Developing standardized methods for breeding preharvest sprouting resistant wheat, challenges and successes in Canadian wheat

Preharvest sprouting (PHS) in spring wheat (Triticum aestivum L.) and durum wheat (T. turgidum L. var durum) causes significant economic losses due to a reduction in grain yield, grain functionality and viability of seed for planting. Average annual estimated losses in Canada are about $100 million. Genetic resistance to PHS reduces these losses. Development of PHS resistant cultivars is complicated by the effects of factors under genetic control, such as spike morphology, seed dormancy, environment, and kernel diseases. Resistance to PHS has been a breeding priority since the late 1960s. Development of RL4137, which is the primary source of PHS resistance in the Canada Western Red Spring market class, has led to cultivar improvements. A white-seeded derivative of RL4137 is the primary source of PHS in the Canada Prairie Spring White and Canada Western Hard White Spring wheat market classes. Procedures to select for PHS resistance vary among breeding programs, market classes and by degree of inbreeding. Methods include artificial sprouting of intact spikes, germination tests, natural weathering in field trials, artificial weathering trials, and indirect assessment of sprouting by measuring Hagberg falling number. Although many genetic loci have been attributed to preharvest sprouting resistance, application of molecular markers is currently limited due to the complex inheritance of the trait. In Canada, cultivars are characterized for their relative level of PHS resistance and the information is made available to producers.     

Methods for characterization of preharvest sprouting resistance in a wheat breeding program

Preharvest sprouting (PHS) can be a serious problem in western Canada resulting in economically important yield and grade losses for wheat producers. Improved PHS resistance is an important breeding objective and wheat breeders require effective and reliable methods to select for PHS resistance. The objective of this study was to evaluate whether differences exist between field and artificial weathering in the evaluation of PHS resistance in breeding lines and to determine whether sprouting scores are a good estimator of PHS resistance when compared to Hagberg falling numbers. Results suggest that both field and artificial weathering can be effectively used to screen for PHS resistance in wheat breeding lines. Although in this study, field weathering resulted in greater sprouting damage than artificial weathering, the ability to control conditions may make artificial weathering the more reliable test for PHS resistance in a breeding program. Mean sprouting scores greater than 7.5 consistently identified lines with low PHS resistance; hence, sprouting scores can be used to discard the most sprouting susceptible lines. However, falling numbers appear to be more reliable to evaluate the PHS resistance of advanced lines. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mapping QTLs for pre-harvest sprouting traits in the spring wheat cross ‘RL4452/AC Domain’

Pre-harvest sprouting (PHS) in spring wheat (Triticum aestivum L.) is a major downgrading factors for grain producers and can significantly reduce end-use quality. PHS resistance is a complex trait influenced by genotype, environment and plant morphological factors. A population of 185 doubled haploid (DH) lines from the spring wheat cross ‘RL4452/AC Domain’ were used as the mapping population to detect quantitative trait loci (QTLs) associated with three PHS traits, germination index (GI), sprouting index (SI) and falling number (FN). Six major QTLs linked with PHS traits were mapped on bread wheat chromosomes 3A, 3D, 4A (2 loci), 4B and 7D. ‘AC Domain’ alleles contributed to PHS resistance on 3A, 4A (locus-2) and 4B, and ‘RL4452’ alleles contributed resistance on 3D, 4A (locus-1) and 7D. QTLs detected on chromosome 4B controlling FN (QFN.crc-4B), GI (QGI.crc-4B) and SI (QSI.crc-4B) were coincident, and explained the largest amount of phenotypic variation in FN (22%), GI (67%) and SI (26%), respectively.     

Three classes of loci controlling preharvest sprouting in rye (Secale cereale L.) discerned by means of bidirectional selective genotyping (BSG)

Bidirectional selective genotyping (BSG) carried out on recombinant inbred lines (RILs) derived from the 541 × Ot1-3 intercross revealed three classes of selection responsive loci underlying preharvest sprouting (PHS) in rye. Ten PHS directional loci (PHSD) located on chromosomes 1RL (3), 3RS (2), 3RL (2), 5RL (2), and 7RS (1) responded significantly to both directions of the disruptive selection and were epistatic to the remaining two classes. Nine PHS resistance loci (PHSR) mapped on chromosomes 1RS, 1RL, 2RS, 3RS, 4RS, 5RS, 5RL, and 6RL (2) responded only to selection for sprouting resistance, being neutral for selection carried out in opposite direction. Eight PHS enhancing loci mapped on chromosomes 2RL, 3RL, 4RL, 5RL, 6RS, and 7RS (3) were affected by selection for sprouting susceptibility and did not respond to selection for sprouting resistance. Map positions of the selection responsive loci coincided with QTLs for PHS and alpha-amylase activity (AA) detected earlier, but BSG coupled with molecular mapping increased precision of PHS dissection in rye. Efficient strategy of marker assisted selection for preharvest sprouting resistance in rye should be based on PHSD and PHSR loci.     

