ACC Deaminase Production Research Articles

Effects of plant growth-promoting rhizobacteria strains producing ACC deaminase on photosynthesis, isoprene emission, ethylene formation and growth of Mucuna pruriens (L.) DC. in response to water deficit

Agricultural sustainability is an increasing need considering the challenges posed by climate change and rapid human population growth. The use of plant growth-promoting rhizobacteria (PGPR) may represent an excellent, new agriculture practice to improve soil quality while promoting growth and yield of important crop species subjected to water stress conditions. In this study, two PGPR strains with 1-Aminocyclopropane-1-Carboxylate (ACC) deaminase activity were co-inoculated in velvet bean plants to verify the physiological, biochemical and molecular responses to progressive water stress. The results of our study show that the total biomass and the water use efficiency of inoculated plants were higher than uninoculated plants at the end of the water stress period. These positive effects may be derived from a lower root ACC content (−45 %) in water-stressed inoculated plants than in uninoculated ones resulting in lower root ethylene emission. Furthermore, the ability of inoculated plants to maintain higher levels of both isoprene emission, a priming compound that may help to protect leaves from oxidative damage, and carbon assimilation during water stress progression may indicate the underlining metabolic processes conferring water stress tolerance. Overall, the experimental results show that co-inoculation with ACC deaminase PGPR positively affects tolerance to water deficit, confirming the potential for biotechnological applications in water-stressed agricultural areas.

Mitigation of Copper Stress in Maize (Zea mays) and Sunflower (Helianthus annuus) Plants by Copper-resistant Pseudomonas Strains.

Use of heavy metal (HM) resistant plant growth-promoting rhizobacteria (PGPR) is among the eco-friendly strategies to increase the resistance of crop plants against the HM stress. In this study, we investigated the effects of two copper (Cu)-resistant PGPR strains (Pseudomonas fluorescens P22 and Pseudomonas sp. Z6) on the growth and nutrition of maize (Zea mays) and sunflower (Helianthus annuus) plants grown in a Cu-contaminated soil under glasshouse conditions. Both PGPR strains significantly increased the plant vegetative parameters including shoot biomass, stem height and diameter, and chlorophyll (SPAD values) index in both crops. In both plants, the PGPR inoculations also significantly elevated the uptake of nutrients including potassium, phosphorus, calcium, magnesium (only by P. fluorescens P22), iron, zinc, manganese, and Cu. Magnitude of the nutritional effects varied between the PGPR strains, e.g., in sunflower, inoculation with P. fluorescens P22 and Pseudomonas sp. Z6 led to an increase in uptake of Zn by 42% and 114%, or Mn by 61% and 88%, respectively, in comparison with control plants. Improved performance of the inoculated plants can be attributed to the plant growth-promoting (e.g., production of auxin and siderophore, phosphate solubilization activities, etc.) and stress removal (e.g., production of ACC-deaminase to drop the ethylene level in stressed plants) properties of the PGPR strains, which were uncovered in our in vitro studies prior to the glasshouse experiment. Beside the plant growth-promoting traits of these PGPR strains, their high resistance to Cu toxicity seemed to be of particular importance for plant fitness improvement under Cu toxicity.

Biofilm Producing Rhizobacteria With Multiple Plant Growth-Promoting Traits Promote Growth of Tomato Under Water-Deficit Stress.

