Novel PCR Assays for the Detection of Biological Agents Responsible for Wheat Rust Diseases: Puccinia triticina and Puccinia striiformis f. sp. tritici

2017 ◽  
Vol 27 (5) ◽  
pp. 299-305 ◽  
Author(s):  
Adam Kuzdraliński ◽  
Anna Kot ◽  
Hubert Szczerba ◽  
Agnieszka Ostrowska ◽  
Michał Nowak ◽  
...  
Keyword(s):  
Puccinia Striiformis ◽  
Infected Plant ◽  
Simultaneous Detection ◽  
Puccinia Triticina ◽  
Biological Agents ◽  
Plant Materials ◽  
Rust Diseases ◽  
Pcr Assays ◽  
Primer Sets ◽  
Wheat Rust

The species <i>Puccinia triticina</i> (Pt) and <i>Puccinia striiformis</i> f. sp. <i>tritici </i>(Pst) are devastating cereal pathogens that cause leaf and stripe rust diseases. We developed PCR assays for the species-specific detection of Pt and Pst, 2 biological agents that cause wheat rust disease. For each pathogen, we validated 3 primer sets that target the second largest subunits of the RNA polymerase II (<i>rpb2</i>) and β-tubulin 1 (<i>tub1</i>) genes. The specificities of the primers were verified using naturally infected plant materials with visual symptoms of disease. All primer sets amplified a single DNA fragment of the expected length. The primer sets LidPr15/16, LidPr1/2, and LidPs13/14 were able to detect small amounts of pure fungal DNA with sensitivities of 0.1, 1, and 10 pg/μL, respectively. A sufficient detection limit (1 pg/μL to 5 ng/μL) was observed for all assays when the sensitivity test was performed with host plant DNA. The study also evaluated the simultaneous detection of both rust pathogens, and the multiplex PCR assay generated amplicons of 240 and 144 bp in length for Pts (LidPs9/10) and Pt (LidPr1/2), respectively.

Advances in understanding the biology and epidemiology of rust diseases of cereals

2021 ◽  
pp. 15-38
Author(s):  
Vanessa Bueno-Sancho ◽  
◽  
Clare M. Lewis ◽  
Diane G. O. Saunders ◽  
◽  
...  
Keyword(s):  
Disease Surveillance ◽  
Crop Production ◽  
Puccinia Striiformis ◽  
Management Strategies ◽  
Puccinia Triticina ◽  
Rust Fungi ◽  
Biological Knowledge ◽  
Parasitic Fungi ◽  
Rust Diseases ◽  
Wheat Rust

Rust fungi (order: Pucciniales) constitute the largest group of plant parasitic fungi and include many species of agricultural importance. This includes the three wheat rust fungi (Puccinia graminis f. sp. tritici, Puccinia striiformis f. sp. tritici and Puccinia triticina) that have posed a threat to crop production throughout history. This chapter provides an overview of the wheat rust pathogen lifecycle that has been critical to the design of effective disease management strategies and discusses recent integration of basic biological knowledge and genomic-led tools within an epidemiological framework. Furthermore, we include a case study on the “field pathogenomics” technique, illustrating the value of genomic-based tools in disease surveillance activities. Bringing together advances in understanding basic pathogen biology, developments in modelling for disease forecasting and identification, alongside genomic-led advances in surveillance and resistance gene cloning, holds great promise for curtailing the threat of these notorious pathogens.

Development and Application of a New PCR Method for Detection of Blumeria graminis f. sp. tritici

2018 ◽  
Vol 28 (3) ◽  
pp. 137-146 ◽  
Author(s):  
Adam Kuzdraliński ◽  
Hubert Szczerba ◽  
Anna Kot ◽  
Agnieszka Ostrowska ◽  
Michał Nowak ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Puccinia Triticina ◽  
Blumeria Graminis ◽  
Pyrenophora Tritici Repentis ◽  
Field Samples ◽  
Expected Length ◽  
Pcr Assays ◽  
Primer Sets ◽  
Wheat Leaves ◽  
Species Specific

