Isoptericola croceus sp. nov., a novel actinobacterium isolated from saline-alkali soil.

A novel actinomycete, designated strain q2T, was isolated from the saline-alkaline soil, collected from Daqing, Heilongjiang province, China. The results of phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain q2T belongs to the genus Isoptericola, and showed the highest sequence similarity to Isoptericola halotolerans KCTC 19046T (98.48%) and Isoptericola chiayiensis KCTC 19740T (98.13%), respectively. The average nucleotide identity values between strain q2T and other members of the genus Isoptericola were lower than 95% recommended for distinguishing novel prokaryotic species. Cells of strain q2T were Gram-staining-positive, aerobic, non-motile, rod-shaped and non-spore-forming. Colonies of strain q2T were golden-yellow pigmented, tidy edged and smooth surfaced. Growth occurred at 15-37 °C (optimum, 29 °C), pH 7.0-10.0 (optimum, pH 8.0). The predominant respiratory quinones were MK-9(H4) and MK-9(H2). The main detected polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, and phosphatidylinositol mannoside. The peptidoglycan compositions were L-alanine, D-aspartic, L-glutamic acid and L-lysine (type A4α). The major cellular fatty acids (> 10%) were anteiso-C15:0, iso-C15:0, and anteiso-C17:0. The G+C content of the genomic DNA was determined to be 69.7%. Based on the phenotypic, physiological, genotypic, and phylogenetic data, strain q2T represents a novel species of the genus Isoptericola, for which the name Isoptericola croceus sp. nov. is proposed. The type strain is q2T (= GDMCC 1.2923T = KCTC 49759T).

Oceanobacillus saliphilus sp. nov., Isolated from Saline-Alkali Soil in Heilongjiang Province, China.

A novel bacterium, designated strain APA_H-1(4)T, was isolated from the saline-alkaline soil, Zhaodong, Heilongjiang Province, China. Phenotypic and chemotaxonomic analyses, and whole-genome sequencing were used to determine the taxonomic position of the strain. Phylogenetic analysis indicated that the isolate belongs to the genus Oceanobacillus, and showed the highest sequence similarity to O. damuensis KCTC 33146T (98.35%, similarity) and 'O. massiliensis' DSM 24644 (98.32%). The average nucleotide identity values between strain APA_H-1(4)T and other members of the genus Oceanobacillus were lower than 82% recommended for distinguishing novel prokaryotic species. The digital DNA-DNA hybridization values of strain APA_H-1(4)T with O. damuensis KCTC 33146T and 'O. massiliensis' DSM 24644 were 13.60 and 17.60%, respectively. Cells of strain APA_H-1(4)T were Gram-staining positive, motile, aerobic, spore-forming rods (0.5-0.7 × 1.8-2.6 µm) with flagella. The growth was found to occur optimally at 37 °C. The whole-cell hydrolysate contained meso-diaminopimelic acid as the diagnostic cell wall diamino acid. The main detected polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unidentified polar lipid. The predominant respiratory quinone was identified as menaquinone-7 (MK-7). The major cellular fatty acid (>10%) was anteiso-C15:0. The G + C content of the genomic DNA was determined to be 38.4% based on the draft genome sequence. Based on the comparative analysis of polyphasic taxonomic data, strain APA_H-1(4)T represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus saliphilus sp. nov. is proposed. The type strain is APA_H-1(4)T (=GDMCC 1.2239T = KCTC 43254T).

Surface display of ACC deaminase on endophytic Enterobacteriaceae strains to increase saline resistance of host rice sprouts by regulating plant ethylene synthesis.