Plant growth-promoting rhizobacteria (PGPR) not only enhance plant growth but also control phytopathogens and mitigate abiotic stresses, including water-deficit stress. In this study, 21 (26.9%) rhizobacterial strains isolated from drought-prone ecosystems of Bangladesh were able to form air–liquid (AL) biofilms in the glass test tubes containing salt-optimized broth plus glycerol (SOBG) medium. Based on 16S rRNA gene sequencing, Pseudomonas chlororaphis (ESR3 and ESR15), P. azotoformans ESR4, P. poae ESR6, P. fluorescens (ESR7 and ESR25), P. gessardii ESR9, P. cedrina (ESR12, ESR16, and ESR23), P. veronii (ESR13 and ESR21), P. parafulva ESB18, Stenotrophomonas maltophilia ESR20, Bacillus cereus (ESD3, ESD21, and ESB22), B. horikoshii ESD16, B. aryabhattai ESB6, B. megaterium ESB9, and Staphylococcus saprophyticus ESD8 were identified. Fourier transform infrared spectroscopy studies showed that the biofilm matrices contain proteins, polysaccharides, nucleic acids, and lipids. Congo red binding results indicated that these bacteria produced curli fimbriae and nanocellulose-rich polysaccharides. Expression of nanocellulose was also confirmed by Calcofluor binding assays and scanning electron microscopy. In vitro studies revealed that all these rhizobacterial strains expressed multiple plant growth-promoting traits including N2 fixation, production of indole-3-acetic acid, solubilization of nutrients (P, K, and Zn), and production of ammonia, siderophores, ACC deaminase, catalases, lipases, cellulases, and proteases. Several bacteria were also tolerant to multifarious stresses such as drought, high temperature, extreme pH, and salinity. Among these rhizobacteria, P. cedrina ESR12, P. chlororaphis ESR15, and B. cereus ESD3 impeded the growth of Xanthomonas campestris pv. campestris ATCC 33913, while P. chlororaphis ESR15 and B. cereus ESD21 prevented the progression of Ralstonia solanacearum ATCC® 11696TM. In a pot experiment, tomato plants inoculated with P. azotoformans ESR4, P. poae ESR6, P. gessardii ESR9, P. cedrina ESR12, P. chlororaphis ESR15, S. maltophilia ESR20, P. veronii ESR21, and B. aryabhattai ESB6 exhibited an increased plant growth compared to the non-inoculated plants under water deficit-stressed conditions. Accordingly, the bacterial-treated plants showed a higher antioxidant defense system and a fewer tissue damages than non-inoculated plants under water-limiting conditions. Therefore, biofilm-producing PGPR can be utilized as plant growth promoters, suppressors of plant pathogens, and alleviators of water-deficit stress.

Environmental Adaptations of an Extremely Plant Beneficial Bacillus subtilis Dcl1 Identified Through the Genomic and Metabolomic Analysis.

Bacterial endophytes ubiquitously colonize the internal tissues of plants and promote the plant growth through diverse mechanisms. The current study describes the mechanistic basis of plant-specific adaptations present in an extremely beneficial endophytic bacterium. Here, the endophytic Bacillus subtilis Dcl1 isolated from the dried rhizome of Curcuma longa was found to have the drought tolerance, IAA and ACC deaminase production and phosphate solubilization properties. The whole genome sequencing and annotation further showed the genome of B. subtilis Dcl1 to have the size of 4,321,654 bp. This also showed the presence of genes for IAA, H2S, acetoin, butanediol, flagella and siderophore production along with phosphate solubilization and biofilm formation for the B. subtilis Dcl1. In addition, the genes responsible for the synthesis of surfactin, iturin, fengycin, bacillibactin, bacillaene, bacilysin, chitinase, chitosanase, protease and glycoside hydrolase could also be annotated from the genome of B. subtilis Dcl1. Identification of genes for the glycine betaine, glutamate and trehalose further indicated the drought stress tolerance features of B. subtilis Dcl1. The presence of the genetic basis to produce the catalase, superoxide dismutase, peroxidases, gamma-glutamyltranspeptidase, glutathione and glycolate oxidase also indicated the plant oxidative stress protective effect of B. subtilis Dcl1. Identification of these properties and the demonstration of its plant probiotic effect in Vigna unguiculata confirmed the applicability of B. subtilis Dcl1 as a biofertilizer, biocontrol and bioremediator agent to enhance the agricultural productivity.

Field evaluation of consortium of bacterial inoculants producing ACC deaminase on growth, nutrients and yield components of rice and wheat

