Development of a PCR-based allele-specific assay from an RFLP probe linked to resistance to cereal cyst nematode in wheat

The RFLP locus Xglk605 identified by the probe Tag605 maps to a proximal position on the long arm of wheat chromosome 2B about 7 cM away from a gene conditioning resistance to cereal cyst nematode in the wheat line AUS10894. The clone Tag605 was partially sequenced and the PCR primer set AWP1 was designed. The 292-bp product, which showed no polymorphism between varieties, was cloned and sequenced. A single base difference was found in the sequence of the AWP1 products amplified and cloned from the wheat lines AUS10894 and 'Spear'. PCR primers were designed with 3′ termini that corresponded to the two alleles. A dual-PCR system was developed in which the primer sets AWP2 and AWP3 produced allele-specific amplification. The concentration of the oligonucleotide primers and the sequence of the primer–template mismatches were critical to the success of discriminatory allele amplification. Key words : Triticum aestivum, STS, cereal cyst nematode, RFLP.

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Investigation of resistance to Pratylenchus penetrans and P. thornei in international wheat lines and its durability when inoculated together with the cereal cyst nematode Heterodera avenae, using qPCR for nematode quantification

European Journal of Plant Pathology ◽

10.1007/s10658-018-1420-0 ◽

2018 ◽

Vol 151 (4) ◽

pp. 875-889 ◽

Cited By ~ 3

Author(s):

Fouad Mokrini ◽

Nicole Viaene ◽

Lieven Waeyenberge ◽

Abdelfattah A. Dababat ◽

Maurice Moens

Keyword(s):

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Heterodera Avenae ◽

Pratylenchus Penetrans ◽

Wheat Lines

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Registration of H192 and H782, White Winter Wheat Lines Resistant to Cereal Cyst Nematode and Powdery Mildew

Journal of Plant Registrations ◽

10.3198/jpr2016.01.0001crg ◽

2016 ◽

Vol 11 (1) ◽

pp. 71-74 ◽

Cited By ~ 1

Author(s):

Dan Qiu ◽

Lei Cui ◽

Yanling Sun ◽

Jingwei Zou ◽

Chaoxing Zheng ◽

...

Keyword(s):

Powdery Mildew ◽

Winter Wheat ◽

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Wheat Lines

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Diagnostic DNA markers for cereal cyst nematode resistance in bread wheat

Australian Journal of Agricultural Research ◽

10.1071/ar01031 ◽

2001 ◽

Vol 52 (12) ◽

pp. 1367 ◽

Cited By ~ 63

Author(s):

F. C. Ogbonnaya ◽

N. C. Subrahmanyam ◽

O. Moullet ◽

J. de Majnik ◽

H. A. Eagles ◽

...

Keyword(s):

Nematode Resistance ◽

Wheat Breeding ◽

Primary Objective ◽

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Aegilops Ventricosa ◽

Breeding Lines ◽

Marker Selection ◽

Efficiency Of Selection ◽

Wheat Lines

The development of cultivars resistant to cereal cyst nematode (CCN) is a primary objective in wheat breeding in the southern wheatbelt of Australia. Nine CCN resistance genes have been identified in wheat and its relatives, some of which confer resistance to the Australian pathotype of CCN (Ha13). Cultivars released in Australia with CCN resistance carry either the Cre1 or CreF gene, with the Cre3 gene present in advanced breeding lines. The biological assay for CCN resistance screening in wheat is time-consuming, not reliable on a single-plant basis, and prone to inconsistencies, thus reducing the efficiency of selection amongst breeding lines. Using gene sequences initially isolated from the Cre3 locus, a DNA-based marker selection system was developed and applied to unambiguously identify wheat lines carrying resistance alleles at theCre1 and/or Cre3 loci in breeding populations derived from diverse genetic backgrounds. Homologues of sequences from the Cre3 locus, located elsewhere in the wheat genome, can also be used to select wheat lines with a newly identified CCN resistance gene (Cre6) introgressed from Aegilops ventricosa. Application of these markers has become an integral part of the southern Australian breeding programs.

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Development and Identification of Wheat Lines H3714 and H4058 Resistant to Cereal Cyst Nematode

ACTA AGRONOMICA SINICA ◽

10.3724/sp.j.1006.2015.00872 ◽

2015 ◽

Vol 41 (6) ◽

pp. 872 ◽

Cited By ~ 2

Author(s):

Xi-Ying SUN ◽

Lei CUI ◽

Lei SUN ◽

Yan-Ling SUN ◽

Dan QIU ◽

...

