Effects of different NS genes of avian influenza viruses and amino acid changes on pathogenicity of recombinant A/Puerto Rico/8/34 viruses

Kim, Il-Hwan,Kwon, Hyuk-Joon,Lee, Su-Hyung,Kim, Dae-Yong,Kim, Jae-Hong Elsevier 2015 Veterinary microbiology Vol.175 No.1

<P><B>Abstract</B></P> <P>To examine the effects of the NS1 and NEP genes of avian influenza viruses (AIVs) on pathogenicity in mice, we generated recombinant PR8 viruses containing 3 different NS genes of AIVs. In contrast to the reverse genetics-generated PR8 (rPR8) strain and other recombinant viruses, the recombinant virus rPR8-NS(0028), which contained the NS gene of A/chicken/KBNP-0028/2000 (H9N2) (0028), was non-pathogenic to mice. The novel single mutations of 0028 NS1 to corresponding amino acid of PR8 NS1, G139D and S151T increased the pathogenicity of rPR8-NS(0028). The replacement of the PL motifs (EPEV or RSEV) of pathogenic recombinant viruses with that of 0028 (GSEV) did not reduce the pathogenicity of the viruses. However, a recombinant virus with an EPEV-grafted 0028 NS gene was more pathogenic than rPR8-NS(0028) but less than rPR8. The lower pathogenicity of rPR8-NS(0028) might be associated with the lower virus titer and IFN-β level in the lungs of infected mice, and be attributed to G139, S151 and GSEV-PL motif of NS1 gene of 0028. In conclusion we defined new amino acid residues of NS1 related to mice pathogenicity and the presence of pathogenic NS genes among low pathogenic AIVs may encourage continuous monitoring of their mammalian pathogenicity.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A mice nonpathogenic NS gene originated from an attenuated H9N2 virus was identified. </LI> <LI> Mice pathogenic NS genes among LPAIVs were identified. </LI> <LI> Novel amino acid changes related to mice pathogenicity were identified. </LI> <LI> A method to generate nonpathogenic PR8-based recombinant virus was suggested. </LI> </UL> </P>

두 개의 새로운 phytotoxin의 Botrytis cinerea 병원성에서의 역할

김금정,윤미영,김흥태,최경자,장경수,최용호,박명수,차병진,김진철 한국식물병리학회 2009 식물병연구 Vol.15 No.2

In the course of study on the roles of phytotoxins in the pathogenicity of Botrytis cinerea, we isolated two novel phytotoxins. They were identified as 3-O-acetyl botcinol and 3-O-acetyl botcinolide. In this study, we investigated correlation between the two phytotoxins and the pathogenicity of B. cinerea. In liquid cultures, the two phytotoxins were not produced by three low pathogenic isolates out of 25 B. cinerea isolates. Among strong or moderate pathogenic isolates, some produced the two phytotoxins, but the others did not. On the other hand, the ethyl acetate extracts of fermentation broths of 10 out of 25 isolates showed phytotoxic activity against various plants tested in a whole plant assay. The phytotoxins were detected in all of the 10 phytotoxic ethyl acetate extracts. In planta, the two phytotoxins were detected in all of the plant tissues infected with strong pathogenic isolates. However, there was no correlation between the ability of B. cinerea isolates to produce the two phytotoxins and their pathogenicities. The two phytotoxins began to detect in tomato plant tissues infected with B. cinerea 2-16 at 3 days after inoculation, increased gradually till 4 days after inoculation, and then decreased. The above results suggest that 3-O-acetyl botcinol and 3-O-acetyl botcinolide are one of pathogenicity factors for B. cinerea, but not a primary determinant of its pathogenicity. In the course of study on the roles of phytotoxins in the pathogenicity of Botrytis cinerea, we isolated two novel phytotoxins. They were identified as 3-O-acetyl botcinol and 3-O-acetyl botcinolide. In this study, we investigated correlation between the two phytotoxins and the pathogenicity of B. cinerea. In liquid cultures, the two phytotoxins were not produced by three low pathogenic isolates out of 25 B. cinerea isolates. Among strong or moderate pathogenic isolates, some produced the two phytotoxins, but the others did not. On the other hand, the ethyl acetate extracts of fermentation broths of 10 out of 25 isolates showed phytotoxic activity against various plants tested in a whole plant assay. The phytotoxins were detected in all of the 10 phytotoxic ethyl acetate extracts. In planta, the two phytotoxins were detected in all of the plant tissues infected with strong pathogenic isolates. However, there was no correlation between the ability of B. cinerea isolates to produce the two phytotoxins and their pathogenicities. The two phytotoxins began to detect in tomato plant tissues infected with B. cinerea 2-16 at 3 days after inoculation, increased gradually till 4 days after inoculation, and then decreased. The above results suggest that 3-O-acetyl botcinol and 3-O-acetyl botcinolide are one of pathogenicity factors for B. cinerea, but not a primary determinant of its pathogenicity.

