezensis (Bv) PGPR strains could increase soybean growth and nodulation by Bradyrhizobium japonicum in greenhouse and field experiments to reduce variability. The treatments included untreated soybean seeds planted in field soil that contained Bv PGPR strains and non-inoculated controls with and without 0.1% ( w/v ) pectin or (1 or 10 mg/200 μL) orange peel (OP) amendment. In greenhouse and field tests, 35 and 55 days after planting (DAP), the plants were removed from pots, washed, and analyzed for treatment effects. In greenhouse trials, the rhizobial inoculant was not added with Bv strains and pectin or OP amendment, but in the field trial, a commercial B. japonicum inoculant was used with Bv strains and pectin amendment. In the greenhouse tests, soybean seeds inoculated with Bv AP193 and pectin had significantly increased soybean shoot length, dry weight, and nodulation by indigenous Bradyrhizobium compared to AP193 without pectin. In the field trial, pectin with Bv AP193 significantly increased the shoot length, dry weight, and nodulation of a commercial Bradyrhizobium japonicum compared to Bv AP193 without pectin. In greenhouse tests, OP amendment with AP193 at 10 mg significantly increased the dry weight of shoots and roots compared to AP193 without OP amendment. The results demonstrate that pectin-rich amendments can enhance Bv-mediated soybean growth promotion and nodulation by indigenous and inoculated B. japonicum .

A Gram-stain-positive, aerobic bacterium, TB-66 T , was isolated from a pile of bat guano in a cave of New Mexico, USA. On the basis of 16S rRNA gene sequence similarity comparisons, strain TB-66 T grouped together with Filibacter limicola showing a 16S rRNA gene sequence similarity of 98.5 % to the type strain. The quinone system of strain TB-66 T consisted predominantly of menaquinone MK-7. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylserine and three unidentified phospholipids. The peptidoglycan type was A4α l-Lys-d-Glu (A11.33). The major fatty acids were C15 : 0 anteiso, C16 : 0, and C16 : 1 ω7c. The G+C content of the genomic DNA was 37.6 (±1.8) mol%. On the basis of the genotypic and phenotypic properties it is clear that strain TB-66 T represents a member of the genus Filibacter, but is distinct from the only other species in the genus, Filibacter limicola DSM 13886 T . We propose a novel species with the name Filibacter tadaridae sp. nov. The type strain is TB-66 T (= CIP 111629 T = LMG 30660 T = CCM 8866 T ).

Bats are known to be reservoirs for a variety of mammalian pathogens, including viruses, fungi, and bacteria. Many of the studies examining the microbial community inhabiting bats have investigated bacterial taxa found within specific bat tissues and isolated bat guano pellets, but relatively few studies have explored bacterial diversity within bat guano piles. In large bat caves, bat guano can accumulate over time, creating piles several meters deep and forming complex interactions with coprophagous organisms in a habitat with low light and oxygen. As the guano decays, the nutrient composition changes, but the bacterial communities deep within the pile have not been characterized. Here, we assess the bacterial communities across varying depths within the guano pile using both culture-independent and culture-dependent methods. We found that although similar taxa are found throughout the guano pile, the relative abundances of taxa within the pile shift, allowing certain taxa to dominate the bacterial community at varying depths. We also identified potential bacterial functions being performed within the bat guano as various depths within the pile and found little variation in terms of the dominant predicted functions, suggesting that although the relative abundances of bacterial taxa are changing, the functions being performed are similar. Additionally, we cultured 15 different bacterial species, including 2 not present in our culture-independent analysis, and discuss the pathogenicity potential of these taxa. This study represents the first characterization of the bacterial community from the extreme environment within a bat guano pile and demonstrates the potential for bat caves as resources for identifying new bacterial species.

The aims of this study were to isolate and characterize N 2 -fixing bacteria from giant reed and switchgrass and evaluate their plant growth promotion and nutrient uptake potential for use as biofertilizers. A total of 190 bacteria were obtained from rhizosphere soil and inside stems and roots of giant reed and switchgrass. All the isolates were confirmed to have nitrogenase activity, 96.9% produced auxin, and 85% produced siderophores. Then the top six strains, including Sphingomonas trueperi NNA-14, Sphingomonas trueperi NNA-19, Sphingomonas trueperi NNA-17, Sphingomonas trueperi NNA-20, Psychrobacillus psychrodurans NP-3, and Enterobacter oryzae NXU-38, based on nitrogenase activity, were inoculated on maize and wheat seeds in greenhouse tests to assess their potential benefits to plants. All the selected strains promoted plant growth by increasing at least one plant growth parameter or increasing the nutrient concentration of maize or wheat plants. NNA-14 outperformed others in promoting early growth and nutrient uptake by maize. Specifically, NNA-14 significantly increased root length, surface area, and fine roots of maize by 14%, 12%, and 17%, respectively, and enhanced N, Ca, S, B, Cu, and Zn in maize. NNA-19 and NXU-38 outperformed others in promoting both early growth and nutrient uptake by wheat. Specifically, NNA-19 significantly increased root dry weight and number of root tips of wheat by 25% and 96%, respectively, and enhanced Ca in wheat. NXU-38 significantly increased root length, surface area, and fine roots of wheat by 21%, 13%, and 26%, respectively, and enhanced levels of Ca and Mg in wheat. It is concluded that switchgrass and giant reed are colonized by N 2 -fixing bacteria that have the potential to contribute to plant growth and nutrient uptake by agricultural crops., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Several studies have shown that mixtures of plant-growth-promoting rhizobacteria (PGPR) could enhance biological control activity for multiple plant diseases through the mechanisms of induced systemic resistance or antagonism. Prior experiments showed that four individual PGPR strains-AP69 (Bacillus altitudinis), AP197 (B. velezensis), AP199 (B. velezensis), and AP298 (B. velezensis)-had broad-spectrum biocontrol activity via antagonism in growth chambers against two foliar bacterial pathogens (Xanthomonas axonopodis pv. vesicatoria and Pseudomonas syringae pv. tomato) and one of two tested soilborne fungal pathogens (Rhizoctonia solani and Pythium ultimum). Based on these findings, the overall hypothesis of this study was that a mixture of two individual PGPR strains would exhibit better overall biocontrol and plant-growth promotion than the individual PGPR strains. Two separate greenhouse experiments were conducted. In each experiment, two individual PGPR strains and their mixtures were tested for biological control of three different diseases and for plant-growth promotion in the presence of the pathogens. The results demonstrated that the two individual PGPR strains and their mixtures exhibited both biological control of multiple plant diseases and plant-growth promotion. Overall, the levels of disease suppression and growth promotion were greater with mixtures than with individual PGPR strains.

