Lysobacter antibioticus HS124 was isolated from rhizosphere soil in previous experiments, which produced lytic enzymes such as chitinase, gelatinase, lipase and protease. In addition, HS124 released an antibiotic compound, 4-hydroxyphenylacetic acid (4-HPAA). When larvae of P. xylostella was treated with HS124 culture broth, its body was destroyed, and degraded with the increase of incubation time, yielding glycine which was detected from HS124 culture broth. When 4-HPAA produced from HS124 was sprayed, larvae mortality increased with increasing concentration of 4-HPAA. When HS124 culture supplemented with Tween 80 was sprayed, its insecticidal activity against larvae was approximately 1.4 times higher compared to the culture without Tween 80. Insecticide (IS), HS124 culture broth (HS124), Magic-pi (MP) and HS124 culture broth+Magic-pi (HS124+MP) were each treated against larvae of P. xylostella to investigate their insecticidal effect where sterile diluted water (SDW) was used as a control. The highest mortality of larvae was found in HS124+MP, followed by IS, MP, HS124 and SDW respectively. Mortality of larvae in HS124 was 31% higher than that in SDW, but 41% lower than that in HS124+MP, meaning that both enzymes and antibiotics produced from HS124 may synergistically act as active agents with plant extract containing neem oil and turmeric in HS124+MP treatment. These results suggested that L. antibioticus HS124 together with plant extract can be one of candidates for biocontrol agents against Plutella xylostella.
Sheath rot and dry rot disease caused by Pseudomonas marginalis and Fusarium oxysporum were serious problems in garlic farmland. In this study, total of 160 indigenous antagonistic bacteria were isolated from 16 farmlands in Yeongcheon, Korea. Among these, 15 strains were able to inhibited P. marginalis and F. oxysporum. The 16s rDNA genes of the selected 15 strains were amplified and sequenced. The strains has strong antagonistic ability against garlic pathogens was achieved Bacillus subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240. The selected 3 strains tested for investigation of antifungal mechanisms further analyses; 3 strains of these validated for production of siderophore, ${\beta}$-glucanase and chitinase using CAS (chrome azurol S) blue agar, CMC-congo red agar and DNS method. The 3 strains were able to utilized insoluble phosphate as dertermined by vanado-molybdate method. The 3 strains verified for production of auxin and gibberellic acid using Salkowski test and holdbrook test. Also, 3 strains showed stimulation germination, stem growth promoting activity on the in vivo test. The 3 strains were able to effectively suppress P. marginalis and F. oxysporum causing sheath rot and dry rot diseases on the in vivo pot test.
Wang, Yiming;Kwon, Soon Jae;Wu, Jingni;Choi, Jaeyoung;Lee, Yong-Hwan;Agrawal, Ganesh Kumar;Tamogami, Shigeru;Rakwal, Randeep;Park, Sang-Ryeol;Kim, Beom-Gi;Jung, Ki-Hong;Kang, Kyu Young;Kim, Sang Gon;Kim, Sun Tae
The Plant Pathology Journal
/
v.30
no.4
/
pp.343-354
/
2014
Rice blast disease caused by Magnaporthe oryzae is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early response genes in rice, we utilized the transcriptome analysis approach using a 300 K tilling microarray to rice leaves infected with compatible and incompatible M. oryzae strains. Prior to the microarray experiment, total RNA was validated by measuring the differential expression of rice defense-related marker genes (chitinase 2, barwin, PBZ1, and PR-10) by RT-PCR, and phytoalexins (sakuranetin and momilactone A) with HPLC. Microarray analysis revealed that 231 genes were up-regulated (>2 fold change, p < 0.05) in the incompatible interaction compared to the compatible one. Highly expressed genes were functionally characterized into metabolic processes and oxidation-reduction categories. The oxidative stress response was induced in both early and later infection stages. Biotic stress overview from MapMan analysis revealed that the phytohormone ethylene as well as signaling molecules jasmonic acid and salicylic acid is important for defense gene regulation. WRKY and Myb transcription factors were also involved in signal transduction processes. Additionally, receptor-like kinases were more likely associated with the defense response, and their expression patterns were validated by RT-PCR. Our results suggest that candidate genes, including receptor-like protein kinases, may play a key role in disease resistance against M. oryzae attack.
