BACKGROUND: This study investigated the effects of rice genetically modified to be resistant against rice blast and rice bacterial blight on the soil microbial community. A comparative analysis of the effects of rice genetically modified rice choline kinase (OsCK1) gene for disease resistance (GM rice) and the Nakdong parental cultivar (non-GM rice) on the soil microbial community at each stage was conducted using rhizosphere soil of the OsCK1 and Nakdong rice. METHODS AND RESULTS: The soil chemistry at each growth stage and the bacterial and fungal population densities were analyzed. Soil DNA was extracted from the samples, and the microbial community structures of the two soils were analyzed by pyrosequencing. No significant differences were observed in the soil chemistry and microbial population density between the two soils. The taxonomic analysis showed that Chloroflexi, Proteobacteria, Firmicutes, Actinobacteria, and Acidobacteria were present in all soils as the major phyla. Although the source tracking analysis per phylogenetic rank revealed that there were differences in the bacteria between the GM and non-GM soil as well as among the cultivation stages, the GM and non-GM soil were grouped according to the growth stages in the UPGMA dendrogram analysis. CONCLUSION: The difference in bacterial distributions between Nakdong and OsCK1 rice soils at each phylogenetic level detected in microbial community analysis by pyrosequencing may be due to the genetic modification done on GM rice or due to heterogeneity of the soil environment. In order to clarify this, it is necessary to analyze changes in root exudates along with the expression of transgene. A more detailed study involving additional multilateral soil analyses is required.
During the rhizoremediation of diesel-contaminated soil, methane (CH4), a representative greenhouse gas, is emitted as a result of anaerobic metabolism of diesel. The application of methantrophs is one of solutions for the mitigation CH4 emissions during the rhizoremediation of diesel-contaminated soil. In this study, CH4-oxidizing rhizobacteria, Methylocystis sp. JHTF4 and Methyloversatilis sp. JHM8, were isolated from rhizosphere soils of tall fescue and maize, respectively. The maximum CH4 oxidation rates for the strains JHTF4 and JHM8 were 65.8 and 33.8 mmol·g-DCW-1·h-1, respectively. The isolates JHTF4 and JHM8 couldn't degrade diesel. The inoculation of the isolate JHTF4 or JHM8 significantly enhanced diesel removal during rhizoremediation of diesel-contaminated soil planted with maize for 63 days. Diesel removal in the tall fescue-planting soil was enhanced by inoculating the isolates until 50 days, while there was no significant difference in removal efficiency regardless of inoculation at day 63. In both the maize and tall fescue planting soils, the CH4 oxidation potentials of the inoculated soils were significantly higher than the potentials of the non-inoculated soils. In addition, the gene copy numbers of pmoA, responsible for CH4 oxidation, in the inoculated soils were significantly higher than those in the non-inoculated soils. The gene copy numbers ratio of pmoA to 16S rDNA (the ratio of methanotrophs to total bacteria) in soil increased during rhizoremediation. These results indicate that the inoculation of Methylocystis sp. JHTF4 and Methyloversatilis sp. JHM8, is a promising strategy to minimize CH4 emissions during the rhizoremediation of diesel-contaminated soil using maize or tall fescue.