A Gram-positive-staining, aerobic, non-endospore-forming bacterial strain (JJ-59 T ), isolated from a field-grown maize plant in Dunbar, Nebraska in 2014 was studied by a polyphasic approach. Based on 16S rRNA gene sequence similarity comparisons, strain JJ-59 T was shown to be a member of the genus Paenibacillus, most closely related to the type strains of Paenibacillus aceris (98.6 % 16S rRNA gene sequence similarity) and Paenibacillus chondroitinus (97.8 %). For all other type strains of species of the genus Paenibacillus lower 16S rRNA gene sequence similarities were obtained. DNA-DNA hybridization values of strain JJ-59 T to the type strains of P. aceris and P. chondroitinus were 26 % (reciprocal, 59 %) and 52 % (reciprocal, 59 %), respectively. Chemotaxonomic characteristics such as the presence of meso-diaminopimelic acid in the peptidoglycan, the major quinone MK-7 and spermidine as the major polyamine were in agreement with the characteristics of the genus Paenibacillus. Strain JJ-59 T shared with its next related species P. aceris the major lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified aminophospholipid, but the presence/absence of certain lipids was clearly distinguishable. Major fatty acids of strain JJ-59 T were anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0, and the genomic G+C content is 47.2 mol%. Physiological and biochemical characteristics of strain JJ-59 T were clearly different from the most closely related species of the genus Paenibacillus. Thus, strain JJ-59 T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus nebraskensis sp. nov. is proposed, with JJ-59 T (=DSM 103623 T =CIP 111179 T =LMG 29764 T ) as the type strain.

A study was designed to screen individual strains of plant growth-promoting rhizobacteria (PGPR) for broad-spectrum disease suppression in vitro and in planta. In a preliminary screen, 28 of 196 strains inhibited eight different tested pathogens in vitro. In a secondary screen, these 28 strains showed broad spectrum antagonistic activity to six different genera of pathogens, and 24 of the 28 strains produced five traits reported to be related to plant growth promotion, including nitrogen fixation, phosphate solubilization, indole-3-acetic acid production, siderophore production, and biofilm formation. In advanced screens, the 28 PGPR strains selected in vitro were tested in planta for biological control of multiple plant diseases including bacterial spot of tomato caused by Xanthomonas axonopodis pv. vesicatoria, bacterial speck of tomato caused by Pseudomonas syringae pv. tomato, damping-off of pepper caused by Rhizoctonia solani, and damping-off of cucumber caused by Pythium ultimum. In all, 5 of the 28 tested strains significantly reduced three of the four tested diseases, and another 19 strains showed biological control to two tested diseases. To understand the observed broad-spectrum biocontrol capacity, antiSMASH was used to predict secondary metabolite clusters of selected strains. Multiple gene clusters encoding for secondary metabolites, e.g., bacillibactin, bacilysin, and microcin, were detected in each strain. In conclusion, selected individual PGPR strains showed broad-spectrum biocontrol activity to multiple plant diseases.

Nonpathogenic soil bacteria can colonize the rhizosphere and induce unique plant phenotypes that may influence plant-insect interactions. However, few studies have considered the influences of bacteria-plant interactions on insect feeding and oviposition. The objective of this study was to determine how rhizobacterial inoculation of bermudagrass affects larval development and ovipositional behaviors of the fall armyworm (Spodoptera frugiperda J.E. Smith). Eight blends of rhizobacteria known to induce root or shoot growth in grasses were applied weekly to hybrid bermudagrass for 5 wk. Oviposition was evaluated in two no-choice trials with bacteria-treated, fertilized, or nontreated grass. Grass blades from these treatments were extracted in polar and nonpolar solvents and assayed for oviposition responses. Another experiment compared the development of fall armyworm larvae on bermudagrass treated with each of the eight rhizobacterial blends for 5 wk to larvae fed nontreated bermudagrass. Females deposited more eggs on nontreated and fertilized grass and ≤34% of eggs on grass treated with rhizobacterial blends. Moths exposed to polar and nonpolar extracts were unable to reproduce these results. Larval and pupal weights at days 10 and 12 and the number of adults to eclose were lower for larvae fed some, but not all, bacteria-treated bermudagrass relative to controls. This is one of the few studies to investigate plant-microbe-insect interactions in an economically important system. Although the effects noted with fall armyworm are limited, induced changes in roots also reported for these bacteria may have greater utility than foliar changes for mediating interactions with biotic or abiotic stresses., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Heterodera glycines, the soybean cyst nematode, is the most economically important plant-parasitic nematode on soybean production in the U.S. The objectives of this study were to evaluate the potential of plant growth-promoting rhizobacteria (PGPR) strains for mortality of H. glycines J2 in vitro and for reducing nematode population density on soybean in greenhouse, microplot, and field trials. The major group causing mortality to H. glycines in vitro was the genus Bacillus that consisted of 92.6% of the total 663 PGPR strains evaluated. The subsequent greenhouse, microplot, and field trials indicated that B. velezensis strain Bve2 consistently reduced H. glycines cyst population density at 60 DAP. Bacillus mojavensis strain Bmo3 suppressed H. glycines cyst and total H. glycines population density under greenhouse conditions. Bacillus safensis strain Bsa27 and Mixture 1 (Bve2 + Bal13) reduced H. glycines cyst population density at 60 DAP in the field trials. Bacillus subtilis subsp. subtilis strains Bsssu2 and Bsssu3, and B. velezensis strain Bve12 increased early soybean growth including plant height and plant biomass in the greenhouse trials. Bacillus altitudinis strain Bal13 increased early plant growth on soybean in the greenhouse and microplot trials. Mixture 2 (Abamectin + Bve2 + Bal13) increased early plant growth in the microplot trials at 60 DAP, and also enhanced soybean yield at harvest in the field trials. These results demonstrated that individual PGPR strains and mixtures can reduce H. glycines population density in the greenhouse, microplot, and field conditions, and increased yield of soybean.