Bacterial isolates Achromobacter, Ochrobactrum Pseudomonas, and Variovorax have often been isolated from plant roots, and most of them have been described as growth enhancing rhizobacteria. These rhizobacteria have been used to boost crops productivity in varied region of agro-climatic regions. To lessen the usage of chemical fertilizers inputs and improve the sustainability of rice–wheat crops, this study was planned to investigate the impacts of bacterial consortium inoculants producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase on plant growth, nutrient content and grain yield in a field trial. Rice and wheat plants subjected to plant growth-promoting rhizobacteria (PGPR) consortia of strain DPC9 (Ochrobactrum anthropi) + DPB13 (Pseudomonas palleroniana) + DPB15 (Pseudomonas fluorescens) + DPB16 (Pseudomonas palleroniana) gave the best results with reference to macronutrient—nitrogen, phosphorus, potassium, calcium and sodium content and yield-related parameters including 1000 grain weight (10.2%, 40.7%), number of grains per panicle/spike (45.5%, 60.6%), and tillers (32.2%, 106.6%). This inoculants significantly increased grain yield (65.6%, 74.4%), straw yield (26.8%, 36.9%) and harvest index (14.9%, 13.8%) of rice–wheat crops, respectively, when compared to their counterpart control plants. Significant positive correlation in parameters which contribute in producing high grain yield in rice–wheat include plant height (r = 0.938, r = 0.852), tillers (r = 0.968, r = 0.881), panicle/spike length (r = 0.844, r = 0.912), number of grains per panicle/spike (r = 0.969, r = 0.815), 1000 grain weight (r = 0.833, r = 0.931), straw yield (r = 0.920, r = 0.918) and harvest index (r = 0.909, r = 0.847) and also surge nutrient enhancement in rice and wheat were observed in consortium treated plants. It can be concluded that consortium of DPC9 + DPB13 + DPB15 + DPB16-producing ACC deaminase can serve as a useful bio-inoculant for sustainable rice–wheat production in diverse agro-ecosystem.

Endophytic bacteria associated with halophyte Seidlitzia rosmarinus Ehrenb. ex Boiss. from saline soil of Uzbekistan and their plant beneficial traits

Endophytic bacteria of halophytic plants play essential roles in salt stress tolerance. Therefore, an understanding of the true nature of plant—microbe interactions under extreme conditions is essential. The current study aimed to identify cultivable endophytic bacteria associated with the roots and shoots of Seidlitzia rosmarinus Ehrenb. ex Boiss. grown in the salt-affected soil in Uzbekistan and to evaluate their plant beneficial traits related to plant growth stimulation and stress tolerance. Bacteria were isolated from the roots and the shoots of S. rosmarinus using culture-dependent techniques and identified by the 16S rRNA gene. RFLP (Restriction Fragment Length Polymorphism) analysis was conducted to eliminate similar isolates. Results showed that the isolates from the roots of S. rosmarinus belonged to the genera Rothia, Kocuria, Pseudomonas, Staphylococcus, Paenibacillus and Brevibacterium. The bacterial isolates from the shoots of S. rosmarinus belonged to the genera Staphylococcus, Rothia, Stenotrophomonas, Brevibacterium, Halomonas, Planococcus, Planomicrobium and Pseudomonas, which differed from those of the roots. Notably, Staphylococcus, Rothia and Brevibacterium were detected in both roots and shoots, indicating possible migration of some species from roots to shoots. The root-associated bacteria showed higher levels of IAA (indole-3-acetic acid) synthesis compared with those isolated from the shoots, as well as the higher production of ACC (1-aminocyclopropane-1-carboxylate) deaminase. Our findings suggest that halophytic plants are valuable sources for the selection of microbes with a potential to improve plant fitness under saline soils.

Seed priming with Pseudomonas putida isolated from rhizosphere triggers innate resistance against Fusarium wilt in tomato through pathogenesis-related protein activation and phenylpropanoid pathway

Pathogenesis-related (PR) proteins are one of the major and preliminary proteins accumulated as a defense against biotic stress. This defense response can be induced by using beneficial rhizobacteria, which has been studied in various host-pathogen interactions. In the present study, eleven Pseudomonas isolates were assessed for their potential to ferment sorbitol, reduce nitrate, and produce mycolytic enzymes, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, phenazine antibiotics, and N-acyl homoserine lactones (AHLs). All isolates were tested against the host-specific pathogen Fusarium oxysporum MTCC1755 in tomato under greenhouse conditions, and shortlisted isolates were tested for their rhizosphere competence. In-vitro test results showed that the isolates were able to produce mycolytic enzymes, including protease, lipase, chitinase, cellulase, and amylase, and the antibiotic phenazine and were negative for pyoluteorin. All the isolates except two were positive for ACC deaminase production. Greenhouse results showed that the isolates M80, M96, and T109 significantly reduced symptoms of Fusarium wilt. Extended greenhouse tests under autoclaved and unautoclaved soil conditions showed that M80, M96, and T109 were excellent rhizosphere competitors and were identified as Pseudomonas putida. In brief, the defense-specific biochemical variations in the host could describe the improved defense against Fusarium wilt occurring in the primed plants. These three Pseudomonas strains could be used as potential biocontrol agents, along with their rhizosphere competence.

