Phosphate Solubilizing Bacillus megaterium mj1212 Regulates Endogenous Plant Carbohydrates and Amino Acids Contents to Promote Mustard Plant Growth.

The current study was conducted to explore the potential of a phosphate solubilizing soil bacterium, Bacillus megaterium mj1212 for enhancing the growth of mustard plants. The newly isolated bacterial strain mj1212 was identified as B. megaterium using phylogenetic analysis and, its phosphate solubilization ability was shown by the clear zone formation on National Botanical Research Institute's Phosphate medium. Moreover, the phosphate solubilization ability of B. megaterium mj1212 was enhanced by optimal culture conditions at pH 7.0 and 35 °C which might be due to the presence of malic and quinic acid in the culture medium. The beneficial effect of B. megaterium mj1212 in mustard plants was determined by an increasing shoot length, root length and fresh weight of plants. In the biochemical analysis revealed that chlorophyll, sucrose, glucose, fructose and amino acids (Asp, Thr, Ser, Glu, Gly, Ala, Cys, Val, Met, Ilu, Leu, Tyr, Phe, Lys, His, Arg and Pro) were higher in B. megaterium mj1212 treated plants, when compared to their control. The result of present study suggests that B. megaterium mj1212 treatment could be act as phosphate biofertilizer to improve the plant growth.

Gibberellin-Producing Bacteria Isolated from Coastal Soil Enhance Seed Germination of Mallow and Broccoli Plants under Saline Conditions.

Salinity hinders plant growth, posing a substantial challenge to sustainable agricultural yield maintenance. The application of plant growth-promoting rhizobacteria (PGPR) offers an emerging strategy to mitigate the detrimental effects of high salinity levels. This study aimed to isolate and identify gibberellin-producing bacteria and their impact on the seed germination of Malva verticillata (mallow) and Brassica oleracea var. italica (broccoli) under salt stress. In this study, seven bacterial isolates (KW01, KW02, KW03, KW04, KW05, KW06, and KW07) were used to assess their capacity for producing various growth-promoting traits and their tolerance to varying amounts of salinity (100 mM and 150 Mm NaCl). The findings revealed that KW05 and KW07 isolates outperformed other isolates in synthesizing indole-3-acetic acid, siderophores, and exopolysaccharides and in solubilizing phosphates. These isolates also enhanced phosphatase activity and antioxidant levels, including superoxide dismutase and catalase. Both KW05 and KW07 isolate highlight the growth-promoting effects of gibberellin by enhancing of growth parameters of Waito-C rice. Further, gas chromatography-mass spectrometry validation confirmed the ability of KW05 and KW07 to produce gibberellins (GAs), including GA1, GA3, GA4, and GA7. Seed germination metrics were enhanced due to the inoculation of KW05 and KW07. Moreover, inoculation with KW05 increased the fresh weight (FW) (7.82%) and total length (38.61%) of mallow under salt stress. Inoculation with KW07 increased the FW (32.04%) and shoot length of mallow under salt stress. A single inoculation of these two isolates increased broccoli plants' FW and shoot length under salt stress. Gibberellin-producing bacteria helps in plant growth promotion by improving salt tolerance by stimulating root elongation and facilitating enhanced absorption of water and nutrient uptake in salty environments. Based on these findings, they can play a role in boosting agricultural yield in salt-affected areas, which would help to ensure the long-term viability of agriculture in coastal regions.

The Combined Inoculation of Curvularia lunata AR11 and Biochar Stimulates Synthetic Silicon and Potassium Phosphate Use Efficiency, and Mitigates Salt and Drought Stresses in Rice.