Journal of the Korean Society of Food Science and Nutrition
/
v.39
no.11
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pp.1684-1690
/
2010
The properties of non-waxy rice, Dongjin 1, cultivated with conventional farming (CF) and environmentally-harmonized farming (EHF) using hairy vetch were compared to determine rice water absorption, physicochemical and pasting properties, antioxidant activities of brown and white rice, and a sensory evaluation of cooked white rice was carried out. EHF was treated with green manure crops such as hairy vetch and chitinase, which produce microorganism culture solution. CF was applied with seed disinfection treatments, fertilizer herbicides, and agricultural chemicals for the control of pests and diseases. The absorption level of EHF rice was higher than that of CF rice grain, regardless of the cultivation methods used. The ash and crude lipid contents were higher, but protein and dietary fiber contents were lower in the CF rice than in the EHF rice. The total starch content, water binding capacity, and swelling power of white rice were higher than those of brown rice, regardless of the cultivation methods used. The DPPH's antioxidant activity was shown as follows: EHF brown rice, EHF white rice and CF rice, in a decreasing order. The initial pasting temperature of EHF rice was lower than that of CF rice, but the peak, cold, and breakdown viscosities exhibited reverse trends. The sensory evaluation showed that the cooked white rice cultivated with EHF was not significantly different from that cultivated with CF (p<0.05). The overall preference of cooked rice did not show significant differences between the two cultivation methods (p<0.05).
In order to produce high-quality fermenting composts, bacteria strains with high activities of extracellular enzymes (cellulase, chitinase, amylase, protease and lipase) were isolated from the soils in 6 provinces of Korea, and characterized by 16S rRNA gene sequence analysis and properties. The selected 7 stains inoculated to livestock manure for 2' fermenting time, and experimental treatment divided into 3 groups, B1, B2 and B3, according to microbial activity and enzyme type. Our results showed that microbe applications (B1, B2 and B3) can increase (p<0.05) both rhizomes (17-38%) and enzyme activities (50-81%) in compost after fermenting time, respectively, compared to non-microbe treatment (control). The microbe application also decreased significantly (p<0.05) the $NH_3$ and $H_2S$ gas contents 13.4 and 27.3% compared with control, and the Propionic acid and Butyric acid gas contents 14.5 and 19.6%, respectively, as compared to the control. The microbial degradation rate (%) of pesticides and heavy metals increased significantly (p<0.05) after fermenting time, respectively, as compared to the control. Especially, microbe applications were more effective in total rhizomes yields and bioactivities than non-microbe treatment. Thus the results of this study could help in development of potential bioinoculants and composting techniques that maybe suitable for crop production, and protectable for earth environment under various conditions.
To investigate protective activity in pepper plants, which were pre-inoculated with arbuscular mycorrhizal (AM) fungi Glomus intra radices (Gi), against pathogenic strain Phytophthora capsici (Pc), pathogenesis-related (PR) proteins and antioxidant enzymes were examined. The growth of root and shoot was the highest in peppers inoculated with G. intraradices, compared with non-inoculated control plants and those challenged by the pathogen with and without mycorrhizae after nine days of infection. Mycorrhizal colonization rate was reduced by about 10% in pathogen-challenged plants, but disease pressure was reduced. The activities of PR proteins, $\beta$-1- 3-glucanase and chitinase, were increased in Pc-treated plants compared to Gi+Pc-treated plants in leaves, but those in roots were suppressed. Superoxide dismutase activity and $H_2O_2${/TEX> content in Gi+Pc and Pc-treated plants were gradually increased in leaves. However, those in roots continuously increased up to 5 days, and then decreased dramatically. Peroxidase activity in leaves and roots increased after P. capsici infection both in plants inoculated with or without G. intraradices. These results suggest that AM fungi, G. intra radices, potentially act as one of the protective agents against plant pathogens. Changes of PR proteins and antioxidative enzymes in mycorrhizae-inoculated pepper appear to be regulated differently in leaves and roots by pathogen infection.