Airborne chemical signals emitted by bacteria influence the behavior of other bacteria and plants. We present an overview of in vitro methods for evaluating bacterial and plant responses to bacterial volatile compounds (BVCs). Three types of equipment have been used to physically separate the bacterial test strains from either other bacterial strains or plants (in our laboratory we use either Arabidopsis or tobacco plant seedlings): a Petri dish containing two compartments (BI Petri dish); two Petri dishes connected with tubing; and a microtiter-based assay. The optimized procedure for the BI Petri dish system is described in this protocol and can be widely used for elucidation of potential function in interactions between diverse microbes and those plant and chemical volatiles emitted by bacteria that are most likely to mediate bacterial or plant responses to BVCs. We also describe a procedure for metabolome-based BVC profiling via dynamic (i.e., continuous airflow) or static headspace sampling using solid-phase microextraction (SPME). Using both these procedures, bacteria-bacteria communications and bacteria-plant interactions mediated by BVCs can be rapidly investigated (within 1-4 weeks).

A Gram-positive-staining, aerobic organism, isolated from the rhizosphere of Zea mays, was investigated in detail. Based on 16S rRNA gene sequence similarity comparisons, strain JJ-247T was grouped into the genus Bacillus, most closely related to Bacillus foraminis (98.4 %). The 16S rRNA gene sequence similarity to the sequences of the type strains of other species of the genus Bacillus was <97.4 %. The fatty acid profile with the major fatty acids, anteiso-C15 : 0, iso-C15 : 0, iso-C14 : 0 and iso-C16 : 0 supported the grouping of the strain to the genus Bacillus. The polar lipid profile contained the major components diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified aminophospholipid. The major quinone was menaquinone MK-7, and the major polyamine was spermidine. The genomic DNA G+C content of strain JJ-247T was 44.5 mol%. DNA-DNA hybridizations with the type strain B. foraminis LMG 23147T resulted in values below 70 %. In addition, physiological and biochemical test results allowed a clear phenotypic differentiation of strain JJ-247T from B. foraminis. As a consequence, JJ-247T represents a novel species of the genus Bacillus, for which we propose the name Bacillus zeae sp. nov., with JJ-247T (=CCM 8726T=LMG 29876T) as the type strain.

In the past decade, increased attention has been placed on biological control of plant-parasitic nematodes using various fungi and bacteria. The objectives of this study were to evaluate the potential of 662 plant growth-promoting rhizobacteria (PGPR) strains for mortality to Meloidogyne incognita J2 in vitro and for nematode management in greenhouse, microplot, and field trials. Results indicated that the mortality of M. incognita J2 by the PGPR strains ranged from 0 to 100% with an average of 39%. Among the PGPR strains examined, 212 of 662 strains (or 33%) caused significantly greater mortality percent of M. incognita J2 than the untreated control. Bacillus was the major genus initiating a greater mortality percentage when compared with the other genera. In subsequent trials, B. velezensis strain Bve2 reduced M. incognita eggs per gram of cotton root in the greenhouse trials at 45 days after planting (DAP) similarly to the commercial standards Abamectin and Clothianidin plus B. firmus I-1582. Bacillus mojavensis strain Bmo3, B. velezensis strain Bve2, B. subtilis subsp. subtilis strain Bsssu3, and the Mixture 2 (Abamectin + Bve2 + B. altitudinis strain Bal13) suppressed M. incognita eggs per gram of root in the microplot at 45 DAP. Bacillus velezensis strains Bve2 and Bve12 also increased seed-cotton yield in the microplot and field trials. Overall, results indicate that B. velezensis strains Bve2 and Bve12, B. mojavensis strain Bmo3, and Mixture 2 have potential to reduce M. incognita population density and to enhance growth of cotton when applied as in-furrow sprays at planting.

A Gram-stain-positive, aerobic, endospore-forming bacterial strain isolated from the rhizosphere of Zea mays was studied to determine its detailed taxonomic position. Based on 16S rRNA gene sequence similarity comparisons, strain JJ-64T was shown to be a member of the genus Paenibacillus, most closely related to the type strains of Paenibacillus silagei (99 %) and Paenibacillus borealis (97.5 %). 16S rRNA gene sequence similarity to all other Paenibacillus species was ≤97.5 %. DNA-DNA hybridization values to the type strains of P. silagei and P. borealis were 51 % (reciprocal 25 %) and 31 % (reciprocal 37 %), respectively. The presence of meso-diaminopimelic acid as the diagnostic diamino acid of the peptidoglycan, the major quinone MK-7 and the polyamine pattern with spermidine as the major component were well in line with the characteristics of the genus Paenibacillus. Furthermore, the polar lipid profile of strain JJ-64T with the predominant lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine and two unidentified aminophospholipids reflected the close phylogenetic relatedness to P. silagei. Major fatty acids were iso- and anteiso-branched components. Physiological and biochemical characteristics allowed the further phenotypic differentiation of strain JJ-64T from the most closely related species. Thus, strain JJ-64T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus rhizoplanae sp. nov. is proposed. The type strain is JJ-64T (=LMG 29875T=CCM 8725T).