Synthetic chemical fertilizers are a fundamental source of nutrition for agricultural crops; however, their limited availability, low plant uptake, and excessive application have caused severe ecological imbalances. In addition, the gravity of environmental stresses, such as salinity and water stress, has already exceeded the threshold limit. Therefore, the optimization of nutrient efficiency in terms of plant uptake is crucial for sustainable agricultural production. To address these challenges, we isolated the rhizospheric fungus Curvularia lunata ARJ2020 (AR11) and screened the optimum doses of biochar, silicon, and potassium phosphate (K2HPO4), and used them-individually or jointly-to treat rice plants subjected to salt (150 mM) and drought stress (20-40% soil moisture). Bioassay analysis revealed that AR11 is a highly halotolerant and drought-resistant strain with an innate ability to produce gibberellin (GA1, GA3, GA4, and GA7) and organic acids (i.e., acetic, succinic, tartaric, and malic acids). In the plant experiment, the co-application of AR11 + Biochar + Si + K2HPO4 significantly improved rice growth under both salt and drought stresses. The plant growth regulator known as abscisic acid, was significantly reduced in co-application-treated rice plants exposed to both drought and salt stress conditions. These plants showed higher Si (80%), P (69%), and K (85%) contents and a markedly low Na+ ion (208%) concentration. The results were further validated by the higher expression of the Si-carrying gene OsLSi1, the salt-tolerant gene OsHKT2, and the OsGRAS23's drought-tolerant transcriptome. Interestingly, the beneficial effect of AR11 was significantly higher than that of the co-application of Biochar + Si + K2HPO4 under drought. Moreover, the proline content of AR11-treated plants decreased significantly, and an enhancement of plant growth-promoting characteristics was observed. These results suggest that the integrated co-application of biochar, chemical fertilizers, and microbiome could mitigate abiotic stresses, stimulate the bioavailability of essential nutrients, relieve phytotoxicity, and ultimately enhance plant growth.

Gibberellin production and phosphate solubilization by newly isolated strain of Acinetobacter calcoaceticus and its effect on plant growth.

Plant growth-promoting rhizobacteria with gibberellins (GA)-producing potential were isolated from soil and screened for plant growth promotion. A new strain, Acinetobacter calcoaceticus SE370, produced extracellular GA and also had phosphate solubilising potential. It produced 10 different gibberellins, including the bioactive GA(1), GA(3) and GA(4) which were at, respectively, 0.45, 6.2 and 2.8 ng/100 ml. The isolate solubilised tricalcium phosphate and lowered pH of the medium during the process. Culture filtrates of the organism after growth on broth promoted growth of cucumber, Chinese cabbage and crown daisy.

Phoma herbarum as a new gibberellin-producing and plant growth-promoting fungus.

Endophytic fungi are known for the production of valuable metabolites, but information on the gibberellin production capacity of this group is limited. We isolated 9 endophytic fungi from the roots of salt-stressed soybean plants and screened them on waito-c rice, in order to identify plant growth promoting fungal strains. The fungal isolate TK- 2-4 gave maximum plant length (20.35 cm) promotion in comparison with wild-type Gibberella fujikuroi (19.5 cm). In a separate experiment, bioassay of TK-2-4 promoted plant length and biomass of soybean cultivar Taegwangkong. The TK-2-4 culture filtrate was analyzed for the presence of gibberellins, and it was found that all physiologically active gibberellins, especially GA(4) and GA(7), were present in higher amounts (GA(1), 0.11 ng/ml; GA(3), 2.91 ng/ml; GA(4), 3.21 ng/ml; and GA(7), 1.4 ng/ml) in conjunction with physiologically inactive GA(9) (0.05 ng/ml), GA(12) (0.23 ng/ ml), GA(15) (0.42 ng/ml), GA(19) (0.53 ng/ml), and GA(20) (0.06 ng/ml). The fungal isolate TK-2-4 was later identified as a new strain of Phoma herbarum, through the phylogenetic analysis of 28S rDNA sequence.

Deletion of xylR gene enhances expression of xylose isomerase in Streptomyces lividans TK24.

Glucose (xylose) isomerases from Streptomyces sp. have been used for the production of high fructose corn syrup for industrial purposes. An 11-kb DNA fragment containing the xyl gene cluster was isolated from Streptomyces lividans TK24 and its nucleotide sequences were analyzed. It was found that the xyl gene cluster contained a putative transcriptional repressor (xylR), xylulokinase (xylB), and xylose isomerase (xylA) genes. The transcriptional directions of the xylB and xylA genes were divergent, which is consistent to those found in other streptomycetes. A gene encoding XylR was located downstream of the xylB gene in the same direction, and its mutant strain produced xylose isomerase regardless of xylose in the media. The enzyme expression level in the mutant was 4.6 times higher than that in the parent strain under xylose-induced condition. Even in the absence of xylose, the mutant strain produce over 60% of enzyme compared with the xylose-induced condition. Gel mobility shift assay showed that XylR was able to bind to the putative xyl promoter, and its binding was inhibited by the addition of xylose in vitro. This result suggested that XylR acts as a repressor in the S. lividans xylose operon.

