Studies on the Isolation of Endophytic Bacteria from Tomato Plants and Their Growth-promoting Activities

[Objective] The study aimed to investigate the growth-promoting activities of endophytic bacteria from tomato plants.[Method]The endophytic bacteria isolated from different tissues of tomato plants were analyzed for the effects of their growth-promoting activities on the germination and growth of tomato plants.The bacteria with growth-promoting activity were preliminarily identified.[Result]Totally 59 endophytic bacterial strains were isolated from roots and stems of tomatoes,of which 4 showed significantly growth-promoting activity to germination and growth of tomato.The results suggest that these strains are endowed with the potential capability of growth-promoting.[Conclusion]The endophytic bacteria with growth-promoting activity were found among the isolates from tomato plants.This provided a good foundation for utilization of these bacteria with growth-promoting activity.

Determination of Growth-promoting and Antagonistic Action of Endophytic Bacteria Strains Itb57 and Itb295 of Tobacco

[ Objective ] The paper was to study the growth-promoting and antagonistic action of endophytic bacteria strains Itb57 and Itb295 of tobacco to explore their functions in biological control. [ Method] The growth-promoting effects of bacterial suspension ~ff endophytic bacteria Itb57 and Itb295 on tobacco seedling un- der different treatment modes were studied using potting method in greenhouse. The antagonistic action of bacterial suspension of endophytic bacteria Itb57 and Itb295 on Phytophthora nicotianae, Alternaria alternata and Botrytis cinerea were measured by duel culture method. [ Result] Bacterial suspensions of enduphytic bacteria Itb57 and Itb295 had certain growth-promoting effects on tobacco seedling, which could significantly increase the fresh weight and dry weight in aerial part; the growth-promoting effect of soaking ＋ spraying and irrigating treatment was the best. Itb57 strain had good antagonistic action against P. nicotianae. A. alterna- ta and B. cinerea, while Itb295 strain only had good antifungal effect against P. nicotianae. [ Conclusion] The results provided basis for the study and application of tobacco endophytic bacteria strains Itb57 and Itb295 in biocontrol of tobacco diseases.

Identification of Growth-promoting Bacteria from Rhizosphere of Pastures and Their Effects on Growth of Lotus corniculatus L.

[Objectives] This study was conducted to explore the interaction between nitrogen-fixing and phosphate-solubilizing strains and the optimal combination of different functional strains,in order to provide a theoretical basis for the development of PGPR compound fertilizers suitable for local environment.[Methods] In this study,16S rDNA gene sequence analysis was used to identify fast-growing and competitive strains from pasture nodules and rhizosphere soils in Guizhou Province,and three representative Rhizobia and phosphorus-solubilizing bacteria were chosen for the test of bacterial combination when reducing 50% of nitrogen and 30% of phosphorus.The effects of different strain combinations on the plant height,root length,aboveground and underground biomass of Lotus corniculatus L.were investigated,and the total nitrogen and total phosphorus contents of the plants were determined.[Results] The mixed bacterial agents could promote the increase of root biomass,and the effects of A1,A3,B3 and C3 were the most obvious.The fresh weight and dry weight of the roots of L.corniculatus increased by 30.35%-168.45% and 26.43%-180.00%,respectively,and A3,B3,B2 and C3 had the best effects.The total phosphorus content of the plants increased by 12.79%-55.25% compared with the CK2;and most of the bacterial agents with significant growth-promoting effects showed decreased total nitrogen contents,while those with non-significant growth-promoting effects showed significantly-increased total nitrogen contents,which were not as much as the CK1.Comprehensively,the most productive combination was C3,namely R27-2 Rhinohizobium fredii and P33-3 Stenotrophomonas rhizophila.[Conclusions] This study can provide a theoretical basis for the production and promotion of bacterial fertilizers.

