Keller B

Krattinger SG

Mishra B

Priyamvada

Saharan MS

Tiwari R

English

Sydney eScholarship

 1Genetic and phenotypic mapping for leaf rust resistance, Lr34 in Indian bread wheat population  Saharan MS1,3, Tiwari R1, Krattinger SG2, Keller B2, Priyamvada1 and Mishra B1 1Directorate of Wheat Research, Karnal, Haryana-132001, INDIA; 2Institute of Plant Biology, University of Zurich, Zollikerstrasse 107, 8008, Zurich, Switzerland 3Corresponding author, E-mail: mssaharan7@yahoo.co.in  ABSTRACT  The Indian spring bread wheat cultivar ‘HD 2189’ shows leaf tip necrosis and resistance to leaf rust pathotypes in India since its release in 1980. We studied 128 single seed descent lines of an ‘HD2189’×’Agra Local’ F6 population to identify and map quantitative trait loci (QTLs) for leaf rust resistance. Percentage of leaf area infected and the response to infection were evaluated in two field trials in 2006-07 crop season at Directorate of Wheat Research, Karnal (India) and were transformed to the area under the disease progress curve (AUDPC). One–hundred and twenty eight F6 lines along with both parental lines were analysed with six polymorphic simple sequence repeats (SSRs) markers (GWM1220, XWM1, SWM5, SWM8, SWM10 & WMC463) and one sequence tagged sites (STS) marker, csLV34. A genetic linkage map based on above markers data was established. The region containing Lr34 was flanked by XSWM8 (proximal) and XWMC463 (distal) and spanned 68.7 cM in ‘HD 2189’×’Agra Local’ population. Marker SWM10 was closely located to Lr34 in present population. By using composite interval mapping and LOD >4.4, QTL analysis of the linkage group representing chromosome 7DS in the HD 2189×Agra Local population defined a QTL  for leaf rust resistance in the interval XGWM1220 - XSWM10 which accounted for 14.8 % of observed phenotypic variation for leaf rust resistance (AUDPC).    INTRODUCTION  Bread wheat (Triticum aestivum L. em Thell) has an allohexaploid genome (AABBDD, 2n = 6x = 42) with seven group of homeologous chromosomes. It is one of the major food crops in the world. Of the total area (approximately 215 million hectares) sown to hexaploid and tetraploid wheat worldwide, 44 per cent (95 million hectares) is in Asia. Of this, 62 million ha is located in China, India and Pakistan (Singh et al., 2004). The rust diseases of wheat pose a constant threat to sustainable wheat production and thus global food security. Leaf rust caused by Puccinia triticina Eriks is a devastating disease of wheat. The leaf rust pathogen is highly variable and has gained additional virulence over a period of time rendering new cultivars grown in India susceptible. The virulence pattern of P. triticina in central, peninsular and southern hill zones underwent a shift during 1980 to 2007. New pathotypes virulent on Lr9, Lr19 and Lr26 have appeared recently (Nayar et al., 2003; Bhardwaj et al., 2005). Although the timely application of fungicides can provide adequate control but their use adds to production costs and they are environment unfriendly. Thus growing resistant varieties is the most effective, environmentally safe and efficient control strategy for wheat rusts.   In contrast to race-specific resistance genes, two adult plant resistance genes (Lr34 and Lr46) have been reported to confer stable resistance to diverse pathotypes of the fungus and are thought to be durable (Singh et al., 1998). Molecular markers are powerful tools for identifying quantitative traits and dissecting these complex traits into Mendelian factors in the form of quantitative trait loci (QTL) as well as for establishing the genomic locations of such genetic loci. Genetic studies using lines from CIMMYT (William et al., 1997), Switzerland (Schnurbusch et al., 2004, Bossolini et al., 2006) or the Indian (Kaur et al., 2000) spring wheat germplasm confirmed the involvement of Lr34 in the expression of durable leaf rust resistance. The objective of present study was to dissect the quantitative and durable leaf rust resistance of the India cultivar ‘HD 2189’ in the ‘HD 2189’ × ‘Agra Local’ population using quantitative trait locus (QTL) analysis.   