Tetty Marta Linda;Jusinta Aliska;Nita Feronika;Ineiga Melisa;Erwina Juliantari
Journal of Microbiology and Biotechnology
/
v.34
no.6
/
pp.1239-1248
/
2024
Peatlands are marginal agricultural lands due to highly acidic soil conditions and poor drainage systems. Drought stress is a big problem in peatlands as it can affect plants through poor root development, so technological innovations are needed to increase the productivity and sustainability of upland rice on peatlands. Rhizobacteria can overcome the effects of drought stress by altering root morphology, regulating stress-responsive genes, and producing exopolysaccharides and indole acetic acid (IAA). This study aimed to determine the ability of rhizobacteria in upland rice to produce exopolysaccharides and IAA, identify potential isolates using molecular markers, and prove the effect of rhizobacteria on viability and vigor index in upland rice. Rhizobacterial isolates were grown on yeast extract mannitol broth (YEMB) medium for exopolysaccharides production testing and Nutrient Broth (NB)+L-tryptophan medium for IAA production testing. The selected isolates identify using sequence 16S rRNA. The variables observed in testing the effect of rhizobacteria were germination ability, vigour index, and growth uniformity. EPS-1 isolate is the best production of exopolysaccharides (41.6 mg/ml) and IAA (60.83 ppm). The isolate EPS-1 was identified as Klebsiella variicola using 16S rRNA sequencing and phylogenetic analysis. The isolate EPS-1 can increase the viability and vigor of upland rice seeds. K. variicola is more adaptive and has several functional properties that can be developed as a potential bioagent or biofertilizer to improve soil nutrition, moisture and enhance plant growth. The use of rhizobacteria can reduce dependence on the use of synthetic materials with sustainable agriculture.
An antagonistic bacteria, Bacillus licheniformis Nl strain which effectively inhibited mycelial growth of gray mold rot pathogen, Botrytis cinerea was isolated from the rhizosphere of perilla crop. Powder soy formulation by B. lichentfomis Nl strain as a biocontrol agent was developed far the first time and estimated its control effect on perilla leaves in this study. First of all, far the mass production of antifungal metabolites of B. lichentfomis Nl strain in flask liquid culture, the most effective carbon and nitrogen source were selected as glucose and tryp-tone, respectively, For the formulation, vegetative biomass of B. licheniformis Nl strain from 5-day-old liquid culture in nutrient broth added glucose and tryptone was mixed with soy flour, rice flour glucose, FeSo$_4$~7$H_2O$, and MnCl$_2$. 4$H_2O$, and dried and pulverized. In plastic house test, powder soy formulation effectually controlled gray mold rot as the control value of 93.1 %, was more effective than chemical fungicide, benomyl showing the control value of 86.1%. Thus, development of powder soy formulation of B. lichentfomis Nl will aid large-scale application of biological control in field trials.
A method for evaluating bacterial chemotactic responses toward several single of combined sugars and sterile mucilage from the different rice cultivars had been tested. Bacterial genus of Azospirillum, Pseudomonas and Agrobacterium were specially identified from the histosphere of different rice cultivars and graminea grasses in saline and reclaimed saline paddy soil. To evaluate chemotaxis of these strains a modification of Fendrik channel method was used. Under this condition Azospirillum lipoferum Ecc 3-1 reacted stereoisomerically fomulating the single migration ring while Agrobacterium radiobacter Ecc 1-1 and Pseudomonas sp Ecc 4-1 did not. Strains specificities of chemotaxis to the single sugar such as D(+)-glucose and D(+)-fructose were less prominent than malic and citric acid. Chemotactic responses to the combined sugar such as D-galacturonic acid and the L-aspartate were found high attracting reaction than other combined sugars. Chemotaxis of associative $N_2$-fixing bacteria to the root exudates of different rice cultivars were differed among bacterial strains and rice cultivars.
You, Young-Hyun;Park, Jong Myong;Lee, Myung-Chul;Kim, Jong-Guk
Korean Journal of Microbiology
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v.51
no.1
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pp.86-95
/
2015
To study the halobacterial diversity at the rhizospheric soil of coastal plant native to Dokdo islands, several host plant were selected and its rhizospheric soil was sampled. Soil sample was diluted serially and pure isolation was done by sub-culture using marine agar media. 26 halophilic strains cultivable at the marine medium containig concentration of 9.0% sodium chloride were selected among total 161 isolates. Their partial 16S rRNA gene sequences extracted from genomic DNA were analyzed and partially identified. Furthermore, to identify their genetic relationship, phylogenetic tree was deduced. Total 26 strains were belongs to Firmicutes (30.8%), Gamma proteobacteria (53.8%), Bacteroidetes (7.7%), Alpha proteobacteria (7.7%), and Actinobacteria (7.7%). These results showed the specific difference from previous researches which has been reported the microbial flora of soil or sea water around the Dokdo islands. Furthermore, 4 among 26 halophilic strains grew at above 12.0% NaCl concentrated marine broth, and 2 strains Idiomarina abyssalis LM4H23 and Halomonas huangheensis AS4H13 grew at 15.0% concentration. These halophilic strains thought to overcoming the severe stress like high salt concentration or variation derived from Dokdo-specific climate and might have unknown, specific relationship with their host coastal plant native to Dokdo islands.
