Soybean cyst nematode: Challenges and opportunities

Shawn M. J. Winter1, Istvan Rajcan, and Barry J. Shelp2

Department of Plant Agriculture, University of Guelph, Guelph, Ontario, Canada N1G 2W1. Received 19 April 2005, accepted 22 September 2005.
Winter, S. M. J., Rajcan, I. and Shelp, B. J. 2006. Soybean cyst nematode: Challenges and opportunities. Can. J. Plant Sci. 86: 2532. Soybean cyst nematode (SCN) is the primary pest responsible for yield losses of Glycine max. Management of SCN remains difficult in commercial soybean production due to the length of its biological cycle, frequent changes in population virulence, and ease of spread via infested soil. Effective management relies on crop rotation in combination with resistant cultivars, which have been derived from a limited germplasm base. Breeding for SCN resistance in soybean is difficult due to the quantitative nature of the trait, genetic variation within SCN populations, time required for phenotyping experimental soybean lines, and environmental factors affecting SCN reproduction. Quantitative trait loci associated with SCN resistance have been identified on 17 of the 20 soybean linkage groups, explaining 191% of the total phenotypic variation. Two major resistance genes, rhg 1 and Rhg 4, have been identified on linkage groups G and A2, respectively. Several minor resistance genes have been identified, but their importance varies with germplasm source and nematode race. Enhancement of SCN resistance in G. max may be achieved by interspecific hybridization with G. soja, the wild ancestor, or by engineering plants with candidate resistance genes such as Hs1pro-1. Key words: Genetic engineering, Glycine soja, soybean cyst nematode, molecular markers, resistance Winter, S. M. J., Rajcan, I. et Shelp, B. J. 2006. Le nmatode kyste du soja : difficults et possibilits. Can. J. Plant Sci. 86: 2532. Le nmatode kyste du soja (NKS) est le principal ravageur responsable des baisses de rendement de Glycine max. La lutte reste complique dans les cultures commerciales cause de la dure du cycle biologique du parasite, des changements frquents de la virulence des populations et de la facilit avec laquelle le NKS se propage dans le sol infest. Pour tre efficace, la lutte repose sur lassolement et lutilisation de cultivars rsistants, issus dun matriel gntique limit. Amliorer le soja pour quil rsiste davantage au NKS nest pas ais en raison de la nature quantitative du caractre gntique, de sa variation au sein des populations de NKS, du temps ncessaire pour valuer le phnotype des lignes exprimentales de soja et des paramtres environnementaux qui affectent la reproduction du NKS. Les locus du caractre quantitatif associ la rsistance au NKS ont t identifis sur 17 des 20 groupes de liaison du soja et expliquent 1 91% de la variation totale du phnotype. On a identifi deux importants gnes codant la rsistance, rhg 1 et Rhg 4, sur les groupes de liaison G et A2, respectivement. Plusieurs autres gnes secondaires ont t dcouverts, mais leur importance varie avec la provenance du matriel gntique et la race du nmatode. On pourrait amliorer la rsistance de G. max au NKS par une hybridation interspcifique avec G. soja, anctre sauvage de la culture, ou en modifiant gntiquement les plants qui portent les gnes susceptibles de coder la rsistance au ravageur tel Hs1pro-l. Mots cls: Gnie gntique, Glycine soja, nmatode kyste du soja, marqueurs molculaires, rsistance

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Soybean [Glycine max (L.) Merr.] is an annual herbaceous legume that is the worlds largest oilseed crop. World soybean production is 197.3 Mt yr1 [United States Department of Agriculture (USDA) 2004], with approximately 98% of this being produced in 10 countries: the United States of America, Brazil, Argentina, China, India, Paraguay, Canada, Indonesia, Bolivia and Italy (Wrather et al. 2001). The United States is the current world leader in soybean production, accounting for about 75.01 Mt or 38% of the global crop (USDA 2004), whereas Canada produces about 2.27 Mt (Statistics Canada 2004). The soybean cyst nematode (Heterodera glycines Ichinohe; SCN) was first observed in the United States in 1954 (Winstead et al. 1955) and in Canada in 1987 (Anderson et al. 1988). Losses in soybean production due to
1Current

SCN are greater than that of any other disease (Wrather et al. 2001), and are estimated at US$1.67B in the United States and US$20.9M in Canada, using a base price of US$220.50 t1. SCN infection commonly results in chlorotic and stunted host plants and yield losses of 1020% under normal conditions; however, it has caused up to 58% yield loss in severely infested regions of Argentina. In this review, we discuss challenges and opportunities associated with SCN. First, we consider the biology of SCN, current SCN management strategies, and virulence of SCN populations. Second, we describe the history of SCN-resistant cultivars and molecular marker technology for identification of SCN-resistant germplasm. Third, we emphasize interspecific breeding with wild soybean germplasm or engineering with natural resistance genes such as Hs1pro1 in order to develop new SCN-resistant cultivars. Abbreviations: FI, female index; QTL, quantitative trait locus or loci; LG, linkage group(s); PI, plant introduction; SCN, soybean cyst nematode
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address: Maizex Seeds Inc., R.R. #2, Tilbury, ON, Canada N0P 2L0. 2To whom correspondence should be addressed (email: bshelp@uoguelph.ca).