Bioprospection of native psychrotolerant plant-growth-promoting rhizobacteria from Peruvian Andean Plateau soils associated with Chenopodium quinoa.

The Peruvian Andean Plateau, one of the main production areas of native varieties of Chenopodium quinoa, is exposed to abrupt decreases in environmental temperature, affecting crop production. Plant-growth-promoting rhizobacteria that tolerate low temperatures could be used as organic biofertilizers in this region. We aimed to bioprospect the native psychrotolerant bacteria of the quinoa rhizosphere in this region that show plant-growth-promoting traits. Fifty-one strains belonging to the quinoa rhizosphere were characterised; 73% of the total could grow at low temperatures (4, 6, and 15 °C), whose genetic diversity based on DNA amplification of interspersed repetitive elements (BOX) showed 12 different profiles. According to the 16S rRNA sequence, bacterial species belonging to the classes Beta- and Gammaproteobacteria were identified. Only three (6%) isolates identified as nonpathogenic bacteria exhibited plant-growth-promoting activities, like IAA production, phosphate solubilization, growth in a nitrogen-free medium, and ACC deaminase production at 6 and 15 °C. ILQ215 (Pseudomonas silesiensis) and JUQ307 (Pseudomonas plecoglossicida) strains showed significantly positive plant growth effects in aerial length (about 50%), radicular length (112% and 79%, respectively), and aerial and radicular mass (above 170% and 210%, respectively) of quinoa plants compared with the control without bacteria. These results indicate the potential of both psychrotolerant strains to be used as potential organic biofertilizers for quinoa in this region.

Isolation and characterization of plant growth-promoting endophytic bacteria Bacillus stratosphericus LW-03 from Lilium wardii.

In the present study, a new strain of Bacillus stratosphericus LW-03 was isolated from the bulbs of Lilium wardii. The isolated endophytic strain LW-03 exhibited excellent antifungal activity against common plant pathogens, such as Fusarium oxysporum, Botryosphaeria dothidea, Botrytis cinerea, and Fusarium fujikuroi. The growth inhibition percentage of Botryosphaeria dothidea was 74.56 ± 2.35%, which was the highest, followed by Botrytis cinerea, Fusarium fujikuroi, and Fusarium oxysporum were 71.91 ± 2.87%, 69.54 ± 2.73%, and 65.13 ± 1.91%, respectively. The ethyl acetate fraction revealed a number of bioactive compounds and several of which were putatively identified as antimicrobial agents, such as 4-hydroxy-2-nonenylquinoline N-oxide, sphingosine ceramides like cer(d18:0/16:0(2OH)), cer(d18:0/16:0), and cer(d18:1/0:0), di-peptides, tri-peptide, cyclopeptides [cyclo(D-Trp-L-Pro)], [cyclo (Pro-Phe)], dehydroabietylamine, oxazepam, 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine like compound (PC(0:0/20:4), phosphatidylethanolamine (PE(18:1/0:0)), 3-Hydroxyoctadecanoic acid, 7.alpha.,27-Dihydroxycholesterol, N-Acetyl-d-mannosamine, p-Hydroxyphenyllactic acid, Phytomonic acid, and 2-undecenyl-quinoloin-4 (1H). The LW-03 strain exhibits multiple plant growth-promoting traits, including the production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, and nitrogen fixation activity. The beneficial effects of the endophytic strain LW-03 on the growth of two lily varieties were further evaluated under greenhouse conditions. Our results revealed plant growth-promoting activity in inoculated plants relative to non-inoculated control plants. The broad-spectrum antifungal activity and multiple plant growth-promoting properties of Bacillus stratosphericus LW-03 make it an important player in the development of biological fertilizers and sustainable agricultural biological control strategies.

Streptomyces sp. CLV45 from Fabaceae rhizosphere benefits growth of soybean plants.

