An improved PCR method for rapid and accurate identification of mating types in the late blight pathogen Phytophthora infestans

2021 ◽  
Author(s):  
Romain Mabon ◽  
Michèle Guibert ◽  
Roselyne Corbiere ◽  
Didier Andrivon
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Multiplex Pcr ◽  
Mating Type ◽  
Mating Types ◽  
Accurate Identification ◽  
Fta Cards ◽  
Multiplex Pcr Assay ◽  
Freeze Dried ◽  
Late Blight Pathogen

Mating type is a critical trait in heterothallic organisms. In plant pathogenic oomycetes, like the late blight pathogen Phytophthora infestans, it is usually identified through pairing between tester and candidate isolates, a method which is both laborious and applicable to live isolates only. Therefore, developing simple and fast PCR tests to reliably identify P. infestans mating types is of great interest for population genetic studies. A multiplex PCR assay combining the amplification of a locus diagnostic for P. infestans and of one diagnostic for the A1 mating type was developed and validated on a collection of 1441 samples, covering the current and past diversity of European P. infestans populations. These samples obtained from either freeze-dried mycelium or from FTA cards on which diseased leaflets had been pressed. The multiplex assay correctly identified mating types in 97.4 % of these samples. The main source of incorrect assignment was the lack of amplification of the A1 diagnostic allele, due to insufficient DNA quality and/or quantity in the reaction mix. This multiplex PCR, applicable to both live and stored material, thus constitutes a useful addition to the set of molecular tools available for population typing in P. infestans.

scholarly journals Complete mitogenome mapping of potato late blight pathogen, Phytophthora infestans A2 mating type

2017 ◽  
Vol 2 (1) ◽  
pp. 90-91
Author(s):  
Virupaksh U. Patil ◽  
G. Vanishree ◽  
Debasis Pattanayak ◽  
Sanjeev Sharma ◽  
Vinay Bhardwaj ◽  
...  
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Mating Type ◽  
Potato Late Blight ◽  
Complete Mitogenome ◽  
Late Blight Pathogen

scholarly journals Genetic Structure of the Phytophthora infestans Population in Morocco

Plant Disease ◽  
2000 ◽  
Vol 84 (2) ◽  
pp. 173-176 ◽  
Author(s):  
M. Sedegui ◽  
R. B. Carroll ◽  
A. L. Morehart ◽  
T. A. Evans ◽  
S. H. Kim ◽  
...  
Keyword(s):  
Genetic Structure ◽  
Late Blight ◽  
Sexual Reproduction ◽  
Phytophthora Infestans ◽  
Mating Type ◽  
Potato Seed ◽  
Mating Types ◽  
Potato Field ◽  
New Genotype

ABSRACT In 1996 to 1998, a late-blight survey was conducted in potato- and tomato-growing regions of Morocco. A total of 149 isolates of Phytophthora infestans were collected and analyzed for the glucose-6-phosphate isomerase (Gpi) and peptidase (Pep) alleles, mating types, and metalaxyl sensitivities. Four genotypes were identified: MO-1 (mating type A1, Gpi 100/100, Pep 92/100), MO-2 (mating type A1, Gpi 86/100, Pep 92/100), MO-3 (mating type A2 Gpi 100/100, Pep 100/100), and MO-4 (mating type A1, Gpi 100/100, Pep 100/100). The potato isolates were MO-1 (1996 & 97), MO-3 (1998), and MO-4 (1998). The frequencies of A1 (MO-4) and A2 (MO-3) mating types in potato fields in 1998 were 26 and 74%, respectively. Potato isolates were pathogenic to both potatoes and tomatoes. The isolates collected from tomatoes in 1997 and 1998 were MO-2. Potato and tomato isolates were insensitive and sensitive to metalaxyl, respectively. The change of genotype population in 1998 was probably caused by migration of a new genotype from Europe associated with importation of potato seed. The detection of A1 and A2 mating types in the same potato field indicates the potential for sexual reproduction of P. infestans in Morocco.