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BIOLOGY OF SOYBEAN CYST NEMATODE Soybean cyst nematodes are round microscopic worms of approximately 480 m in length (Ichinohe 1952). They live in the soil and are parasitic, infecting and reproducing in the roots of specific hosts in the Fabaceae family. Legumes such as soybean, pea (Pisum sativum L.), birdsfoot trefoil (Lotus corniculatus L.) and common bean (Phaseolus vulgaris L.) are efficient hosts for SCN. The life cycle of SCN is approximately 21 d at 25C (Lauritis et al. 1983), resulting in two to five generations per year. Embryos develop into juveniles within the eggs, then become dormant or hatch out and become infectious. Root diffusate from the host plant stimulates hatching and emergence (Tefft and Bone 1985; Schmitt and Riggs 1991). Depending on the origin of the egg, the juveniles may also have to emerge through the cyst or female body. These hatched juveniles, which are invasive for only 611 d (Robinson et al. 1987), migrate toward host roots in the water film between soil particles (Wallace 1963); migration is influenced by gradients in pH, amino acids, sugars or heat (Sharma and Sharma 1998). Upon arrival at a host plant the juveniles travel along the root, repeatedly pressing their lips against its surface and initiating stylet probing in search of a suitable entry point (Doncaster and Seymour 1973). Identification of a successful entry point increases probing frequency until the cuticle of the root is penetrated. Sites of invasion attract other individuals, initiating colony formation within the root. Juveniles preferentially migrate to zones of elongation and commonly penetrate the vascular tissues of the root (Atkinson and Harris 1989). Once migration slows, a change in stylet activity occurs and a specialized feeding structure known as the syncytium is induced. The syncytium is a large distinctive metabolically active group of plant cells that surround the nematodes head and provides a food source of plant nutrients. Once the feeding process is initiated, the juveniles become immobile (Sharma and Sharma 1998) and then enlarge and differentiate sexually into female or male adults (Mankau and Linford 1960; Mller et al. 1981). Adult males, which become mobile and move out of the root to fertilize the females, are unable to feed on plant tissue, resulting in a lifespan of only a few weeks (Zunke and Eisenback 1998). Adult female nematodes do not become mobile, and remain receptive for at least 2 mo (Triantaphyllou and Hirschmann 1962). Upon fertilization, their bodies swell with egg development, resulting in the rupture of the cortex and epidermis of the root while still attached to the feeding site. The initial egg production results in the deposition of 50 to 200 eggs in an external gelatinous matrix, which provides antimicrobial and desiccation protection (Riggs and Niblack 1999). Upon degeneration of the vaginal muscles, a further 50 to 200 eggs are deposited within the body cavity until the adult female dies. The body cavity enlarges to a lemon shape and turns from white to yellow to brown with development. Egg dormancy is initiated from mid-summer to October and is critical for winter survival (Yen et al. 1995). The life cycle begins as dormancy is broken and hatching occurs usually from April to June. Dormancy is

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dependent on several factors, including host phenology and soil temperature (Hill and Schmitt 1989). A unique feature of cyst nematodes is the ability of the female to turn her body into a protective case or cyst, in which her progeny may remain viable for more than 12 yr (Riggs and Niblack 1999). Cysts of SCN can spread easily via water, wind, soil and machinery. Soil environment influences the biological success of SCN. For examples, the duration of the complete life cycle is 18, 22 and 37 d at constant temperatures of 31, 24 and 17C, respectively (Alston and Schmitt 1988), temperatures of 20 and 30C result in cyst counts that are 30% less than those at 27C (Palmateer et al. 2000), non-irrigated soils tend to have higher SCN populations than irrigated soils (Koenning and Barker 1995), and cyst counts are positively correlated with percent sand (Avendao et al. 2004). MANAGEMENT OF SOYBEAN CYST NEMATODE Management of SCN is complicated by the prolonged viability of eggs within unique protective cysts (Riggs and Niblack 1999), combined with population variability (Riggs et al. 1981; Young 1984; Leudders 1989; Colgrove et al. 2002) and the numerous methods by which eggs are dispersed (Riggs and Niblack 1999). In addition, high input conditions that promote soybean growth also enhance the ability of SCN to reproduce (Ishibashi et al. 1973). The narrow host range of SCN, however, is a characteristic that can be exploited. The use of resistant soybean cultivars is the primary method for managing SCN in commercial production. They are effective tools, but their success depends on the genetic variability of the SCN population. In infested soils, resistant cultivars can yield 1050% higher than susceptible cultivars (Epps et al. 1981; Hartwig 1981; Young and Hartwig 1992). The presence of identifiable SCN races facilitates the effective use of resistant cultivars. Crop rotation is also a valuable tool in managing SCN populations. In Arkansas, a 3-yr rotation consisting of 1 yr of a non-host crop, 1 yr of a resistant soybean cultivar, and 1 yr of a susceptible soybean cultivar is effective (Slack et al. 1981). More northerly areas of soybean production often require longer rotations for equally effective results (Niblack 1993). The use of non-host crops places minimal selection pressure on nematode populations and stabilizes population growth. Many growers resist long crop rotations because non-host crops might be less-profitable and might require additional equipment (Riggs and Schuster 1998). Chemicals are costly, but have been effective in controlling SCN in the past. The efficacy of nematicides can be variable due to environmental conditions and improper application (Smith et al. 1991). Recent environmental and health concerns have resulted in the discontinuation of many highly successful nematicides (Roberts 1993), and control of SCN using these chemicals is no longer widely recommended (Riggs and Schuster 1998). Biological control theoretically provides an additional SCN management tool. An unknown fungus, Arkansas Fungus 18 (ARF18), develops on cysts, eggs and juveniles of H. glycines and causes 2060% reduction in nematode

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numbers in infested soils (Kim and Riggs 1991). Currently, the amount required for field application is not economical or practical for commercial use. VIRULENCE OF SOYBEAN CYST NEMATODE POPULATIONS Variation in the virulence of SCN populations was identified a few years after the development of the first commercially available SCN-resistant soybean cultivars (Ross 1962). This variation together with the biological diversity within populations resulted in the need for a classification scheme. To characterize the heterogeneous populations of SCN, a race scheme was developed (Golden et al. 1970) and later expanded (Riggs and Schmitt 1988). A female index (FI), defined as the ratio of the number of females that develop on differential lines (different sources of resistance) to those that develop on the standard susceptible cultivar Lee, multiplied by 100, is used to determine the race identity of SCN populations; an arbitrary value of < 10% is designated as resistant (Golden et al. 1970). The race characterization scheme describes SCN populations in broad terms, and permits intra-race variability (Riggs et al. 1981; Riggs and Schmitt 1988; Riggs and Schmitt 1991; Riggs et al. 1991; Rao-Arelli et al. 1992). Several concerns have been expressed with the race classification scheme. Ambiguity about race designation of a SCN population is common and may be attributed to the use of differential host plants from different sources, variability in inoculum preparation, and the inability to completely recover all cysts (Riggs et al. 1988). Other factors such as the time of sampling and the temperature during testing greatly influence race designations (Palmateer et al. 2000; Colgove et al. 2002). Directional selection studies using resistant cultivars revealed unpredictable frequency changes in virulent individuals within SCN populations (Riggs et al. 1981; Young 1984; Leudders 1989; Anand et al. 1995; Noel and Edwards 1996; Colgrove et al. 2002). The race scheme only considers the average phenotype and not the genetic diversity of the population (Niblack et al. 2002). The HG (named after Heterodera glycines) Type test is a classification scheme with enhanced flexibility to describe SCN population variation (Niblack et al. 2002). The ability of a population to infect seven different indicator lines (i.e., 1. PI 548402, 2. PI 88788, 3. PI 90763, 4. PI 437654, 5. PI 309332, 6. PI 89772 and 7. PI 548316), as well as the standard susceptible line Lee 74, is evaluated under standardized bioassay conditions. If fewer than 100 females are observed on Lee 