Sa-Youl Ghim

Genome Sequence of the Polymyxin-Producing Plant-Probiotic Rhizobacterium Paenibacillus polymyxa E681

Paenibacillus polymyxa E681, a spore-forming, low-G+C, Gram-positive bacterium isolated from the rhizosphere of winter barley grown in South Korea, has great potential for agricultural applications due to its ability to promote plant growth and suppress plant diseases. Here we present the complete genome sequence of P. polymyxa E681. Its 5.4-Mb genome encodes functions specialized to the plant-associated lifestyle and characteristics that are beneficial to plants, such as the production of a plant growth hormone, antibiotics, and hydrolytic enzymes.

Biological Control and Plant Growth Promoting Capacity of Rhizobacteria on Pepper under Greenhouse and Field Conditions

Plant growth promoting rhizobacteria Ochrobactrum lupini KUDC1013 and Novosphingobium pentaromativorans KUDC1065 isolated from Dokdo Island, S. Korea are capable of eliciting induced systemic resistance (ISR) in pepper against bacterial spot disease. The present study aimed to determine whether plant growth-promoting rhizobacteria (PGPR) strains including strain KUDC1013, strain KUDC1065, and Paenibacillus polymyxa E681 either singly or in combinations were evaluated to have the capacity for potential biological control and plant growth promotion effect in the field trials. Under greenhouse conditions, the induced systemic resistance (ISR) effect of treatment with strains KUDC1013 and KUDC1065 differed according to pepper growth stages. Drenching of 3-week-old pepper seedlings with the KUDC-1013 strain significantly reduced the disease symptoms. In contrast, treatment with the KUDC1065 strain significantly protected 5-week-old pepper seedlings. Under field conditions, peppers treated with PGPR mixtures containing E681 and KUDC1013, either in a two-way combination, were showed greater effect on plant growth than those treated with an individual treatment. Collectively, the application of mixtures of PGPR strains on pepper might be considered as a potential biological control under greenhouse and field conditions.

Impact of a Bacterial Volatile 2,3-Butanediol on Bacillus subtilis Rhizosphere Robustness

Volatile compounds, such as short chain alcohols, acetoin, and 2,3-butanediol, produced by certain strains of root-associated bacteria (rhizobacteria) elicit induced systemic resistance in plants. The effects of bacterial volatile compounds (BVCs) on plant and fungal growth have been extensively studied; however, the impact of bacterial BVCs on bacterial growth remains poorly understood. In this study the effects of a well-characterized bacterial volatile, 2,3-butanediol, produced by the rhizobacterium Bacillus subtilis, were examined in the rhizosphere. The nature of 2,3-butanediol on bacterial cells was assessed, and the effect of the molecule on root colonization was also determined. Pepper roots were inoculated with three B. subtilis strains: the wild type, a 2,3-butanediol overexpressor, and a 2,3-butanediol null mutant. The B. subtilis null strain was the first to be eliminated in the rhizosphere, followed by the wild-type strain. The overexpressor mutant was maintained at roots for the duration of the experiment. Rhizosphere colonization by a saprophytic fungus declined from 14 days post-inoculation in roots treated with the B. subtilis overexpressor strain. Next, exudates from roots exposed to 2,3-butanediol were assessed for their impact on fungal and bacterial growth in vitro. Exudates from plant roots pre-treated with the 2,3-butanediol overexpressor were used to challenge various microorganisms. Growth was inhibited in a saprophytic fungus (Trichoderma sp.), the 2,3-butanediol null B. subtilis strain, and a soil-borne pathogen, Ralstonia solanacearum. Direct application of 2,3-butanediol to pepper roots, followed by exposure to R. solanacearum, induced expression of Pathogenesis-Related (PR) genes such as CaPR2, CaSAR8.2, and CaPAL. These results indicate that 2,3-butanediol triggers the secretion of root exudates that modulate soil fungi and rhizosphere bacteria. These data broaden our knowledge regarding bacterial volatiles in the rhizosphere and their roles in bacterial fitness and as important inducers of plant defenses.