Forty-nine pigments were extracted from the collections of 106 pigment producing bacteria from the plant rhizosphere soil. Antibacterial activity test was performed in the subjects of the extracted pigments with plant pathogenic bacteria including Xanthomonas axonopodis and Xanthomonas campestris, and with plant pathogenic fungi including Botrytis cinerea, Colletotrichum acutatum, and Fusarium oxysporum. The yellow pigment by Chryseobacterium sp. RBR9 and the red pigment by of Methylobacterium sp. RI13 showed the antibacterial activities against Xanthomonas axonopodis and Xanthomonas campestris. The violet pigment by Collimonas sp. DEC-B5 showed the antibacterial activity as well as the antifungal activities against Botrytis cinerea and Fusarium oxysporum. Especially, the violet pigment inhibited the growth of Botrytis cinerea more than 65% at MIC $20{\mu}M$. Upon the HPLC analysis result for the isolation of pigment with antifungal activity, violacein (91.6%) and deoxyviolacein (8.4%) were isolated for the pigment by Collimonas sp. DEC-B5. The production amount of the pigment was increased more than 10 times higher when D-mannitol 1.5% and yeast extract 0.2% were added as the nitrogen source to SCB medium. This study suggests that produced violacein by Collimonas sp. DEC-B5 will be effective to control strawberry gray-mold rot fungi by its preventive activity.
Lee, Min-Kyoung;Lee, Gil-Seung;Yim, Woo-Jong;Hong, In-Soo;Palaniappan, Pitchai;Siddikee, Md. Ashaduzzaman;Boruah, Hari P. Deka;Madhaiyan, Munusamy;Ahn, Ki-Sup;Sa, Tongmin
Korean Journal of Soil Science and Fertilizer
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v.42
no.4
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pp.266-273
/
2009
Use of plant growth promoting symbiotic and non-symbiotic free-living beneficial bacteria as external source of nitrogen is a major research concern for sustainable crop production in the $21^{st}$ century. In view of this, an experiment was conducted under controlled conditions to determine the effects of inoculation with Methylobacterium suomiense CBMB120, a plant growth promoting (PGP) root and shoot colonizer on red pepper, for the purpose of reducing external chemical nitrogen fertilization. Amendments with organic fertilizer and chemical fertilizer in the form of NPK were made at dosages of 50%, 75% and 100%, at 425 and $115kg/ha^{-1}$ measurements. The soil type used was loam, with a pH of 5.13. The growth responses were measured as plant height at 19, 36 and 166 days after transplantation and final biomass production after 166 days. It was found that inoculation with M. suomiense CBMB120 promotes plant height increase during the active growth phase at 19 and 36 days by 14.17% and 10.03%, respectively. Thereafter, the bacteria inoculated plantlets showed canopy size increment. A highly significant inoculation effect on plant height at p<0.01 level was found for 100% level of organic matter and chemical amendment in red pepper plantlets after 36 days and 19 days from transplantation. Furthermore, there was a significantly higher (10.30% and 6.84%) dry biomass accumulation in M. suomiense CBMB120 inoculated plants compared to un-inoculated ones. A 25% reduction in the application of chemical nitrogen can be inferred with inoculation of M. suomiense CBMB120 at with comparable results to that of 100% chemical fertilization alone. Enumeration of total bacteria in rhizosphere soil confirms that the introduced bacteria can multiply along ther hizosphere soil. Large scale field study may lead to the development of M. suomiense CBMB120 as an efficient biofertilizer.