MATERIALS AND METHODS  A population of 128 F6 derived recombinant inbred lines (RILs) of  cross between the two Indian spring bread wheat cultivars ‘HD 2189’ (resistant) and ‘Agra Local’ (susceptible) was phenotyped for leaf rust resistance at two locations at the Directorate of Wheat Research, Karnal (Haryana) India. Leaf rust epidemic was initiated by inoculating 3 week old plants of spreader rows with urediniospore-water-tween 20 suspension having equal proportions of predominant four leaf rust pathotypes (12-2, 77-2, 77-5 and 104-2). The infection types TR, R, TMS, MS, TS and S) were recorded by following McNeal et al., (1971). Area under disease progress curve (AUDPC) was calculated using a computer package developed at CIMMYT, Mexico (CIMMYT. 1988) based on three leaf rust scorings recorded at 10 days equal interval.    Genomic DNA of 128 RILs and parents, ‘HD 2189’ and ‘Agra Local’ was extracted from fresh leaves using cetyltrimethylammonium bromide (CTAB) method. One–hundred and twenty eight F6 lines alongwith both parental lines were analysed with six polymorphic simple sequence repeats (SSRs) markers (GWM1220, XWM1, SWM5, SWM8, SWM10 & WMC463) and one  2 sequence tagged sites (STS) marker, csLV34. Microsatellite primers were used for amplification as described in original research papers (Bossolini et al., 2006; Lagudah et al., 2006; Seah et al., 1998). PCR products of SSR markers were resolved on polyacrylamide gel (PAGE) separated using LiCor DNA sequencer 4200. Visualization of the amplified PCR products of STS marker, csLV34 was by using a 2.5 per cent high-resolution agarose gel coupled with ethidium bromide staining. The linkage map was constructed using MAPMAKER 3.0b for MS-DOS (Lander et al., 1987). QTL analysis was performed with genotypic data of seven markers and phenotypic data for leaf rust i. e. the Area Under Disease Progress Curve (AUDPC). Interval QTL analysis was carried out with the composite interval mapping (CIM) program, PLABQTL, version 1.1 (Utz and Melchinger, 2000).    RESULTS AND DISCUSSION  One–hundred and twenty eight F6 lines along with both parental lines were phenotyped for leaf rust under artificially inoculated conditions at two locations in Karnal (India) during 2006-07. AUDPC values ranged from 0 to1082.50 on RILs. AUDPC values of 300 and 820 were calculated on parents ‘HD 2189’ and ‘Agra Local’, respectively. One hundred twenty eight lines (F6) from the cross between ‘HD 2189’ and ‘Agra Local’ were genotyped with the six SSR markers most closely linked to Lr34, alongwith the STS marker csLV34. We found that the region containing Lr34 and flanked by XSWM8 (proximal) and XWMC463 (distal) spanned 68.7 cM in “HD 2189 × ‘Agra Local’ population. In this population, QTL analysis defined a QTL for leaf rust resistance in the interval XGWM1220 – XSWM10. QTL analysis (composite interval mapping) of the linkage group representing chromosome 7DS in the ‘HD 2189’ × ‘Agra Local’ population revealed a QTL at position 20 with XSWM10, which accounted for 14.8 % of observed phenotypic variation for leaf rust resistance (AUDPC) with corresponding LOD 4.4. This QTL is considered to be due to the effects of the known slow-rusting gene, Lr34. These data indicate the usefulness of SWM10 for marker-assisted breeding programs for Lr34/Yr18.  Our finding suggest that leaf resistance locus on 7DS corresponds to the Lr34 gene. Leaf rust resistance loci were also earlier detected in other wheat populations (Messmer et al., 2000, Suenaga et al., 2003, Schnurbusch et al, 2004). Lr13 had also been postulated in cultivar ‘HD 2189’ (Nayar et al., 1999). Lr34 in combination with Lr13 confers high level of adult plant resistance which is durable in nature in cultivars Frontana and Era (Roelfs, 1988). In an earlier study, XSWM10 mapped 0.7cM proximal to Lr34 in ‘Arina’ × ‘Forno’ population (Bossolini et al., 2006).  