Nitrous oxide (N 2 O) emissions are increasing at an unprecedented rate owing to the increased use of nitrogen (N) fertilizers. Thus, new innovative management tools are needed to reduce emissions. One potential approach is the use of microbial inoculants in agricultural production. In a previous incubation study, we observed reductions in N 2 O emissions when microbial-based inoculants were added to soil (no plants present) with N fertilizers under laboratory incubations. This present study evaluated the effects of microbial-based inoculants on N 2 O and carbon dioxide (CO 2 ) emissions when applied to soil planted with corn (Zea mays L.) under controlled greenhouse conditions. Inoculant treatments consisted of (i) SoilBuilder (SB), (ii) a metabolite extract of SoilBuilder (SBF), and (iii) a mixture of 4 strains of plant-growth-promoting Bacillus spp. (BM). Experiments included an unfertilized control and 3 N fertilizers: urea, urea - ammonium nitrate with 32% N (UAN-32), and calcium - ammonium nitrate with 17% N (CAN-17). Cumulative N 2 O fluxes from pots 41 days after planting showed significant reductions in N 2 O of 15% (SB), 41% (BM), and 28% (SBF) with CAN-17 fertilizer. When UAN-32 was used, reductions of 34% (SB), 35% (SBF), and 49% (BM) were obtained. However, no reductions in N 2 O emissions occurred with urea. Microbial-based inoculants did not affect total CO 2 emissions from any of the fertilized treatments or the unfertilized control. N uptake was increased by an average of 56% with microbial inoculants compared with the control (nonmicrobial-based treatments). Significant increases in plant height, SPAD chlorophyll readings, and fresh and dry shoot mass were also observed when the microbial-based treatments were applied (with and without N). Overall, results demonstrate that microbial inoculants can reduce N 2 O emissions following fertilizer application depending on the N fertilizer type used and can enhance N uptake and plant growth. Future studies are planned to evaluate the effectiveness of these microbial inoculants in field-based trials and determine the mechanisms involved in N 2 O reduction.

A Gram-stain-positive, aerobic organism, showing an irregular cell morphology, was isolated from the root tissue of cucumber (Cucumis sativus) and investigated in detail for its taxonomic position. On the basis of the 16S rRNA gene sequence analysis, strain AP-38T was shown to be most closely related to Isoptericola variabilis (99.1 %) and Isoptericola nanjingensis (98.9 %). The 16S rRNA gene sequence similarity to all other species of the genus Isoptericola was ≤98.5 %. DNA-DNA relatedness to Isoptericola variablis DSM 10177T and Isoptericola nanjingensis DSM 24300T was 31(reciprocal 41 %) and 34 (reciprocal 34 %), respectively. The diagnostic diamino acid of the peptidoglycan was l-lysine. The quinone system contained predominantly menaquinones MK-9(H4) and MK-9(H2). In the polar lipid profile, major compounds were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and two phosphatidylinositol mannosides. The polyamine pattern contained the major components spermidine and spermine and significant amounts of tyramine. In the fatty acid profile, anteiso-C15 : 0 and iso-C15 : 0 were present in major amounts. These data support the allocation of the strain to the genus Isoptericola. The results of physiological and biochemical characterization additionally provide phenotypic differentiation of strain AP-38T from I. variabilis and I. nanjingensis. AP-38T represents a novel species of the genus Isoptericola, for which we propose the name Isoptericola cucumis sp. nov., with AP-38T (= LMG 29223T=CCM 8653T) as the type strain.

A Gram-positive-staining, aerobic, endospore-forming bacterial strain, isolated from the stem of a cucumber plant, was studied in detail for its taxonomic position. Based on 16S rRNA gene sequence similarity comparisons, strain AP-115 T was grouped into the genus Paenibacillus, most closely related to Paenibacillus amylolyticus (98.8 %), Paenibacillus tundrae and Paenibacillus barcinonensis (both 98.4 %). The 16S rRNA gene sequence similarity to other species of the genus Paenibacillus was ≤98.4 %. Chemotaxonomic characterization supported allocation of the strain to the genus Paenibacillus. The quinone system contained exclusively menaquinone MK-7, and in the polar lipid profile diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylserine were predominating. The major component in the polyamine pattern was spermidine, and the diagnostic diamino acid of the peptidoglycan was meso-diaminopimelic acid. The major fatty acids were iso- and anteiso-branched fatty acids. The results of physiological and biochemical tests allowed phenotypic differentiation of strain AP-115 T from closely related species. Thus, AP-115 T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus cucumis sp. nov. is proposed, with AP-115 T (=LMG 29222 T =CCM 8653 T ) as the type strain.

In commercial agriculture, populations and interactions of rhizosphere microflora are potentially affected by the use of specific agrichemicals, possibly by affecting gene expression in these organisms. To investigate this, we examined changes in bacterial gene expression within the rhizosphere of glyphosate-tolerant corn (Zea mays) and soybean (Glycine max) in response to long-term glyphosate (PowerMAX™, Monsanto Company, MO, USA) treatment. A long-term glyphosate application study was carried out using rhizoboxes under greenhouse conditions with soil previously having no history of glyphosate exposure. Rhizosphere soil was collected from the rhizoboxes after four growing periods. Soil microbial community composition was analyzed using microbial phospholipid fatty acid (PLFA) analysis. Total RNA was extracted from rhizosphere soil, and samples were analyzed using RNA-Seq analysis. A total of 20-28 million bacterial sequences were obtained for each sample. Transcript abundance was compared between control and glyphosate-treated samples using edgeR. Overall rhizosphere bacterial metatranscriptomes were dominated by transcripts related to RNA and carbohydrate metabolism. We identified 67 differentially expressed bacterial transcripts from the rhizosphere. Transcripts downregulated following glyphosate treatment involved carbohydrate and amino acid metabolism, and upregulated transcripts involved protein metabolism and respiration. Additionally, bacterial transcripts involving nutrients, including iron, nitrogen, phosphorus, and potassium, were also affected by long-term glyphosate application. Overall, most bacterial and all fungal PLFA biomarkers decreased after glyphosate treatment compared to the control. These results demonstrate that long-term glyphosate use can affect rhizosphere bacterial activities and potentially shift bacterial community composition favoring more glyphosate-tolerant bacteria., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

Gossypol is an important allelochemical produced by the subepidermal glands of some cotton varieties and important for their ability to respond to changing biotic stress by exhibiting antibiosis against some cotton pests. Plant growth-promoting rhizobacteria (PGPR) are root-colonizing bacteria that increase plant growth and often elicit defence against plant pathogens and insect pests. Little is known about the effect of PGPR on cotton plant-insect interactions and the potential biochemical and molecular mechanisms by which PGPR enhance cotton plant defence. Here, we report that PGPR (Bacillus spp.) treated cotton plants showed significantly higher levels of gossypol compared with untreated plants. Similarly, the transcript levels of the genes (i.e. (+)-δ-cadinene synthase gene family) involved in the biosynthesis of gossypol were higher in PGPR-treated plants than in untreated plants. Furthermore, the levels of jasmonic acid, an octadecanoid-derived defence-related phytohormone and the transcript level of jasmonic acid responsive genes were higher in PGPR-treated plants than in untreated plants. Most intriguingly, Spodoptera exigua showed reduced larval feeding and development on PGPR-treated plants. These findings demonstrate that treatment of plants with rhizobacteria may induce significant biochemical and molecular changes with potential ramifications for plant-insect interactions., (© 2015 John Wiley & Sons Ltd.)