We developed new PCR assays that target beta-tubulin (<i>TUB2</i>) and 14 alpha-demethylase (<i>CYP51</i>) genes and used them for the species-specific detection of <i>Blumeria graminis</i> f. sp. <i>tritici</i> (<i>Bgt</i>). Based on fungi DNA sequences available in the NCBI (National Center for Biotechnology Information) GenBank database we developed simplex and duplex PCR assays. The specificities of the primer sets were evaluated using environmental samples of wheat leaves collected during the 2015/2016 growing season across Poland. Primer sets<i></i> LidBg17/18 and LidBg21/22 strongly amplified fragments of the expected length for all 67 tested samples. Primer specificity was confirmed using field samples of <i>Zymoseptoria tri­tici</i>, <i>Puccinia triticina</i> (syn.<i> P. recondita</i> f. sp.<i> tritici</i>), <i>P. striiformis</i> f. sp.<i> tritici</i>, and <i>Pyrenophora tritici-repentis</i>.

scholarly journals Accomplishments in wheat rust research in South Africa

2020 ◽  
Vol 116 (11/12) ◽  
Author(s):  
Zacharias A. Pretorius van der Walt ◽  
Renée Prins ◽  
Elsabet Wessels ◽  
Cornel M. Bender ◽  
Botma Visser ◽  
...  
Keyword(s):  
South Africa ◽  
South African ◽  
New Technologies ◽  
Puccinia Triticina ◽  
Sources Of Resistance ◽  
Breeding Lines ◽  
Rust Diseases ◽  
African Populations ◽  
Pathogen Populations ◽  
Wheat Rust

Rust diseases, although seasonal, have been severe constraints in wheat production in South Africa for almost 300 years. Rust research gained momentum with the institution of annual surveys in the 1980s, followed by race identification, an understanding of rust epidemiology, and eventually a focused collaboration amongst pathologists, breeders and geneticists. Diversity in South African populations of Puccinia triticina, P. graminis f. sp. tritici and P. striiformis f. sp. tritici has been described and isolates are available to accurately phenotype wheat germplasm and study pathogen populations at national, regional and global levels. Sources of resistance have been, and still are, methodically analysed and molecular marker systems were developed to incorporate, stack and verify complex resistance gene combinations in breeding lines and cultivars. Vigilance, capacity, new technologies, collaboration and sustained funding are critical for maintaining and improving the current research impetus for future management of these important diseases.

SCREENING OF WHEAT LR-GENES FOR RESISTANCE TO PUCCINIA TRITICINA IN THE NORTH CAUCASUS REGION CONDITIONS

1970 ◽  
pp. 54-56
Author(s):  
G. V. Volkova ◽  
O. A. Kudinova ◽  
O. F. Vaganova
Keyword(s):  
Winter Wheat ◽  
Leaf Rust ◽  
Puccinia Triticina ◽  
North Caucasus ◽  
Breeding Programs ◽  
Lr Genes ◽  
Caucasus Region ◽  
Highly Efficient ◽  
The North ◽  
Wheat Rust

Currently, more than 70 wheat rust resistance genes are known, but few of them are effective. The purpose of this work is to screen lines of Lr gene carriers for resistance to leaf rust under conditions of the North Caucasus region. Investigations were carried out in 2016-2018 at the infectious site of VNIIBZR. Research material was 49 near isogenic lines of winter wheat cultivar Thatcher. Infectious material was the combined populations of P. triticina, obtained as a result of route surveys of industrial and breeding crops of winter wheat in the areas of the Krasnodar, Stavropol Territories and the Rostov Region, conducted in 2016-2018. According to the assessment, the genes are ranked as follows: - highly efficient genes (plants with no signs of damage): Lr9, Lr42, Lr43 + 24 and Lr50; effective (1R-5R) Lr genes: 19, 24, 29, 36, 37, 38, 45, 47; moderately effective (10MR-20MR) Lr genes: 17, 18, 21, 22a, 28, 32, 41, 52. The remaining Lr-lines were susceptible to P. triticina (25 MR - 90S) to varying degrees. Highly efficient and effective genes Lr9, Lr19, Lr24, Lr29, Lr38, Lr42, Lr43 + 24, Lr47 and Lr50 showed resistance in the seedling phase and can be recommended for inclusion in breeding programs to protect wheat from leaf rust in different phases of plant ontogenesis in the North Caucasus region.