BACKGROUND: Most endophytic bacteria in consortia, which provide robust and broad metabolic capacity, are attractive for applications in plant metabolic engineering. The aim of this study was to investigate the effects of engineered endophytic bacterial strains on rice sprout ethylene level and growth under saline stress. A protocol was developed to synthesize engineered strains by expressing bacterial 1-aminocyclopropane-1-carboxylate (ACC) deaminase gene on cells of endophytic Enterobacter sp. E5 and Kosakonia sp. S1 (denoted as E5P and S1P, respectively). RESULTS: Results showed that ACC deaminase activities of the engineered strains E5P and S1P were significantly higher than those of the wild strains E5 and S1. About 32-41% deaminase was expressed on the surface of the engineered strains. Compared with the controls without inoculation, inoculation with the wild and engineered strains increased the deaminase activities of sprouts. Inoculation with the engineered strains increased 15-21% more deaminase activities of sprouts than with the wild strains, and reduced the ethylene concentrations of sprouts more significantly than with wild strains (P < 0.05). Inoculation with the wild and engineered strains promoted the growth of sprouts, while the promoting effects were more profound with the engineered strains than with the wild strains. The engineered strains improved saline resistance of sprouts under salt concentrations from 10 to 25 g L-1. The engineered strains promoted longer roots and shoots than the wild strains under the salt stresses, indicating that the ACC deaminases on the endophytic bacterial cells could result in plant-produced ACC degradation and inhibit plant ethylene formation. CONCLUSIONS: The protocols of expressing enzymes on endophytic bacterial cells showed greater potentials than those of plant over-expressed enzymes to increase the efficiency of plant metabolic pathways.

Nitrogen removal from synthetic wastewater using single and mixed culture systems of denitrifying fungi, bacteria, and actinobacteria.

The aim of this study was to investigate the effects of single and mixed culture of denitrifying fungi, bacteria, and actinobacteria on nitrogen removal and N2O emission in treatment of wastewater. Denitrifying endophytes of Pseudomonas sp. B2, Streptomyces sp. A9, and Fusarium sp. F3 isolated from rice plants were utilized for treatment of synthetic wastewater containing nitrate and nitrite. Experiments were conducted under shaking and static conditions. Results showed that under the static condition, more than 97 % of nitrate removal efficiencies were reached in all the treatments containing B2. The nitrate removal rates within the first 12 h in the treatments of B2, B2+A9, B2+F3, and B2+A9+F3 were 7.3, 9.8, 11, and 11 mg L-1 h-1, respectively. Under the shaking condition, 100 % of nitrite was removed in all the treatments containing B2. The presence of A9 and F3 with B2 increased the nitrite removal rates under both the shaking and static conditions. Compared to the B2 system, the mixed systems of B2+A9, B2+F3, and B2+A9+F3 reduced N2O emission (78.4 vs. 19.4, 1.80, and 0.03 µM in 4 weeks, respectively). Our results suggested that B2 is an important strain that enhances nitrogen removal from wastewater. Mixed cultures of B2 with A9 and F3 can remove more nitrate and nitrite from wastewater and reduce nitrite accumulation and N2O emission in the denitrification process.

Kocuria subflava sp. nov., isolated from marine sediment from the Indian Ocean.

A novel Gram-staining positive, catalase-positive, oxidase-negative, aerobic, non-motile coccus, designated strain YIM 13062(T), was isolated from a marine sediment sample collected from the Indian Ocean. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 13062(T) belongs to the genus Kocuria, and is closely related to Kocuria polaris NBRC 103063(T) (97.8 % similarity), Kocuria rosea NBRC 3768(T) (97.6 % similarity) and Kocuria carniphila JCM 14118(T) (97.4 % similarity). The strain grew optimally at 28 °C, pH 8.0 and in the presence of 2-4 % (w/v) NaCl. Cell-wall peptidoglycan type was Lys-Ala3 (type A3α). The major isoprenoid quinones were MK-6(H2) and MK-7(H2). The polar lipids of strain YIM 13062(T) consisted of diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), one unidentified phospholipid (PL), one unidentified aminophospholipid (APL), two unidentified aminolipids (AL) and four unidentified lipids (L). Major fatty acids of the novel isolate were anteiso-C15:0, iso-C14:0 and C18:1 2OH. The genomic DNA G+C content of strain YIM 13062(T) was 68.0 mol%. The level of DNA-DNA relatedness between strain YIM 13062(T) and K. polaris NBRC 103063(T), K. rosea NBRC 3768(T), K. carniphila JCM 14118(T) were 53.2, 48.8 and 42.6 %, respectively. On the basis of genotypic and phenotypic data, it is apparent that strain YIM 13062(T) represents a novel species of the genus Kocuria, for which the name Kocuria subflava sp. nov. is proposed. The type strain is YIM 13062(T) (=CGMCC 4.7252(T)=KCTC 39547(T)).