Plant growth-promoting properties and anti-fungal activity of endophytic bacterial strains isolated from Thymus altaicus and Salvia deserta in arid lands

Endophytes,as crucial components of plant microbial communities,significantly contribute to enhancing the absorption of nutrients such as nitrogen and phosphorus by their hosts,promote plant growth,and degrade pathogenic fungal mycelia.In this study,an experiment was conducted in August 2022 to explore the growth-promoting potential of endophytic bacterial strains isolated from two medical plant species,Thymus altaicus and Salvia deserta,using a series of screening media.Plant samples of Thymus altaicus and Salvia deserta were collected from Zhaosu County and Habahe County in Xinjiang Uygur Autonomous Region,China,in July 2021.Additionally,the inhibitory effects of endophytic bacterial strains on the four pathogenic fungi(Fusarium oxysporum,Fulvia fulva,Alternaria solani,and Valsa mali)were determined through the plate confrontation method.A total of 80 endophytic bacterial strains were isolated from Thymus altaicus,while a total of 60 endophytic bacterial strains were isolated from Salvia deserta.The endophytic bacterial strains from both Thymus altaicus and Salvia deserta exhibited plant growth-promoting properties.Specifically,the strains of Bacillus sp.TR002,Bacillus sp.TR005,Microbacterium sp.TSB5,and Rhodococcus sp.TR013 demonstrated strong cellulase-producing activity,siderophore-producing activity,phosphate solubilization activity,and nitrogen-fixing activity,respectively.Out of 140 endophytic bacterial strains isolated from Thymus altaicus and Salvia deserta,104 strains displayed anti-fungal activity against Fulvia fulva,Alternaria solani,Fusarium oxysporum,and Valsa mali.Furthermore,the strains of Bacillus sp.TR005,Bacillus sp.TS003,and Bacillus sp.TSB7 exhibited robust inhibition rates against all the four pathogenic fungi.In conclusion,the endophytic bacterial strains from Thymus altaicus and Salvia deserta possess both plant growth-promoting and anti-fungal properties,making them promising candidates for future development as growth-promoting agents and biocontrol tools for plant diseases.

Application Progress of Plant Growth-promoting Bacteria in Crops

Plant growth-promoting bacteria(PGPBs)can promote plant growth and improve crop yield.They can induce plant systemic resistance to resist biotic and abiotic stresses.In recent years,with the development of green ecological agriculture,new biological fertilizers such as microbial inocula and microbial fertilizers based on PGPBs have been gradually applied in crop planting.Based on plant growth promotion and disease control,the application progress of PGPBs in crops from the aspects of growth promotion mechanism,growth promotion effect,resistance to biological and abiotic stresses were discussed,aiming to provide reference for the relevant research and application of PGPBs in crops.

Rhizobacteria facilitate physiological and biochemical drought tolerance of Halimodendron halodendron (Pall.) Voss

Growth-promoting bacteria(GPB)have shown promising effects on serving plants against environmental constraints such as drought.Nevertheless,simultaneous effects of different GPB have less been considered for arid land plants and under field conditions.We investigated the effects of single and combined application of GPB,including free-living nitrogen-fixing bacteria(NFB),phosphate solubilizing bacteria(PSB),potassium solubilizing bacteria(KSB),a combination of NFB,PSB,and KSB(NPK),and control,at three drought stress treatments.In order to better understand the interactions between drought and GPB,we measured the morphological,biochemical,and physiological plant traits.The target plant was salt tree(Halimodendron Halodendron(Pall.)Voss),a legume shrub native to arid lands of Central and West Asia.All biofertilizer treatments enhanced the growth,physiology,and biochemistry of salt tree seedlings,and there were significant differences among the treatments.KSB and PSB treatments increased photosynthetic pigments,but KSB treatment was more efficient in transpiration rate and stomatal regulation and increased the soluble carbohydrates.PSB treatment had the highest effect on root traits,such as taproot length,root volume,cumulative root length,and the ratio of root to shoot.NFB treatment enhanced root diameter and induced biomass translocation between root systems.However,only the application of mixed biofertilizer(i.e.,NPK treatment)was the most significant treatment to improve all plant morphological and physiological characteristics of salt tree under drought stress.Therefore,our results provided improvement of some specific plant traits simultaneous with application of three biofertilizers to increase growth and establishment of salt tree seedlings in the degraded arid lands.

