Biological Control of Oomycete Soilborne Diseases Caused by Phytophthora capsici, Phytophthora infestans, and Phytophthora nicotianae in Solanaceous Crops

Elena Volynchikova,Ki Deok Kim 한국균학회 2022 Mycobiology Vol.50 No.5

Oomycete pathogens that belong to the genus Phytophthora cause devastating diseases in solanaceous crops such as pepper, potato, and tobacco, resulting in crop production losses worldwide. Although the application of fungicides efficiently controls these diseases, it has been shown to trigger negative side effects such as environmental pollution, phytotoxicity, and fungicide resistance in plant pathogens. Therefore, biological control of Phytophthora- induced diseases was proposed as an environmentally sound alternative to conventional chemical control. In this review, progress on biological control of the soilborne oomycete plant pathogens, Phytophthora capsici, Phytophthora infestans, and Phytophthora nicotianae, infecting pepper, potato, and tobacco is described. Bacterial (e.g., Acinetobacter, Bacillus, Chryseobacterium, Paenibacillus, Pseudomonas, and Streptomyces) and fungal (e.g., Trichoderma and arbuscular mycorrhizal fungi) agents, and yeasts (e.g., Aureobasidium, Curvibasidium,andMetschnikowia) have been reported as successful biocontrol agents of Phytophthora pathogens. These microorganisms antagonize Phytophthora spp. via antimicro- bial compounds with inhibitory activities against mycelial growth, sporulation, and zoospore germination. They also trigger plant immunity-inducing systemic resistance via several path- ways, resulting in enhanced defense responses in their hosts. Along with plant protection, some of the microorganisms promote plant growth, thereby enhancing their beneficial rela- tions with host plants. Although the beneficial effects of the biocontrol microorganisms are acceptable, single applications of antagonistic microorganisms tend to lack consistent effi- cacy compared with chemical analogues. Therefore, strategies to improve the biocontrol per- formance of these prominent antagonists are also discussed in this review.

Genetic DNA Marker for A2 mating type in Phytophthora infestans

Kim, Kwon-Jong,Lee, Youn-Su The Microbiological Society of Korea 2002 The journal of microbiology Vol.40 No.4

The Phytophthora infestans requires two mating types for sexual reproduction. Amplified fragment length polymorphism (AFLP) was used to specifically detect different mating types of P. infestans. The AFLP primers E+AA (5'-GACTGCGTACCAATTCAA-3') and M+CAA (5'-GATGAGTCCTGAG-TAAC AA-3') detected a fragment that is specific in the A2 mating type of P. infestans. This fragment was cloned and sequenced. Based on the sequence data, PHYB-1 and PHYB-2 primer were designed to detect the A2 mating type of P. infestans. A single 347 bp segment was observed in the A2 mating type of P. infestans, but not in the A1 mating type of P. infestans or other Phytophthora spp. Identification of mating type was performed with phenotype (sexual reproduction) and genotype (CAPs marker) methods. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using mating type-specific primers, a unique band was obtained within annealing temperatures of 57$^{\circ}C$-62$^{\circ}C$ and DNA levels of 10pg-100 ng (data not shown).

단일적응 감자 유전자원들의 역병저항성 및 주요 농업형질 평가

박영은(Young Eun Park),조현묵(Hyun Mook Cho),조지홍(Ji-Hong Cho),조광수(Kwang Soo Cho),김현준(Hyun Jun Kim),후안 란데오(Juan Landeo) 한국원예학회 2011 원예과학기술지 Vol.29 No.5