In vivo Pathogenicity Test of Oak Wilt Fungus (Raffaelea quercus-mongolicae) on Oriental Chestnut Oak (Quercus acutissima)

Su Hee Yi,Jin Heung Lee,Sang Tae Seo,Jong Kyu Lee 강원대학교 산림과학연구소 2017 Journal of Forest Science Vol.33 No.4

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

In vivo Pathogenicity Test of Oak Wilt Fungus (Raffaelea quercus-mongolicae) on Oriental Chestnut Oak (Quercus acutissima)

Yi, Su Hee,Lee, Jin Heung,Seo, Sang Tae,Lee, Jong Kyu Institute of Forest Science 2017 Journal of Forest Science Vol.33 No.4

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

In vivo Pathogenicity Test of Oak Wilt Fungus (Raffaelea quercus-mongolicae) on Oriental Chestnut Oak (Quercus acutissima)

이수희,이진흥,서상태,이종규 강원대학교 산림과학연구소 2017 Journal of Forest Science Vol.33 No.4

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

Novel mutations in avian PA in combination with an adaptive mutation in PR8 NP exacerbate the virulence of PR8-derived recombinant influenza A viruses in mice

Lee, Chung-Young,An, Se-Hee,Kim, Ilhwan,Choi, Jun-Gu,Lee, Youn-Jeong,Kim, Jae-Hong,Kwon, Hyuk-Joon Elsevier 2018 Veterinary microbiology Vol.221 No.-

<P><B>Abstract</B></P> <P>The polymerase complex of the low-pathogenic avian influenza virus [A/chicken/Korea/KBNP-0028/2000] (0028) has previously been characterized, and novel amino acid residues present in the polymerase acidic protein (PA) that likely contribute to pathogenicity toward mammals have been identified. In the present study, our aims were to generate A/Puerto Rico/8/34 (PR8)-derived recombinant viruses containing the <I>0028-PA</I> gene with a single amino acid mutation and to test their pathogenicity and replication ability. We found that the recombinant viruses acquired additional single mutations in the nucleoprotein (NP). Because the additional mutations in NP did not affect viral pathogenicity, but rather attenuated viral replication and polymerase activity, the incompatibility of the avian <I>PA</I> gene within the PR8 backbone may have induced an adaptive mutation in <I>NP</I>. To minimize the differences due to NP mutations, we generated 0028-PA mutants with an E375G mutation, not affecting viral replication and pathogenicity, in the <I>NP</I> gene. The PR8-PA(0028)-E684G mutant showed significantly higher viral replication in mammalian cells as compared to PR8-PA(0028) and led to 100% mortality in mice, with significantly increased interferon β expression. Thus, the E684G mutation in the <I>PA</I> gene may play an important role in viral pathogenicity in mice by increasing viral replication and the host immune response.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The incompatibility of avian PA within the PR8 backbone induced mutations in NP. </LI> <LI> The E684G PA mutation increased viral replication in ECEs and MDCK cells. </LI> <LI> The E684G PA mutation increased viral replication and pathogenicity in mice. </LI> <LI> E684G mutation in PA may be a potential pathogenicity-related mutation. </LI> </UL> </P>

Benzylideneacetone suppresses both cellular and humoral immune responses of Spodoptera exigua and enhances fungal pathogenicity

Jung-A Park,김용균 한국응용곤충학회 2011 Journal of Asia-Pacific Entomology Vol.14 No.4

An entomopathogenic fungus, Beauveria bassiana, had significant insecticidal activity against the beet armyworm, Spodoptera exigua. However, it took almost one week to cause significant mortality. This study used a mixture treatment with an immunosuppressant to enhance the fungal pathogenicity. A bacterial metabolite, benzylideneacetone (BZA), had a significant synergistic effect on the fungal pathogenicity against S. exigua, although it had little insecticidal activity by itself. The mixture treatment shortened median lethal time of B. bassiana by approximately 2 days. The synergistic activity of BZA on the pathogenicity of B. bassiana was induced by its immunosuppressive effects on both cellular and humoral antifungal responses of S. exigua. In response to B. bassiana, S. exigua larvae can form hemocytic nodules. Nodules were significantly suppressed by BZA treatment. Moreover, BZA inhibited expression of some antimicrobial peptide genes of S. exigua in response to fungal challenge. The immunosuppressive condition induced by BZA allowed B. bassiana to easily colonize and multiply in the hemocoel of treated larvae, which resulted in significant enhancement of the pathogenicity of B. bassiana.