74, the test is discarded and repeated. The population type is then based on the identifier numbers of the indicator lines that exhibit a FI 10. For example, a population that produces FI 10 on PI 548402, PI 88788 and PI 89772 is an HG Type 1.2.6. The HG Type test does not identify genotypes within a population, and SCN populations with the same HG designation may not behave in the same way because they can easily differ in characteristics not measured by the test. HG Type testing does permit more accurate management recommendations than the Race scheme described above because of the additional resistant germplasm used in the test, and the test is easily expanded as new soybean germplasm is released and deployed.

SOYBEAN CYST NEMATODE RESISTANT SOYBEAN CULTIVARS The first SCN-resistant soybean sources identified in the mid 1950s were plant introductions PI 90763 and PI 84751 and cultivars Ilsoy and Peking (Ross and Brim 1957). These sources of resistant germplasm were undesirable for commercial use due to the presence of a black seed coat. A major resistance gene (Rhg4) was later identified as closely linked with the seed coat colour gene (Matson and Williams 1965). This linkage made it difficult for breeders to develop SCN-resistant cultivars with a desirable yellow seed coat. Peking was chosen as the initial parental source of resistance due to its agronomic characteristics, and after one cycle of breeding, three SCN-resistant cultivars, Custer, Pickett and Dyer were released in 1965, 1967 and 1968, respectively (Brim and Ross 1966; Hartwig and Epps 1968; Leudders et al. 1968). These cultivars were not widely grown due to their relatively low yields, but they provided the parentage base for future cultivars resistant to Races 1 and 3 (Anand et al. 1998). In the 1970s, a second cycle of breeding resulted in three more cultivars from each of Custer, Pickett and Dyer. Shortly after their release a new SCN race, Race 4, was identified (Hartwig 1981), prompting another germplasm evaluation, which identified resistance to Race 4 in PI 88788, PI 89772, PI 87631-1, Cloud, Columbia, Peking, PI 84751 and PI 90763 (Epps and Hartwig 1972). Subsequently, breeding with PI 88788 as a resistant parent resulted in the release of the first cultivar, Bedford, with resistance to SCN Race 4 (Hartwig and Epps 1978). Later revisions to the race classification scheme classified PI 88788 as resistant to Race 14. Forrest, the most widely grown cultivar in the southern United States in the 1970s, derived SCN resistance to Races 1 and 3 from Dyer, and was estimated to prevent US$405M in crop losses from 1975 to 1980 (Bradley and Duffy 1982). However, repeated use of resistant germplasm sources resulted in the development of new races of SCN (Riggs et al. 1981). An additional germplasm screen identified PI 437654 as resistant to all SCN races (Anand et al. 1988; Diers et al. 1997) and it was used in the development of the resistant cultivar Hartwig (Anand 1992). Cyst X, the most recent addition to the list of SCNresistant cultivars, was derived from a cross between Williams 82 and Hartwig (Vierling et al. 2000). It was made available to public and private breeding programs in 1997 and patent protected in 2000. The locus responsible for Cyst X resistance is on linkage group (LG) B1 and explains more than 90% of the total phenotypic variation to Race 3 SCN (Vierling et al. 1996). Peking, PI 88788 and PI 209332 were used extensively as sources for SCN resistance in breeding programs, resulting in a narrow resistant germplasm base (Diers and Arelli 1999). In the mid-western United States, 80% of public SCN-resistant cultivars released during the 1990s and 93% of private industry cultivars available in 1998 derived SCN resistance from PI 88788 (Diers and Arelli 1999), whereas in Ontario, Canada, all cultivars available in 2005 derived resistance from PI 88788 (Ontario Oil and Protein Seed