A facultative anaerobic, Gram-positive staining, endospore-forming bacterium, isolated from the rhizosphere of cucumber ( Cucumis sativus ), was taxonomically investigated. Based on 16S rRNA gene sequence similarity comparisons, strain AP-6 T clustered together with other species of the genus Bacillus and showed highest similarities with Bacillus drentensis LMG 21831 T (99.1 %), Bacillus vireti LMG 21834 T (98.7 %) and Bacillus soli LMG 21838 T (98.5 %). The 16S rRNA gene sequence similarity to the sequences of the type strains of other species of the genus Bacillus was 98.5 % or less. Chemotaxonomic features supported the grouping of the strain in the genus Bacillus ; for example, the major fatty acids were anteiso-C 15 : 0 , iso-C 15 : 0 and C 16 : 0 , the polar lipid profile contained the major components diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified glycolipid, the major quinone was menaquinone MK-7 and the major compound in the polyamine pattern was spermidine. Additionally, DNA-DNA hybridization with B. drentensis LMG 21831 T , B. vireti LMG 21834 T and B. soli LMG 21838 T resulted in relatedness values that were clearly below 70 %. Physiological and biochemical test results were also different from those of the most closely related species. As a consequence, AP-6 T represents a novel species of the genus Bacillus , for which the name Bacillus cucumis sp. nov. is proposed, with AP-6 T ( = CIP 110974 T  = CCM 8651 T ) as the type strain.

Glyphosate is one of the most widely used herbicides in agriculture with predictions that 1.35 million metric tons will be used annually by 2017. With the advent of glyphosate tolerant (GT) cropping more than 10 years ago, there is now concern for non-target effects on soil microbial communities that has potential to negatively affect soil functions, plant health, and crop productivity. Although extensive research has been done on short-term response to glyphosate, relatively little information is available on long-term effects. Therefore, the overall objective was to investigate shifts in the rhizosphere bacterial community following long-term glyphosate application on GT corn and soybean in the greenhouse. In this study, rhizosphere soil was sampled from rhizoboxes following 4 growth periods, and bacterial community composition was compared between glyphosate treated and untreated rhizospheres using next-generation barcoded sequencing. In the presence or absence of glyphosate, corn and soybean rhizospheres were dominated by members of the phyla Proteobacteria, Acidobacteria, and Actinobacteria. Proteobacteria (particularly gammaproteobacteria) increased in relative abundance for both crops following glyphosate exposure, and the relative abundance of Acidobacteria decreased in response to glyphosate exposure. Given that some members of the Acidobacteria are involved in biogeochemical processes, a decrease in their abundance could lead to significant changes in nutrient status of the rhizosphere. Our results also highlight the need for applying culture-independent approaches in studying the effects of pesticides on the soil and rhizosphere microbial community., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)

Certain bacterial species associate with plant roots in soil. The plant growth-promoting rhizobacteria (PGPR) stimulate plant growth and yield in greenhouse and field. Here, we examined whether application of known bacilli PGPR strains stimulated growth and asexual reproduction in the succulent plant Kalanchoe daigremontiana. Four PGPR strains B. amyloliquefaciens IN937a, B. cereus BS107, B. pumilus INR7, and B. subtilis GB03 were applied to young plantlets by soil-drenching, and plant growth and development was monitored for three months. Aerial growth was significantly stimulated in PGPR-inoculated plants, which was observed as increases in plant height, shoot weight, and stem width. The stimulated growth influenced plant development by increasing the total number of leaves per plant. Treatment with bacilli also increased the total root biomass compared with that of control plants, and led to a 2-fold increase in asexual reproduction and plantlet formation on the leaf. Collectively, our results firstly demonstrate that Bacillus spp. promote vegetative development of K. daigremontiana, and the enhanced growth stimulates asexual reproduction and plantlet formation.

To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR) strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group) are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32 to 90%, with 2839 genes within the core genome of B. amyloliquefaciens subsp. plantarum. Comparative genomic analyses of B. amyloliquefaciens strains identified genes that are linked with biological control and colonization of roots and/or leaves, including 73 genes uniquely associated with subsp. plantarum strains that have predicted functions related to signaling, transportation, secondary metabolite production, and carbon source utilization. Although B. amyloliquefaciens subsp. plantarum strains contain gene clusters that encode many different secondary metabolites, only polyketide biosynthetic clusters that encode difficidin and macrolactin are conserved within this subspecies. To evaluate their role in plant pathogen biocontrol, genes involved in secondary metabolite biosynthesis were deleted in a B. amyloliquefaciens subsp. plantarum strain, revealing that difficidin expression is critical in reducing the severity of disease, caused by Xanthomonas axonopodis pv. vesicatoria in tomato plants. This study defines genomic features of PGPR strains and links them with biocontrol activity and with host colonization.

The rhizobacterium Serratia marcescens strain 90-166 elicits induced systemic resistance against plant pathogens and herbivores and promotes plant growth under greenhouse and field conditions. Strain 90-166 secretes volatile compounds, siderophores, salicylic acid, and quorum-sensing autoinducers as bacterial determinants toward plant health. Herein, we present its draft genome sequence., (Copyright © 2015 Jeong et al.)

Certain pathovars of Pseudomonas amygdali, which is newly reclassified from Pseudomonas syringae by DNA-DNA hybridization and ribotyping, cause many serious diseases of major crop plants. Herein, we present draft genome sequences of P. amygdali pv. tabaci strain ATCC 11528 and P. amygdali pv. lachrymans strain 98A-744., (Copyright © 2015 Jeong et al.)