Inhibition of protein kinase CKII activity by resveratrol, a natural compound in red wine and grapes

Resveratrol is a phytoalexin found in grapes and other foods that has been shown to have anticancer and anti-inflammatory effects. Because protein kinase CKII is involved in cell proliferation and oncogenesis, we examined whether resveratrol could modulate CKII activity. Resveratrol was shown to inhibit the phosphotransferase activity of CKII with IC(50) of about 10 microM. Steady state studies revealed that resveratrol acted as a competitive inhibitor with respect to the substrate ATP. A value of 1.2 microM was obtained for the apparent K(i). Resveratrol also inhibited the catalytic reaction of CKII with GTP as substrate. Furthermore, resveratrol inhibits endogenous CKII activity on protein substrates in HeLa cell lysates. These results suggest that resveratrol is likely to function by inhibiting oncogenic disease, at least in part, through the inhibition of CKII activity.

Determinants of Plant Growth-promoting Ochrobactrum lupini KUDC1013 Involved in Induction of Systemic Resistance against Pectobacterium carotovorum subsp. carotovorum in Tobacco Leaves

The plant growth-promoting rhizobacterium Ochrobactrum lupini KUDC1013 elicited induced systemic resistance (ISR) in tobacco against soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum. We investigated of its factors involved in ISR elicitation. To characterize the ISR determinants, KUDC1013 cell suspension, heat-treated cells, supernatant from a culture medium, crude bacterial lipopolysaccharide (LPS) and flagella were tested for their ISR activities. Both LPS and flagella from KUDC1013 were effective in ISR elicitation. Crude cell free supernatant elicited ISR and factors with the highest ISR activity were retained in the n-butanol fraction. Analysis of the ISR-active fraction revealed the metabolites, phenylacetic acid (PAA), 1-hexadecene and linoleic acid (LA), as elicitors of ISR. Treatment of tobacco with these compounds significantly decreased the soft rot disease symptoms. This is the first report on the ISR determinants by plant growth-promoting rhizobacteria (PGPR) KUDC1013 and identifying PAA, 1-hexadecene and LA as ISR-related compounds. This study shows that KUDC1013 has a great potential as biological control agent because of its multiple factors involved in induction of systemic resistance against phytopathogens.

Shewanella dokdonensis sp. nov., isolated from seawater.

A novel bacterial strain, designated UDC329(T), was isolated from a sample of seawater collected at Dong-do, on the coast of Dokdo Island, in the East Sea of the Republic of Korea. The Gram-staining-negative, motile, facultatively anaerobic, non-spore-forming rods of the strain developed into dark orange-yellow colonies. The strain grew optimally between 25 and 30 °C, with 1% (w/v) NaCl and at pH 7. It grew in the absence of NaCl, but not with NaCl at >7% (w/v). The predominant menaquinone was MK-7, the predominant ubiquinones were Q-7 and Q-8, and the major fatty acids were iso-C(15:0) (33.52%) and C(17:1)ω8c (11.73%). The genomic DNA G+C content of strain UDC329(T) was 50.2 mol%. In phylogenetic analyses based on 16S rRNA and gyrB gene sequences, strain UDC329(T) was grouped with members of the genus Shewanella and appeared most closely related to Shewanella fodinae JC15(T) (97.9% 16S rRNA gene sequence similarity), Shewanella indica KJW27(T) (95.0%), Shewanella algae ATCC 51192(T) (94.8%), Shewanella haliotis DW01(T) (94.5%) and Shewanella chilikensis JC5(T) (93.9%). The level of DNA-DNA relatedness between strain UDC329(T) and S. fodinae JC15(T) was, however, only 27.4%. On the basis of phenotypic, genotypic and DNA-DNA relatedness data, strain UDC329(T) represents a novel species in the genus Shewanella, for which the name Shewanella dokdonensis sp. nov. is proposed. The type strain is UDC329(T) (=KCTC 22898(T)=DSM 23626(T)).

A cry for help from leaf to root: above ground insect feeding leads to the recruitment of rhizosphere microbes for plant self-protection against subsequent diverse attacks.

Plants have evolved general and specific defense mechanisms to protect themselves from diverse enemies, including herbivores and pathogens. To maintain fitness in the presence of enemies, plant defense mechanisms are aimed at inducing systemic resistance: in response to the attack of pathogens or herbivores, plants initiate extensive changes in gene expression to activate “systemic acquired resistance” against pathogens and “indirect defense” against herbivores. Recent work revealed that leaf infestation by whiteflies, stimulated systemic defenses against both an airborne pathogen and a soil-borne pathogen, which was confirmed by the detection of the systemic expression of pathogenesis-related genes in response to salicylic acid and jasmonic acid-signaling pathway activation. Further investigation revealed that plants use self protection mechanisms against subsequent herbivore attacks by recruiting beneficial microorganisms called plant growth-promoting rhizobacteria/fungi, which are capable of reducing whitefly populations. Our results provide new evidence that plant-mediated aboveground to belowground communication and vice versa are more common than expected.