Engineering the robustness of Saccharomyces cerevisiae by introducing bifunctional glutathione synthase gene.

Robust, high-yielding Saccharomyces cerevisiae is highly desirable for cost-effective cellulosic ethanol production. In this study, the bifunctional glutathione (GSH) synthetase genes GCSGS at high copy number was integrated into ribosomal DNA of S. cerevisiae by Cre-LoxP system. Threefold higher GSH contents (54.9 µmol/g dry weight) accumulated in the engineered strain BY-G compared to the reference strain. Tolerance of BY-G to H2O2 (3 mM), temperature (40 °C), furfural (10 mM), hydroxymethylfurfural (HMF, 10 mM) and 0.5 mM Cd(2+) increased compared to reference strain. Twofold higher ethanol concentration was obtained by BY-G in simultaneous saccharification and fermentation of corn stover compared to the reference strain. The results showed that intracellular GSH content of S. cerevisiae has an influence on robustness. The strategy is used to engineer S. cerevisiae strains adaptive to a combination of tolerance to inhibitors and raised temperature that may occur in high solid simultaneous saccharification and fermentation of lignocellulosic feedstocks.

Determination of ionophore antibiotics nactins produced by fecal Streptomyces from sheep.

To investigate the correlation between fecal actinobacteria and host animals, Streptomyces was isolated from fresh faeces of healthy sheep and secondary metabolites were analyzed. The most frequently isolated strain S161 with antibiotic activity against bacteria and fungi were analyzed. The S161 showed the highest 99 % similarity to Streptomyces canus DSB17 based on the 16S rRNA gene sequence analysis. Metabolite analysis based on MS and NMR spectra showed that S161 produces nactins, cyclotetralactones derived from nonactic acid and homononactic acid as building units of ionophoretic character. Due to ionophores are antimicrobial compounds that are commonly fed to ruminant animals to improve feed efficiency, stable beneficial interactions between Streptomyces bacteria and vertebrates have been demonstrated.

Siderophore production by actinobacteria.

Produced by bacteria, fungi and plants, siderophores are low-molecular-weight chelating agents (200-2,000 Da) to facilitate uptake of iron (Fe). They play an important role in extracellular Fe solubilization from minerals to make it available to microorganisms. Siderophores have various chemical structures and form a family of at least 500 different compounds. Some antibiotics (i.e., albomycins, ferrimycins, danomycins, salmycins, and tetracyclines) can bind Fe and some siderophores showed diverse biological activities. Functions and applications of siderophores derived from actinobacteria were reviewed to better understand the diverse metabolites.

Removal of Cr(VI) from aqueous solutions by fruiting bodies of the jelly fungus (Auricularia polytricha).

The aim of this study was to investigate the potential to remove chromium (Cr) from aqueous solutions using the fruiting body of Auricularia polytricha. Batch experiments were conducted under various conditions, and different models were used to characterize the biosorption process. Results showed that, for both fresh and dried fruiting bodies of A. polytricha, removal efficiencies of Cr(VI) and total Cr reached maximum values at pH values of 1 and 2, respectively. The process of Cr(VI) removal by A. polytricha included the sorption process as well as the reduction of Cr(VI) to Cr(III). Spectra of X-ray photoelectron spectroscopy of the biosorbent revealed that most of the Cr loaded on the biomass surface was in the trivalent form. The Freundlich model fitted the isotherm process better than the Langmuir model in the concentration range examined. The pseudo-second-order model well described the adsorption process of Cr onto the biomass. The biosorption capacity of Cr(VI) by fruiting bodies was much higher than that by most of other biosorbents reported. The results suggest that the fruiting bodies of A. polytricha should be a promising biomaterial for Cr removal from water contaminated by the heavy metal.