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Crop Committee 2005). Genetic diversity studies suggested that many sources of resistance are genetically similar and cluster into two major groups (Diers et al. 1997). However, a more recent study identified that the resistant differentials used in the race test are indeed genetically diverse (Zhang et al. 1999), a result that could be explained by specific allelic composition at the resistant loci, as described recently for Rhg1 (Brucker et al. 2005). The presence of such allelic differences at resistance loci provides impetus for using Glycine soja Sieb. and Zucc. as a source of novel resistance genes and alleles since allelic differences between G. soja and G. max are common and well established (Maughan et al. 1995; Powell et al. 1996). MOLECULAR MARKERS ASSOCIATED WITH SOYBEAN CYST NEMATODE RESISTANCE Molecular markers allow traits of interest to be identified, mapped and identified in subsequent generations. Linkage groups (LG) reflect the nature of the relationships among molecular markers within and among individual chromosomes. The use of molecular markers in combination with phenotypic data enables regions on the chromosome to be associated with a particular trait. If many genes control the trait of interest, the associated region identified is termed a quantitative trait locus (QTL). A recent genetic linkage map of soybean, which has over 1000 SSR markers saturating 20 linkage groups (Song et al. 2004), allows researchers to accurately assign specific genetic markers associated with traits of interest or QTL to LGs. Breeding for SCN resistance using either the classical approach or molecular markers has been complicated by the length of time for screening plant material and control of the trait by multiple genes. Previous research indicates that SCN resistance QTL are located on 17 of the 20 LG within the soybean genome, and explain 191% of the total phenotypic variation [for a review, see Concibido et al. (2004)]. A major QTL, designated as rhg1, is located on LG G associated with the RFLP marker C006V (Concibido et al. 1994) and explains 54% of the total phenotypic variation in disease response in PI 209332 to Race 6, 50% to Race 3 and 35% to Race 1 (Concibido et al. 1996). This multiple-race (non-race specific) response suggests that the gene-for-gene theory does not apply to SCN resistance, and that despite the common use of the term resistance, it is more likely a case of genes/loci carrying either partial resistance or tolerance to SCN, which are terms rarely used in the SCN literature. Separate studies indicated that Peking (Chang et al. 1997; Concibido et al. 1997; Meksem et al. 2001), PI 90763 (Concibido et al. 1997), PI 88788 (Concibido et al. 1997), PI 209332 (Concibido et al. 1994), PI 89772 (Yue et al. 2001b), PI 437654 (Webb et al. 1995), Peking + PI 437654 (Prabhu et al. 1999) and Peking + PI 88788 + PI 90763 (Heer et al. 1998) have the major QTL on LG G near rhg1. The resistance conferred by rhg1 to multiple SCN races led to extensive mapping of this region, with the SSR, Satt309, being identified only 0.4 cM from the rhg1 gene (Cregan et al. 1999). A second major QTL for SCN resistance on LG A2 has been designated as the Rhg4 gene. It is closely linked to the

i locus controlling seed coat colour (Matson and Williams 1965). This region was the first SCN-resistant locus to be mapped using molecular markers (Weisemann et al. 1992). In PI 209332, LG A2 explains 15% of the total phenotypic variation to SCN Race 3 (Concibido et al. 1994) and a significant portion of the variation in Peking (Mahalingam and Skorupska 1995; Chang et al. 1997; Meksem et al. 2001), PI 437654 (Webb et al. 1995) and Peking + PI 88788 + PI 90763 (Heer et al. 1998). Although two additional recessive genes, rhg2 and rgh3, have been named and hypothesised as part of a three recessive gene model (Caldwell et al. 1960), they are yet to be assigned to a linkage group on the soybean genome map. It is conceivable that some of the identified QTL are located at or near one of these two genes. Many minor QTL have been identified on LG A1, B1, B2, C1, C2, D1a, D2, E, F, G, H, I, J, L, M and N (Concibido et al. 2004). The total number of QTL per LG range from one (e.g., LG H and I) to four (LG G) (Concibido et al. 2004). The position of each QTL on the soybean genetic map, however, can be considered tentative at best due to differences in size and structure of mapping populations, DNA marker platforms and limited number of DNA markers at the time of publication of each report (Concibido et al. 2004). Perhaps not surprisingly, the greatest number of resistant QTL are identified in Peking (nine independent QTL) as the oldest and most frequently studied source of SCN resistance, whereas low numbers are identified in recent sources such as PI 88788 or PI 90763 (one and two QTL, respectively) (Concibido et al. 2004). The allelic diversity found at the minor QTL is similar to that found at major QTL, but the amount of variation explained tends to be smaller for the minor QTL. The existence of most minor QTL has often been reported only once and requires independent confirmation; this is a common deficiency with QTL mapping. The most consistent reports are