Multiplex PCR assay for simultaneous identification of slow rust resistance genes Lr34, Lr46 and Lr68 in wheat (Triticum aestivum L.)

Currently, production of wheat cultivars (Triticum aestivum L.) that show durable field resistance against fungal pathogens is a priority of many breeding programs. This type of resistance involves race-nonspecific mechanisms and can be identified at adult-plant stages. Until now, seven genes (Lr34/Yr18, Lr46/Yr29, Lr67/Yr46, Lr68, Lr75, Lr77 and Lr78) conferring durable types of resistance against multiple fungal pathogens have been identified in the wheat gene pool. In this study we showed a multiplex Polymerase Chain Reaction (multiplex PCR) assay, which was developed for detection of slow rusting resistance genes Lr34, Lr46, Lr68, using molecular markers: csLV34, Xwmc44 and csGS, respectively. Identification of molecular markers was performed on 40 selected wheat genotypes which are the sources of slow rusting genes according to literature reports. Multiplex PCR is an important tool to reduce the time and cost of analysis. This multiplex PCR protocol can be applicable for genotyping processes and marker assisted resistance breeding of wheat.

[1]  J. Nawracała,et al.  Development of multiplex PCR to detect slow rust resistance genes Lr34 and Lr46 in wheat , 2019, Journal of Applied Genetics.

[2]  J. Dubcovsky,et al.  A High‐Resolution Map of Wheat QYr.ucw‐1BL, an Adult Plant Stripe Rust Resistance Locus in the Same Chromosomal Region as Yr29 , 2019, The plant genome.

[3]  D. Kurasiak-Popowska,et al.  Identification of Leaf Rust Resistance Genes in Selected Wheat Cultivars and Development of Multiplex PCR , 2019, Open life sciences.

[4]  J. Nawracała,et al.  Identification of Powdery Mildew Blumeria graminis f. sp. tritici Resistance Genes in Selected Wheat Varieties and Development of Multiplex PCR , 2019, Open Chemistry.

[5]  J. Arnemann.  Multiplex-PCR , 2019, Springer Reference Medizin.

[6]  B. Mccallum,et al.  Quantitative Trait Loci Conferring Leaf Rust Resistance in Hexaploid Wheat. , 2018, Phytopathology.

[7]  G. Bai,et al.  Mapping and characterization of the new adult plant leaf rust resistance gene Lr77 derived from Santa Fe winter wheat , 2018, Theoretical and Applied Genetics.

[8]  A. Hanzalová,et al.  Multiplex PCR assay to detect rust resistance genes Lr26 and Lr37 in wheat , 2018 .

[9]  G. Bai,et al.  Adult Plant Leaf Rust Resistance Derived from Toropi Wheat is Conditioned by Lr78 and Three Minor QTL. , 2018, Phytopathology.

[10]  R. Singh,et al.  Identification and Mapping of Adult Plant Resistance Loci to Leaf Rust and Stripe Rust in Common Wheat Cultivar Kundan. , 2017, Plant disease.

[11]  E. Lagudah,et al.  Starving the enemy , 2016, Science.

[12]  T. Wicker,et al.  Characterization of Lr75: a partial, broad-spectrum leaf rust resistance gene in wheat , 2016, Theoretical and Applied Genetics.

[13]  J. Patrick,et al.  A recently evolved hexose transporter variant confers resistance to multiple pathogens in wheat , 2015, Nature Genetics.

[14]  R. Singh,et al.  Progress towards genetics and breeding for minor genes based resistance to Ug99 and other rusts in CIMMYT high yielding spring wheat , 2014 .

[15]  Z. Dar,et al.  Status and strategies in breeding for rust resistance in wheat , 2013 .

[16]  M. Lillemo,et al.  QTL for spot blotch resistance in bread wheat line Saar co-locate to the biotrophic disease resistance loci Lr34 and Lr46 , 2012, Theoretical and Applied Genetics.

[17]  E. Lagudah,et al.  Lr68: a new gene conferring slow rusting resistance to leaf rust in wheat , 2012, Theoretical and Applied Genetics.

[18]  D. Somers,et al.  Allelic variation for the rust resistance gene Lr34/Yr18 in Canadian wheat cultivars , 2011, Euphytica.

[19]  S. Chao,et al.  Mapping and validation of QTL which confer partial resistance to broadly virulent post-2000 North American races of stripe rust in hexaploid wheat , 2011, Theoretical and Applied Genetics.

[20]  G. Ash.  Wheat Rusts: An Atlas of Resistance Genes. , 1996, Australasian Plant Pathology.

[21]  S. Chakrabortya,et al.  Climate change , plant diseases and food security : an overview , 2011 .

[22]  E. Lagudah.  Molecular genetics of race non-specific rust resistance in wheat , 2011, Euphytica.

[23]  Zhonghu He,et al.  Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.) , 2010 .

[24]  B. Keller,et al.  Gene-specific markers for the wheat gene Lr34/Yr18/Pm38 which confers resistance to multiple fungal pathogens , 2009, Theoretical and Applied Genetics.

[25]  R. Tiwari,et al.  STS marker based tracking of slow rusting Lr34 gene in Indian wheat genotypes , 2009 .

[26]  B. Keller,et al.  A Putative ABC Transporter Confers Durable Resistance to Multiple Fungal Pathogens in Wheat , 2009, Science.

