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result(s) for
"Phytophthora - isolation "
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A giant NLR gene confers broad-spectrum resistance to Phytophthora sojae in soybean
2021
Phytophthora
root and stem rot caused by
P. sojae
is a destructive soybean soil-borne disease found worldwide. Discovery of genes conferring broad-spectrum resistance to the pathogen is a need to prevent the outbreak of the disease. Here, we show that soybean
Rps11
is a 27.7-kb nucleotide-binding site-leucine-rich repeat (NBS-LRR or NLR) gene conferring broad-spectrum resistance to the pathogen.
Rps11
is located in a genomic region harboring a cluster of large NLR genes of a single origin in soybean, and is derived from rounds of unequal recombination. Such events result in promoter fusion and LRR expansion that may contribute to the broad resistance spectrum. The NLR gene cluster exhibits drastic structural diversification among phylogenetically representative varieties, including gene copy number variation ranging from five to 23 copies, and absence of allelic copies of
Rps11
in any of the non-
Rps11
-donor varieties examined, exemplifying innovative evolution of NLR genes and NLR gene clusters.
While multiple resistance-to-
Phytophthora sojae
loci/alleles have been mapped in soybean, many of them have become ineffective to newly evolved isolates. Here, the authors show that a 27.7-kb nucleotide-binding site-leucine-rich repeat gene confers broad-spectrum resistance to
P. sojae
in soybean.
Journal Article
A LAMP at the end of the tunnel: A rapid, field deployable assay for the kauri dieback pathogen, Phytophthora agathidicida
by
Nelson, Briana C. W.
,
McLenachan, Patricia A.
,
Winkworth, Richard C.
in
Analysis
,
Araucariaceae - genetics
,
Araucariaceae - microbiology
2020
Notes how the root rot causing oomycete, Phytophthora agathidicida, threatens the long-term survival of the iconic New Zealand kauri tree. Describes a loop-mediated isothermal amplification (LAMP) assay for the detection of P. agathidicida that targets a portion of the mitochondrial apocytochrome b coding sequence. Discusses the various ways such an assay can be used in the communities affected. Source: National Library of New Zealand Te Puna Matauranga o Aotearoa, licensed by the Department of Internal Affairs for re-use under the Creative Commons Attribution 3.0 New Zealand Licence.
Journal Article
Oomycete diversity and ecology in declining alder stands in Switzerland
by
Lygis, Vaidotas
,
Mizerienė, Goda
,
Prospero, Simone
in
Alnus - microbiology
,
Alnus - parasitology
,
Alnus glutinosa
2025
In this study, we assessed the occurrence and diversity of four oomycete genera ( Phytophthora , Phytopythium , Pythium , and Globisporangium ) in 13 declining alder ( Alnus glutinosa and A. incana ) stands in Switzerland. For this, we sampled and analyzed soil from tree rhizosphere, water from streams and rivers along which the stands were located, and symptomatic alder bark. The overall isolation rate was 47.2%, with a total of 400 oomycete isolates recovered at all 13 sites. The highest incidence of oomycete isolates was in soil samples (baiting, 82.5% isolation rate), followed by water (baiting, 14.7%), and bark (direct isolation, 2.7%). Of all recovered oomycete isolates, 90.3% could be successfully assigned to a known species, for a total of 23 species identified, including both preferential saprotrophs and pathogens. Among all genera, Phytophthora was the most abundant with 273 isolates (75.6%), followed by Phytopythium , Pythium , and Globisporangium . Oomycete species diversity showed a significant variation among substrates. Only one species— Phytophthora lacustris —was abundant in all substrates, while 16 species were restricted to a specific substrate, mainly soil. The rhizosphere of symptomatic alder trees harbored the most diverse oomycete community, highlighting once again the importance of soil as a reservoir for these microorganisms. Only two Phytophthora species were isolated from alder bark lesions, namely, P. × alni , the known causal agent of alder decline, and P. lacustris . The low recovery rate of P. × alni might be due to attempts to isolate it from old, inactive lesions, but may also suggest that alder decline might be caused by other oomycetes infecting the root system of the trees.
Journal Article
A qPCR assay for the detection of Phytophthora abietivora, an emerging pathogen on fir species cultivated as Christmas trees
by
Tanguay, Philippe
,
Bilodeau, Guillaume J.
