Combining wheat rust and fusarium head blight resistance genes and QTL using marker-assisted selection

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Date
2014-01
Authors
Senoko, Katleho Joyce
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University of the Free State
Abstract
English: Wheat (Triticum aestivum) is one of the most important cereal crops worldwide and cultivated in a wide range of environments. The production of wheat, based on consumption and other demands, needs to be increased to meet the annual requirements for its domesticated uses. Wheat productivity is influenced by biotic and abiotic stresses. Therefore, appropriate, efficient and environmentally friendly control measures to lessen such stresses need to be applied. Fungal diseases of wheat, including leaf rust, stripe rust, stem rust, powdery mildew and Fusarium head blight (FHB) can cause yield losses. The diseases result in a reduction in grain yield and affect grain quality negatively. The use of resistant cultivars is an effective, economical and environmentally safe approach to lower fungicide dependence and to reduce production losses due to foliar diseases. In this study, we combined rust and FHB resistance genes and/or quantitative trait loci (QTL) and molecular marker analyses were applied using polymorphic microsatellites (simple sequence repeats, SSR). The aim of the study was to combine five rust resistance genes (against leaf, stem and stripe rust) and five FHB resistance genes/QTL for type I and II resistance into a single wheat genotype with the aid of both marker-assisted and phenotypic selection. Molecular markers used in the study are already being used in commercial breeding programmes in South Africa. The study was divided into two phases. Phase one was aimed at combining rust resistance genes/QTL into a single genotype using parental lines AvocetYrSp, Blade, Kariega and CSLr19-149-299 and to select the best lines based on the number and combination of homozygous genes/QTL present. The genes Sr2, Sr26, Lr19, Lr34/Yr18/Sr57 and QTL QYr.sgi-2B were sourced from parental lines and combined in a double cross breeding scheme followed by several rounds of selfing. Selection was done using marker-assisted selection (MAS). The two best rust resistant lines were selected. Line S16(7.3) contained QYr.sgi-2B.1, Lr34/Yr18/Sr57 and Lr19 in a homozygous state while line S726(3.2) contained Lr34/Yr18/Sr57 and Lr19 in a homozygous and Sr26 in a heterozygous state. Phenotypic screening furthermore indicated the presence of YrSp in line S726(3.2). FHB resistance QTL for the second phase of the project were sourced from Frontana (type I resistance) and CM-82036 (type II resistance). The FHB resistant lines were crossed with the two best rust resistant lines in a double cross breeding scheme and selection was done using molecular markers. The study was conducted over a period of three years. A total of 954 individuals of a double cross population were screened with five markers associated with rust resistance genes/QTL and five markers associated with FHB resistance QTL. The following markers were used to screen for accumulated rust resistance genes/QTL: cssfr5 (Lr34/Yr18/Sr57), Gwm148 and Gwm501 (QYr.sgi-2B.1), STSLr19130 (Lr19) and Sr26#43 (Sr26). The markers DuPw227 (3A), Barc133 (3B), Gwm156, Gwm293 and Barc197.2 (5A), Gwm133 and Gwm644 (6B) and Gwm233 (7A) were used to identify individuals containing FHB resistance QTL. The number of resistance markers in the double cross family ranged between one to nine out of the 10 tested markers. The two best lines, 116.3 and 123.1, contained all markers except the marker linked to the minor FHB QTL on chromosome 7A. All tested lines contained Lr34/Yr18/Sr57 in a homozygous state while all other markers were segregating. The best lines will be self-pollinated for use in future breeding programmes.