Molecular cloning and expression of a thermostable xylose (glucose) isomerase gene, xylA, from Streptomyces chibaensis J-59.

In the present study, the xylA gene encoding a thermostable xylose (glucose) isomerase was cloned from Streptomyces chibaensis J-59. The open reading frame of xylA (1167 bp) encoded a protein of 388 amino acids with a calculated molecular mass of about 43 kDa. The XylA showed high sequence homology (92% identity) with that of S. olivochromogenes. The xylose (glucose) isomerase was expressed in Escherichia coli and purified. The purified recombinant XylA had an apparent molecular mass of 45 kDa, which corresponds to the molecular mass calculated from the deduced amino acid and that of the purified wild-type enzyme. The N-terminal sequences (14 amino acid residues) of the purified protein revealed that the sequences were identical to that deduced from the DNA sequence of the xylA gene. The optimum temperature of the purified enzyme was 85 degrees C and the enzyme exhibited a high level of heat stability.

Gibberellins-producing rhizobacteria increase endogenous gibberellins content and promote growth of red peppers.

The growth of red pepper plants was enhanced by treatment with the rhizobacterium, Bacillus cereus MJ-1. Red pepper shoots showed a 1.38-fold increase in fresh weight (fw) and roots showed a 1.28-fold fw gain. This plant growth-promoting rhizobacterium (PGPR) has been reported to produce gibberellins (GAs). Other GAs-producing rhizobacteria, Bacillus macroides CJ-29 and Bacillus pumilus CJ-69, also enhanced the fw of the plants. They were less effective than B. cereus MJ-1, though. The endogenous GAs content of pepper shoots inoculated with MJ-1 was also higher than in shoots inoculated with CJ-29 or CJ-69. When inoculated with MJ-1, bacterial colonization rate of the roots was higher than that of roots inoculated with CJ-29 or CJ-69. These results support the idea that the plant growth-promoting effect of the bacteria also positively related with the efficiency of root colonization by the bacteria. In addition, we identified the major endogenous GAs of the red pepper as originating from both the early C-13 hydroxylation and the early non C-13 hydroxylation pathways, with the latter being the predominant pathway of GA biosynthesis in red pepper shoots.

Enterococcus faecium LKE12 Cell-Free Extract Accelerates Host Plant Growth via Gibberellin and Indole-3-Acetic Acid Secretion.

The use of microbial extracts containing plant hormones is a promising technique to improve crop growth. Little is known about the effect of bacterial cell-free extracts on plant growth promotion. This study, based on phytohormonal analyses, aimed at exploring the potential mechanisms by which Enterococcus faecium LKE12 enhances plant growth in oriental melon. A bacterial strain, LKE12, was isolated from soil, and further identified as E. faecium by 16S rDNA sequencing and phylogenetic analysis. The plant growth-promoting ability of an LKE12 bacterial culture was tested in a gibberellin (GA)-deficient rice dwarf mutant (waito-C) and a normal GA biosynthesis rice cultivar (Hwayongbyeo). E. faecium LKE12 significantly improved the length and biomass of rice shoots in both normal and dwarf cultivars through the secretion of an array of gibberellins (GA1, GA3, GA7, GA8, GA9, GA12, GA19, GA20, GA24, and GA53), as well as indole-3-acetic acid (IAA). To the best of our knowledge, this is the first study indicating that E. faecium can produce GAs. Increases in shoot and root lengths, plant fresh weight, and chlorophyll content promoted by E. faecium LKE12 and its cell-free extract inoculated in oriental melon plants revealed a favorable interaction of E. faecium LKE12 with plants. Higher plant growth rates and nutrient contents of magnesium, calcium, sodium, iron, manganese, silicon, zinc, and nitrogen were found in cell-free extract-treated plants than in control plants. The results of the current study suggest that E. faecium LKE12 promotes plant growth by producing GAs and IAA; interestingly, the exogenous application of its cell-free culture extract can be a potential strategy to accelerate plant growth.