,
Gauthier, Marie-Krystel
in
Abies - microbiology
,
Abies - parasitology
,
Assaying
2025
Emerging species of the Phytophthora genus are among the most important threats to global plant biodiversity, horticulture and trade. For instance, Phytophthora root rot (PRR) of Christmas trees, mainly Fraser ( Abies fraseri ) and balsam ( Abies balsamea ) firs, is responsible for an average of 10% of the observed diseased trees in plantations. Diagnosing PRR involves isolation followed by morphological and molecular identification of the causal agents. However, these methods are rarely adapted to larger scale monitoring such as in situ detection. For these applications, molecular detection of environmental DNA (eDNA) provides the fast and high-throughput results needed. Phytophthora abietivora was associated with PRR in firs ( Abies spp.) cultivated as Christmas trees in the province of Québec (Canada). This study focused on developing a sensitive and specific qPCR assay targeting P. abietivora and validating its efficiency on DNA extracted from soil and roots. A set of primers and probe was designed for this assay, and parameters such as the limit of detection (LOD 95% ) and limit of quantification (LOQ) were measured. The assay was tested on DNA obtained from healthy-looking and PRR symptomatic Fraser and balsam firs. The assay was shown to be semi-specific because it cross-reacted with P. europaea and three other phylogenetically close species, but deemed specific in the context of PRR of firs. The LOD 95% was estimated at 10 copies per reaction (C q of 35.7) and the LOQ to 33 P. abietivora oospores per gram of soil. Out of 488 DNA samples from soil, 68 were positive for P. abietivora from which 42 (61.7%) were from PRR symptomatic trees. Only a slight overlap (3 out of 7 samples) was observed with previously obtained baiting results . This assay will be useful for rapid diagnostics of P. abietivora infected trees and as a prospecting tool to better characterize the natural distribution and dissemination of the disease.
Journal Article
Microbiota Characterization of Compost Using Omics Approaches Opens New Perspectives for Phytophthora Root Rot Control
by
Ros, Margarita
,
Blaya, Josefa
,
Marhuenda, Frutos C.
in
Abundance
,
Agricultural wastes
,
Analysis
2016
Phytophthora root rot caused by Phytophthora nicotianae is an economically important disease in pepper crops. The use of suppressive composts is a low environmental impact method for its control. Although attempts have been made to reveal the relationship between microbiota and compost suppressiveness, little is known about the microorganisms associated with disease suppression. Here, an Ion Torrent platform was used to assess the microbial composition of composts made of different agro-industrial waste and with different levels of suppressiveness against P. nicotianae. Both bacterial and fungal populations responded differently depending on the chemical heterogeneity of materials used during the composting process. High proportions (67-75%) of vineyard pruning waste were used in the most suppressive composts, COM-A and COM-B. This material may have promoted the presence of higher relative abundance of Ascomycota as well as higher microbial activity, which have proved to be essential for controlling the disease. Although no unique fungi or bacteria have been detected in neither suppressive nor conducive composts, relatively high abundance of Fusarium and Zopfiella were found in compost COM-B and COM-A, respectively. To the best of our knowledge, this is the first work that studies compost metabolome. Surprisingly, composts and peat clustered together in principal component analysis of the metabolic data according to their levels of suppressiveness achieved. This study demonstrated the need for combining the information provided by different techniques, including metagenomics and metametabolomics, to better understand the ability of compost to control plant diseases.
Journal Article
eDNA from roots: a robust tool for determining Phytophthora communities in natural ecosystems
2018
Abstract
Proper isolation and identification of Phytophthora species is critical due to their broad distribution and huge impact on natural ecosystems throughout the world. In this study, five different sites were sampled and seven methods were compared to determine the Phytophthora community. Three traditional isolation methods were conducted (i) soil baiting, (ii) filtering of the bait water and (iii) isolation from field roots using Granny Smith apples. These were compared to four sources of eDNA used for metabarcoding using Phytophthora-specific primers on (i) sieved field soil, (ii) roots from field, (iii) filtered baiting water and (iv) roots from bait plants grown in the glasshouse in soil collected from these sites. Six Phytophthora species each were recovered by soil baiting using bait leaves and from the filtered bait water. No Phytophthora species were recovered from Granny Smith apples. eDNA extracted from field roots detected the highest number of Phytophthora species (25). These were followed by direct DNA isolation from filters (24), isolation from roots from bait plants grown in the glasshouse (19), and DNA extraction from field soil (13). Therefore, roots were determined to be the best substrate for detecting Phytophthora communities using eDNA.
This study provides a comparison of traditional isolation methods to metabarcoding techniques to determine Phytophthora species diversity.