Lutibacter oricola sp. nov., a marine bacterium isolated from seawater

A bacterial strain, UDC377(T), was isolated from seawater samples collected at Seo-do on the coast of Dokdo island in the East Sea, and was subjected to taxonomic study using a polyphasic approach. Strain UDC377(T) was pale-yellow, Gram-staining-negative, non-motile, rod-shaped and aerobic. The strain grew optimally at 25-28 °C, in the presence of 2 % (w/v) NaCl and at pH 7.0-7.5. Strain UDC377(T) produced carotenoid pigments; however, it did not produce flexirubin-type pigments. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain UDC377(T) clustered with members of the genus Lutibacter and appeared most closely related to Lutibacter agarilyticus KYW566(T) (96.0 % 16S rRNA gene sequence similarity) followed by L. aestuarii MA-My1(T) (95.0 %), L. litoralis CL-TF09(T) (94.9 %), L. maritimus S7-2(T) (94.1 %) and L. flavus IMCC1507(T) (94.0 %). The DNA G+C content of strain UDC377(T) was 30.8 mol%. Strain UDC377(T) contained MK-6 as the predominant menaquinone, iso-C15 : 0 3-OH, iso-C15 : 0 and iso-C16 : 0 3-OH as the major fatty acids, and phosphatidylethanolamine, two unknown aminolipids and six unknown lipids as the major polar lipids. Based on phenotypic properties and phylogenetic data presented, strain UDC377(T) is considered to represent a novel species of the genus Lutibacter, for which the name Lutibacter oricola sp. nov. is proposed. The type strain is UDC377(T) ( = DSM 24956(T) = KCTC 23668(T)).

Paenibacillus dongdonensis sp. nov., isolated from rhizospheric soil of Elymus tsukushiensis

A Gram-staining-positive, endospore-forming and rod-shaped bacterial strain, designated KUDC0114(T), was isolated from rhizospheric soil of Elymus tsukushiensis from Dongdo Island, one of the largest of the Dokdo Islands, South Korea. The strain displayed optimal growth at 37 °C, pH 8.5 in the absence of NaCl. Based on phylogenetic analysis of 16S rRNA gene sequences, strain KUDC0114(T) represented a member of the genus Paenibacillus and was most closely related to Paenibacillus taichungensis BCRC 17757(T) (98.46%). The cell-wall peptidoglycan was A1γ type, and the predominant quinone was menaquinone-7 (MK-7). The major cellular fatty acids were anteiso-C(15 : 0) and C(16 : 0). The DNA-DNA hybridization of strain KUDC0114(T) with nine other strains indicated less than 23% relatedness, and its DNA G+C content was 44.30 mol%. Based on genomic, phenotypic and phylogenetic analyses, KUDC0114(T) should be classified as representing novel species within the genus Paenibacillus. The name Paenibacillus dongdonensis sp. nov. is proposed. The type strain is KUDC0114(T) ( = DSM27607(T) = KCTC33221(T)).

Linoleic acid-induced expression of defense genes and enzymes in tobacco.

Linoleic acid (LA) is a naturally occurring fatty acid (FA) found to elicit induced systemic resistance (ISR) of tobacco against the bacterial soft rot pathogen, Pectobacterium carotovorum subsp. carotovorum (PCC). In this study, we examined effects of six doses of exogenous LA on the induction of defense genes and enzymes. The optimum ISR activity was observed in plants treated with 0.1mM LA where the effect of LA on membrane permeability was minimal. The application of LA as a root drench enhanced the activity of defense enzymes such as phenylalanine ammonia-lyase (PAL), peroxidase (POD), and polyphenol oxidase (PPO) and induced the expression of β-glucuronidase (GUS). PAL and POD activities were increased in a concentration dependent manner while the maximum PPO activity was observed after treatment with 0.01mM LA. An RT-PCR analysis of the defense-related genes, Coi1, NPR1, PR-1a and PR-1b, of tobacco plants treated with 0.1mM LA revealed an association of LA with elicitation of ISR in tobacco.