Plant growth-promoting bacteria such as Streptomyces are an attractive alternative for increasing the sustainability of agricultural systems. In this study, Streptomyces isolates obtained from rhizosphere soil of plants in the family Fabaceae were characterized for their plant growth-promoting traits, including the production of siderophores, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, indole-3-acetic acid (IAA), and phenazines. Soybean seeds were bacterized with selected isolates to test growth promotion. All isolates produced IAA, and the isolate CLV45 was the most efficient, reaching 398.53mg of IAA per gram of cells. CLV41, CLV45, and CLV46 showed high activity for ACC deaminase whereas CLV42, CLV44, and CLV46 were efficient in siderophore production. Pyocyanin was detected in all isolates; CLV41, CLV43, and CLV45 produced phenazine-carboxylic acid as well. Selected for IAA and ACC deaminase production combined with production of siderophores and phenazines, CLV42, CLV44, and CLV45 were tested for their growth promotion potential. Seed bacterization with CLV45 resulted in plants with increased shoot growth (36.63%) and dry mass (17.97%) compared to control plants. Results suggest that moderate or high levels of auxin and ACC deaminase production by the isolate CLV45 positively affected the growth of soybean plants, making it a strong candidate for further studies on biofertilizer formulation.

The Endophytic Bacteria Bacillus velezensis Lle-9, Isolated from Lilium leucanthum, Harbors Antifungal Activity and Plant Growth-Promoting Effects.

Bacillus velezensis is an important plant growth-promoting rhizobacterium with immense potential in agriculture development. In the present study, Bacillus velezensis Lle-9 was isolated from the bulbs of Lilium leucanthum. The isolated strain showed antifungal activities against plant pathogens like Botryosphaeria dothidea, Fusarium oxysporum, Botrytis cinerea and Fusarium fujikuroi. The highest percentage of growth inhibition i.e., 68.56±2.35% was observed against Fusarium oxysporum followed by 63.12 ± 2.83%, 61.67 ± 3.39% and 55.82 ± 2.76% against Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi, respectively. The ethyl acetate fraction revealed a number of bioactive compounds and several were identified as antimicrobial agents such as diketopiperazines, cyclo-peptides, linear peptides, latrunculin A, 5α-hydroxy-6-ketocholesterol, (R)-S-lactoylglutathione, triamterene, rubiadin, moxifloxacin, 9-hydroxy-5Z,7E,11Z,14Zeicosatetraenoic acid, D-erythro-C18-Sphingosine, citrinin, and 2- arachidonoyllysophosphatidylcholine. The presence of these antimicrobial compounds in the bacterial culture might have contributed to the antifungal activities of the isolated B. velezensis Lle- 9. The strain showed plant growth-promoting traits such as production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, and nitrogen fixation and phosphate solubilization. IAA production was accelerated with application of exogenous tryptophan concentrations in the medium. Further, the lily plants upon inoculation with Lle-9 exhibited improved vegetative growth, more flowering shoots and longer roots than control plants under greenhouse condition. The isolated B. velezensis strain Lle-9 possessed broad-spectrum antifungal activities and multiple plant growth-promoting traits and thus may play an important role in promoting sustainable agriculture. This strain could be developed and applied in field experiments in order to promote plant growth and control disease pathogens.

Streptomyces lydicus M01 Regulates Soil Microbial Community and Alleviates Foliar Disease Caused by Alternaria alternata on Cucumbers.

Due to the adverse effect on the environment caused by excessive use of chemical fertilizers, the development of sustainable agriculture attracts a growing demand of biological based fertilizers composed of living microorganisms. In this study, an Actinobacteria Streptomyces lydicus M01 was isolated from the rhizosphere soil of Pyrus calleryana. This strain effectively promoted the plant growth and suppressed a foliar disease caused by Alternaria alternata on cucumbers. S. lydicus M01 exhibited growth promoting characteristics such as phosphate solubilization, IAA secretion, siderophore and ACC deaminase production. Through Illumina sequencing of the 16S rRNA gene and ITS gene of the soil microbes, we found that the application of S. lydicus M01 altered the composition of the microbial community by promoting beneficial groups, including bacteria genera Pseudarthrobacter, Sphingomonas, Rhodanobacter, and Pseudomonas, fungi genera Fusicolla, Humicola, Solicoccozyma, and Paraphaeosphaeria. Most of these bacteria and eukaryotes exhibit positive effects on growth promotion, such as nutrient accumulation, auxin secretion, abiotic stress alleviation, biological control, or bioremediation. Furthermore, studies on the reactive oxygen species (ROS) level and antioxidants of cucumber leaves revealed that S. lydicus M01 treatment reduced the ROS accumulation and increased the activities of antioxidases related with ROS scavenging, which indicated an enhanced disease resistance of cucumbers under biotic stress. Thus, our results suggest that the application of S. lydicus M01 can systemically affect plant microbiome interactions and represent a promising sustainable solution to improve agricultural production instead of chemical fertilizers.