Journal Article
Distinct regions of the Phytophthora essential effector Avh238 determine its function in cell death activation and plant immunity suppression
2017
Phytophthora pathogens secrete effectors to manipulate host innate immunity, thus facilitating infection. Among the RXLR effectors highly induced during Phytophthora sojae infection, Avh238 not only contributes to pathogen virulence but also triggers plant cell death. However, the detailed molecular basis of Avh238 functions remains largely unknown.
We mapped the regions responsible for Avh238 functions in pathogen virulence and plant cell death induction using a strategy that combines investigation of natural variation and large-scale mutagenesis assays. The correlation between cellular localization and Avh238 functions was also evaluated.
We found that the 79th residue (histidine or leucine) of Avh238 determined its cell deathinducing activity, and that the 53 amino acids in its C-terminal region are responsible for promoting Phytophthora infection. Transient expression of Avh238 in Nicotiana benthamiana revealed that nuclear localization is essential for triggering cell death, while Avh238-mediated suppression of INF1-triggered cell death requires cytoplasmic localization.
Our results demonstrate that a representative example of an essential Phytophthora RXLR effector can evolve to escape recognition by the host by mutating one nucleotide site, and can also retain plant immunosuppressive activity to enhance pathogen virulence in planta.
Journal Article
A Novel Phytophthora sojae Resistance Rps12 Gene Mapped to a Genomic Region That Contains Several Rps Genes
by
Robertson, Alison E.
,
Abeysekara, Nilwala S.
,
Bhattacharyya, Madan K.
in
Base Sequence
,
Binding sites
,
Biology and Life Sciences
2017
Phytophthora sojae Kaufmann and Gerdemann, which causes Phytophthora root rot, is a widespread pathogen that limits soybean production worldwide. Development of Phytophthora resistant cultivars carrying Phytophthora resistance Rps genes is a cost-effective approach in controlling this disease. For this mapping study of a novel Rps gene, 290 recombinant inbred lines (RILs) (F7 families) were developed by crossing the P. sojae resistant cultivar PI399036 with the P. sojae susceptible AR2 line, and were phenotyped for responses to a mixture of three P. sojae isolates that overcome most of the known Rps genes. Of these 290 RILs, 130 were homozygous resistant, 12 heterzygous and segregating for Phytophthora resistance, and 148 were recessive homozygous and susceptible. From this population, 59 RILs homozygous for Phytophthora sojae resistance and 61 susceptible to a mixture of P. sojae isolates R17 and Val12-11 or P7074 that overcome resistance encoded by known Rps genes mapped to Chromosome 18 were selected for mapping novel Rps gene. A single gene accounted for the 1:1 segregation of resistance and susceptibility among the RILs. The gene encoding the Phytophthora resistance mapped to a 5.8 cM interval between the SSR markers BARCSOYSSR_18_1840 and Sat_064 located in the lower arm of Chromosome 18. The gene is mapped 2.2 cM proximal to the NBSRps4/6-like sequence that was reported to co-segregate with the Phytophthora resistance genes Rps4 and Rps6. The gene is mapped to a highly recombinogenic, gene-rich genomic region carrying several nucleotide binding site-leucine rich repeat (NBS-LRR)-like genes. We named this novel gene as Rps12, which is expected to be an invaluable resource in breeding soybeans for Phytophthora resistance.
Journal Article
Tightly linked Rps12 and Rps13 genes provide broad-spectrum Phytophthora resistance in soybean
2021
The
Phytophtora
root and stem rot is a serious disease in soybean. It is caused by the oomycete pathogen
Phytophthora sojae
. Growing
Phytophthora
resistant cultivars is the major method of controlling this disease. Resistance is race- or gene-specific; a single gene confers immunity against only a subset of the
P. sojae
isolates. Unfortunately, rapid evolution of new
Phytophthora sojae
virulent pathotypes limits the effectiveness of an
Rps
(“resistance to
Phytophthora sojae
”) gene to 8–15 years. The current study was designed to investigate the effectiveness of
Rps12
against a set of
P. sojae
isolates using recombinant inbred lines (RILs) that contain recombination break points in the
Rps12
region. Our study revealed a unique
Rps
gene linked to the
Rps12
locus. We named this novel gene as
Rps13
that confers resistance against
P. sojae
isolate V13, which is virulent to recombinants that contains
Rps12
but lack
Rps13
. The genetic distance between the two
Rps
genes is 4 cM. Our study revealed that two tightly linked functional
Rps
genes with distinct race-specificity provide broad-spectrum resistance in soybean. We report here the molecular markers for incorporating the broad-spectrum
Phytophthora
resistance conferred by the two
Rps
genes in commercial soybean cultivars.
Journal Article