Bioassay-guided fractionation of the organic extracts of a Bacillus amyloliquefaciens strain (AP183) led to the discovery of a new macrocyclic polyene antibiotic, bacillusin A (1). Its structure was assigned by interpretation of NMR and MS spectroscopic data as a novel macrodiolide composed of dimeric 4-hydroxy-2-methoxy-6-alkenylbenzoic acid lactones with conjugated pentaene-hexahydroxy polyketide chains. Compound 1 showed potent antibacterial activities against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium with minimum inhibitory concentrations in a range of 0.6 to 1.2 μg/mL. The biosynthetic significance of this unique class of antibiotic compounds is briefly discussed.

Bacillus amyloliquefaciens GB03 has been used as a representative commercialized strain of the bacilli for biological control against a broad spectrum of plant pathogens and as a bio-fertilizer to promote growth and yield of field crops for more than two decades. Herein, we present the genome sequence and a brief analysis of strain GB03., (Copyright © 2014 Choi et al.)

Bacillus pumilus INR7 is an endophytic bacterium that has been commercialized as a biological control product against soilborne pathogens as well as foliar pathogens by direct antagonism and induction of systemic resistance. In the current study, we provide the genome sequence and a possible explanation of the function of strain INR7., (Copyright © 2014 Jeong et al.)

Due to increased restrictions on the use of chemical nematicides, alternative nematode management strategies, including biocontrol, are needed. The objectives of this study were to evaluate the potential of Bacillus firmus GB-126 and Paecilomyces lilacinus 251 in commercial formulations applied separately or concomitantly to manage Rotylenchulus reniformis in cotton grown under greenhouse, microplot, and field conditions. In the greenhouse, seed treated with B. firmus (1.4 × 10 7 CFU/seed), an application of P. lilacinus (0.3% vol/vol of water), or the combination of B. firmus and P. lilacinus reduced the number of females, eggs, and vermiforms of R. reniformis (P ≤ 0.02) and increased populations of free-living nematodes (P ≤ 0.01). In microplots and field conditions, populations of R. reniformis vermiforms decreased when exposed to B. firmus and P. lilacinus biocontrol agents at midseason (P ≤ 0.04). Furthermore, stem diameter and free-living nematode numbers increased (P ≤ 0.01) with the combination of B. firmus and P. lilacinus. In the field, numbers of females, eggs, and vermiform life stages at the end of the growing season decreased in the presence of the biocontrol agents applied individually or concomitantly (P ≤ 0.01). Cotton yields from the application of B. firmus GB-126 and P. lilacinus 251 were similar to those from aldicarb, the chemical nematicide standard.

Bacteria and plant derived volatile organic compounds have been reported as the chemical triggers that elicit induced resistance in plants. Previously, volatile organic compounds (VOCs), including acetoin and 2,3-butanediol, were found to be emitted from plant growth-promoting rhizobacteria (PGPR) Bacillus subtilis GB03, which had been shown to elicit ISR and plant growth promotion. More recently, we reported data that stronger induced resistance could be elicited against Pseudomonas syringae pv maculicola ES4326 in plants exposed to C13 VOC from another PGPR Paenibacillus polymyxa E681 compared with that of strain GB03. Here, we assessed whether another long hydrocarbon C16 hexadecane (HD) conferred protection to Arabidopsis from infection of a biotrophic pathogen, P. syringae pv maculicola and a necrotrophic pathogen, Pectobacterium carotovorum subsp carotovorum. Collectively, long-chain VOCs can be linked to a plant resistance activator for protecting plants against both biotrophic and necrotrophic pathogens at the same time.

Numerous root-associated bacteria (rhizobacteria) are known to elicit induced systemic resistance (ISR) in plants. Bacterial cell-density-dependent quorum sensing (QS) is thought to be important for ISR. Here, we investigated the role of QS in the ISR elicited by the rhizobacterium, Serratia marcescens strain 90-166, in tobacco. Since S. marcescens 90-166 produces at least three QS signals, QS-mediated ISR in strain 90-166 has been difficult to understand. Therefore, we investigated the ISR capacity of two transgenic tobacco (Nicotiana tabacum) plants that contained either bacterial acylhomoserine lactone-producing (AHL) or -degrading (AiiA) genes in conjunction with S. marcescens 90-166 to induce resistance against bacterial and viral pathogens. Root application of S. marcescens 90-166 increased ISR to the bacterial pathogens, Pectobacterium carotovorum subsp. carotovorum and Pseudomonas syringae pv. tabaci, in AHL plants and decreased ISR in AiiA plants. In contrast, ISR to Cucumber mosaic virus was reduced in AHL plants treated with S. marcescens 90-166 but enhanced in AiiA plants. Taken together, these data indicate that QS-dependent ISR is elicited by S. marcescens 90-166 in a pathogen-dependent manner. This study provides insight into QS-dependent ISR in tobacco elicited by S. marcescens 90-166.

There is currently much interest in developing crop management practices that will decrease NO emissions from agricultural soils. Many different approaches are being investigated, but to date, no studies have been published on how microbial inoculants affect NO emissions. This study was conducted to test the hypothesis that microbial-based inoculants known to promote root growth and nutrient uptake can reduce NO emissions in the presence of N fertilizers under controlled conditions. Carbon dioxide and CH fluxes were also measured to evaluate microbial respiration and determine the aerobic and anaerobic conditions of the incubated soil. The microbial-based treatments investigated were SoilBuilder (SB), a metabolite extract of SoilBuilder (SBF), and a mixture of four strains of plant growth-promoting spp. Experiments included two different N fertilizer treatments, urea and urea-NHNO 32% N (UAN), and an unfertilized control. Emissions of NO and CO were determined from soil incubations and analyzed with gas chromatography. After 29 d of incubation, cumulative NO emissions were reduced 80% by SB and 44% by SBF in soils fertilized with UAN. Treatment with spp. significantly reduced NO production on Days 1 and 2 of the incubation in soils fertilized with UAN. In the unfertilized treatment, cumulative emissions of NO were significantly reduced 92% by SBF. Microbial-based treatments did not reduce NO emissions associated with urea application. Microbial-based treatments increased CO emissions from soils fertilized with UAN, suggesting a possible increase in microbial activity. Overall, the results demonstrated that microbial-based inoculants can reduce NO emissions associated with N fertilizer application, and this response varies with the type of microbial-based inoculant and fertilizer., (Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.)