Effect of Inoculation with Three Phytohormone Producers Phytobacteria with ACC Deaminase Activity on Root Length of Lens esculenta Seedlings

Plant-associated bacteria that inhabit the rhizosphere may influence the plant growth by their contribution to the endogenous pool of phytohormones and by the activity of ACC deaminase to decrease the ethylene concentration. The aim of this study was to analyse the root length growth by the promoting effect of indole acetic acid producers phytobacteria with ACC deaminase activity, on inoculated seeds of Lens esculenta as synergistic effect on root elongation. In this study, although the roots of L. esculenta seedlings do not show a significant promotion, these phytobacteria could be recommended to treat plants analyzing their added inoculum to increase plant biomass and retard the effect of ethylene on cultures supplied with Tryptophan and ACC.

Potential of Novel Halotolerant and Psychrotrophic Strain of Planococcus sp. for Water Treatment

To screen multifunctional bacteria for water treatment,a chemical oxygen demand( COD)degradation and flocculation strain S2 A15 was obtained from the offshore sewage outfall at Weihai International Beach. Based on the phylogenetic characteristics,a novel strain was identified as a Planococcus species. Strain S2 A15 was determined to have the ability of flocculation and COD degradation. A series of experiments showed that the strain S2 A15 could be used for the treatment of four types of wastewater,including domestic wastewater( 400 mg/L and 800 mg/L) and high salt domestic wastewater( 400 mg/L and 800 mg/L). Among them,the best effect was exerted by the strain that reduced by 76.9% in domestic wastewater with 400 mg/L COD. The flocculation ratio reached 60.19%. The optimal treatment conditions are also discussed. We confirmed that the strain S2 A15 had salt tolerance and low temperature resistance. The best growth of S2 A15 at salt concentration of 6% and further confirmed that the strain could degrade COD at a low temperature.

Selection and validation of reference genes for quantitative real-time polymerase chain reaction analyses of Serratia ureilytica DW2

Background:Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C.pilosula;nonetheless,until now,no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction(RT–qPCR)data.Methods:To screen stable reference genes of S.ureilytica DW2,the expression of its eight candidate reference genes(16S rRNA,ftsZ,ftsA,mreB,recA,slyD,thiC,and zipA)under different treatment conditions(pH,temperature,culture time,and salt content)was assayed by RT–qPCR.The expression stability of these genes was analyzed using different algorithms(geNorm,NormFinder,and BestKeeper).To verify the reliability of the data,the expression of the glucose dehydrogenase(gdh)gene under different soluble phosphate levels was quantified using the most stably expressed reference gene.Results:The results showed that the zipA and 16S rRNA genes were the most stable reference genes,and the least stable genes were thiC and recA.The expression of gdh was consistent with the phosphate solubilization ability on plates containing the National Botanical Research Institute phosphate growth medium.Conclusion:Therefore,this study provides a stable and reliable reference gene of Serratia for the accurate quantification of functional gene expression in future studies.

Isolation,Identification,and Characterization of an Aluminum-Tolerant Bacterium Burkholderia sp.SB1 from an Acidic Red Soil

Aluminum(Al) toxicity usually occurs in acidic soils worldwide, which is detrimental to the growth of organisms. An Al-tolerant bacterium, SB1, was isolated from an acidic red soil of Chingkang Mountain, located in Jiangxi Province of China. Polyphasic analysis,including a 16 S rDNA phylogenetic tree as well as morphological and physicochemical properties, revealed that the isolate was a gramnegative, rod-shaped bacterium, which was recognized as Burkholderia sp. SB1 and had extreme acidity tolerance(pH 2.2) and excellent Al resistance(270 mg L^(-1) Al^(3+)). It could remove Al by up to 97.7% at a concentration of 54 mg L^(-1) Al^(3+). SB1 behavior under different temperatures and antibiotics was also examined. SB1 preferred moderate temperature conditions, ranging from 25 to37?C, and exhibited notable resistance to multiple antibiotics(including ampicillin, streptomycin, and tetracycline), except for being sensitive to chloramphenicol. Therefore, as the first reported bacterium to possess favorable Al resistance and excellent Al removal,Burkholderia sp. SB1 can potentially be used as an agent for bioremediation of Al-contaminated acidic red soils.

