Mee Kyung Sang

The volatile‐producing Flavobacterium johnsoniae strain GSE09 shows biocontrol activity against Phytophthora capsici in pepper

Aims:  Previously, we selected a bacterial strain (GSE09) antagonistic to Phytophthora capsici on pepper, which produced a volatile compound (2,4‐di‐tert‐butylphenol), inhibiting the pathogen. In this study, we identified strain GSE09 and characterized some of the biological traits of this strain in relation to its antagonistic properties against P. capsici. In addition, we examined bacterial colonization on the root surface or in rhizosphere soil and the effect of various concentrations of the volatile compound and strain GSE09 on pathogen development and radicle infection as well as radicle growth.

Biocontrol Activity and Induction of Systemic Resistance in Pepper by Compost Water Extracts Against Phytophthora capsici

We investigated the effects of water extracts of composts (CWE) from commercial compost facilities for controlling root and foliar infection of pepper plants by Phytophthora capsici. Among 47 CWE tested, CWE from composts Iljuk-3, Iljuk-7, Shinong-8, and Shinong-9 significantly (P < 0.05) inhibited zoospore germination, germ tube elongation, mycelial growth, and population of P. capsici. All selected CWE significantly (P < 0.05) reduced the disease incidence and severity in the seedling and plant assays compared with the controls. However, there were no significant differences in zoospore germination, disease incidence, and disease severity between treatments of untreated, autoclaved, and filtered CWE. In addition, CWE significantly (P < 0.05) suppressed leaf infection of P. capsici through induced systemic resistance (ISR) in plants root-drenched with CWE. The tested CWE enhanced the expression of the pathogenesis-related genes, CABPR1, CABGLU, CAChi2, CaPR-4, CAPO1, or CaPR-10 as well as beta-1,3-glucanase, chitinase, and peroxidase activities, which resulted in enhanced plant defense against P. capsici in pepper plants. Moreover, the CWE enhanced the chemical and structural defenses of the plants, including H(2)O(2) generation in the leaves and lignin accumulation in the stems. The CWE could also suppress other fungal pathogens (Colletotrichum coccodes in pepper leaves and C. orbiculare in cucumber leaves) through ISR; however, it failed to inhibit other bacterial pathogens (Xanthomonas campestris pv. vesicatoria in pepper leaves and Pseudomonas syringae pv. lachrymans in cucumber leaves). These results suggest that a heat-stable chemical(s) in the CWE can suppress root and foliar infection by P. capsici in pepper plants. In addition, these suppressions might result from direct inhibition of development and population of P. capsici for root infection, as well as indirect inhibition of foliar infection through ISR with broad-spectrum protection.

Root Treatment with Rhizobacteria Antagonistic to Phytophthora Blight Affects Anthracnose Occurrence, Ripening, and Yield of Pepper Fruit in the Plastic House and Field

We previously selected rhizobacterial strains CCR04, CCR80, GSE09, ISE13, and ISE14, which were antagonistic to Phytophthora blight of pepper. In this study, we investigated the effects of root treatment of rhizobacteria on anthracnose occurrence, ripening, and yield of pepper fruit in the plastic house and field in 2008 and 2009. We also examined the effects of volatiles produced by the strains on fruit ripening and on mycelial growth and spore development of Colletotrichum acutatum and Phytophthora capsici in the laboratory, identifying the volatile compounds by gas chromatography-mass spectrometry (GC-MS). In the house tests, all strains significantly (P < 0.05) reduced anthracnose incidence on pepper fruit; strains GSE09 and ISE14 consistently produced higher numbers of pepper fruit or increased the fresh weight of red fruit more than the controls in both years. In the field tests, all strains significantly (P < 0.05) reduced anthracnose occurrence on either green or red pepper fruit; strain ISE14 consistently produced higher numbers or increased fresh weights of red fruit more than the controls in both years. In the laboratory tests, volatiles produced by strains GSE09 and ISE13 only stimulated maturation of pepper fruit from green (unripe) to red (ripe) fruit; the volatiles of certain strains inhibited the growth and development of C. acutatum and P. capsici. On the other hand, GC-MS analysis of volatiles of strains GSE09 and ISE13 revealed 17 distinct compounds in both strains, including decane, dodecane, 1,3-di-tert-butylbenzene, tetradecane, 2,4-di-tert-butylphenol, and hexadecane. Among these compounds, 2,4-di-tert-butylphenol only stimulated fruit ripening and inhibited growth and development of the pathogens. Taken together, strains GSE09 and ISE14 effectively reduced anthracnose occurrence and stimulated pepper fruit ripening and yield, possibly via bacterial volatiles. Therefore, these two strains could be potential agents for controlling Phytophthora blight and anthracnose, and for increasing fruit ripening and yield. To our knowledge, this is the first report of volatiles such as 2,4-di-tert-butylphenol produced by rhizobacteria being related to both fruit ripening and pathogen inhibition.