Keyword(s):

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Wheat Lines

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Comparison of PCR Primers for Analyzing Denitrifying Microorganisms in the Hyporheic Zone

Applied Sciences ◽

10.3390/app10124172 ◽

2020 ◽

Vol 10 (12) ◽

pp. 4172 ◽

Cited By ~ 1

Author(s):

Heejung Kim

Keyword(s):

Hyporheic Zone ◽

Stream Water ◽

Pcr Primers ◽

Polymerase Chain Reaction Primer ◽

Aqueous Environments ◽

Specific Amplification ◽

Specific Band ◽

Primer Sets ◽

Denitrifying Microorganisms ◽

Pcr Primer

In this study, the specific amplifications of six denitrification-associated genes using PCR(Polymerase Chain Reaction) primer sets were compared. Thereafter, the PCR primer sets that were determined to be suitable for each denitrification-associated gene were used to test samples from sixteen aqueous environments (three from groundwater, three from stream water, and ten from hyporheic zone water). The specific amplification was determined using PCR primer sets for denitrification-associated genes and nucleic acids from eleven types of strains. NosZ was the most frequently amplified gene from the nucleic acid of type, with a specific band seen in all eleven strains. The specific band amplification and PCR time of the strains were analyzed to select one PCR primer set for each gene. The selected PCR primer sets were used to analyze sixteen samples from the aqueous environments in which denitrifying microorganisms were expected to be present. Specific bands of narG, nirS, and nosZ were most frequently observed in the hyporheic water samples. The results showed that microorganisms containing nirG (involved in the reduction of nitrate to nitrite), nirS (involved in the reduction of nitrite to nitric oxide), and nosZ (involved in the reduction of nitrous oxide to nitrogen gas) were the most abundant in the hyporheic zone samples used in this study.

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Analysis of ascorbate peroxidase genes expressed in resistant and susceptible wheat lines infected by the cereal cyst nematode, Heterodera avenae

Plant Cell Reports ◽

10.1007/s00299-010-0903-z ◽

2010 ◽

Vol 29 (10) ◽

pp. 1169-1178 ◽

Cited By ~ 23

Author(s):

Ester Simonetti ◽

Eva Alba ◽

María Jesús Montes ◽

Ángeles Delibes ◽

Isidoro López-Braña

Keyword(s):

Ascorbate Peroxidase ◽

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Heterodera Avenae ◽

Wheat Lines

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Identification of microsatellite markers associated with the cereal cyst nematode resistance gene Cre3 in wheat

Australian Journal of Agricultural Research ◽

10.1071/ar04085 ◽

2004 ◽

Vol 55 (12) ◽

pp. 1205 ◽

Cited By ~ 8

Author(s):

E. M. Martin ◽

R. F. Eastwood ◽

F. C. Ogbonnaya

Keyword(s):

Microsatellite Markers ◽

Microsatellite Marker ◽

Wheat Chromosome ◽

Triticum Aestivum L ◽

Wheat Breeding ◽

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Heterodera Avenae ◽

Pcr Marker ◽

Diverse Range

Cereal cyst nematode (CCN) is a root disease caused by the pathogen Heterodera avenae Woll. that significantly reduces wheat (Triticum aestivum L.) grain yields in temperate countries. The Cre3 gene, located on chromosome 2DL, provides high levels of resistance to the Australian pathotype and isolates from Syria and Algeria, and has become available to wheat breeders. Selection for lines carrying the Cre3 gene in Australian wheat breeding programs is currently based on a dominant PCR marker (Cre3spf/r) diagnostic for the Cre3 gene. However, this marker has limitations that increase the cost and reduce selection efficiency in screening early-generation breeding lines. Such limitations would be minimised by the identification of a microsatellite marker linked to the Cre3 gene. We have constructed 2 genetic linkage maps of wheat chromosome 2DL and identified microsatellite markers mapping closely to the diagnostic Cre3spf/r marker. These closely linked markers were validated in a diverse range of germplasm, and one microsatellite marker, Xgwm301, which mapped 4 cM from Cre3spf/r, was shown to be highly associated with the presence of the Cre3 gene. Amplification conditions for the Xgwm301 locus were optimised, and its use in marker-assisted selection to identify Cre3 CCN-resistant wheat in the Australian Grain Technologies breeding program is demonstrated.

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Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650362 ◽

1996 ◽

Vol 75 (05) ◽

pp. 757-759 ◽

Cited By ~ 11

Author(s):

Rainer Blasczyk ◽

Markus Ritter ◽

Christian Thiede ◽

Jenny Wehling ◽

Günter Hintz ◽

...

Keyword(s):

Direct Sequencing ◽

Activated Protein C ◽

Factor V Leiden ◽

Pcr Amplification ◽

Factor V ◽

Specific Pcr ◽

Pcr Rflp ◽

Allele Specific ◽

Specific Amplification ◽

Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

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