Benzylideneacetone suppresses both cellular and humoral immune responses of Spodoptera exigua and enhances fungal pathogenicity

Park, Jung-A,Kim, Yong-Gyun 한국응용곤충학회 2011 Journal of Asia-Pacific Entomology Vol.14 No.4

An entomopathogenic fungus, Beauveria bassiana, had significant insecticidal activity against the beet armyworm, Spodoptera exigua. However, it took almost one week to cause significant mortality. This study used a mixture treatment with an immunosuppressant to enhance the fungal pathogenicity. A bacterial metabolite, benzylideneacetone (BZA), had a significant synergistic effect on the fungal pathogenicity against S. exigua, although it had little insecticidal activity by itself. The mixture treatment shortened median lethal time of B. bassiana by approximately 2 days. The synergistic activity of BZA on the pathogenicity of B. bassiana was induced by its immunosuppressive effects on both cellular and humoral antifungal responses of S. exigua. In response to B. bassiana, S. exigua larvae can form hemocytic nodules. Nodules were significantly suppressed by BZA treatment. Moreover, BZA inhibited expression of some antimicrobial peptide genes of S. exigua in response to fungal challenge. The immunosuppressive condition induced by BZA allowed B. bassiana to easily colonize and multiply in the hemocoel of treated larvae, which resulted in significant enhancement of the pathogenicity of B. bassiana.

사과흰날개무늬병균의 Cytochalasin E 독소 생산과 병원성

이동혁,최경희,엄재열 한국식물병리학회 2009 식물병연구 Vol.15 No.1

Cytochalasin E (CE) is a secondary metabolite secreted by Rosellinia necatrix, caused by white root rot, and has toxicity to apple as a toxin during disease progress. This study was conducted to demonstrate the relationship between the production of CE and its pathogenicity. CE producing isolates and non-producing isolates of R. nectatrix were isolated from the mycerial mat of diseased roots and was detected on that using a TLC and HPLC analysis and in vivo pathogenicity test. CE non-producing isolates were not pathogenic to apple roots and not detected CE by TLC and HPLC analysis. It was shown that the production of CE was related to the pathogenicity of R. nectatrix. Cytochalasin E (CE) is a secondary metabolite secreted by Rosellinia necatrix, caused by white root rot, and has toxicity to apple as a toxin during disease progress. This study was conducted to demonstrate the relationship between the production of CE and its pathogenicity. CE producing isolates and non-producing isolates of R. nectatrix were isolated from the mycerial mat of diseased roots and was detected on that using a TLC and HPLC analysis and in vivo pathogenicity test. CE non-producing isolates were not pathogenic to apple roots and not detected CE by TLC and HPLC analysis. It was shown that the production of CE was related to the pathogenicity of R. nectatrix.

보문 : 병원성 검정 및 RAPD 분석에 의한 국내 인삼뿌리썩음병균(Cylindrocarpon destructans)의 유전적 다양성

서문원 ( Mun Won Seo ),김선익 ( Sun Ick Kim ),송정영 ( Jeong Young Song ),김홍기 ( Hong Gi Kim ) 한국균학회 2011 韓國菌學會誌 Vol.39 No.1

Ginseng root rot caused by Cylindrocarpon destructans is one of the most destructive diseases of ginseng (Panax ginseng). We analyzed the features of the species through pathogenicity test and genetic diversity analysis of C. destructans in Korea, for its application as basic data to attempt for effective control. C. destructans isolated from rotted ginseng roots exhibited a variety of colonial colors on media. It was assumed that there may exist genetic diversity in the population by the diversity of pathogenicity among isolates observed when artificially inoculated into ginseng roots. Pathogenicity tests using ex vivo wound inoculation with agar mixture inoculation on ginseng roots were performed similar results as were observed appear to be useful for rapid pathogen inspection. According to RAPD analysis results, Korean C. destructans isolates formed a single genetic group which can be distinguished readily from closely related other fungi. C. destructans group was divided into two small groups. Therefore, we were able to confirm pathogenicity and genetic difference between the isolates in each of the groups of the pathogen.