Chlorothalonil-biotransformation by glutathione S-transferase of Escherichia coli.

It has recently been reported that one of the most important factors of yeast resistance to the fungicide chlorothalonil is the glutathione contents and the catalytic efficiency of glutathione S-transferase (GST) (Shin et al, 2003). GST is known to catalyze the conjugation of glutathione to a wide variety of xenobiotics, resulting in detoxification. In an attempt to elucidate the relation between chlorothalonil-detoxification and GST, the GST of Escherichia coli was expressed and purified. The drug-hypersensitive E. coli KAM3 cells harboring a plasmid for the overexpression of the GST gene can grow in the presence of chlorothalonil. The purified GST showed chlorothalonil-biotransformation activity in the presence of glutathione. Thus, chlorothalonil is detoxified by the mechanism of glutathione conjugation catalyzed by GST.

Glutathione-dependent biotransformation of the fungicide chlorothalonil.

A gene responsible for the chlorothalonil biotransformation was cloned from the chromosomal DNA of Ochrobactrum anthropi SH35B, capable of efficiently dissipating the chlorothalonil. The gene encoding glutathione S-transferase (GST) of O. anthropi SH35B was expressed in Escherichia coli, and the GST was subsequently purified by affinity chromatography. The fungicide chlorothalonil was rapidly transformed by the GST in the presence of glutathione. LC-MS analysis supported the formation of mono-, di-, and triglutathione conjugates of chlorothalonil by the GST. The monoglutathione conjugate was observed as an intermediate in the enzymatic reaction. The triglutathione conjugate has not been previously reported and seems to be the final metabolite in the biotransformation of chlorothalonil. The glutathione-dependent biotransformation of chlorothalonil catalyzed by the bacterial GST is reported.

Gibberellin-producing Promicromonospora sp. SE188 improves Solanum lycopersicum plant growth and influences endogenous plant hormones.

Plant growth-promoting rhizobacteria (PGPR) producing gibberellins (GAs) can be beneficial to plant growth and development. In the present study, we isolated and screened a new strain of Promicromonospora sp., SE188, isolated from soil. Promicromonospora sp. SE188 secreted GAs into its growth medium and exhibited phosphate solubilization potential. The PGPR produced physiologically active (GA(1) and GA(4)) and inactive (GA(9), GA(12), GA(19), GA(20), GA(24), GA(34), and GA(53)) GAs in various quantities detected by GC/MS-SIM. Solanum lycopersicum (tomato) plants inoculated with Promicromonospora sp. SE188 showed a significantly higher shoot length and biomass as compared to controls where PGPR-free nutrient broth (NB) and distilled water (DW) were applied to plants. The presence of Promicromonospora sp. SE188 significantly up-regulated the non C-13 hydroxylation GA biosynthesis pathway (GA(12)→GA(24)→GA(9)→GA(4)→ GA(34)) in the tomato plants as compared to the NB and DW control plants. Abscisic acid, a plant stress hormone, was significantly down-regulated in the presence of Promicromonospora sp. SE188. Contrarily, salicylic acid was significantly higher in the tomato plant after Promicromonospora sp. SE188 inoculation as compared to the controls. Promicromonospora sp. SE188 showed promising stimulation of tomato plant growth. From the results it appears that Promicromonospora sp. SE188 has potential as a bio-fertilizer and should be more broadly tested in field trials for higher crop production in eco-friendly farming systems.

Burkholderia sp. KCTC 11096BP as a newly isolated gibberellin producing bacterium.

We isolated 864 bacteria from 553 soil samples and bioassayed them on cucumber and crown daisy for plant growth promotion. A new bacterial strain, Burkholderia sp. KCTC 11096BP gave maximum growth promotion and was selected for further investigations. The culture filtrate of this bacterium was thus analyzed for the presence of gibberellins and we found physiologically active gibberellins were found (GA(1), 0.23 ng/100 ml; GA(3), 5.11 ng/100 ml and GA(4), 2.65 ng/100 ml) along with physiologically inactive GA(9), GA(12), GA(15), GA(20), and GA(24). The bacterial isolate also solubilised tricalcium phosphate and lowered the pH of the medium during the process. The isolate was identified as a new strain of Burkholderia through phylogenetic analysis of 16S rDNA sequence. Gibberellin production capacity of genus Burkholderia is reported for the first time in current study.