those for the rhg1 and Rhg4 loci (Concibido et al. 2004). Crosses have revealed significant resistance not identified in either parent. For example, in the cross Peking by PI 437654, two resistant QTL to SCN Race 3 were located on LG B1 (Vierling et al. 1996). These QTL on LG B1, identifiable by RFLP markers A006 and A567, explained 91 and 1% of the total phenotypic variation, respectively. If the sources of resistance are considered independently, a resistant QTL is only identified on LG B1 at A567 in PI437654, and it explains 27% of the total phenotypic variation (Webb 2003). The QTL on LG B1 is not derived from Peking (Concibido et al. 2004). However, it is known to be present in resistant source PI 89772, but it only explains 717% of the resistance to SCN Races 1, 2 and 5 (Yue et al. 2001b). On the other hand, the number of races showing significant response at a QTL ranges from one to five (LG G, rhg1 locus for races 1, 2, 3, 5 and 14) per locus (Concibido et al., 2004). Resistant QTL can be found in several regions on an individual linkage group. For example, on LG G, three loci are associated with SCN resistance in addition to the major gene, rhg1 (Concibido et al. 2004). The SCN resistant sources Peking (Concibido et al. 1997), PI 438489B (Yue et al. 2001a) and PI 468916 (Wang et al. 2001) have resistant

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QTL at other locations on LG G. Of the three additional regions of SCN resistance on LG G, all are associated with resistance to Race 3 SCN, suggesting a possible rearrangement of the resistant genes on LG G. Multiple QTL for SCN resistance have been identified on LGs A1, B1, B2, C2, D1a, D2, E, G and M [for a review, see Concibido et al. (2004)]. Minor loci for SCN resistance vary with regards to race specificity. For example, a minor locus for resistance to SCN Race 3 occurs on LG J in resistant sources PI 209332 (Concibido et al. 1996) and PI 90763 (Concibido et al. 1997). A minor QTL is also present on LG E in P438489B and PI 468916 and associated with resistance to SCN Races 2 and 14, and 3, respectively (Wang et al. 2001;Yue et al. 2001a). Significant interaction between major and minor markers associated with SCN resistance has been previously identified (Mahalingam and Skorupska 1995; Webb et al. 1995; Chang et al. 1997; Kilo et al. 1997; Heer et al. 1998; Prabhu et al. 1999; Meksem et al. 2001; Wang et al. 2001; Yue et al. 2001b). For example, single factor ANOVA does not significantly associate either Rhg4 or rhg1 with SCN resistance to Race 3 in a Flyer Hartwig population; however, their interaction accounts for 16% of the total phenotypic variation (Prabhu et al. 1999). Interaction between significant markers has been attributed to epistasis (Heer et al. 1998; Yue et al. 2001a; Concibido et al. 2004). INTERSPECIFIC BREEDING WITH GLYCINE SOJA Recently, soybean breeders and geneticists began to use G. soja as an alternative source of SCN resistance for the improvement of elite cultivars. This wild ancestor of the domesticated soybean enables many common interspecific obstacles such as infertility and pod abortion to be circumvented. In G. max G. soja populations, three backcrosses to G. max are required to attain a reasonable number of agronomically suitable lines (Junyi et al. 1982; Ertl and Fehr 1985; Carpenter and Fehr 1986). Loci associated with SCN resistance have been documented in a cross between G. max (A81-356022) and G. soja (PI 468916) (Wang et al. 2001). Significant QTL, which derive resistance from G. soja, are located on LG G and E, and respectively, explain 27 and 23% of the total phenotypic variation to SCN Race 3. The region on LG G is not believed to be a major SCN resistance gene as the QTL is located 230 cM away from the rhg1 gene. Therefore, this QTL may be unique as it has not previously been associated with SCN resistance. By contrast, the QTL on LG E may or may not be unique to G. soja as this region has previously been associated with SCN resistance in G. max (Yue et al. 2001a). G. soja-specific QTL could represent unique genes for broadening SCN resistance in G. max cultivars, but little information on their use is available. The development and release of the first cultivars incorporating such QTL may take some years due to the backcrossing and linkage drag issues. In addition to the preliminary efforts described above, it is reasonable to believe that the more than 1000 plant introductions of G. soja available at the USDA soybean germplasm collection in Urbana-Champaign, IL, carry unutilized genes and alleles for SCN resistance.