Bacterial and fungal taxon changes in soil microbial community composition induced by short-term biochar amendment in red oxidized loam soil.

To take full advantage of biochar as a soil amendment, the objective of this study was to investigate the effects of biochar addition on soil bacterial and fungal diversity and community composition. Incubation experiments with a forest soil (a red oxidized loam soil) with and without biochar amendment were conducted for 96 days. The culture-independent molecular method was utilized to analyze soil bacterial and fungal species after the incubation experiments. Results showed that bacteria and fungi responded differently to the biochar addition during the short-term soil incubation. Twenty four and 18 bacterial genara were observed in the biochar amended and unamended soils, respectively, whereas 11 and 8 fungal genera were observed in the biochar amended and unamended soils, respectively. Microbial taxa analysis indicated that the biochar amendment resulted in significant shifts in both bacterial and fungal taxa during the incubation period. The shift for bacteria occurred at the genus and phylum levels, while for fungi only at the genus level. Specific taxa, such as Actinobacteria of bacteria and Trichoderma and Paecilomyces of fungi, were enriched in the biochar amended soil. The results reveal a pronounced impact of biochar on soil microbial community composition and an enrichment of key bacterial and fungal taxa in the soil during the short time period.

Ruminant feces harbor diverse uncultured symbiotic actinobacteria.

To isolate actinobacteria from ruminant feces and elucidate their correlations with ruminants, the actinobacterial community in sheep (Ovis aries) and cattle (Bos taurus) feces was determined by cultivation and clone library methods. Most of actinobacteria isolated belonged to Streptomyces, Amycolatopsis, Micromonospora, and Cellulosimicrobium genera. The strains showed above 99 % similarity with the type strains, respectively. All the strains isolated could grow on media containing pectin, cellulose, or xylan as the sole carbon sources. However, most antibacterial and antifungal activities were found in Streptomyces species. Clone library analysis revealed that the genera Mycobacterium, Aeromicrobium, Rhodococcus, Cellulomonas were present in cattle and sheep feces. In contrast, the 16S rRNA genes showed less than 98 % similarity with the type strains. The analysis of actinobacterial community in ruminant feces by clone library and cultivation yielded a total of 10 actinobacterial genera and three uncultured actinobacterial taxa. The ruminant feces harbored diverse actinobacterial community. Ruminants may represent an underexplored reservoir of novel actinomycetes of potential interest for probiotics and drug discovery.

Endosphere mycobiome in mature rice roots originate from both seedlings and soils.

Plant-fungus symbioses have functional relevance during plant growth and development. However, it is still unknown whether the endosphere fungi in mature plants originated from soils or seeds. To elucidate the origination of endosphere fungi in mature rice roots, the fungal communities in surface sterilized roots and shoots of mature rice plants germinated in soils, rhizosphere soils and seedlings germinated under sterile conditions were analyzed by Illumina-based sequencing and compared. Total 62 fungal OTUs shared in the seedlings, shoots and roots, 126 OTUs shared in the rhizosphere soils, shoots and roots. Fungal OTUs coexisted in the four types of samples belonged to genera of Rhizophagus, Trichoderma, Fusarium, Atractiella, Myrmecridium, Sporothrix, Microdochium, Massariosphaeria, and Phialemonium. The principle component analysis (PCA) and NMDS plot suggested that the fungal community structure in rhizosphere soils was different from that in seedlings significantly. Rhizosphere soil, shoot and root contained more similar fungal community. The fungal community in seedling was similar to that in shoot and root of mature plants. The results suggested that endophytic fungal communities in mature rice plants originated from both seedlings and rhizosphere soils, and more fungal taxa originated from rhizosphere soils. Mature rice plants contain mycobiome transmitted vertically from seeds, which suggests that inoculation of endophytic fungi isolated from seedlings might be an effective way to introduce beneficial fungal inoculants into rice plants successfully.