Production of an anti-fungal substance for biological control of Phytophthora capsici causing phytophthora blight in red-peppers by Streptomyces halstedii.

The culture broth of Streptomyces halstedii AJ-7 suppressed the growth of Phytophthora capsici, which causes phytophthora blight in red-peppers, with less than 1% survival of the pathogen after 12 h of treatment. The low molecular fraction (< or = 10 kDa) of the culture broth retained anti-fungal activity against P. capsici after being held at 100 degrees C for 6 h.

Purification and characterization of an extracellular chitinase from the antifungal biocontrol agent Streptomyces halstedii.

An extracellular chitinase from Streptomyces halstedii AJ-7, a broad spectrum antifungal biocontrol agent, was characterized and purified. The apparent molecular weight of the purified protein was 55 kDa, Km value and Vmax of the protein for colloidal chitin were 3.2 mg ml(-1) and 118 micromol h(-1), respectively. The growth and chitinase activity of S. halstedii AJ-7 were enhanced by adding of 0.1% killed mycelium of Fusarium oxysporium in a medium containing 0.2% colloidal chitin.

Growth promotion of red pepper plug seedlings and the production of gibberellins by Bacillus cereus, Bacillus macroides and Bacillus pumilus.

The growth of red pepper plug seedlings was promoted by Bacillus cereus MJ-1, B. macroides CJ-29, and B. pumilus CJ-69 isolated from the rhizosphere. Gibberellins (GAs), a well-known plant growth-promoting hormone, were detected in the culture broth of their rhizobacteria. Among the GAs, the contents of GA1, GA3, GA4, and GA7, physiologically active GAs, were comparatively higher than those of others, suggesting that the growth promoting effect was originated from the GAs. This isthe first report on the production of GA5, GA8, GA34, GA44, and GA53 by bacteria.

Effects of green tea catechin on polymorphonuclear leukocyte 5'-lipoxygenase activity, leukotriene B4 synthesis, and renal damage in diabetic rats.

The purpose of this study was to investigate the effects of green tea catechin on polymorphonuclear leukocyte 5'-lipoxygenase activity, leukotriene B4 synthesis, and renal damage in diabetic rats. Male Sprague-Dawley rats weighing 100 +/- 10 g were randomly assigned to 1 normal group and 3 diabetic groups given a catechin-free diet (DM-0C group), 0.25% catechin diet (DM-0.25C group), or 0.5% catechin diet (DM-0.5C group), respectively. 5'-Lipoxygenase activity in the polymorphonuclear leukocytes significantly increased by 54% in the DM-0C group compared to the normal group, while the level in the DM-0.5C group remained the same as in the normal group. The leukotriene B4 content in the polymorphonuclear leukocytes increased 55% in the DM-0C group compared to the normal group, whereas the DM-0.25C and DM-0.5C groups exhibited the same level as the normal group. The superoxide radical content in the kidney microsomes increased 116% in the DM-0C group when compared to the normal group, yet decreased 29% in the DM-0.25C group and 50% in the DM-0.5C group compared to DM-0C group. The lipofuscin content was 197 and 136% higher in the DM-0C and DM-025C groups, respectively, than in the normal group, whereas the DM-0.5C group exhibited the same content as in the normal group. The carbonyl value increased 118% in the DM-0C group compared to the normal group, and the DM-0.25C and DM-0.5C groups were not significantly different from the DM-0C group. Accordingly, these results indicate that dietary catechin inhibited the generation of superoxide radicals, oxidized protein, and lipid peroxide in the kidney of streptozotocin-induced diabetic rats. Furthermore, green tea catechin supplementation in diabetic rats also appeared to inhibit the production of leukotriene B4 based on regulating the activity of 5'-lipoxygenase, thereby potentially reducing renal oxidative damage and inflammatory reactions.