scholarly journals Development of PCR Assays for the Identification of Species and Pathotypes of Elsinoë Causing Scab on Citrus

Plant Disease ◽  
2007 ◽  
Vol 91 (7) ◽  
pp. 865-870 ◽  
Author(s):  
J. W. Hyun ◽  
N. A. Peres ◽  
S.-Y. Yi ◽  
L. W. Timmer ◽  
K. S. Kim ◽  
...  
Keyword(s):  
Internal Transcribed Spacer ◽  
Sweet Orange ◽  
Random Amplified Polymorphic Dna ◽  
Its Region ◽  
The United States ◽  
Specific Primer ◽  
Identification Of Species ◽  
Orange Fruit ◽  
Pcr Assays ◽  
Primer Sets

Two scab pathogens of citrus, Elsinoë fawcettii and E. australis, cause citrus scab and sweet orange scab, respectively, and pathotypes of each species have been described. The two species cannot be readily distinguished by morphological or cultural characteristics and can be distinguished only by host range and the sequence of the internal transcribed spacer (ITS) region. In this study, random amplified polymorphic DNA (RAPD) assays clearly distinguished E. fawcettii and E. australis, and the sweet orange and natsudaidai pathotypes within E. australis also could be differentiated. We developed specific primer sets, Efaw-1 for E. fawcettii; Eaut-1, Eaut-2, Eaut-3, and Eaut-4 for E. australis; and EaNat-1 and EaNat-2 for the natsudaidai pathotype within E. australis using RAPD products unique to each species or pathotype. Other primer sets, Efaw-2 and Eaut-5, which were specific for E. fawcettii and E. australis, respectively, were designed from previously determined ITS sequences. The Efaw-1 and Efaw-2 primer sets successfully identified E. fawcettii isolates from Korea, Australia, and the United States (Florida) and the Eaut-1 to Eaut-5 primer sets identified both the sweet orange pathotype isolates of E. australis from Argentina and the natsudaidai pathotype isolates from Korea. The EaNat-1 and EaNat-2 primer sets were specific for isolates of the natsudaidai pathotype. The Efaw-1 and Efaw-2 primer sets successfully detected E. fawcettii from lesions on diseased leaves and fruit from Korea and primer pairs Eaut-1, Eaut-2, Eaut-3, Eaut-4, and Eaut-5 detected E. australis from lesions on sweet orange fruit from Brazil.

scholarly journals Virulence surveillance of wheat black stem rust fungus

2014 ◽  
Vol 11 (2) ◽  
pp. 803-812
Author(s):  
Baghdad Science Journal
Keyword(s):  
Leaf Rust ◽  
Disease Severity ◽  
Stem Rust ◽  
Yellow Rust ◽  
Rust Fungus ◽  
The North ◽  
Rust Diseases ◽  
Northern Regions ◽  
Infection Disease ◽  
Wheat Rust

General survey for wheat rust diseases in Iraqi fields was done during the seasons of 2010, 2011 and 2012. The survey covered different fields in southern, middle and northern regions. Results of the first season indicated that most of Iraqi cultivars such as Tmmoze2, IPA 99 and Mexipak showed different types of susceptibility to both yellow and leaf rust infection. Disease severity increased when the conditions were favorable for infections with using susceptible cultivars. The severity of leaf rust was less in the north region comparing with the middle and south regions. Most of the introduced cultivars such as Sham6 and Cimmyto showed susceptible reaction to yellow and leaf rust. Yellow rust was in epiphytotic form at the Iraqi-Syrian-Turkish triangle where the disease severity was 100%. Low disease severity of stem rust was observed on some cultivars (1-5%), except for the cultivar Mexipak which showed 40%S in Najaf. Rusts at season of 2011 were restricted mostly in Baghdad and the yellow rust was dominant. The AUDPC of 15 wheat cultivars showed that Sawa and Sali were highly susceptible to the three types of rusts while Babil113 and Tamoze2 were resistant. No rusts were detected at season 2012.