Drought-Tolerant Pseudomonas spp. Improve the Growth Performance of Finger Millet(Eleusine coracana(L.) Gaertn.)Under Non-Stressed and Drought-Stressed Conditions

Application of plant growth-promoting bacteria(PGPB)is an environmentally sustainable option to reduce the effects of abiotic and biotic stresses on plant growth and productivity.Three 1-aminocyclopropane-1-carboxylic acid(ACC)deaminase-producing drought-tolerant bacteria were isolated from a rain-fed agriculture field in the Central Himalaya of Kumaun region,Uttarakhand,India and evaluated for their efficiency in improving finger millet(Eleusine coracana(L.)Gaertn.)plant growth under non-stressed and drought-stressed conditions.These bacteria withstood a substrate metric potential of -1.0 MPa(30% polyethylene glycol 8000)and therefore were considered drought-tolerant.These strains were identified as Pseudomonas spp.by fatty acid methyl ester analysis and 16S rRNA gene sequencing.The ACC deaminase activity of these strains was characterized at the biochemical level,and the presence of acd S gene,the structural gene for ACC deaminase,was confirmed by the polymerase chain reaction.Two sets of pot trials in glass house were set up,one for normal(non-stressed)and the other for drought-stressed conditions.After 5 weeks,one set of plants was subjected to drought stress for 5 d,while the other set continued to be watered.The same growth parameters were recorded for both sets of plants after 40 d of plant growth.The results of pot trials showed that treatments inoculated with ACC deaminase-producing bacterial strains significantly improved the growth performance of finger millet plants and foliar nutrient content as compared to uninoculated treatments under both non-stressed and drought-stressed conditions.In addition,a significant increase in antioxidant activity was observed,wherein bacterial stain inoculation improved plant fitness by protecting it from oxidative damage induced by drought.

A heterotrophic nitrification-aerobic denitrification bacterium Halomonas venusta TJPU05 suitable for nitrogen removal from high-salinity wastewater

A novel salt-tolerant heterotrophic nitrification and aerobic denitrification(HN-AD)bacterium was isolated and identified as Halomonas venusta TJPU05(H.venusta TJPU05).The nitrogen removal performance of H.venusta TJPU05 in simulated water(SW)with sole or mixed nitrogen sources and in actual wastewater(AW)with high concentration of salt and nitrogen was investigated.The results showed that 86.12%of NH_(4)^(+)-N,95.68%of NO_(3)^(-)-N,100%of NO_(2)^(-)-N and 84.57%of total nitrogen(TN)could be removed from SW with sole nitrogen sources within 24 h at the utmost.H.venusta TJPU05 could maximally remove 84.06%of NH_(4)^(+)-N,92.33%of NO_(3)^(-)-N,92.9%of NO_(2)^(-)-N and 77.73%of TN from SW with mixed nitrogen source when the salinity was above 8%.The application of H.venusta TJPU05 in treating AW with high salt and high ammonia nitrogen led to removal efficiencies of 50.96%,47.28%and 43.19%for NH_(4)^(+)-N,NO_(3)^(-)-N and TN respectively without any optimization.Furthermore,the activities of nitrogen removal–related enzymes of the strain were also investigated.The successful detection of high level activities of ammonia oxygenase(AMO),hydroxylamine oxidase(HAO),nitrate reductase(NAR)and nitrite reductase(NIR)enzymes under high salinity condition further proved the HN-AD and salt-tolerance capacity of H.venusta TJPU05.These results demonstrated that the H.venusta TJPU05 has great potential in treating high-salinity nitrogenous wastewater.

Biosurfactant-assisted phytoremediation of potentially toxic elements in soil:Green technology for meeting the United Nations Sustainable Development Goals

Biosurfactants are biomolecules produced by microorganisms, low in toxicity, biodegradable, and relatively easy to synthesize using renewable waste substrates. Biosurfactants are of great importance with a wide and versatile range of applications, including the bioremediation of contaminated sites. Plants may accumulate soil potentially toxic elements(PTEs), and the accumulation efficacy may be further enhanced by the biosurfactants produced by rhizospheric microorganisms. Occasionally, the growth of bacteria slows down in adverse conditions, such as highly contaminated soils with PTEs. In this context,the plant's phytoextraction capacity could be improved by the addition of metal-tolerant bacteria that produce biosurfactants. Several sources, categories,and bioavailability of PTEs in soil are reported in this article, with the focus on the cost-effective and sustainable soil remediation technologies, where biosurfactants are used as a remediation method. How rhizobacterial biosurfactants can improve PTE recovery capabilities of plants is discussed, and the molecular mechanisms in bacterial genomes that support the production of important biosurfactants are listed. The status and cost of commercial biosurfactant production in the international market are also presented.