Biosorption behavior and mechanism of heavy metals by the fruiting body of jelly fungus (Auricularia polytricha) from aqueous solutions.

The aim of this study was to investigate the biosorption characteristics of Cd(2+), Cu(2+), and Pb(2+) by the fruiting body of jelly fungus Auricularia polytricha. Batch experiments were conducted to characterize the kinetics, equilibrium, and mechanisms of the biosorption process. Optimum values of pH 5, biomass dosage 4 g L(-1), and contact time 60 min provided maximum biosorption capacities of A. polytricha for Cd(2+), Cu(2+), and Pb(2+) of 63.3, 73.7, and 221 mg g(-1), respectively. The maximum desorption was achieved using 0.05 mol L(-1) HNO(3) as an elute. The fruiting body was reusable at least for six cycles of operations. The pseudo-second-order model was the best to describe the biosorption processes among the three kinetic models tested. Freundlich and Dubinin-Radushkevich models fitted the equilibrium data well, indicating a heterogeneous biosorbent surface and the favorable chemisorption nature of the biosorption process. A Fourier transform infrared spectroscopy analysis indicated that carboxyl, amine/hydroxyl, amino, phosphoryl, and C-N-C were the main functional groups to affect the biosorption process. Synergistic ion exchange and surface complexation were the dominant mechanisms in the biosorption process. The present work revealed the potential of jelly fungus (fruiting body of A. polytricha) to remove toxic heavy metals from contaminated water.

Bacteroides fragilis Supplementation Deteriorated Metabolic Dysfunction, Inflammation, and Aorta Atherosclerosis by Inducing Gut Microbiota Dysbiosis in Animal Model.

BACKGROUND: The gut microbial ecosystem is an important factor that regulates host health and the onset of chronic diseases, such as inflammatory bowel diseases, obesity, hyperlipidemia, and diabetes mellitus, which are important risk factors for atherosclerosis. However, the links among diet, microbiota composition, and atherosclerotic progression are unclear. METHODS AND RESULTS: Four-week-old mice (-/- mice, C57Bl/6) were randomly divided into two groups, namely, supplementation with culture medium (control, CTR) and Bacteroides fragilis (BFS), and were fed a high-fat diet. The gut microbiota abundance in feces was evaluated using the 16S rDNA cloning library construction, sequencing, and bioinformatics analysis. The atherosclerotic lesion was estimated using Oil Red O staining. Levels of CD36, a scavenger receptor implicated in atherosclerosis, and F4/80, a macrophage marker in small intestine, were quantified by quantitative real-time PCR. Compared with the CTR group, the BFS group showed increased food intake, fasting blood glucose level, body weight, low-density lipoprotein level, and aortic atherosclerotic lesions. BFS dramatically reduced Lactobacillaceae (LAC) abundance and increased Desulfovibrionaceae (DSV) abundance. The mRNA expression levels of CD36 and F4/80 in small intestine and aorta tissue in the BFS group were significantly higher than those in the CTR group. CONCLUSIONS: gut microbiota dysbiosis was induced by BFS. It was characterized by reduced LAC and increased DSV abundance and led to the deterioration of glucose/lipid metabolic dysfunction and inflammatory response, which likely promoted aorta plaque formation and the progression of atherosclerosis.

Tryptophan was metabolized into beneficial metabolites against coronary heart disease or prevented from producing harmful metabolites by the in vitro drug screening model based on Clostridium sporogenes.