Deciphering metal toxicity responses of flax (Linum usitatissimum L.) with exopolysaccharide and ACC-deaminase producing bacteria in industrially contaminated soils

Rapid industrialization is the main reason of heavy metals contamination of soil colloids and water reservoirs. Heavy metals are persistent inorganic pollutants; deleterious to plants, animals and human beings because of accumulation in food chain. The aim of the current work was to evaluate the role of indole acetic acid (IAA), exopolysaccharide (EPS) and ACC-deaminase producing plant growth promoting rhizobacteria (PGPR) i.e .B. gibsonii PM11 and B. xiamenensis PM14 in metal phytoremediation of metals, their survival and plant growth promotion potential in metal polluted environment as well as alterations in physio-biochemical responses of inoculated L. usitatissimum plants towards heavy metal toxicity. Two bacterial strains Bacillus gibsonii (PM11) and Bacillus xiamenensis (PM14), previously isolated from sugarcane's rhizosphere, were screened for metal tolerance (50 mg/l to 1000 mg/l) and plant growth promoting traits like IAA, ACC-deaminase, EPS production and nitrogen fixing ability under metal stress. The response of flax plant (Linum usitatissimum L.) was analyzed in a pot experiment containing both industrially contaminated and non-contaminated soils. Experiment was comprised of six different treatments, each with three replicates. At the end of the experiment, role of metal tolerant plant growth promoting bacterial inoculation was elucidated by analyzing the plant growth parameters, chlorophyll contents, antioxidative enzymes, and metal uptake both under standard and metal contaminated rhizospheres. Results revealed that root and shoot length, plant's fresh and dry weight, proline content, chlorophyll content, antioxidant enzymatic activity was increased in plants inoculated with plant growth promoting bacteria as compared to non-inoculated ones both in non-contaminated and industrial contaminated soils. In current study, inoculation of IAA, EPS and ACC-deaminase producing bacteria enhances plant growth and nutrient availability by minimizing metal-induced stressed conditions. Moreover, elevated phytoextraction of multi-metals from industrial contaminated soils by PGPR inoculated L. usitatissimum plants reveal that these strains could be used as sweepers in heavy metals polluted environment.

The ACC-Deaminase Producing Bacterium Variovorax sp. CT7.15 as a Tool for Improving Calicotome villosa Nodulation and Growth in Arid Regions of Tunisia.

Calicotome villosa is a spontaneous Mediterranean legume that can be a good candidate as pioneer plants to limit regression of vegetation cover and loss of biodiversity in Tunisian arid soils. In order to grow legumes in such soils, pairing rhizobia and nodule associated bacteria (NAB) might provide numerous advantages. In this work, cultivable biodiversity of rhizobial symbionts and NAB in nodules of C. villosa plants growing in five arid regions of south Tunisia was characterized. Phylogenetic analysis using 16S rDNA gene, dnak, recA and nodD sequences separated nodule-forming bacteria in six clades associated to genera Ensifer, Neorhizobium, Phyllobacterium and Rhizobium. Among NAB, the strain Variovorax sp. CT7.15 was selected due to its capacity to solubilise phosphate and, more interestingly, its high level of aminocyclopropane-1-carboxylate deaminase (ACC deaminase) activity. C. villosa plants were inoculated with representative rhizobia of each phylogenetic group and co-inoculated with the same rhizobia and strain CT7.15. Compared with single rhizobia inoculation, co-inoculation significantly improved plant growth and nodulation, ameliorated plant physiological state and increased nitrogen content in the plants, independently of the rhizobia used. These results support the benefits of pairing rhizobia and selected NAB to promote legume growth in arid or degraded soils.

ACC-deaminase producing plant growth promoting rhizobacteria and biochar mitigate adverse effects of drought stress on maize growth.