Afrikaans: Koring (Triticum aestivum) is een van die belangrikste graangewasse wêreldwyd en word oor ‘n wye reeks omgewings geproduseer. Die produksie van koring, gebaseer op verbruik en aanvraag, moet verhoog word om in die toenemende jaarlikse behoeftes van plaaslike gebruik te voorsien. Koringproduktiwiteit word deur biotiese en abiotiese stres beïnvloed. Toepaslike, doeltreffende en omgewings-vriendelike beheermaatreëls moet dus toegepas word om hierdie stresfaktore te verlig. Verskeie swamsiektes van koring, insluitend blaarroes, streeproes, stamroes, poeieragtige meeldou en Fusarium-aarskroei (FAS) kan oesverliese veroorsaak en kwaliteit negatief beïnvloed. Die gebruik van weerstandbiedende kultivars is ‘n effektiewe, ekonomiese en veilige benadering tot minder swamdodertoedienings en ook laer verliese a.g.v. blaarsiektes. In hierdie studie is roes- en FAS weerstandsgene en/of kwantitatiewe eienskaploki (“QTL”) gekombineer en molekulêre merkeranalise toegepas d.m.v. polimorfiese mikrosatelliete (“SSR”). Die doel van die studie was om vyf roesweerstandsgene (teen blaar-, stam- en streeproes) en vyf FAS weerstandsgene/QTL vir tipe I en II weerstand m.b.v. merker-ondersteunde en fenotipiese seleksie in ‘n enkel koringgenotipe te kombineer. Die molekulêre merkers wat in hierdie studie gebruik is word reeds in kommersiële teelprogramme in Suid-Afrika toegepas. Die studie is in twee fases verdeel. In fase een is roesweerstandsgene/QTL vanaf die skenkerlyne AvocetYrSp, Blade, Kariega en CSLr19-149-299 in ‘n enkel genotipe gekombineer. Die beste lyne op grond van die aantal en kombinasie van homosigotiese gene/QTL teenwoordig is geselekteer. Die Sr2, Sr26, Lr19, Lr34/Yr18/Sr57 gene en QYr.sgi-2B QTL is vanaf die skenkerlyne verkry en in ‘n dubbelkruising teelskema gekombineer gevolg deur verskeie rondtes van selfbestuiwing. Seleksie is m.b.v. merker-ondersteunde seleksie gedoen. Die twee beste roesweerstandbiedende lyne is geselekteer. QYr.sgi-2B.1, Lr34/Yr18/Sr57 en Lr19 was homogeen in lyn S16(7.3) terwyl Lr34/Yr18/Sr57 en Lr19 homogeen en Sr26 heterogeen was in lyn S726(3.2). Fenotipiese toetse het verder aangetoon dat die YrSp geen in lyn S726(3.2) teenwoordig was. FAS weerstandsgene/QTL wat gedurende die tweede fase van die projek gebruik is, is vanaf Frontana (tipe I weerstand) en CM-82036 (tipe II weerstand) verkry. Die FAS weerstandbiedende lyne is met die twee roesweerstandbiedende lyne in ‘n dubbelkruising teelskema gekruis en seleksie is m.b.v. molekulêre merkers gedoen. Die studie is oor ‘n tydperk van drie jaar uitgevoer. ‘n Totaal van 954 individue van die dubbelkruispopulasie is met vyf merkers gekoppel aan roesweerstandsgene en vyf merkers gekoppel aan FAS weerstandsgene/QTL getoets. Die volgende merkers is gebruik om vir die gekombineerde roesweerstandsgene/QTL te toets: cssfr5 (Lr34/Yr18/Sr57), Gwm148 en Gwm501 (QYr.sgi-2B.1), STSLr19130 (Lr19) en Sr26#43 (Sr26). Die merkers DuPw227 (3A), Barc133 (3B), Gwm156, Gwm293 en Barc197.2 (5A), Gwm133 en Gwm644 (6B) en Gwm233 (7A) is gebruik om individue wat FAS weerstandsgene/QTL bevat te identifiseer. Die aantal merkers gekoppel aan weerstand in die dubbelkruispopulasie het gewissel tussen een en nege uit die 10 getoetste merkers. Die twee beste lyne, 116.3 en 123.1, het al die merkers behalwe die merker gekoppel aan die kleinere QTL op chromosoom 7A bevat. Lr34/Yr18/Sr57 was homosigoties in al die lyne wat getoets is terwyl al die ander merkers nog steeds gesegregeer het. Die beste lyne sal selfbestuif word vir gebruik in toekomstige teelprogramme.
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Keywords
Simple sequence repeats (SSR), Rust, Quantitative trait loci (QTL), Phenotypic screening, Fusarium head blight (FHB), Marker-assisted selection (MAS), Wheat rusts, Wheat -- Disease and pest resistance, Genetic markers, Fusarium diseases of plants, Dissertation (M.Sc.Agric. (Plant Sciences: Plant Breeding))--University of the Free State, 2014
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