Phytophthora infestans에 의해 유발되는 감자역병은 전세계의 모든 감자 재배지대에서 발생하는 가장 치명적인 병 중의 하나로서 감자역병에 대한 저항성 품종 육성은 매우 중요한 육종 목표로 인식되고 있다. B3C1집단은 국제감자연구소로부터 도입되었으며 감자역병에 대하여 수평저항성을 가지고 있다. 그러나 이 집단은 페루의 저온단일 환경에서 선발된 계통들로서 고온장일의 국내 환경에 대한 적응성 검정은 이루어진 바 없었다. 따라서 본 연구는 국내의 고온 장일 환경 조건에서 B3C1 계통의 역병저항성과 주요 농업 관련 형질들을 평가하여 국내 재배 가능성을 검토해 보고, 앞으로 역병저항성 품종 육성을 위한 육종 소재로써의 가치를 평가하고자 수행되었다. 무방제 포장에서 역병저항성을 평가한 결과 모든 B3C1 계통들은 매우 높은 역병저항성을 나타내어 역병저항성 자원으로 가치가 높은 것으로 확인되었다. 그러나 모든 B3C1 계통에서 공통적으로 나타나는 극도의 만숙성 괴경 탈립성 불량 괴경 비대 불량 그리고 지상부 및 땅속 줄기의 과번무 현상 등은 반드시 개량되어야 할 것으로 판단되었다. B3C1 계통들은 강릉 지역 보다는 대관령 지역에서 더 높은 적응성을 나타내었으며 최종적으로 20 계통 중 LB-8(CIP393077.159)과 LB-11(CIP393371.159) 2 계통을 선발하였다. 역병에 강하고 국내의 고온장일 환경에 적응성이 높은 계통을 육성하기 위하여 B3C1 계통들은 국내 육성 계통들과 교배를 실시하였으며 지속적인 계통 선발이 진행되고 있다. Potato late blight caused by the fungus Phytophthora infestans is one of the most vital diseases damaging the potato plant. It is for this reason that breeding potato cultivars resistant to late blight is now becoming a major concern around the world. The B3C1 clones has been introduced by the Highland Agriculture Research Center RDA. The clones which came from International Potato Center in 2005 have a durable resistance to late blight. The clones were bred under a short-day condition in Peru. However there was still no report on the adaptability of these clones to the long-day condition in Korea. Therefore this study was conducted to evaluate the late blight resistance and major agronomic characteristics of B3C1 clones under Korea's long-day condition. This study was also done to generate genetic resources for developing new varieties resistant to late blight. In this study it was found out that in naturally infested field with P. infestans AUDPC (area under disease progress curve) values of all B3C1 clones were significantly lower than those of the control varieties,'Superior','Atlantic' and'Haryeong'. It was found out that B3C1 clones had a high level of resistance to late blight and that they could be used as genetic resources to breed potato varieties with late blight resistance. However several undesirable characteristics such as extremely late maturity excessive growth of stems and stolons and production of tubers that cannot easily be removed from the stolons were also observed. Among the twenty B3C1 clones two clones LB-8 (CIP393077.159) and LB-11 (CIP393371.159) were selected for cultivating at the highland area of Korea. Two B3C1 clones were crossed with Korean breeding lines and clonal selection for the progenies is still in progress.

Rapid and Accurate Species-Specific Detection of Phytophthora infestans Through Analysis of ITS Regions in Its rDNA

Kim, KYoung Su,Lee, Youn Su 한국미생물 · 생명공학회 2000 Journal of microbiology and biotechnology Vol.10 No.5

Polymerase chain reaction (PCR) was used to specifically detect Phytophthora infestans by analyzing the sequences of the ribosomal internal transcribed spacer regions (ITS) in the rDNA of the Phytophthora species. Based on the sequence data, PISP-1 together with the ITS3 primer were used to detect P. infestans. A single ca. 450bp segment was observed in P infestans, but not in the other fungal or bacterial isolates. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using these species-specific primers, a unique band was obtained within annealing temperatures of 55℃-61℃ and at template DNA levels of 10pg-100ng.

감자역병균 Phytophthora infestans의 특이적 SCAR Marker 개발

이상표 ( S. P. Lee ),김권종 ( K. J. Kim ),라숭도 ( S. D. Ra ),엄숭희 ( S. H. Eum ),이윤수 ( Y. S. Lee ) 강원대학교 농업생명과학연구원 2002 강원 농업생명환경연구 Vol.13 No.-

<i>Nicotiana benthamiana Matrix Metalloprotease</i> 1 (<i>NMMP</i>1) gene confers disease resistance to <i>Phytophthora infestans</i> in tobacco and potato plants

Ha, Jang Ho,Jang, Hyun A.,Moon, Ki-Beom,Baek, Kwang Hyun,Choi, Gyung Ja,Choi, Doil,Cho, Hye Sun,Kwon, Suk Yun,Jeon, Jae-Heung,Oh, Sang-Keun,Kim, Hyun-Soon G. Fischer 2017 Journal of plant physiology Vol. No.