Incidence of the potato late blight pathogen, Phytophthora infestans, in Syria and its mating type

2017 ◽  
Vol 124 (6) ◽  
pp. 533-537 ◽  
Author(s):  
Walid Naffaa ◽  
Soulima Ibrahim ◽  
Taissir Abou Alfadil ◽  
Antonious Al-Daoude
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Mating Type ◽  
Potato Late Blight ◽  
Late Blight Pathogen

scholarly journals First Report of A1 and A2 Mating Types of Phytophthora infestans on Potato and Tomato in Nepal

Plant Disease ◽  
1998 ◽  
Vol 82 (9) ◽  
pp. 1064-1064 ◽  
Author(s):  
S. K. Shrestha ◽  
K. Shrestha ◽  
K. Kobayashi ◽  
N. Kondo ◽  
R. Nishimura ◽  
...  
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Mating Type ◽  
Simultaneous Occurrence ◽  
Mating Types ◽  
Isozyme Loci ◽  
Oospore Formation ◽  
The Hill ◽  
Pathogenic Diversity ◽  
Important Disease

Late blight caused by Phytophthora infestans (Mont.) de Bary is an important disease of potato and tomato that occurs annually in the hills and occasionally in the terai (plain) of Nepal. In 1996 and 1997, each year, 50 samples of late blight-infected potato and tomato leaves were collected from the hill and terai areas. The pathogen was cultured on Rye A agar. Each isolate was paired on clear V8 agar with reference isolates DN111 (A1 mating type) and DN107 (A2 mating type) received from Hokkaido University, Japan, and examined for oospore formation after 10 to 15 days of incubation at 20°C. The proportion of A2 isolates was 6% in 1996 and 42% in 1997. The A2 isolates were mainly from the high hills (2,000 to 2,500 m) where local and Andean types of potatoes are grown. Analysis of genotypes of isolates at the glucosephosphate isomerase (GPI-1), malic enzyme (ME), and peptidase (PEP-1) (1,2) isozyme loci revealed genetic diversity between A1 and A2 isolates. A1 isolates from potato were either homozygous (100/100) or heterozygous (86/100) for GPI-1, whereas all A1 isolates from tomato were heterozygous (86/100). All A1 isolates were homozygous (100/100) at the ME locus and heterozygous (92/100) at the PEP-1 locus. A2 isolates were homozygous (100/100) at all isozyme loci. The results show that both A1 and A2 mating types of P. infestans are present in Nepal, and that they display different isozyme genotypes. It is speculated that the A1 type may have migrated with potatoes from Europe while the A2 type may have been introduced with Andean potatoes from Latin America more recently. The simultaneous occurrence of both mating types may allow the fungus to increase its pathogenic diversity and to survive by means of oospores. References: (1) A. A. Mosa et al. Plant Pathol. 42:26, 1993. (2) P. W. Tooley et al. J. Hered. 76:431, 1985.

scholarly journals Phenotypic Characterization of Recent Clonal Lineages of Phytophthora infestans in the United States

Plant Disease ◽  
2013 ◽  
Vol 97 (7) ◽  
pp. 873-881 ◽  
Author(s):  
G. Danies ◽  
I. M. Small ◽  
K. Myers ◽  
R. Childers ◽  
W. E. Fry
Keyword(s):  
United States ◽  
Late Blight ◽  
Phytophthora Infestans ◽  
The United States ◽  
Phenotypic Characterization ◽  
Phenotypic Traits ◽  
Mating Types ◽  
Late Blight Disease ◽  
Blight Disease

Phytophthora infestans, the causal agent of late blight disease, has been reported in the United States and Canada since the mid-nineteenth century. Due to the lack of or very limited sexual reproduction, the populations of P. infestans in the United States are primarily reproducing asexually and, thus, show a simple genetic structure. The emergence of new clonal lineages of P. infestans (US-22, US-23, and US-24) responsible for the late blight epidemics in the northeastern region of the United States in the summers of 2009 and 2010 stimulated an investigation into phenotypic traits associated with these genotypes. Mating type, differences in sensitivity to mefenoxam, differences in pathogenicity on potato and tomato, and differences in rate of germination were studied for clonal lineages US-8, US-22, US-23, and US-24. Both A1 and A2 mating types were detected. Lineages US-22, US-23, and US-24 were generally sensitive to mefenoxam while US-8 was resistant. US-8 and US-24 were primarily pathogenic on potato while US-22 and US-23 were pathogenic on both potato and tomato. Indirect germination was favored at lower temperatures (5 and 10°C) whereas direct germination, though uncommon, was favored at higher temperatures (20 and 25°C). Sporangia of US-24 released zoospores more rapidly than did sporangia of US-22 and US-23. The association of characteristic phenotypic traits with genotype enables the prediction of phenotypic traits from rapid genotypic analyses for improved disease management.

scholarly journals EIN2-mediated signaling is involved in pre-invasion defense in Nicotiana benthamiana against potato late blight pathogen, Phytophthora infestans