A better understanding of the genetic basis of leaf rust resistance in present population would greatly improve the breeding efficiency of this trait for pyramiding of rust resistance genes.   ACKNOWLEDGEMENTS  Senior author is grateful to Indo Swiss Collaboration in Biotechnology, Department of Biotechnology (Govt. of India), Prof. Beat Keller Group, Institute of Plant Biology (University of Zurich), Switzerland and Directorate of Wheat Research, Karnal (ICAR) for providing the facilities for carrying out the research work.   REFERENCES  Bhardwaj, S.C., Prashar, M., Kumar, S., Jain, S.K. and Datta, D. 2005. Lr19 resistance in wheat becomes susceptible to Puccinia triticina in India. Plant Dis. 89:1360. Bossolini, E., Krattinger S.G. and Keller, B. 2006. Development of simple sequence repeat markers specific for the Lr34 resistance region of wheat using sequence information from rice and Aegilops tauschii. Theor. Appl. Genet. 113:1049-1062. CIMMYT. 1988. A programme for calculation of AUDPC. CIMMYT, Mexico DF: a software package. Kaur, M., Saini, R.G. and Preet, K. 2000. Adult plant leaf rust resistance from 111 wheat (Triticum aestivum L.) cultivars. Euphytica 113: 235-243.  Lagudah, E.S., McFadden, H., Singh, R. P., Huerta-Espino, J., Bariana, H. S. and Spielmeyer, W. 2006. Molecular genetic characterization of the Lr34/Yr18 slow rusting resistance gene region in wheat. Theor. Appl. Genet. 114:21-30. Lander, E., Green, P., Abrahamson, J., Barlow, A., Daly, M.J., Lincoln, S.E. and Newburg, L. 1987. MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174-181.  McNeal, F.H., Conzak, C.F., Smith, E.P., Tate, W.S. and Russel, T.S. 1971. A uniform system for recording and processing cereal research data. USDA ARS pp.  42. Messmer, M.M., Seyfarth, R., Keller, M., Schachermayr, G., Winzeler, M., Zanetti, S., Feuillet, C. and Keller, B. 2000. Genetic analysis of durable leaf rust resistance in winter  wheat. Theor. Appl. Genet. 100: 419-431.  Nayar, S.K., Jain, S.K., Prashar, M., Bhardwaj, S.C., Kumar, S. and Menon, M.K. 2003. Appearance of new pathotype of Puccinia recondita tritici virulent on Lr9 in India. Indian Phytopathol. 56:196-198. Roelfs, A.P. 1988. Resistance to leaf and stem rusts in wheat. Pages 10-12 in: Breeding Strategies for Resistance to Rusts of Wheat. Simmonds, N.W. and RajaRam, S. eds. CIMMYT, Mexico, D. F. Schnurbusch, T., Paillard, S., Schori, A., Messmer, M., Schachermayr, G., Winzeler, M. and Keller, B. 2004: Dissection of quantitative and durable leaf rust resistance in Swiss winter wheat reveals a major resistance QTL in the Lr34 chromosomal region. Theor. Appl. Genet. 108: 477-484.  3Seah, S. 1998. Cloning and characterization of a family of disease resistance analogs from wheat and barley. Theor. Appl. Genet. 97:737-945. Suenaga, K., Singh, R.P., Huerta-Espino, J. and William, H.M. 2003. Microsatellite markers for genes Lr34/Yr18 and other quantitative trait loci for leaf rust and stripe rust resistance in bread wheat. Phytopathology 93: 881-890. Singh, R.P., Mujeeb-Kazi, A. and Huerta-Espino, J. 1998. Lr46: A gene conferring slow-rusting resistance to leaf rust in wheat. Phytopathology 88: 890-894.  Singh, R.P., William, H.M., Huerta-Espino, J. and Rosewarne, G. 2004. Wheat rust in Asia: meeting with old and new technologies. Pages 1-13 in:  Utz, H.F. and Melchinger, A.E. 2000. PLABQTL: A computer program to map QTL. Version 1.1. Institute of Plant Breeding, Seed Science, and Population Genetics, University of Hohenheim, Germany. William, H. M., Hoisington, D., Singh, R.P. and Gonzalez de Leon., D. 1997. Detection of quantitative trait loci associated with leaf rust resistance in bread wheat. Genome 40:253-260.    

Genetic and phenotypic mapping for leaf rust resistance; Lr34 in Indian bread wheat  population

https://ses.library.usyd.edu.au/bitstream/2123/3223/1/P156.pdf

Genetic and phenotypic mapping for leaf rust resistance; Lr34 in Indian bread wheat population

Abstract

Similar works

Full text

Available Versions

Sydney eScholarship