<P><B>Abstract</B></P> <P>We previously isolated <I>Nicotiana benthamiana</I> matrix metalloprotease 1 (<I>NMMP1</I>) from tobacco leaves. The <I>NMMP1</I> gene encodes a highly conserved, Zn-containing catalytic protease domain that functions as a factor in the plant’s defense against bacterial pathogens. Expression of <I>NMMP1</I> was strongly induced during interactions between tobacco and one of its pathogens, <I>Phytophthora infestans</I>. To elucidate the role of the <I>NMMP1</I> in defense of <I>N. benthamiana</I> against fungal pathogens, we performed gain-of-function and loss-of-function studies. <I>NMMP1</I>-overexpressing plants had stronger resistance responses against <I>P. infestans</I> infections than control plants, while silencing of <I>NMMP1</I> resulted in greater susceptibility of the plants to the pathogen. This greater susceptibility correlated with fewer <I>NMMP1</I> transcripts than the non-silenced control. We also examined cell death as a measure of disease. The amount of cell death induced by the necrosis-inducing <I>P. infestans</I> protein 1, PiNPP1, was dependent on <I>NMMP1</I> in <I>N. benthamiana</I>. Potato plants overexpressing <I>NMMP1</I> also had enhanced disease resistance against <I>P. infestans.</I> RT-PCR analysis of these transgenic potato plants revealed constitutive up-regulation of the potato defense gene <I>NbPR5. NMMP1</I>-overexpressing potato plants were taller and produced heavier tubers than control plants. We suggest a role for <I>NMMP1</I>in pathogen defense and development.</P>

강원지역 감자 역병균 Phytophthora infestans의 교배형 및 약제저항성 변화

박경훈,윤종철,김병섭,차병진,류경열,정규식,김점순,권민 한국식물병리학회 2010 식물병연구 Vol.16 No.3

Potato late blight caused by Phytophthora infestans was the most constrain disease at potato cultivation areas. The mating type distribution and fungicides response of P. infestans were investigated to elucidate the changes of pathogen from Gangwon province. On the fungal isolates in 2006, 58.7% were A1 mating type and 41.3%were A2 mating type. In 2007, A1 mating type isolates increased to 93.3% and A2 mating type isolates were collected from Jinbu areas as much as 6.7%. About 234 isolates analysed for metalaxyl response, the results was resistance 73.7%, intermediate 18.8% and sensitive 7.5% in 2006. And it was resistance 59.4%,intermediate 4.0% and sensitive 36.6% in 2007. It meant that mating type distribution and fungicide response were very different over the collection sites. Minimal inhibition concentration (MIC) of dimethomorph examined with 126 isolates in 2006 and 106 isolates in 2007. MIC over 1.0 μg/ml was 56.3% in 2006 and it was 3.8% in 2007. The average EC50 value of P. infestans was 0.37 μg/ml in 2006, but it decreased to 0.12 μg/ml in 2007. Fungicides response and pathogenesis of P. infestans should be monitored continuously to enhance the chemical efficacy at potato fields. 강원지역의 감자포장에서 2006년과 2007년에 역병균을 분리하여, 교배형, metalaxyl과 dimethomorph 반응을 조사하였다. 각 지역에서 분리한 역병균의 교배형을 조사한 결과, 2006년 분리된 조사 균주는 155 균주 중 A1형이 58.7%, A2형이 41.3%로 확인 되었으며, 2007년 분리된 균주에서 A2 교배형은 6.7%로 발생 비율이 매우 낮았다. metalaxyl에 대한 반응성은 2006년 133균주에서 저항성 균주가 73.75%, 중도 저항성 균주가 18.8%, 감수성 균주가 7.5%로 나타났으며, 2007년 조사한 101 균주 중에서 저항성이 56.4%, 중도저항성 균이 4%, 감수성 균이 36.6%로 분리되었다. 강원도의 대부분 지역에서 A2 교배형 발생과 metalaxyl에 저항성을 갖는 균주를 확인할 수 있었는데, 이러한 결과는 병원균이 지속적으로 기주 및 환경요인에 따라 변화되고 있음을 의미한다. Dimethomorph약제 반응 조사결과 MIC 값은 0.6미만, 06~1.0, 1.0이상인 세 개의 그룹으로 분류 되었으며, 2006년에는 1.0 μg/ml이상인 균주는 56.3%이었으나 2007년에는 1.0 μg/ml이상인 균주가 4%였다. EC50 값은 2007년에 조사한 균주들의 평균값이 0.12 μg/ml로 2006년 평균값 0.37 μg/ml 보다 감소하였다. 감자 역병균들의 MIC 값과 EC50 값을 고려한 결과, 2007년에 조사한 병원균의 약제 반응정도가 2006년에 분리한 균주보다 감소하는 것으로 확인되었다.