Biocontrol Activity and Primed Systemic Resistance by Compost Water Extracts Against Anthracnoses of Pepper and Cucumber

We investigated direct and indirect effects of compost water extracts (CWEs) from Iljuk-3, Iljuk-7, Shinong-8, and Shinong-9 for the control of anthracnoses caused by Colletotrichum coccodes on pepper and C. orbiculare on cucumber. All tested CWEs significantly (P < 0.05) inhibited in vitro conidial germination and appressorium formation of the fungal pathogens; however, DL-β-amino-n-butyric acid (BABA) failed to inhibit the conidial development of the pathogens. Direct treatments of the CWEs and BABA on pepper and cucumber leaves at 1 and 3 days before or after inoculation significantly (P < 0.05) reduced anthracnose severities; Iljuk-3, Shinong-9, and BABA for pepper and Iljuk-7 for cucumber had more protective activities than curative activities. In addition, root treatment of CWEs suppressed anthracnoses on the plants by the pathogens; however, CWE treatment on lower leaves failed to reduce the diseases on the upper leaves of the plants. The CWE root treatments enhanced not only the expression of the pathogenesis-related (PR) genes CABPR1, CABGLU, CAChi2, CaPR-4, CAPO1, and CaPR-10 in pepper and PR1-1a, PR-2, PR-3, and APOX in cucumber but also the activity of β-1,3-glucanase, chitinase, and peroxidase and the generation of hydrogen peroxide in pepper and cucumber under pathogen-inoculated conditions. However, the CWE treatments failed to induce the plant responses under pathogen-free conditions. These results indicated that the CWEs had direct effects, reducing anthracnoses by C. coccodes on pepper leaves and C. orbiculare on cucumber leaves through protective and curative effects. In addition, CWE root treatments could induce systemic resistance in the primed state against pathogens on plant leaves that enhanced PR gene expression, defense-related enzyme production, and hydrogen peroxide generation rapidly and effectively immediately after pathogen infection. Thus, the CWEs might suppress anthracnoses on leaves of both pepper and cucumber through primed (priming-mediated) systemic resistance.

Chryseobacterium kwangjuense sp. nov., isolated from pepper (Capsicum annuum L.) root.

The yellow-pigmented, Gram-stain-negative, rod-shaped bacterium KJ1R5(T) was isolated from the root of a pepper plant grown in a field in Kwangju, Korea. Strain KJ1R5(T) was characterized by physiological, biochemical, and molecular genetic analyses. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain KJ1R5(T) was most closely related to members of the genus Chryseobacterium, and that the strain exhibited the highest similarities with type strains of Chryseobacterium vrystaatense (97.0 %) and Chryseobacterium rhizosphaerae (97.1 %). DNA-DNA hybridization reassociation values between strain KJ1R5(T) and type strains of C. vrystaatense KACC 11675(T) and C. rhizosphaerae KACC 14918(T) were 46.9 and 38.4 %, respectively. The DNA G+C content of KJ1R5(T) is 40.2 mol%. The predominant respiratory quinone of KJ1R5(T) was menaquinone MK-6; major cellular fatty acids were iso-C15 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C17 : 1ω9c, and iso-C17 : 0 3-OH. On the basis of these phenotypic and genotypic characteristics, the strain significantly differed from representative strains belonging to the genus Chryseobacterium. Thus, we propose that strain KJ1R5(T) represents a novel species of the genus Chryseobacterium, named Chryseobacterium kwangjuense sp. nov. The type strain is KJ1R5(T) (= KACC 13029(T) = JCM 15904(T)).

Application of Volatile Antifungal Plant Essential Oils for Controlling Pepper Fruit Anthracnose by Colletotrichum gloeosporioides

Anthracnose caused by Colletotrichum gloeosporioides has been destructive during pepper fruit production in outdoor fields in Korea. In vitro antifungal activities of 15 different plant essential oils or its components were evaluated during conidial germination and mycelial growth of C. gloeosporioides. In vitro conidial germination was most drastically inhibited by vapour treatments with carvacrol, cinnamon oil, trans-cinnamaldehyde, citral, p-cymene and linalool. Inhibition of the mycelial growth by indirect vapour treatment with essential oils was also demonstrated compared with untreated control. Carvacrol, cinnamon oil, trans-cinnamaldehyde, citral and eugenol were among the most inhibitory plant essential oils by the indirect antifungal efficacies. Plant protection efficacies of the plant essential oils were demonstrated by reduced lesion diameter on the C. gloeosporioides-inoculated immature green pepper fruits compared to the inoculated control fruits without any plant essential oil treatment. In planta test showed that all plant essential oils tested in this study demonstrated plant protection efficacies against pepper fruit anthracnose with similar levels. Thus, application of different plant essential oils can be used for eco-friendly disease management of anthracnose during pepper fruit production.