Background: Fern Distortion Syndrome (FDS) is a serious disease of Leatherleaf fern (Rumohra adiantiformis). The main symptom of FDS is distortion of fronds, making them unmarketable. Additional symptoms include stunting, irregular sporulation, decreased rhizome diameter, and internal discoloration of rhizomes. We previously reported an association of symptoms with increased endophytic rhizome populations of fluorescent pseudomonads (FPs). The aim of the current study was to determine if FPs from ferns in Costa Rica with typical FDS symptoms would recreate symptoms of FDS., Methodology and Findings: Greenhouse tests were conducted over a 29-month period. Micro-propagated ferns derived from tissue culture were first grown one year to produce rhizomes. Then, using an 8×9 randomized complete block experimental design, 8 replicate rhizomes were inoculated by dipping into 9 different treatments before planting. Treatments included water without bacteria (control), and four different groups of FPs, each at a two concentrations. The four groups of FPs included one group from healthy ferns without symptoms (another control treatment), two groups isolated from inside rhizomes of symptomatic ferns, and one group isolated from inside roots of symptomatic ferns. Symptoms were assessed 12 and 17 months later, and populations of FPs inside newly formed rhizomes were determined after 17 months. Results showed that inoculation with mixtures of FPs from ferns with FDS symptoms, but not from healthy ferns, recreated the primary symptom of frond deformities and also the secondary symptoms of irregular sporulation, decreased rhizome diameter, and internal discoloration of rhizomes., Conclusions: These results suggest a model of causation of FDS in which symptoms result from latent infections by multiple species of opportunistic endophytic bacteria containing virulence genes that are expressed when populations inside the plant reach a minimum level.

Bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against Edwardsiella ictaluri and Aeromonas hydrophila, the causative agents of enteric septicemia of catfish (ESC) and motile aeromonad septicaemia (MAS), respectively. Twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. Each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×10(7) CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (P<0.05). A similar challenge experiment conducted in Vietnam with four of the five Bacillus strains also showed protective effects against E. ictaluri in striped catfish. Safety of the four strains exhibiting the strongest biological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture.

Background: Some strains of plant growth-promoting rhizobacteria (PGPR) elicit induced systemic resistance (ISR) by emission of volatile organic compounds (VOCs) including short chain alcohols, acetoin, and 2,3-butanediol. The objective of this study was to evaluate whether species-specific VOCs from PGPR strain Paenibacillus polymyxa E681 can promote growth and induce resistance in Arabidopsis., Methodology/principal Findings: The efficacy of induction was strain-specific, with stronger protection against Pseudomonas syringae pv. maculicola ES4326 in plants exposed to VOCs from P. polymyxa E681 versus Arabidopsis plants exposed to VOCs from a reference strain Bacillus subtilis GB03, which was previously shown to elicit ISR and plant growth promotion. VOC emissions released from E681 primed transcriptional expression of the salicylic acid, jasmonic acid, and ethylene signaling marker genes PR1, ChiB, and VSP2, respectively. In addition, strain E681 produced more than thirty low molecular-weight VOCs, of which tridecane was only produced by E681 and not found in GB03 or IN937a volatile blends. These strain-specific VOCs induced PR1 and VSP2 genes., Conclusions/significance: These results provide new insight into the existence of a long chain VOC signaling molecule produced by P. polymyxa that can serve as a bacterial trigger of induced systemic resistance in planta.

A syndrome has been recognized on leatherleaf fern (Rumohra adiantiformis) in Costa Rica for many years that causes widespread damage but has not been described in the literature. A full description of the syndrome, termed fern distortion syndrome (FDS), is reported here, along with evidence that FDS is a new disease and that it is associated with endophytic fluorescent pseudomonads but not with any other major groups of pathogens or pests. The main aboveground symptoms of FDS are twisting and distortions of fronds, which make the fronds unmarketable. In advanced cases of FDS, fronds are often thickened, new frond growth ceases or slows dramatically, and uneven sporulation is apparent on the underside of fronds. Symptoms of FDS belowground are reduced diameter of rhizomes and reduced overall root mass. The incidence of FDS in Costa Rica was typically over 80%, and severity typically ranged from 1.26 to 2.48 using a 0 to 3 rating scale in fields propagated vegetatively with rhizomes from fields with FDS. In contrast, in three fields planted 1.5 to 4 years previously with rhizomes derived from tissue culture, incidence and severity were markedly lower: 23 to 34% and 0.24 to 0.36, respectively. Paired sampling of symptomatic and asymptomatic plants revealed significantly greater populations of fluorescent pseudomonads inside rhizomes of symptomatic plants.

The search for microorganisms that improve soil fertility and enhance plant nutrition has continued to attract attention due to the increasing cost of fertilizers and some of their negative environmental impacts. The objectives of this greenhouse study with tomato were to determine (1) if reduced rates of inorganic fertilizer coupled with microbial inoculants will produce plant growth, yield, and nutrient uptake levels equivalent to those with full rates of the fertilizer and (2) the minimum level to which fertilizer could be reduced when inoculants were used. The microbial inoculants used in the study were a mixture of plant growth-promoting rhizobacteria (PGPR) strains Bacillus amyloliquefaciens IN937a and Bacillus pumilus T4, a formulated PGPR product, and the arbuscular mycorrhiza fungus (AMF), Glomus intraradices. Results showed that supplementing 75% of the recommended fertilizer rate with inoculants produced plant growth, yield, and nutrient (nitrogen and phosphorus) uptake that were statistically equivalent to the full fertilizer rate without inoculants. When inoculants were used with rates of fertilizer below 75% of the recommended rate, the beneficial effects were usually not consistent; however, inoculation with the mixture of PGPR and AMF at 70% fertility consistently produced the same yield as the full fertility rate without inoculants. Without inoculants, use of fertilizer rates lower than the recommended resulted in significantly less plant growth, yield, and nutrient uptake or inconsistent impacts. The results suggest that PGPR-based inoculants can be used and should be further evaluated as components of integrated nutrient management strategies.

