• Research in Plant Disease
• /
• v.18 no.4
• /
• pp.268-276
• /
• 2012

Broad bean wilt virus 2 (BBWV2), genus Fabavirus, subfamily Comovirinae, family Secoviridae, causes damage in many economically important horticultural and ornamental crops. Sequence alignments showed several conserved sequences in 5' non-coding regions (5' NCRs) of RNA 1 and RNA 2 in all BBWV2 strains characterized so far. Based on this observation, we generated a hpRNA construct (pIR-BBWV2) harboring an inverted repeat containing a 210 bp cDNA fragment homologous to 5' NCR portion of BBWV2 RNA 1 to investigate the silencing potential for its ability to interfere with a rapidly replicating BBWV2. Agrobacterium-mediated transient expression of the IR-BBWV2 had a detrimental effect on BBWV2 infection, showing no distinct symptoms in non-inoculated leaves of the agroinfiltrated Nicotiana benthamiana plants. BBWV2 genomic RNAs were not detected by RT-PCR from tissues of both the inoculated leaves and upper leaves of the agroinfiltrated plants. Accumulation of virus-derived small interfering RNAs was detected in the inoculated leaf tissues of N. benthamiana plants elicited by transient expression of IR-BBWV2 indicating that RNA silencing is responsible for the resistance to BBWV2.

• The Plant Pathology Journal
• /
• v.34 no.6
• /
• pp.514-531
• /
• 2018

Tomato spotted wilt virus (TSWV; Genus Orthotospovirus: Family Tospoviridae) is one of the most destructive viruses affecting a wide range of horticultural crops on a worldwide basis. In 2015 and 2016, 171 leaf and fruit samples from tomato (Solanum lycopersicum) plants with viral symptoms were collected from the fields in various regions of Iran. ELISA test revealed that the samples were infected by TSWV. The results of RT-PCR showed that the expected DNA fragments of about 819 bp in length were amplified using a pair of universal primer corresponding to the RNA polymerase gene and DNA fragments of ca 777 bp and 724 bp in length were amplified using specific primers that have been designed based on the nucleocapsid (N) and non-structural (NSs) genes, respectively. The amplified fragments were cloned into pTG19-T and sequenced. Sequence comparisons with those available in the GenBank showed that the sequences belong to TSWV. The high nucleotide identity and similarities of new sequences based on the L, N, and NSs genes showed that minor evolutionary differences exist amongst the isolates. The phylogenetic tree grouped all isolates six clades based on N and NSs genes. Phylogenetic analysis showed that the Iranian isolates were composed a new distinct clade based on a part of polymerase, N and NSs genes. To our knowledge, this is the first detailed study on molecular characterization and genetic diversity of TSWV isolates from tomato in Iran that could be known as new clade of TSWV isolates.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.3
• /
• pp.474-480
• /
• 2007

Fusarium wilt of cucumbers was effectively controlled by Escherichia coli expressing an endochitinase gene (chiA), and the rate was as effective (60.0%) as the wild-type strain S. proteamaculans 3095 (55.0%) where the gene was cloned. However, live cells of soil inoculated E. coli host harboring the chiA gene did not proliferate but declined 100-fold from $10^8$ CFU during the first week and showed less than 10 cells after day 14, suggesting that E. coli was able to express and produce the chitinase enzyme to the soil even as the population was gradually decreasing. Because the majority of the strains was alive for only a short period of time and the Fusarium-affected seedlings showed symptoms of wilting within 7-10 days, it seems that the pathogen control was decided early after the introduction of the biocontrol agent, eliminating the survival of the antagonist. These results indicated that soil inoculated E. coli could sufficiently express and produce the recombinant protein to control the pathogen, and root or soil colonization of the antagonist might not be a significant factor in determining the efficacy of biological control.