Agronomic evaluation of Herbaspirillum seropedicae strain ZAE94 as an inoculant to improve maize yield in Brazil

Diazotrophic bacteria applied as a seed inoculant can improve the grain yield of several crops including maize. The current study aimed to test the agronomic efficiency and contribution of biological nitrogen fixation(BNF) of the endophytic diazotroph Herbaspirillum seropedicae strain ZAE94 to maize under field conditions. Eighteen field assays were conducted in four different locations during consecutive years on two hybrids and two varieties of maize in a random block design with four replicates using a peat-based inoculant. The inoculant containing the ZAE94 strain was applied without nitrogen(N)fertilization or with 40 kg N ha^(-1) and was compared to the application of 40 and 80 kg N ha^(-1) without inoculation. Crop productivity and N accumulation in the grain were evaluated in addition to ^(15)N natural abundance(δ^(15)N) to evaluate BNF in the treatments without N fertilization. Fertilization at 40 kg N ha^(-1) plus bacterial inoculation produced crop yields similar to the treatment with 80 kg N ha^(-1) and increased grain N content, especially in the off-season with 40 kg N ha^(-1). The inoculation treatments showed lower δ^(15)N values than the non-inoculated treatments, which was most evident in the off-season. The BNF contributed about 30% of N accumulated in plants inoculated with ZAE94. On average, 64% of the N fertilized plots showed an increase of the parameters evaluated in the inoculated treatments, compared with the control. Inoculation also increased root length, root volume, and leaf area, and these parameters were positively correlated with plant weight using a hydroponic assay. This study revealed that the application of H. seropedicae inoculant increased the amount of N in plants owing to BNF, and there is a better chance of yield response to inoculation under low N fertilizer application in the off-season.

Co-inoculation of indigenous Pseudomonas oryzihabitans and Bradyrhizobium sp.modulates the growth,symbiotic efficacy,nutrient acquisition,and grain yield of soybean

Soil microbes play a vital role in improving plant growth,crop productivity,and soil health through solubilization of essential nutrients.Present investigation was conducted to access the efficacy of Bradyrhizobium sp.LSBR-3 and the indigenous phosphate-solubilizing Pseudomonas oryzihabitans LSE-3 in improving the symbiosis,nutrient accumulation,and yield of soybean.The isolate LSE-3,selected on the basis of phosphate solubilization,was screened for beneficial traits,antagonistic activities,and pathogenicity.The levels of indole acetic acid production(50.34±2.35μg mL^(-1)),phosphate solubilization(184.4±7.4 mg L^(-1)),biofilm formation(optical density at 560 mm,1.3896±0.04),siderophore production(121.46±1.61μg mL^(-1)),and 1-aminocyclopropane-1-carboxylate deaminase activity(0.51±0.07 mmolα-ketobutyrateμg^(-1) protein h^(-1))were significantly higher with the dual inoculants(LSBR-3 and LSE-3)than with the single inoculant LSBR-3.The plant growth-promoting traits of single and dual inoculants were evaluated for the synergistic effects on soybean under field conditions.Soybean plots treated with LSBR-3+LSE-3 exhibited improvement in seed germination,plant height,plant biomass,and chlorophyll content compared with the uninoculated control.Dual inoculant treatments resulted in significantly higher symbiotic efficacy evidenced by increased nodulation(40.0±0.75 plant^(-1)),nodule biomass(188.52±6.29 mg plant^(-1)),and leghemoglobin content(11.02±0.83 mg g^(-1) fresh nodule),and significantly increased activities of phosphatase(75.16±3.17 and 58.77±6.08μg p-nitrophenol g^(-1) h^(-1) for alkaline and acid phosphatase,respectively)and dehydrogenase(32.66±1.92μg triphenylformazan g^(-1) h^(-1))compared with the control.Dual inoculation with LSBR-3 and LSE-3 enhanced the uptake of macro-and micronutrients,reduced Na content in shoots,and resulted in 10.85%higher grain yield and ca.US$96.80 ha^(-1) higher profit compared with the control.This is the first report on the effectiveness of combined inoculation of LSE-3 and LSBR-3 in promoting the growth,symbiotic efficacy,and yield of soybean for sustainable agriculture.