[27]  D. Garvin,et al.  Analysis of the Lr34/Yr18 Rust Resistance Region in Wheat Germplasm , 2008 .

[28]  P. K. Gupta,et al.  Wheat Genomics: Present Status and Future Prospects , 2008, International journal of plant genomics.

[29]  Zhonghu He,et al.  Development of two multiplex PCR assays targeting improvement of bread-making and noodle qualities in common wheat , 2008 .

[30]  D. Somers,et al.  Molecular marker analysis of Lr34 in Canada Western Red Spring wheat cultivars , 2008 .

[31]  B. Mccallum,et al.  Inheritance and genetic mapping of leaf rust resistance genes in the wheat cultivar Buck Manantial , 2008 .

[32]  R. Singh,et al.  Molecular genetic characterization of the Lr34/Yr18 slow rusting resistance gene region in wheat , 2006, Theoretical and Applied Genetics.

[33]  M. Côté,et al.  Identification of Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma on Inoculated and Naturally Infected Fruit Using Multiplex PCR. , 2004, Plant disease.

[34]  M. Kaur,et al.  Adult plant leaf rust resistance from 111 wheat (Triticum aestivum L.) cultivars , 2000, Euphytica.

[35]  G. Bryan,et al.  The potential of microsatellites for high throughput genetic diversity assessment in wheat and barley , 1998, Genetic Resources and Crop Evolution.

[36]  A. Ballabio,et al.  Screening for steroid sulfatase (STS) gene deletions by multiplex DNA amplification , 1990, Human Genetics.

[37]  C. Mundt,et al.  Associations and genetics of three components of slow rusting in leaf rust of wheat , 2004, Euphytica.

[38]  R. Singh,et al.  Microsatellite markers for genes lr34/yr18 and other quantitative trait Loci for leaf rust and stripe rust resistance in bread wheat. , 2003, Phytopathology.

[39]  B. Salmanowicz,et al.  Multiplex PCR identification of wheat HMW glutenin subunit genes by allele-specific markers. , 2003, Journal of applied genetics.

[40]  R. Singh,et al.  Characterization of Lr46, a gene conferring partial resistance to wheat leaf rust. , 2004, Hereditas.

[41]  N. U. Ahmed,et al.  Mapping of durable adult plant and seedling resistances to stripe rust and stem rust diseases in wheat , 2001 .

[42]  M. Sorrells,et al.  Mapping Yr28 and Other Genes for Resistance to Stripe Rust in Wheat , 2000 .

[43]  C. Morris,et al.  Catalogue of gene symbols for wheat: 2000 supplement. , 2000 .

[44]  M. Ganal,et al.  A microsatellite map of wheat. , 1998, Genetics.

[45]  R. Singh,et al.  Lr46: a gene conferring slow-rusting resistance to leaf rust in wheat. , 1998, Phytopathology.

[46]  F. Salamini,et al.  Catalogue of gene symbols for wheat , 1998 .

[47]  N. Heerema,et al.  Multiplex PCR: critical parameters and step-by-step protocol. , 1997, BioTechniques.

[48]  D. Rubiales,et al.  Characterization of Lr34, a major gene conferring nonhypersensitive resistance to wheat leaf rust. , 1995 .

[49]  C. Lengauer,et al.  Rapid screening of the Y chromosome in idiopathic sterile men, diagnostic for deletions in AZF, a genetic Y factor expressed during spermatogenesis , 2009, Andrologia.

[50]  R. Stern,et al.  Efficient 12-mutation testing in the CFTR gene: a general model for complex mutation analysis. , 1993, Human molecular genetics.

[51]  D. Ledbetter,et al.  Multiplex PCR of three dinucleotide repeats in the Prader-Willi/Angelman critical region (15q11-q13): molecular diagnosis and mechanism of uniparental disomy. , 1993, Human molecular genetics.

[52]  R. Singh,et al.  Genetics of adult-plant resistance of leaf rust in 'Frontana' and three CIMMYT wheats , 1992 .

[53]  R. Singh.  Genetic association of leaf rust resistance gene Lr34 with adult plant resistance to stripe rust in bread wheat , 1992 .

[54]  D. van Beers,et al.  Fast Multiplex polymerase chain reaction on boiled clinical samples for rapid viral diagnosis. , 1990, Journal of virological methods.

[55]  P. Dyck.  THE INHERITANCE OF LEAF RUST RESISTANCE IN WHEAT CULTIVARS KENYON AND BUCK MANANTIAL , 1989 .

[56]  R A Gibbs,et al.  Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. , 1988, Nucleic acids research.

[57]  J. S. Heslop-Harrison,et al.  Nuclear dna amounts in angiosperms. , 1976, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[58]  R. D. Wilcoxson.  Genetics of slow rusting in cereals. , 1981 .

[59]  P. Dyck,et al.  ADULT-PLANT LEAF RUST RESISTANCE IN PI 250413, AN INTRODUCTION OF COMMON WHEAT , 1979 .

[60]  H. Ohm.  Three Components of Slow Leaf-Rusting at Different Growth Stages in Wheat , 1976 .

[61]  C. N. Law,et al.  Genetic control of durable resistance to yellow rust (Puccinia striiformis) in the wheat cultivar Hybride de Bersee. , 1975 .

[62]  L. H. Shebeski,et al.  Glenlea red spring wheat , 1972 .

[63]  N. Borlaug.  Wheat breeding and its impact on world food supply , 1968 .