In our previous study of 2,130 Chinese patients with coronary heart disease (CHD), we found that tryptophan (TRP) metabolites contributed to elevated risks of death. Many TRP-derived metabolites require the participation of intestinal bacteria to produce, and they play an important role in the pathogenesis of metabolic diseases such as CHD. So it is necessary to metabolize TRP into beneficial metabolites against CHD or prevent the production of harmful metabolites through external intervention. Indole-3-butyric acid (IBA) may be a key point of gut microbiota that causes TRP metabolism disorder and affects major adverse cardiovascular events in CHD. Therefore, this study aimed to develop a method based on in vitro culture bacteria to evaluate the effects of IBA on specific microbial metabolites quickly. We detected the concentrations of TRP and its metabolites in 11 bacterial strains isolated from feces using liquid chromatography-mass spectrometry, and selected Clostridium sporogenes as the model strain. Then, IBA was used in our model to explore its effect on TRP metabolism. Results demonstrated that the optimal culture conditions of C. sporogenes were as follows: initial pH, 6.8; culture temperature, 37°C; and inoculum amount, 2%. Furthermore, we found that IBA increases the production of TRP and 5-HIAA by intervening TRP metabolism, and inhibits the production of KYNA. This new bacteria-specific in vitro model provides a flexible, reproducible, and cost-effective tool for identifying harmful agents that can decrease the levels of beneficial TRP metabolites. It will be helpful for researchers when developing innovative strategies for studying gut microbiota.

Seasonal and spatial diversity of microbial communities in marine sediments of the South China Sea.

This study was conducted to characterize the diversity of microbial communities in marine sediments of the South China Sea by means of 16S rRNA gene clone libraries. The results revealed that the sediment samples collected in summer harboured a more diverse microbial community than that collected in winter, Deltaproteobacteria dominated 16S rRNA gene clone libraries from both seasons, followed by Gammaproteobacteria, Acidobacteria, Nitrospirae, Planctomycetes, Firmicutes. Archaea phylotypes were also found. The majority of clone sequences shared greatest similarity to uncultured organisms, mainly from hydrothermal sediments and cold seep sediments. In addition, the sedimentary microbial communities in the coastal sea appears to be much more diverse than that of the open sea. A spatial pattern in the sediment samples was observed that the sediment samples collected from the coastal sea and the open sea clustered separately, a novel microbial community dominated the open sea. The data indicate that changes in environmental conditions are accompanied by significant variations in diversity of microbial communities at the South China Sea.

Biosorption of Cd, Cu, Pb, and Zn from aqueous solutions by the fruiting bodies of jelly fungi (Tremella fuciformis and Auricularia polytricha).

In this study, dried and humid fruiting bodies of Tremella fuciformis and Auricularia polytricha were examined as cost-effective biosorbents in treatment of heavy metals (Cd(2+), Cu(2+), Pb(2+), and Zn(2+)) in aqueous solution. The humid T. fuciformis showed the highest capacity to adsorb the four metals in the multi-metal solutions. The Pb(2+) adsorption rates were 85.5%, 97.8%, 84.8%, and 91.0% by dried T. fuciformis, humid T. fuciformis, dried A. polytricha, and humid A. polytricha, respectively. The adsorption amount of Pb(2+) by dried and humid T. fuciformis in Cd(2+) + Pb(2+), Cu(2+) + Pb(2+), Pb(2+) + Zn(2+), Cd(2+) + Cu(2+) + Pb(2+), and Cd(2+) + Zn(2+) + Pb(2+) solutions were not lower than that in Pb(2+) solutions. The results suggested that in humid T. fuciformis, Cd(2+), Cu(2+), and Zn(2+) promoted the Pb(2+) adsorption by the biomass. In the multi-metal solutions of Cd(2+) + Cu(2+) + Pb(2+) + Zn(2+), the adsorption amount and rates of the metals by all the test biosorbents were in the order of Pb(2+) > Cu(2+) > Zn(2+) > Cd(2+). Compared with the pseudo first-order model, the pseudo second-order model described the adsorption kinetics much better, indicating a two-step biosorption process. The present study confirmed that fruiting bodies of the jelly fungi should be useful for the treatment of wastewater containing Cd(2+), Cu(2+), Pb(2+), and Zn(2+).

Characterization of a deep-sea sediment metagenomic clone that produces water-soluble melanin in Escherichia coli.