The continued use of chemical fertilizers and manures for enhanced soil fertility and crop productivity often results in unexpected harmful environmental effects, including leaching of nitrate into ground water, surface run-off of phosphorus and nitrogen run-off, and eutrophication of aquatic ecosystems. Integrated nutrient management systems are needed to maintain agricultural productivity and protect the environment. Microbial inoculants are promising components of such management systems. This review is a critical summary of the efforts in using microbial inoculants, including plant growth-promoting rhizobacteria and arbuscular mycorrhizal fungi for increasing the use efficiency of fertilizers. Studies with microbial inoculants and nutrients have demonstrated that some inoculants can improve plant uptake of nutrients and thereby increase the use efficiency of applied chemical fertilizers and manures. These proofs of concept studies will serve as the basis for vigorous future research into integrated nutrient management in agriculture.

Plant-growth-promoting rhizobacteria (PGPR) are associated with plant roots and augment plant productivity and immunity; however, recent work by several groups shows that PGPR also elicit so-called 'induced systemic tolerance' to salt and drought. As we discuss here, PGPR might also increase nutrient uptake from soils, thus reducing the need for fertilizers and preventing the accumulation of nitrates and phosphates in agricultural soils. A reduction in fertilizer use would lessen the effects of water contamination from fertilizer run-off and lead to savings for farmers.

A 3 year field study was conducted with field corn from 2005 to 2007 to test the hypothesis that microbial inoculants that increase plant growth and yield can enhance nutrient uptake, and thereby remove more nutrients, especially N, P, and K from the field as part of an integrated nutrient management system. The field trial evaluated microbial inoculants, which include a commercially available plant growth-promoting rhizobacteria (PGPR), arbuscular mycorrhiza fungi (AMF), and their combination across 2 tillage systems (no-till and conventional till) and 2 fertilization regimes (poultry litter and ammonium nitrate). Data were collected on plant height, yield (dry mass of ears and silage), and nutrient content of corn grain and silage. In addition, nutrient content of soil was determined, and bioavailability of soil nutrient was measured with plant root simulator probes. Results showed that inoculants promoted plant growth and yield. For example, grain yields (kg.ha(-1)) in 2007 for inoculants were 7717 for AMF, 7260 for PGPR+AMF, 7313 for PGPR, 5725 for the control group, and for fertilizer were 7470 for poultry litter and 6537 for NH4NO3. Nitrogen content per gram of grain tissues was significantly enhanced in 2006 by inoculant, fertilizer, and their interactions. Significantly higher amounts of N, P, and K were removed from the plots with inoculants, based on total nutrient content of grain per plot. These results supported the overall hypothesis and indicate that application of inoculants can lead to reduction in the build up of N, P, and K in agricultural soils. Further studies should be conducted to combine microbial inoculants with reduced rates of fertilizer.

The role of exopolysaccharides (EPSs) from a plant growth-promoting rhizobacterium, Burkholderia gladioli IN26, on elicitation of induced systemic resistance was investigated. A purified EPS induced expression of PR- 1a::GUS on tobacco and elicited induced resistance against Colletotrichum orbiculare on cucumber. The maximum level of disease protection was noted when seeds were soaked in 200 ppm of the EPS. Our results indicate that EPS from specific rhizobacteria can elicit induced resistance and suggest that bacterial EPS might be a useful elicitor of resistance under field conditions.

We evaluated a commercial biopreparation of plant growth-promoting rhizobacteria (PGPR) strains Bacillus subtilis GB03 and B. amyloliquefaciens IN937a formulated with the carrier chitosan (BioYield) for its capacity to elicit growth promotion and induced systemic resistance against infection by Cucumber Mosaic Virus (CMV) and Pseudomonas syringae pv. tomato DC3000 in Arabidopsis thaliana. The biopreparation promoted plant growth of Arabidopsis hormonal mutants, which included auxin, gibberellic acid, ethylene, jasmonate, salicylic acid, and brassinosteroid insensitive lines as well as each wild-type. The biopreparation protected plants against CMV based on disease severity in wild-type plants. However, virus titre was not lower in control plants and those treated with biopreparation, suggesting that the biopreparation induced tolerance rather than resistance against CMV. Interestingly, the biopreparation induced resistance against CMV in NahG plants, as evidenced by both reduced disease severity and virus titer. The biopreparation also elicited induced resistance against P. syringae pv. tomato in the wild-type but not in NahG transgenic plants, which degrade endogenous salicylic acid, indicating the involvement of salicylic acid signaling. Our results indicate that some PGPR strains can elicit plant growth promotion by mechanisms that are different from known hormonal signaling pathways. In addition, the mechanism for elicitation of induced resistance by PGPR may be pathogen-dependent. Collectively, the two-Bacilli strain mixture can be utilized as a biological inoculant for both protection of plant against bacterial and viral pathogens and enhancement of plant growth.

For several years, we have noticed that plant growth-promoting rhizobacteria (PGPR), which consistently promote plant growth in greenhouse tests during spring, summer, and fall, fail to elicit plant growth promotion during the midwinter under ambient light conditions. This report tests the hypothesis that photoperiod regulates elicitation of growth promotion and induced systemic resistance (ISR) by PGPR. A commercially available formulation of PGPR strains Bacillus subtilis GB03 and Bacillus amyloliquefaciens IN937a (BioYield) was used to grow tomato and pepper transplants under short-day (8 h of light) (SD) and long-day (12 h of light) (LD) conditions. Results of many experiments indicated that under LD conditions, BioYield consistently elicited significant increases in root and shoot mass as well as in several parameters of root architecture. However, under SD conditions, such increases were not elicited. Differential root colonization of plants grown under LD and SD conditions and changes in leachate quality partially account for these results. BioYield elicited ISR in tomato and pepper under both LD and SD conditions, indicating that although growth promotion was not elicited under SD conditions, induced resistance was. Overall, the results indicate that PGPR-mediated growth promotion is regulated by photoperiod, while ISR is not.