CLONING OF CANDIDATE SOYBEAN CYST NEMATODE RESISTANCE GENES In recent years, the ability to genetically engineer plants has revolutionized the possibilities for nematode control (Williamson 1999; Atkinson et al. 2003; McLean et al. 2003). This approach typically involves the manipulation and incorporation of various genes that could directly interfere with feeding cells or the nematode. However, this could also involve genes of uncertain function. For example, there are claims that candidate genes for rhg1 and Rhg4 have been cloned, and that both genes are receptor kinases (see Concibido et al. 2004). However, it is unclear if these candidate genes are associated with SCN resistance, and complementation studies to confirm the identity of the genes have not been reported. Another gene known as Hs1pro-1 reportedly provides resistance against the beet cyst nematode, H. schachtii Schmidt (Cai et al. 1997), and interspecific hybridizations between H. schachtii and H. glycines are fertile, with the cross being successfully taken through to the F2 generation (Potter and Fox 1965). This lack of a reproductive barrier between these two species of cyst nematode demonstrates an extremely high genetic relatedness, and although different plant hosts are parasitized, the molecular mechanisms employed are likely similar. Thus, the mechanism of resistance provided by the Hs1pro-1 gene against H. schachtii may also be effective against H. glycines. Interestingly, the presence of an Hs1pro-1 homologue in soybean fails to distinguish between susceptible and tolerant germplasm, but complementation studies to confirm the identity of the gene have not been conducted (see Concibido et al. 2004). Further analysis of these candidate resistance genes is necessary to provide insights into the genetic mechanisms involved. CONCLUSIONS The SCN is a serious pest of the soybean crop. The reproduction of SCN is very dependent on soil environment, and soil variables such as temperature need to be considered in research on SCN resistance. Management of SCN is primarily dependent upon a combination of crop rotation and resistant cultivars, which minimize selection pressure on SCN populations and reduce soil populations. It has been difficult to separate SCN populations that differ in reproductive ability on known sources of resistance. The Race scheme and HG Type test are common methods of characterizing populations with respect to known resistant soybean germplasm and can be used as tools in the management of SCN. SCN populations often display broad variation in virulence, and can rapidly overcome resistance in commercially available soybean cultivars, which are derived from a limited genetic base. A number of different resistance genes in a single cultivar may be necessary to ensure long-term resistance. Recently, molecular markers have become important for identifying QTL for SCN resistance and predicting plant phenotype within experimental populations. This ability, combined with marker-assisted selection, is an invaluable tool for plant breeders. Wild soybean germplasm represents an alternative source of SCN resistance for breeding into elite cultivars, but its use is often avoided because of possi-

