Recombinase Polymerase Amplification (RPA) for the Rapid Isothermal Detection of Spongospora subterranea f. sp. subterranea and Potato Mop-Top Virus

2019 ◽  
Vol 96 (6) ◽  
pp. 617-624
Author(s):  
Joseph B. DeShields ◽  
Natalia Moroz ◽  
Lauren E. Braley ◽  
Guadalupe Arlene Mora-Romero ◽  
Kiwamu Tanaka
Keyword(s):  
Recombinase Polymerase Amplification ◽  
Spongospora Subterranea ◽  
Potato Mop Top Virus

scholarly journals Perunan maltokaariviruksen leviäminen perunakasvin eri osiin

2008 ◽  
pp. 1-6
Author(s):  
Johanna Aura ◽  
Jari P.T. Valkonen
Keyword(s):  
Spongospora Subterranea ◽  
Potato Mop Top Virus

Perunan maltokaarivirus (Potato mop-top virus, PMTV) on Pohjoismaissa laajalle levinnyt ja vaikeastitorjuttava maalevintäinen taudinaiheuttaja. Sen levittäjänä pellolla toimii sienenkaltainen mikrobi,Spongospora subterranea, joka aiheuttaa mukuloiden pintaan kuorirokkoa. PMTV aiheuttaamukuloihin kuoliota, maltokaarioireita, ja tuottaa siten laatutappioita.PMTV tartuttaa mukuloita ja perunan juuristoa, mutta se havaitaan harvoin maanpäällisissäosissa kuten lehdissä. Siten PMTV-tartunnan havaitseminen perunakasvustossa kasvukauden aikanatai testaaminen idätettyjen mukuloiden versoista ei ole mahdollista samaan tapaan kuin muidenperunanvirusten osalta. Havaintojemme mukaan PMTV kuitenkin leviää mukulasta ituihin varastooloissa(5ºC, pimeä). Tämä viittaa siihen, että virus kykenisi sinänsä leviämään kehittyviin versoihin,mutta leviäminen estyy vielä tuntemattoman mekanismin vaikutuksesta. Tutkimuksemme tavoitteenaoli selvittää tätä mekanismia.Kasvatushuonekokeissa PMTV:n havaittiin leviävän infektoituneesta siemenmukulasta kaikkiinperunan maanalaisiin osiin: juuriin, maavarsiin sekä niihin muodostuviin mukuloihin. Lehdissä virustaoli vain hyvin vähäisiä määriä, jotka olivat niin pieniä, että ne voitiin havaita vain testaamallavirusgenomin (RNA) läsnäoloa kvantitatiivisen PCR-menetelmän avulla, mutta ei virustestaukseennormaalisti käytettävän, vasta-aineisiin perustuvan ELISA-menetelmän avulla. Itävien mukuloidenaltistaminen valolle ja korkeammalle lämpötilalle johti virusmäärien vähentymiseen iduissa. Näidentulosten pohjalta tarkastellaan lähemmin virus-RNA:han kohdentuvaa hajotusta, ns. RNA-hiljennystä,mahdollisena mekanismina, joka estää PMTV:n kerääntymistä perunan yhteyttäviin osiin.

Development of High Resolution DNA Melting Analysis for Simultaneous Detection of Potato Mop-Top Virus and Its Vector, Spongospora subterranea, in Soil

Plant Disease ◽  
2020 ◽  
Author(s):  
Xianzhou Nie ◽  
Mathuresh Singh ◽  
Dahu Chen ◽  
Cassandra Gilchrist ◽  
Yasmine Soqrat ◽  
...  
Keyword(s):  
High Resolution ◽  
Simultaneous Detection ◽  
Soil Samples ◽  
Dna Melting ◽  
Spongospora Subterranea ◽  
Rt Pcr ◽  
Efficient Detection ◽  
Melting Analysis ◽  
Potato Mop Top Virus ◽  
Negative Detection

In this study, a set of duplex reverse transcription (RT)-PCR-mediated high resolution DNA melting (HRM) analyses for simultaneous detection of potato mop-virus (PMTV) and its protist vector, Spongospora subterranea f.sp. subterranea (Sss), was developed. The infestation of soil by PMTV was detected by using a tobacco-based baiting system. Total RNA extracted from the soil led to successful RT-PCR gel-electrophoresis detection of both PMTV and Sss. To facilitate more efficient detection, newly designed primer pairs for PMTV RNA species (i.e., RNA-Rep, -CP, and -TGB) were analyzed together with the existing Sss primers using real-time RT-PCR. The resulting amplicons exhibited melting profiles that could be readily differentiated. Under duplex RT-PCR format, all PMTV and Sss primer combinations led to successful detection of respective PMTV RNA species and Sss in the samples by high resolution DNA melting (HRM) analyses. When the duplex HRM assay was applied to soil samples collected from six fields at four different sites in New Brunswick, Canada, positive detection of PMTV and/or Sss was found in 63-100% samples collected from fields in which PMTV-infected tubers had been observed. In contrast, the samples from fields where neither PMTV- nor Sss-infected tubers had been observed resulted in negative detection by the assay. Bait tobacco bioassay for PMTV and Sss produced similar results. Between 63%-83% and 100% of the soil samples collected from PMTV-infested fields led to PMTV and Sss infections in the bait tobacco plants, respectively; whereas no PMTV or Sss infected plants were obtained from soil samples collected from PMTV/Sss-free fields.

scholarly journals Initial infection of roots and leaves reveals different resistance phenotypes associated with coat protein gene-mediated resistance to Potato mop-top virus