Characterization of Bacillus luciferensis Strain KJ2C12 from Pepper Root, a Biocontrol Agent of Phytophthora Blight of Pepper

In this study, we characterized the bacterial strain KJ2C12 in relation with its biocontrol activity against Phytophthora capsici on pepper, and identified this strain using morphological, physiological, biochemical, fatty acid methyl ester, and 16S rRNA gene sequence analyses. Strain KJ2C12 significantly (P

Biocontrol of Phytophthora Blight and Anthracnose in Pepper by Sequentially Selected Antagonistic Rhizobacteria against Phytophthora capsici.

We previously developed a sequential screening procedure to select antagonistic bacterial strains against Phytophthora capsici in pepper plants. In this study, we used a modified screening procedure to select effective biocontrol strains against P. capsici; we evaluated the effect of selected strains on Phytophthora blight and anthracnose occurrence and fruit yield in pepper plants under field and plastic house conditions from 2007 to 2009. We selected four potential biocontrol strains (Pseudomonas otitidis YJR27, P. putida YJR92, Tsukamurella tyrosinosolvens YJR102, and Novosphingobium capsulatum YJR107) among 239 bacterial strains. In the 3-year field tests, all the selected strains significantly (P < 0.05) reduced Phytophthora blight without influencing rhizosphere microbial populations; they showed similar or better levels of disease suppressions than in metalaxyl treatment in the 2007 and 2009 tests, but not in the 2008 test. In the 2-year plastic house tests, all the selected strains significantly (P < 0.05) reduced anthracnose incidence in at least one of the test years, but their biocontrol activities were variable. In addition, strains YJR27, YJR92, and YJR102, in certain harvests, increased pepper fruit numbers in field tests and red fruit weights in plastic house tests. Taken together, these results indicate that the screening procedure is rapid and reliable for the selection of potential biocontrol strains against P. capsici in pepper plants. In addition, these selected strains exhibited biocontrol activities against anthracnose, and some of the strains showed plant growth-promotion activities on pepper fruit.

Biocontrol of Korean Ginseng Root Rot Caused by Phytophthora cactorum Using Antagonistic Bacterial Strains ISE13 and KJ1R5

In this biocontrol research, we evaluated disease suppressive effects of antagonistic bacterial strains ISE13 and KJ1R5 against Korean ginseng root rot caused by P. eaetorum. We also examined the effects of nutrient solution in the hydroponic culture system for Korean ginseng on biological activity of the bacterial strains. As results of dual culture tests of the bacterial strains on juice agar, the strain ISE13 showed antifungal activity against P. eaetorum and other plant pathogenic fungi, but the strain KJ1R5 did not. When their inhibitory effects against infection of P. eaetorum on the roots grown in either nutrient solution or water were tested, the strains ISE13 and KJ1R5 inhibited the disease severity of Korean ginseng roots only grown with water, compared to buffer-treated, inoculated controls. However, the nutrient solution used for hydroponic cultures of ginseng in pots caused higher levels of disease severity by the strains ISE13 and KJ1R5 from to and from to , respectively. In this study, the bacterial strains ISE13 and KJ1R5 could be potentially biocontrol agents to suppress Korean ginseng root rot caused by P. eaetorum. However, more attention using nutrient solution in hydroponic cultures for Korean ginseng production should be applied in biocontrol of plant diseases using the antagonistic microorganisms.

Microbial Population, Aflatoxin Contamination and Predominant Aspergillus Species in Korean Stored Rice

We evaluated microbial populations and aflatoxin production in unhulled and white rice from rice processing complexes of the National Agricultural Cooperative Federation in five regions in Korea and identified three predominant Aspergillus species. Fungal and bacterial populations in rice samples were significantly different between regions in 2007. Aflatoxins were also detected and varied at the levels of 2.45 - 3.43 ng per g unhulled rice grain and 1.29 - 2.09 ng per g white rice grain. Unhulled rice generally detected higher level of aflatoxins than white rice regardless of sampling regions; however, no significant differences were found in Anseong and Cheonan in 2005 and Cheonan and Gimpo in 2007. Aflatoxin production between sampling regions was not different regardless of rice type and sampling year. Although the fungal diversity was highly distinct from region to region, three Aspergillus isolates were predominant in the rice samples; thus, representative isolates AC317, AF57, and AF8 were selected and identified based on their morphological and molecular characteristics. Consequently, isolates AC317, AF57, and AF8 were identified as A. candidus, A. flavus, and A. fumigatus, respectively. These fungi can produce mycotoxins that are harmful for consumers and thus it is important to detect and reduce the population of storage fungi in rice.