To access to the microbial genetic resources of deep-sea sediment by a culture-independent approach, the sediment DNA was extracted and cloned into fosmid vector (pCC1FOS) generating a library of 39,600 clones with inserts of 24-45 kb. The clone fss6 producing red-brown pigment was isolated and characterized. The pigment was identified as melanin according to its physico-chemical characteristics. Subcloning and sequences analyses of fss6 demonstrated that one open reading frame (ORF2) was responsible for the pigment production. The deduced protein from ORF2 revealed significant amino acid similarity to the 4-hydroxyphenylpyruvate dioxygenase (HPPD) from deep-sea bacteria Idiomarina loihiensis. Further study demonstrated that the production of melanin was correlated with homogentistic acid (HGA). The p-hydroxyphenylpyruvate produced by the Escherichia coli host was converted to HGA through the oxidation reaction of introduced HPPD. The results demonstrate that expression of DNA extracted directly from the environment might generate applicable microbial gene products. The construction and analysis of the metagenomic library from deep-sea sediment contributed to our understanding for the reservoir of unexploited deep-sea microorganisms.

Isolation of phytase-producing yeasts from rice seedlings for prospective probiotic applications.

The yeasts transmitted from seeds to sprouts might be used as probiotics for host plants. To investigate the inheritable yeasts of rice plants for probiotics, the fungal internal transcribed spacer (ITS) regions (ITS1 and ITS2) in rice sprouts were analyzed by Illumina-based sequencing. The fungal genera Candida, Mortierella, Alternaria, Penicillium, and Tomentella were revealed by both ITS1 and ITS2 sequence analysis. The endophytic yeasts were isolated from rice sprouts by yeast selective medium. Compared with the negative controls, inoculation of isolate Y3 released 2.2 folds higher concentration of free phosphate in soybean meal broth. Most of the phytase activities were located in the yeast cell interiors. The shoot lengths, shoot fresh weights, and root fresh weights of inoculated seedlings increased by 35%, 80%, and 60% compared with the control seedlings, respectively. The results suggested that the rice sprouts contained diverse phytase-producing yeasts transmitted from seeds. These yeasts might be adopted as prospective probiotics to improve rice growth by increasing phosphate utilization efficacy.

Engineering banana endosphere microbiome to improve Fusarium wilt resistance in banana.

BACKGROUND: Plant microbiome highlights the importance of endosphere microbiome for growth and health of the host plant. Microbial community analysis represents an elegant way to identify keystone microbial species that have a more central position in the community. The aim of this study was to access the interactions between the keystone bacterial species and plants during banana Fusarium wilt process, by comparing the endophytic bacterial and fungal community in banana roots and shoot tips during growth and wilting processes. The keystone bacterial species were isolated and further engineered to improve banana wilt resistance. RESULTS: Banana endosphere microbiome structure varied during plant growth and wilting processes. Bacterial and fungal diversity in the shoot tips and roots increased with the development of the banana plantlets. The bacterial groups belonging to the Enterobacteriaceae family with different relative abundances were detected in all the samples. The Klebsiella spp. might be the keystone bacteria during the growth of banana plantlets. The relative abundance of Fusarium associated with the wilt disease did not increase during the wilting process. The endophytic Enterobacteriaceae strains Enterobacter sp. E5, Kosakonia sp. S1, and Klebsiella sp. Kb were isolated on Enterobacteriaceae selective medium and further engineered by expressing 1-aminocyclopropane-1-carboxylate (ACC) deaminase on the bacterial cell walls (designated as E5P, S1P, and KbP, respectively). Pot experiments suggested that plants inoculated with strains E5, E5P, S1, and S1P increased resistance to the Fusarium wilt disease compared with the controls without inoculation, whereas the Klebsiella inoculation (Kb and KbP) did not increase the wilt resistance. Compared with the inoculation with the wild strains E5 and S1, the inoculation with engineered strains E5P and S1P significantly increased wilt resistance and promoted plant growth, respectively. The results illustrated that the keystone species in the banana microbiome may not be dominant in numbers and the functional role of keystone species should be involved in the wilt resistance. CONCLUSION: The ACC deaminase activity of engineered bacteria was essential to the Fusarium wilt resistance and growth promotion of banana plants. Engineering keystone bacteria in plant microbiome with ACC deaminase on the cell walls should be a promising method to improve plant growth and disease resistance.