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CANADIAN JOURNAL OF PLANT SCIENCE Chang, S. J. C., Doubler, T. W., Kilo, V. Y., Abu-Thredeih, J., Prabhu, R., Freire, V., Suttner, R., Klein, J., Schmidt, M. E., Gibson, P. T. and Lightfoot, D. A. 1997. Association of loci underlying field resistance to soybean sudden death syndrome (SDS) and cyst nematode (SCN) race 3. Crop Sci. 37: 965971. Colgrove, A. L., Smith, G. S., Wrather, J. A., Heinz, R. D. and Niblack, T. L. 2002. Lack of predictable race shift in Heterodera glycines Infested field plots. Plant Dis. 86: 11011108. Concibido, V. C., Denny, R. L., Boutin, S. R., Hautea, R., Orf, J. H. and Young, N. D. 1994. DNA marker analysis of loci underlying resistance to soybean cyst nematode (Heterodera glycines Ichinohe). Crop Sci. 34: 240246. Concibido, V. C., Denny, R. L., Lange, D. A., Orf, J. H. and Young, N. D. 1996. RFLP mapping and marker-assisted selection of soybean cyst nematode resistance in PI 209332. Crop Sci. 36: 16431650. Concibido, V. C., Diers, B. W. and Arelli, P. R. 2004. A decade of QTL mapping for cyst nematode resistance in soybean. Crop Sci. 44: 11211131. Concibido, V. C., Lange, D. A., Denny, R. L., Orf, J. H. and Young, N. D. 1997. Genome mapping of soybean cyst nematode resistance genes in Peking, PI 90763, and PI 88788 using DNA markers. Crop Sci. 37: 258264. Cregan, P. B., Mudge, J., Fickus, E. W., Danesh, D., Denny, R. and Young, N. D. 1999. Two simple sequence repeat markers to select for soybean cyst nematode resistance coditioned by the rhg1 locus. Theor. Appl. Genet. 99: 811818. Diers, B. W. and Arelli, P. R. 1999. Management of parasitic nematodes of soybean through genetic resistance. Pages 300306 in H. E. Kauffman, ed. Proc. of the World Soybean Research Conference. 6th. Chicago, Illinois. Superior Printing, Champaign, IL. Diers, B. W., Skorupska, H. T., Rao-Arelli, A. P. and Cianzio, S. R. 1997. Genetic relationships among soybean plant introductions with resistance to soybean cyst nematodes. Crop Sci. 37: 19661972. Doncaster, C. C. and Seymour, M. K. 1973. Exploration and selection of penetration site by Tylenchida. Nematologica 19: 137145. Epps, J. M. and Hartwig, E. E. 1972. Reaction of soybean varieties and strains to race 4 of the soybean cyst nematode. J. Nematol. 4: 222 (Abstr.). Epps, J. M., Young, L. D. and Hartwig, E. E. 1981. Evaluation of nematicides and resistant cultivar for control of soybean cyst nematode race 4. Plant Dis. 65: 665666. Ertl, D. S. and Fehr, W. R. 1985. Agronomic performance of soybean genotypes from Glycine max ( Glycine soja crosses. Crop Sci. 25: 589592. Golden, A. M., Epps, J. M., Riggs, R. D., Duclos, L. A., Fox, J. A. and Bernard, R. L. 1970. Terminology and identity of infraspecific forms of the soybean cyst nematode (Heterodera glycines). Plant Dis. Rep. 54: 544546. Hartwig, E. E. 1981. Breeding productive soybean cultivars resistant to the soybean cyst nematode for the southern United States. Plant Dis. 65: 303307. Hartwig, E. E. and Epps, J. M. 1968. Dyer soybeans. Crop Sci. 8: 402. Hartwig, E. E. and Epps, J. M. 1978. Registration of Bedford soybeans. Crop Sci. 18: 915. Heer, J. A., Knap, H. T., Mahalingam, R., Shipe, E. R., Arelli, P. R. and Matthews, B. F. 1998. Molecular markers for resistance to Heterodera glycines in advanced soybean germplasm. Mol. Breed. 4: 359367. Hill, N. S. and Schmitt, D. P. 1989. Influence of temperature and soybean phenology on dormancy induction of Heterodera glycines. J. Nematol. 21: 361369.

ble linkage drag of deleterious traits. The development and use of molecular markers for identifying desirable DNA regions has made, and will continue to make, the use of wild germplasm more efficient. Several candidate SCN resistance genes have reportedly been cloned, but further analysis is necessary to provide insights into the mechanisms of genetic resistance. ACKNOWLEDGEMENTS S.M.J.W. was supported by funds to I.R. and B.J.S. from the Food Systems Biotechnology Center at the University of Guelph, the Ontario Soybean Growers, and the Ontario Ministry of Agriculture and Food.
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