The fermented manure derivative known as Preparation 500 is traditionally used as a field spray in biodynamic agriculture for maintaining and increasing soil fertility. This work aimed at characterizing the product from a microbiological standpoint and at assaying its bioactive properties. The approach involved molecular taxonomical characterization of the culturable microbial community; ARISA fingerprints of the total bacteria and fungal communities; chemical elemental macronutrient analysis via a combustion analyzer; activity assays for six key enzymes; bioassays for bacterial quorum sensing and chitolipooligosaccharide production; and plant hormone-like activity. The material was found to harbor a bacterial community of $2.38{\times}10^8$ CFU/g dw dominated by Gram-positives with minor instances of Actinobacteria and Gammaproteobacteria. ARISA showed a coherence of bacterial assemblages in different preparation lots of the same year in spite of geographic origin. Enzymatic activities showed elevated values of ${\beta}$-glucosidase, alkaline phosphatase, chitinase, and esterase. The preparation had no quorum sensing-detectable signal, and no rhizobial nod gene-inducing properties, but displayed a strong auxin-like effect on plants. Enzymatic analyses indicated a bioactive potential in the fertility and nutrient cycling contexts. The IAA activity and microbial degradation products qualify for a possible activity as soil biostimulants. Quantitative details and possible modes of action are discussed.
Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.
Inflammation is a protective response to infection or injury. However, prolonged inflammation can contribute to the pathogenesis of many diseases, such as cancer, diabetes, arthritis, atherosclerosis, and Alzheimer's disease. Recent studies have shown that activated macrophages, inflammatory effector cells, can react to tissue insults in a polarized manner, in which their phenotypes are polarized into two major subtypes, categorized as M1 or M2. Classical M1 activation involves the production of pro-inflammatory cytokines, such as interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$, and free radicals, while M2 or alternative activation is an anti-inflammatory phenotype involved in homeostatic processes, such as wound healing, debris scavenging, and the dampening of inflammation via the production of very low levels of pro-inflammatory cytokines and high levels of anti-inflammatory mediators, including IL-10. As part of our ongoing effort to isolate anti-inflammatory compounds from seaweeds, we investigated the effects of phlorotannins isolated from the brown alga Ecklonia stolonifera on macrophage polarization. Mouse peritoneal macrophages were treated with various concentrations of the extracts, and real-time RT-PCR analyses were performed to examine the expression of polarization markers: IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ for M1 and arginase-1, peroxisome proliferator-activated receptor (PPAR)-${\gamma}$, found inflammatory zone-1 (Fizz-1), chitinase 3-like 3 (Ym1), and$Kr{\ddot{u}}ppel$-like factor 4 (Klf-4) for M2. The pretreatment of cells with eckol, dieckol, and phlorofucofuroeckol-A (PFF-A), isolated from the ethyl acetate fraction of E. stolonifera ethanolic extract, potentiated the anti-inflammatory M2 phenotype of the macrophages. These results indicate that phlorotannins derived from E. stolonifera can be used to enrich macrophages with markers of the M2 anti-inflammatory state.
Seed germination is the important stage to express many genes for regulation of energy metabolism, starch degradation and cell division from seed dormancy state. For the functional analysis of seed germination mechanisms, we were analyzed the rice cDNA clones (Oryzasativa cultivar Ilpum) obtained from seed imbibition during 48 hours. Total number of 18,101 Expressed Sequence Tags (ESTs) were clustered using SeqMan program. Among them, 8,836 clones were identified as unique clones. We identified the chitinase gene specifically expressed in seed germination and amylase gene involved to starch degradation from the full length cDNA analysis, and several genes were registered to NCBI GeneBank. To analyzed the commonly expressed genes between inmature seed and germinated seed, 25,66 inmature ESTs and 18,101 germinated ESTs were clustered using SeqMan program and identified 2,514 clones as commonly expressed unigene. Among them, alpha-glubulin and alcohol dehydrogenase I were supposed to LEA genes only expressed in the immature and germinated seed stages. For the clustering of orthologous group genes, we further analyzed the 8,836 EST clones from germinating seeds using NCBI clusters of orthologous groups database. Among the clones, 5,076 clones were categorized into information storage and processing, cellular processes and signaling, metabolism and poorly characterized genes, proportioning 783 (14.29%), 1,484 (27%), 1,363 (24.8%) and 1,869 (34%) clones to the previous four categories, respectively.
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