Anti-oomycete Activity of Furanocoumarins from Seeds of Psoralea corylifolia against Phytophthora infestans

Shim, Sang-Hee,Kim, Jin-Cheol,Jang, Kyoung-Soo,Choi, Gyung-Ja The Korean Society of Plant Pathology 2009 Plant Pathology Journal Vol.25 No.1

In the course of a searching natural antifungal compounds from plant seeds, we found that the methanol extract of Psoralea corylifolia seeds showed potent control efficacy against tomato late blight caused by Phytophthora infestans and wheat leaf rust Puccinia recondita. Under bioassay-guided purification, we isolated two furanocoumarins, psoralen and isopsoralen, with anti-oomycete activity against P. infestans. By 1-day protective application, both compounds strongly reduced the disease development of P. infestans on tomato seedlings, but hardly controlled development of leaf rust on wheat seedlings. This is the first report on the anti-oomycete activity of P. corylifolia as well as that of psoralen and isopsoralen.

Multiple recognition of RXLR effectors is associated with nonhost resistance of pepper against <i>Phytophthora infestans</i>

Lee, Hyun-Ah,Kim, Shin-Young,Oh, Sang-Keun,Yeom, Seon-In,Kim, Saet-Byul,Kim, Myung-Shin,Kamoun, Sophien,Choi, Doil Blackwell Publishing Ltd 2014 The New phytologist Vol.203 No.3

<P><P>Nonhost resistance (NHR) is a plant immune response to resist most pathogens. The molecular basis of NHR is poorly understood, but recognition of pathogen effectors by immune receptors, a response known as effector-triggered immunity, has been proposed as a component of NHR.</P><P>We performed transient expression of 54 <I>Phytophthora infestans</I>RXLR effectors in pepper (<I>Capsicum annuum</I>) accessions. We used optimized heterologous expression methods and analyzed the inheritance of effector-induced cell death in an F<SUB>2</SUB> population derived from a cross between two pepper accessions.</P><P>Pepper showed a localized cell death response upon inoculation with <I>P. infestans</I>, suggesting that recognition of effectors may contribute to NHR in this system. Pepper accessions recognized as many as 36 effectors. Among the effectors, PexRD8 and Avrblb2 induced cell death in a broad range of pepper accessions. Segregation of effector-induced cell death in an F<SUB>2</SUB> population derived from a cross between two pepper accessions fit 15 : 1, 9 : 7 or 3 : 1 ratios, depending on the effector.</P><P>Our genetic data suggest that a single or two independent/complementary dominant genes are involved in the recognition of RXLR effectors. Multiple loci recognizing a series of effectors may underpin NHR of pepper to <I>P. infestans</I> and confer resistance durability.</P></P>

Bikaverin and fusaric acid from <i>Fusarium oxysporum</i> show antioomycete activity against <i>Phytophthora infestans</i>

Son, S.W.,Kim, H.Y.,Choi, G.J.,Lim, H.K.,Jang, K.S.,Lee, S.O.,Lee, S.,Sung, N.D.,Kim, J.-C. Blackwell Publishing Ltd 2008 Journal of applied microbiology Vol.104 No.3

<P>Abstract</P><P>Aims: </P><P>To isolate and identify antioomycete substances from <I>Fusarium oxysporum</I> EF119 against <I>Phytophthora infestans</I> and to investigate their antimicrobial activities against various plant pathogenic bacteria, oomycetes and true fungi.</P><P>Methods and Results: </P><P>Two antioomycete substances were isolated from liquid cultures of <I>F. oxysporum</I> EF119, which shows a potent disease control efficacy against tomato late blight caused by <I>P. infestans</I>. They were identified as bikaverin and fusaric acid by mass and nuclear magnetic resonance spectral analyses. They inhibited the mycelial growth of plant pathogenic oomycetes and fungi. Fusaric acid also effectively suppressed the cell growth of various plant pathogenic bacteria, but bikaverin was virtually inactive. Treatment with bikaverin at 300 <I>&mgr;</I>g ml<SUP>−1</SUP> suppressed the development of tomato late blight by 71%. Fusaric acid provided effective control against tomato late blight and wheat leaf rust over 67% at concentrations more than 100 <I>&mgr;</I>g ml<SUP>−1</SUP>.</P><P>Conclusions: </P><P>Both bikaverin and fusaric acid showed <I>in vitro</I> and <I>in vivo</I> antioomycete activity against <I>P. infestans</I>.</P><P>Significance and Impact of the Study: </P><P><I>Fusarium oxysporum</I> EF119 producing both bikaverin and fusaric acid may be used as a biocontrol agent against tomato late blight caused by <I>P. infestans</I>.</P>