• Research in Plant Disease
• /
• v.18 no.2
• /
• pp.101-108
• /
• 2012

In 2006, pinewood nematode, Bursaphelenchus xylophilus, was isolated from about 50 years old trees of Pinus densiflora and P. koraiensis showing leaf-wilt and -drying symptoms in Gwangju, Gyeonggi-do and Chuncheon, Gangwon-do. Isolation of pinewood nematodes from sapwood of infected pine trees showed no difference in population density between tree species and among the sampling heights on the main stem. Migration of pinewood nematodes in the host tree were investigated by inoculation of red pine (P. densiflora, 3 years old) seedlings with B. xylophilus. The nematodes seemed to move in red pine seedlings prior to multiplication and it might have taken about 20 days to start multiplication and expression of symptoms including wilt and dieback. In initial time after inoculation, nematodes started migration through the cortical resin canal from inoculated site and further showed upward and downward movements. More nematodes were observed in cortical resin canal during early period of inoculation and later in resin canal of xylem and tracheid also while, the pith still remained free from nematode. The density of B. xylophilus was higher in seedlings of low-vigor with poor root growth than in seedlings of normal root growth. Seedlings showing high density of B. xylophilus exhibited stem discoloration and secondary infection by fungus at the inoculation site.

• The Plant Pathology Journal
• /
• v.39 no.1
• /
• pp.108-122
• /
• 2023

Fusarium oxysporum f. sp. lycopersici (Fol) and Fusarium oxysporum f. sp. cubense (Foc), are the causal agent of Fusarium wilt disease of tomato and banana, respectively, and cause significant yield losses worldwide. A cost-effective measure, such as biological control agents, was used as an alternative method to control these pathogens. Therefore, in this study, six isolates of the Streptomyces-like colony were isolated from soils and their antagonistic activity against phytopathogenic fungi and plant growth-promoting (PGP) activity were assessed. The results showed that these isolates could inhibit the mycelial growth of Fol and Foc. Among them, isolate STRM304 showed the highest percentage of mycelial growth reduction and broad-spectrum antagonistic activity against all tested fungi. In the pot experiment study, the culture filtrate of isolates STRM103 and STRM104 significantly decreased disease severity and symptoms in Fol inoculated plants. Similarly, the culture filtrate of the STRM304 isolate significantly reduced the severity of the disease and symptoms of the disease in Foc inoculated plants. The PGP activity test presents PGP activities, such as indole acetic acid production, phosphate solubilization, starch hydrolysis, lignin hydrolysis, and cellulase activity. Interestingly, the application of the culture filtrate from all isolates increased the percentage of tomato seed germination and stimulated the growth of tomato plants and banana seedlings, increasing the elongation of the shoot and the root and shoot and root weight compared to the control treatment. Therefore, the isolate STRM103 and STRM104, and STRM304 could be used as biocontrol and PGP agents for tomato and banana, respectively, in sustainable agriculture.

• The Korean Journal of Pesticide Science
• /
• v.10 no.4
• /
• pp.313-319
• /
• 2006

Two hundred bacterial strains were isolated from the soil around healthy tomato plants in a polyvinyl house, where most of the other plants showed bacterial wilt symptoms. The strains were screened in vitro for their antibacterial activity. Among them, a strain, KPB3 showed strong bactericidal activity against bacterial wilt pathogen, Ralstonia solanacearum. The strain KPB3 was identified using physiological and biochemical tests, and 16S rRNA analyses. Based on these tests, the strain was found to be closer to genus Paenibacillus. To control the bacterial wilt caused by R. solanacearum, greenhouse experiments were conducted to determine the effectiveness of the Paenibacillus strain KPB3. Drench application of this strain ($4{\times}10^8$ CFU $mL^{-1}$) into the pots containing tomato plants, post-inoculated with the pathogen, R. solanacearum could drastically reduce the disease severity, compared to the non-treated plants. To evaluate effectiveness of this strain under field conditions, experiments were carried out in polyvinyl houses infested with R. solanacearum, during spring and autumn of the year 2006. It was observed that, during spring, bacterial wilt was more prevalent compared to the autumn. During spring, 50.9% disease incidences occurred in non-treated controls, while, Paenibacillus strain KPB3 treated plants showed 24.6% disease incidences. Similarly, during autumn, around 17.2% plants were infected with bacterial wilt in non- treated polyvinyl houses, compared to the Paenibacillus strain KPB3 treated plants, which showed 7.0% disease incidences. These results demonstrated that, Paenibacillus strain KPB3 is a potential biological control agent against bacterial wilt caused by R. solanacearum, effective under greenhouse as well as field conditions. This is the first report showing biocontrol of R. solanacearum using a Paenibacillus spp. under field conditions.