Availability of good quality irrigation water is a big challenge in arid and semi arid regions of the world. Drought stress results in poor plant growth and low yield; however, the rhizobacteria, capable of producing 1-aminocyclopropane-1-carboxylate (ACC)-deaminase are likely to improve crop growth and productivity under drought stress. Similarly, biochar could also ameliorate the negative impacts of drought stress. Therefore, this pot experiment was conducted to evaluate the role of ACC-deaminase producing plant growth promoting rhizobacteria (PGPR) alone and in combinations with timber-waste biochar in improving maize growth under drought stress. The ACC-deaminase producing rhizobacteria, Pseudomonas aeruginosa, Enterobacter cloacae, Achromobacter xylosoxidans and Leclercia adecarboxylata were studied along with two rates (0.75 and 1.50% of the soil weight) of biochar under three moisture levels i.e., normal moisture, mild drought stress and severe drought stress. The E. cloacae in conjunction with higher rate of biochar produced a significant improvement i.e., up to 60, 73, 43, 69, 76 and 42% respectively, in grain yield plant-1, photosynthetic rate, stomatal conductance, chlorophyll a, total chlorophyll and carotenoids contents of maize as compared to the control under mild drought stress. Similarly, A. xylosoxidans with higher rate of biochar also enhanced grain yield plant-1, photosynthetic rate, stomatal conductance, chlorophyll a, total chlorophyll and carotenoids contents of maize up to 200, 213, 113, 152, 148 and 284%, respectively over control under severe drought stress. In conclusion, combination of ACC-deaminase containing PGPR, A. xylosoxidans and biochar (0.75%) proved an effective technique to improve maize growth and productivity under drought stress.

Isolation and Characterization of Plant Growth-Promoting Endophytic Bacteria Paenibacillus polymyxa SK1 from Lilium lancifolium.

Paenibacillus polymyxa is a plant growth-promoting rhizobacterium that has immense potential to be used as an environmentally friendly replacement of chemical fertilizers and pesticides. In the present study, Paenibacillus polymyxa SK1 was isolated from bulbs of Lilium lancifolium. The isolated endophytic strain showed antifungal activities against important plant pathogens like Botryosphaeria dothidea, Fusarium oxysporum, Botrytis cinerea, and Fusarium fujikuroi. The highest percentage of growth inhibition, i.e., 66.67 ± 2.23%, was observed for SK1 against Botryosphaeria dothidea followed by 61.19 ± 3.12%, 60.71 ± 3.53%, and 55.54 ± 2.89% against Botrytis cinerea, Fusarium fujikuroi, and Fusarium oxysporum, respectively. The metabolite profiling of ethyl acetate fraction was assessed through the UHPLC-LTQ-IT-MS/MS analysis, and putative identification was done with the aid of the GNPS molecular networking workflow. A total of 29 compounds were putatively identified which included dipeptides, tripeptides, cyclopeptides (cyclo-(Leu-Leu), cyclo(Pro-Phe)), 2-heptyl-3-hydroxy 4-quinolone, 6-oxocativic acid, anhydrobrazilic acid, 1-(5-methoxy-1H-indol-3-yl)-2-piperidin-1-ylethane-1,2-dione, octadecenoic acid, pyochelin, 15-hydroxy-5Z,8Z,11Z, 13E-eicosatetraenoic acid, (Z)-7-[(2R,3S)-3-[(2Z,5E)-Undeca-2,5-dienyl]oxiran-2-yl]hept-5-enoic acid, arginylasparagine, cholic acid, sphinganine, elaidic acid, gossypin, L-carnosine, tetrodotoxin, and ursodiol. The high antifungal activity of SK1 might be attributed to the presence of these bioactive compounds. The isolated strain SK1 showed plant growth-promoting traits such as the production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, nitrogen fixation, and phosphate solubilization. IAA production was strongly correlated with the application of exogenous tryptophan concentrations in the medium. Furthermore, inoculation of SK1 enhanced plant growth of two Lilium varieties, Tresor and White Heaven, under greenhouse condition. In the light of these findings, the P. polymyxa SK1 may be utilized as a source of plant growth promotion and disease control in sustainable agriculture.

Endophytic bacteria promote growth of the medicinal legume Clitoria ternatea L. by chemotactic activity.

Part of the native root nodule endophytic microflora referring to members of the genera Proteobacteria and Sphingobacteria were used to test their bioefficacy as seed biopriming. These were quantified for their plant growth promoting (PGP) attributes such as IAA production, P and K-solubilization and ACC deaminase production. Results showed that significantly highest IAA was produced by E. hormaechi RCT10. The highest P-solubilization was observed with S. maltophila RCT31 it was solubilizing all the substrate tri-calcium phosphate, di-calcium phosphate, and zinc phosphate. Significantly highest K-solubilization was observed with S. maltophila RCT31 followed by E. turicensis RCT5. However, the maximum zinc solubilization was reported with S. maltophila RCT31 followed by E. turicensis RCT5. The maximum ACC deaminase was quantified in the bacterium. Results revealed that the E. hormaechi RCT10 utilized seed leechates most effectively while root exudates were maximally utilized by S. maltophila RCT31. The pots experiment proves that S. maltophila RCT31 was the most effective bacterium and it was replication vis-à-vis field experiment. In particular, S. maltophila RCT31 holds strong potential to be possibly used as a bioformulation for the medicinal legume, as an economical and eco-friendly alternative to agrochemicals.