2017 ◽  
Vol 12 (4) ◽  
pp. e1300733 ◽  
Author(s):  
Soriya Rin ◽  
Yuri Mizuno ◽  
Yusuke Shibata ◽  
Mayuka Fushimi ◽  
Shinpei Katou ◽  
...  
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Nicotiana Benthamiana ◽  
Potato Late Blight ◽  
Late Blight Pathogen

scholarly journals Phytophthora infestans Mating Types on Tomato (Solanum lycopersicum) in Southern Spain

Plant Disease ◽  
2007 ◽  
Vol 91 (1) ◽  
pp. 109-109 ◽  
Author(s):  
J. M. Segura ◽  
M. de Cara ◽  
M. Santos ◽  
J. Tello
Keyword(s):  
Phytophthora Infestans ◽  
Mating Type ◽  
Agar Medium ◽  
The Other ◽  
Southern Spain ◽  
White Mold ◽  
Mating Types ◽  
Tomato Plants ◽  
Cornell University ◽  
Tomato Seeds

During 2004, an unusual spread of Phytophthora infestans on tomato plants in greenhouses located in Almería and Granada provinces, southern Spain, was observed. Infected plants had water-soaked, brown spots on leaves and stems and necrotic areas with white mold on the surface of fruits. Three isolates were obtained by plating diseased tissue on V8 juice agar medium and maintained on rye agar at 18°C. These isolates were analyzed for the mating type. Crosses were carried out using V8 juice agar and rye agar. The two parental isolates US1 (A1) and US8 (A2) were both provided by W. E. Fry, Cornell University, Ithaca, NY. Two of the Spanish isolates were homothallic and the other isolate belonged to the uncommon mating type A1A2. To confirm the occurrence of the two mating types, 43 single-sporangium progeny were produced and analyzed from the A1A2 mating type. Thirty eight isolates were A1, two were A2, one was A1A2 mating type, and two were sterile. Assessment of five single-sporangium progeny from the homothallic type resulted in two A1, two homothallic, and one sterile isolate. A1A2 isolates produced oospores when crossed with either A1 or A2, but not when self-crossed. Previously, the A1A2 mating type has been found in Israel in the field and was obtained from oospores produced on tomato seeds (2,3). Since 2003, mating types of P. infestans isolates recovered from potato (60) and tomato (8) in southern Spain have been characterized. Seventy-five percent of the isolates recovered from potato were A1 and 25% were A2 mating types. Isolates recovered from tomato were 50% A1 and 50% A2 (1). To our knowledge, this is the first report of the occurrence of the A1A2 mating type and homothallic P. infestans isolates on tomato in Spain. References: (1) E. Andujar et al. Congr. Sociedad Española de Fitopatol. 12:244, 2004. (2) E. Rubin and Y. Cohen. Phytoparasitica 32:237, 2004. (3) E. Rubin and Y. Cohen. Plant Dis. 90:741, 2006.

Genetic mapping and non-Mendelian segregation of mating type loci in the oomycete, Phytophthora infestans.

Genetics ◽  
1995 ◽  
Vol 141 (2) ◽  
pp. 503-512 ◽  
Author(s):  
H S Judelson ◽  
L J Spielman ◽  
R C Shattock
Keyword(s):  
Phytophthora Infestans ◽  
Mating Type ◽  
Dna Markers ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Mating Types ◽  
Mendelian Segregation ◽  
Range Restriction ◽  
Type Locus ◽  
Mating Type Locus

Abstract DNA markers linked to the determinants of mating type in the oomycete, Phytophthora infestans, were identified and used to address the genetic basis of heterothallism in the normally diploid fungus. Thirteen loci linked to the A1 and A2 mating types were initially identified by bulked segregant analysis using random amplified polymorphic DNA markers (RAPDs) and subsequently scored in three crosses polymorphisms (SSCP), cleaved amplified polymorphisms (CAPS), or allele-specific polymerase chain reaction markers (AS-PCR). All DNA markers mapped to a single region, consistent with a single locus determining both mating types. Long-range restriction mapping also demonstrated the linkage of the markers to one region and delimited the mating type locus to a 100-kb region. The interval containing the mating type locus displayed non-Mendelian segregation as only two of the four expected genotypes were detected in progeny. This is consistent with a system of balance lethal loci near the mating type locus. A model for mating type determination is presented in which the balanced lethals exclude form progeny those with potentially conflicting combinations of mating type alleles, such as those simultaneously expressing A1 and A2 functions.

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