scholarly journals Detection and Enumeration of Aromatic Oxygenase Genes by Multiplex and Real-Time PCR

2003 ◽  
Vol 69 (6) ◽  
pp. 3350-3358 ◽  
Author(s):  
Brett R. Baldwin ◽  
Cindy H. Nakatsu ◽  
Loring Nies
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Gene Copy Number ◽  
Pcr Amplification ◽  
Gene Copy ◽  
Hydrocarbon Biodegradation ◽  
Polymerization Temperature ◽  
Primer Sets ◽  
The Impact

ABSTRACT Our abilities to detect and enumerate pollutant-biodegrading microorganisms in the environment are rapidly advancing with the development of molecular genetic techniques. Techniques based on multiplex and real-time PCR amplification of aromatic oxygenase genes were developed to detect and quantify aromatic catabolic pathways, respectively. PCR primer sets were identified for the large subunits of aromatic oxygenases from alignments of known gene sequences and tested with genetically well-characterized strains. In all, primer sets which allowed amplification of naphthalene dioxygenase, biphenyl dioxygenase, toluene dioxygenase, xylene monooxygenase, phenol monooxygenase, and ring-hydroxylating toluene monooxygenase genes were identified. For each primer set, the length of the observed amplification product matched the length predicted from published sequences, and specificity was confirmed by hybridization. Primer sets were grouped according to the annealing temperature for multiplex PCR permitting simultaneous detection of various genotypes responsible for aromatic hydrocarbon biodegradation. Real-time PCR using SYBR green I was employed with the individual primer sets to determine the gene copy number. Optimum polymerization temperatures for real-time PCR were determined on the basis of the observed melting temperatures of the desired products. When a polymerization temperature of 4 to 5�C below the melting temperature was used, background fluorescence signals were greatly reduced, allowing detection limits of 2 � 102 copies per reaction mixture. Improved in situ microbial characterization will provide more accurate assessment of pollutant biodegradation, enhance studies of the ecology of contaminated sites, and facilitate assessment of the impact of remediation technologies on indigenous microbial populations.

scholarly journals Efficacy of plant resistance inducers on cereal crops against a complex of pathogens

2020 ◽  
Vol 21 ◽  
pp. 00029
Author(s):  
Irina Matveeva ◽  
Anastasia Danilova ◽  
Yuriy Kim ◽  
Olesya Miroshnichenko ◽  
Alexander Grigoriev
Keyword(s):  
Grain Yield ◽  
Plant Resistance ◽  
Leaf Spot ◽  
Puccinia Striiformis ◽  
Puccinia Triticina ◽  
Puccinia Hordei ◽  
Cereal Crops ◽  
Yellow Leaf ◽  
Biological Efficacy ◽  
Resistance Inducers

The article describes the benefits of using plant resistance inducers on cereal crops and the basic principles of their impact on pests development (Pyrenophora tritici-repentis, Puccinia triticina, Puccinia striiformis, Puccinia graminis, Puccinia hordei). The assessment results of the biological and economic effectiveness of L-479 and DL-59 preparations are presented in comparison with the chemical Kolosal,CE and biological Albit, which were used as standards for reducing the development of yellow leaf spot and rust diseases. L-479 product was found to be the most effective on wheat against brown, yellow and stem rust, and yellow leaf spot. Its biological efficacy ranged from 48.3 % to 57.0 % depending on the pathogen, and the increase in grain yield ranged from 3.7 % to 22.1 %. DL-59 product showed the best biological efficacy against dwarf leaf rust of barley – 89.8 %, the increase in grain yield was 3.5 %.

scholarly journals Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses

2013 ◽  
Vol 158 (8) ◽  
pp. 1743-1753 ◽  
Author(s):  
Ilona Stefańska ◽  
Tomasz Dzieciatkowski ◽  
Lidia B. Brydak ◽  
Magdalena Romanowska
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Influenza Viruses ◽  
Avian Influenza Viruses ◽  
Pcr Assays
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