• Research in Plant Disease
• /
• v.18 no.4
• /
• pp.361-369
• /
• 2012

A population density model for bacterial wilt, which is caused by Ralstonia solanacearum, in hot pepper was developed to estimate the primary infection date after overwintering in the field. We developed the model mechansitically to predict reproduction of the pathogen and pathogensis on seedlings of the host. The model estimates the pathogen's populations both in the soil and in the host. In order to quantify environmental infection factors, various temperatures and initial population densities were determined for wilt symptoms on the seedlings of hot pepper in a chamber. Once, the pathogens living in soil multiply up to 400 cells/g of soil, they can infect successfully in the host. Primary infection in a host was supposed to be started when the population of the pathogen were over $10^9$ cells/g of root tissue. The estimated primary infection dates of bacterial wilt in 2011 in Korea were mostly mid-July or late-July which were 10-15 days earlier than those in 2010. Two kinds of meterological data, synoptic observation and field measurements from paddy field and orchard in Kyunggi, were operated the model for comparing the result dates. About 1-3 days were earlier from field data than from synoptic observation.

• Research in Plant Disease
• /
• v.19 no.4
• /
• pp.273-280
• /
• 2013

Tomato spotted wilt virus (TSWV) was occurred at 8 areas including Naju, Suncheon, Younggwang, Youngam, and Shinan in Jeonnam province and the crops of Younggwang were severely damaged by TSWV. The hot pepper (Capsicum annuum), bell pepper (Capsicum annuum v ar. angulosum) and tomato (Solanum lycopersicon) in greenhouse and hot pepper in open field were infected by TSWV. Especially, hot pepper was severely damaged by TSWV infection. The survey data indicated that 1.1-30% in the nursery field at Naju, Suncheon, and Jangheung were infected by TSWV. Plants were infected by TSWV from early June to August. However, TSWV-infected seedlings from nursery fields showed the disease symptoms from May after transplanting. In pepper greenhouses, Frankliniella occidentalis was more dominant insect vector than Frankliniella intonsa. But in open field, the population of insect vector was opposed to greenhouse. In addition, the removal of weeds was able to delay the incidence of TSWV via side-window of greenhouse in Winter. Taken together, the control of weed and insect vector nearby side-window of greenhouse is important to prevent TSWV infection of plants.

• Research in Plant Disease
• /
• v.12 no.3
• /
• pp.221-225
• /
• 2006

Cucumber mosaic virus(CMV) and Broad bean wilt virus 2(BBWV2) were isolated from Gentiana scabra plants showing typical mosaic and yellowing symptoms. When the inoculum of mosaic symptom propagated in Nicotiana benthamiana was inoculated to primary leaves of Vigna sinensis, the local lesions of different types was developed. Type one produced a small necrotic spot(SNS) of pinpoint type, while the other one showed a large necrotic spot(LNS) of halo type. LNS on primary leaves of V. sinensis was also induced by inoculum from yellowing symptom on G scabra. Single lesion from SNS induced a typical mosaic symptom on N. Benthamiana. On the other hand, LNS produced a chlorotic ring symptom on inoculated leaves and mosaic plus necrotic ringspot on upper leaves of N. benthamiana. An isolate of CMV from SNS and BBWV2 from LNS were detected by using dsRNA analysis, RT-PCR and agar gel double-diffusion test. Thus, our results should provide a tool of a simple method for discrimination from mixed infected plants by CMV and BBWV2.

• Research in Plant Disease
• /
• v.26 no.4
• /
• pp.256-263
• /
• 2020

Incidence of Fusarium wilt was surveyed in fields of summer radish in Gangwon province in Korea in 2018 and 2019. The disease started in early July and spread rapidly in hot summer of late July and August and in severe case, reached up to 80% in a field in Gangneung area. Symptoms in the seedling stage include poor growth and browning of internal tissue of root. During mid-growth, the leaves of diseased plant turned yellow over time, the surface of the roots changed from white to blackish, and the vascular tissues turned brown. A total of 23 isolates was obtained from the diseased plants and identified as Fusarium oxysporum f. sp. raphani by elongation factor-1α and intergenic spacer sequence analysis. Pathogenicity of the isolates was tested by artificial inoculation to the radish and other plants. All the isolates tested were pathogenic to radish plant, although there were differences in virulence on radish 11 cultivars. However, the isolates were not virulent to other plants except some cruciferous vegetables including Brussels sprouts, rocket, stock, and turnip. The results of pathogenicity test showed that it is necessary to rotate with crops other than cruciferous vegetables in order to prevent Fusarium wilt from radish fields.