2002 ◽  
Vol 83 (5) ◽  
pp. 1201-1209 ◽  
Author(s):  
Anna Germundsson ◽  
Maria Sandgren ◽  
Hugh Barker ◽  
Eugene I. Savenkov ◽  
Jari P. T. Valkonen
Keyword(s):  
Coat Protein ◽  
Nicotiana Benthamiana ◽  
Protein Gene ◽  
Resistance Mechanism ◽  
Spongospora Subterranea ◽  
Mechanical Inoculation ◽  
Systemic Movement ◽  
Cp Gene ◽  
Transgenic Potatoes ◽  
Potato Mop Top Virus

Resistance to the pomovirus Potato mop-top virus (PMTV) was studied in potato (Solanum tuberosum cv. Saturna) and Nicotiana benthamiana transformed with the coat protein (CP) gene of PMTV. The incidence of PMTV infections was reduced in tubers of the CP-transgenic potatoes grown in the field in soil infested with the viruliferous vector, Spongospora subterranea. However, in those tubers that were infected, all three virus RNAs were detected and virus titres were high. The CP-transgenic N. benthamiana plants were inoculated with PMTV using two methods. Following mechanical inoculation of leaves, no RNA 3 (the CP-encoding RNA homologous to the transgene) was detected in leaves, but in some plants low amounts of RNA 3 were detected in roots; RNA 2 was readily detected in leaves and roots of several plants. Inoculation of roots using viruliferous S. subterranea resulted in infection of roots in all plants and the three PMTV RNAs were detected. However, no systemic movement of PMTV from roots to the above-ground parts was observed, indicating a novel expression of resistance. These data indicate that the CP gene-mediated resistance to PMTV specifically restricts accumulation of PMTV RNA 3, and is more effective in leaves than roots. Furthermore, expression of resistance is different depending on whether leaves or roots are inoculated. Data do not exclude the possibility that both a protein-mediated and an RNA-mediated resistance mechanism are involved.

scholarly journals Evaluation of Effects of Chemical Soil Treatments and Potato Cultivars on Spongospora subterranea Soil Inoculum and Incidence of Powdery Scab and Potato Mop-Top Virus in Potato

Plant Disease ◽  
2020 ◽  
Vol 104 (11) ◽  
pp. 2807-2816
Author(s):  
Yuan Zeng ◽  
Ana Cristina Fulladolsa ◽  
Andrew M. Cordova ◽  
Patrick O’Neill ◽  
Stewart M. Gray ◽  
...  
Keyword(s):  
Field Experiments ◽  
Potato Cultivar ◽  
Potato Cultivars ◽  
Powdery Scab ◽  
Infested Soil ◽  
Spongospora Subterranea ◽  
Pathogen Population ◽  
Chemical Treatments ◽  
Soil Inoculum ◽  
Potato Mop Top Virus

Spongospora subterranea is a soilborne plasmodiophorid that causes powdery scab in potato. It also transmits potato mop-top virus (PMTV), which causes necrotic arcs (spraing) in potato tubers. Three field experiments were conducted in naturally S. subterranea-infested soil to investigate the effects of two chemicals, Omega 500F (fluazinam) and FOLI-R-PLUS RIDEZ (biological extract), on powdery scab, PMTV, and changes in S. subterranea inoculum with six different potato cultivars. The efficacy of soil treatment with these two chemicals on tuber lesions, root galling, and pathogen population was also assessed in greenhouse trials. The chemical treatments did not reduce powdery scab, root gall formation, or S. subterranea inoculum in the field or greenhouse trials. Postharvest S. subterranea soil inoculum in fields varied across farms and among potato cultivars but the pathogen population consistently increased by the end of the growing season. The evaluated russet cultivars were more tolerant to powdery scab than the yellow- or red-skinned cultivars but all were susceptible to PMTV. In the field, powdery scab indices and soil inoculum changes were positively correlated, while postharvest S. subterranea inoculum was positively correlated with root galling in both greenhouse trials. Powdery scab and PMTV occurred in noninoculated potting mix, indicating that peat-based potting mix is a source for both pathogens. These results demonstrate that chemical management methods currently used by farmers are ineffective, that S. subterranea and PMTV in potting mix can cause severe epidemics in greenhouses, and that potato cultivar choices impact inoculum increases in soil.

scholarly journals Draft Genome Resource for the Potato Powdery Scab Pathogen Spongospora subterranea

2018 ◽  
Vol 31 (12) ◽  
pp. 1227-1229 ◽  
Author(s):  
Stefan Ciaghi ◽  
Sigrid Neuhauser ◽  
Arne Schwelm
Keyword(s):  
Functional Genomics ◽  
Illumina Sequencing ◽  
Genome Assembly ◽  
Draft Genome ◽  
Potato Industry ◽  
Powdery Scab ◽  
Spongospora Subterranea ◽  
Potato Mop Top Virus ◽  
Single Tuber

The Plasmodiophorida (Phytomyxea, Rhizaria) are a group of protists that infect plants. Of this group, Spongospora subterranea causes major problems for the potato industry by causing powdery scab and root galling of potatoes and as vector for the Potato mop-top virus (PMTV) (genus Pomovirus, family Virgaviridae). A single tuber isolate (SSUBK13) of this uncultivable protist was used to generate DNA for Illumina sequencing. The data were assembled to a draft genome of 28.08 Mb consisting of 2,340 contigs and an L50 of 280. A total of 10,778 genes were predicted and 93% of the BUSCO genes were detected. The presented genome assembly is only the second genome of a plasmodiophorid. The data will accelerate functional genomics to study poorly understood interaction of plasmodiophorids and their hosts.

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