This experiment was conducted to develop fertilizer which promotes plant growth as well as suppressing pathogenic fungi. The fertilizer was made from the mixture of Ju-Back (Korean rice wine cake) and indigenous rhizosphere-bacterium. The main ingredients of Ju-Back were investigated as 6.04% total nitrogen, 42.59% total carbohydrate, 1.01% available phosphate, 73.42% organic matter, 7.72% potassium oxide, 1.35% calcium oxide, 0.53% magnesium oxide. The enzyme activities of Ju-Back were estimated to be 980 units/g for ${\alpha}-amylase$, 300 units/g for glucoamylase, and 1800 units/g for acid pretense. Indigenous rhizosphere bacteria which produced antifungal agent were isolated from soil, and was selected KMU-13 strain which can antagonize against various plant pathogenic fungi (Botrytis cinerea KACC 40573, Sclerotinia sclerotiorum KACC 41065, Fusairum oxysporum KACC 40052, Pythium aphanidermatum KACC 40156, Phytophthora capsici KACC 40476 and Glomerella cingulata KACC 40299). KMU-13 strain was identified as Bacillus subtilis KMU-13 by biochemical and 16s rDNA analysis. The organic fertilizer was made as prototype which was composed 20% Ju-Back, 70% carrier, 9.7% microorganism cultivated solution, 0.3% trace-element. We also investigated an application of fertilizer using Ju-Back for cultivating lettuce (Lactuca sativar) which were grown in three soil conditions that had chemical fertilizer, barnyard manure, lime power, urea, potassium chloride and superphosphate as a control, the whole quantity (80 kg/10a) of posted fertilizer with the control and the half quantity (40 kg/10a) with the control. The growth characteristics were examined and analysed with several weeks interval from 3 weeks to 8 weeks on head length (cm), head width (cm/head), number of leaf and fresh weight (g/plant). The results are summarized as follows. The head width and fresh weight of lettuce were the highest at posted fertilizer 1 (whole quantity) was applied chemical, organic matter (Ju-Back) and carrier. The head length was the highest at posted fertilizer 2 (whole quantity) was applied Ju-Back only.
One of the physiologically important ginseng diseases is red-colored phenomena (RCP) that is caused by accumulation of red-colored substances on the epidermis of ginseng roots. Although RCP severely deteriorates the quality of ginseng products, there has been little information on what red-colored substance is and how RCP occurs. Therefore, the heavy losses of cultivators and ginseng industry are suffering by RCP, For this reason, we have investigated with the morphochernical characteristics of RCP to find out main cause of it. The red-colored substances (RS) on the epidermis of red-colored ginseng (RCG) were examined using inverted light microscope, confocal laser scanning microscope (CLSM)and furier transform infrared (FT/IR) spectrometer. Red brown substances were accumulated in the cell wall of the epidermis from early stage to late stage of RCC. Especially, cell wall of the late stage of RCG was covered with the sub-stances with 80~ 130 fm thick. Therefore, the cell wall of RCG cannot protect the ginseng root cells from the mechanical damages, bacteria and fungi. To analyse red substances of roots, RS were isolated from epidermis of RCG and extracted using various solvents. RS is strongly insoluble but it was bleached by oxidizing agents including 12% (v/v) NaOCl. Therefore, RS was Presumed to make up of high chelation power. The proriles of FT/IR spectra or both healthy ginseng (HEG) and RCG showed a significant difference at two wavelength,2857 cm$\^$-1/(C-H) and 1032 cm$\^$-1/(S=O), respectively. Furthermore, absorption peak of 2857cm$\^$-l/ appears on the only epidermis of RCG. The other peak is shown lower absorption rate on the epidermis of RCG than that of healthy ginseng. Also, FT/IR spectra of the mixture of carboxym-ethylcellulose (CMC) and iron (Fe$\^$3+/) were very similar to RCG spectrum profiles. One of a interesting fact is that the contents of phenolic compounds at the epidermis of healthy ginseng were highest. The results of these experiments sup-port the RCP was closely related with the chemical interaction between inorganic elements (Fe) of rhizosphere and organic matters (cellulose, cellobiose, cell sap, etc.) of ginseng roots.
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