Rhizobacteria producing ACC deaminase mitigate water-stress response in finger millet (Eleusine coracana (L.) Gaertn.).

The aim of the study was to examine the influence of single and consortia treatments of drought tolerant rhizobacteria producing ACC deaminase together with additional plant growth promoting (PGP) characteristics on finger millet growth, antioxidant and nutrient concentration under water-stressed and irrigated (no stress) conditions. These rhizobacteria belong to the Variovorax sp. Achromobacter spp. Pseudomonas spp. and Ochrobactrum sp. The single inoculant of RAA3 (Variovorax paradoxus) and a consortium inoculant of four bacteria, i.e., DPC9 (Ochrobactrum anthropi), DPB13 (Pseudomonas palleroniana), DPB15 (Pseudomonas fluorescens) and DPB16 (Pseudomonas palleroniana), significantly boosted the overall growth parameters and nutrient concentrations in leaves of finger millet. Moreover, elevated levels of the reactive oxygen species scavenging enzymes-superoxide dismutase (17.3%, 11.6%), guaiacol peroxidase (38.7%, 22.2%), catalase (33.7%, 21.3%) and ascorbate peroxidase (18.2%, 10.0%); cellular osmolytes-proline (41.5%, 25.0%), phenol (44.5%, 37.5%); higher leaf chlorophyll (64.4%, 30.8%) and a reduced level of hydrogen peroxide (50.7%, 59.5%) and malondialdehyde (48.4%,72.5%) were noted, respectively, after single inoculation of RAA3 and a consortium treatment by strains DPC9 + DPB13 + DPB15 + DPB16, in contrast with non-treated plants mainly under water-stressed conditions. This finding clearly illustrates that PGPB that express ACC deaminase along with additional PGP traits could be an efficient approach for improving plant health in environments, where agricultural practices are reliant on rain for water.

Multi-strain Inoculation with PGPR Producing ACC Deaminase is More Effective Than Single-strain Inoculation to Improve Wheat (Triticum aestivum) Growth and Yield

Multi-strain Inoculation with PGPR Producing ACC Deaminase is More Effective Than Single-strain Inoculation to Improve Wheat (Triticum aestivum) Growth and Yield

Phylogenetic analysis of halophyte-associated rhizobacteria and effect of halotolerant and halophilic phosphate-solubilizing biofertilizers on maize growth under salinity stress conditions.

The main objective of the present work was to evaluate plant growth-promoting abilities of bacterial strains from the rhizosphere of halophytes and their effect on maize growth under salinity stress. Halophilic bacteria were identified using 16S rRNA sequence analysis and their plant growth-promoting abilities were characterized. Phylogenetic analysis showed that bacterial strains belonging to Bacillus, Halobacillus and Pseudomonas were dominant in the rhizosphere of halophytes. More than 93% strains showed P-solubilization activity and IAA production. About 54% strains were able to produce ACC deaminase, 29% strains showed positive results for nitrogen fixation, 41 and 21% strains showed siderophores and HCN production ability respectively. More than 90% strains showed antifungal activity against more than two fungal pathogens and production of different hydrolytic enzymes. To study the plant growth-promoting effect on maize, five bacterial strains Bacillus safensis HL1HP11 and Bacillus pumilus HL3RS14, Kocuria rosea HL1RP8, Enterobacter aerogenes AT1HP4 and Aeromonas veronii AT1RP10 were used as inoculants; in the form of seed coat and enriched soil-based phosphate biofertilizers. All bacterial strains positively affected the maize growth as compared to non-inoculated control+NaCl plants. Plants inoculated with Bacillus HL3RS14-based soil biofertilizers showed maximum increase in dry weights of root (48-124%) and shoot (52-131%) as compared to control+NaCl (soil+rock phosphate, no inoculum). PGPR inoculations under salinity stress conditions showed high concentrations of proline, glycine betaine and malondialdehyde. These results indicated that under saline soil conditions, halophilic PGPR strains combined with carrier materials are promising candidates as biofertilizers.