Characterization of DFR allelic variations and their associations with pre-harvest sprouting resistance in a set of red-grained Chinese wheat germplasm

Pre-harvest sprouting (PHS) of wheat greatly reduces the quality and economic value of grain, and PHS resistance is one of the most important traits in wheat breeding. Red-grained wheat varieties are generally more resistant to PHS than white-grained ones; however, some are still susceptible. The red pigment of red-grained wheat is synthesized through the flavonoid biosynthetic pathway, in which the dihydroflavonol-4-reductase gene (DFR) is one of the genes involved in anthocyanin synthesis. In this study, a set of 120 red-grained Chinese wheat cultivars and lines with distinct PHS resistance were used to characterize TaDFR genotype variations and their association with PHS resistance. Whereas no variation or functional variation of TaDFR genes was detected on chromosomes 3A and 3D, a novel TaDFR allele, designated TaDFR-Bb, was explored on chromosome 3B. Compared with TaDFR-Ba, an 8 bp insertion (CTCTAGGA) was identified in the promoter region of TaDFR-B in most of the PHS resistant red-grained wheat varieties and advanced lines. Based on this, a CAPS marker was designed and validated with a set of Chinese red-grained wheat cultivars and lines with distinct PHS resistance. In most cases, TaDFR-Bb was associated with higher PHS resistance. An association study indicated that wheat varieties with the 8 bp insertion (average seed germination index 23.6 %) were significantly more resistant (P < 0.01) to PHS than those without the insertion (average seed germination index 69.5 %). Further study on gene expression demonstrated that the insertion led to increased TaDFR-B expression in cultivars with PHS resistance. Transient expression of TaDFR-B in coleoptiles of wheat cv. Chinese Spring revealed that increasing TaDFR gene expression did not induce the synthesis of anthocyanins.     

Integration of marker-assisted selection for resistance to pre-harvest sprouting with selection for grain-filling rate in breeding of white-kernelled wheat for the Chinese environment

Few Chinese high yielding white-grained wheat cultivars possess sufficient dormancy to avoid pre-harvest sprouting (PHS). Because the field evaluation of PHS is difficult, the identification of informative molecular markers is a priority for improving the level of dormancy. In this report, the effectiveness of phenotypic and genotypic selection was compared. Four microsatellite loci Xbarc57, Xbarc294, Xbarc310 and Xbarc321, mapped on the short arm of chromosome 3A, were used for selection in white-grained wheat F₅ lines which were also selected on the basis of their grain filling rate (GFR). One of these (later designated cv. Zhongmai911) was further selected on the basis of its allelic constitution at the four SSR loci. This cultivar combines a high level of PHS resistance with high grain yield. The results suggested that rapid GFR and PHS resistance can be bred simultaneously.     

Mapping quantitative trait loci controlling pre-harvest sprouting resistance in a red × white seeded spring wheat cross

Hard white wheat (Triticum aestivum L.) is a value-added product because of its processing advantages over red wheat; however, white wheat tends to be more susceptible to pre-harvest sprouting (PHS). To identify quantitative trait loci (QTLs) associated with PHS tolerance, we developed a doubled haploid (DH) mapping population from the cross AC Domain (red seeded) × White-RL4137 (white seeded). A genetic map was constructed using microsatellite markers located on chromosome groups 3, 4, 5 and 6. A population of 174 DH lines was characterized for important aspects of PHS including sprouting index, germination index, Hagberg falling number and seed coat colour. A total of 11 QTLs were identified on group 3 chromosomes and on chromosome 5D. Seven QTLs associated with the PHS traits were found to be co-incident with seed coat colour on chromosomes 3A, 3B and 3D. The 5D PHS QTL was notable because it is independent of seed coat colour.     

Increased grain dormancy in white-grained wheat by introgression of preharvest sprouting tolerance QTLs

White-grained wheat cultivars have long been recognized to be less resistant to preharvest sprouting (PHS) than the red-grained ones. Previously two QTLs for grain dormancy, QPhs.ocs-3A.1 (QPhs-3AS) and QPhs.ocs-4A.1 (QPhs-4AL) were identified in a highly dormant Japanese red wheat, Zenkoujikomugi (Zen). Aiming at improvement of PHS tolerance in white-grained wheat, the introgression effect of these two QTLs in a white-grained population consisting of 40 recombinant inbred lines (RILs) developed from a cross between Zen and white-grained Spica was examined here. Random 20 RILs with red grains were also developed from the same cross and used as a control population. The RILs were grown in the field and in the glasshouse to evaluate the grain dormancy by germination test. Several SSR markers closely linked to the QPhs-3AS and QPhs-4AL were used to estimate the alleles at the QTLs. Dormancy variation in the RILs was significantly associated with the differences for grain color and the alleles at QPhs-3AS over several years. Although allelic variation was detected in a SSR marker closely linked to QPhs-4AL there was no difference in germination data between the Zen-allele and the Spica-allele groups. As expected, the red-grained RILs with the Zen allele at QPhs-3AS were the most dormant. Some white-grained RILs with the Zen allele at QPhs-3AS showed higher dormancy compared to the red-grained RILs with the alternative allele. These results demonstrated that introgression of the QPhs-3AS gene could contribute to the increased grain dormancy in white-grained wheat.     

Simultaneous selection for yield-related traits and susceptibility to Fusarium head blight in spring wheat RIL population

Fusarium head blight (FHB), caused by the fungal plant pathogen Fusarium, is a fungal disease that occurs in wheat and can cause significant yield and grain quality losses. The present paper examines variation in the resistance of spring wheat lines derived from a cross between Zebra and Saar cultivars. Experiments covering 198 lines and parental cultivars were conducted in three years, in which inoculation with Fusarium culmorum was applied. Resistance levels were estimated by scoring disease symptoms on kernels. In spite of a similar reaction of parents to F. culmorum infection, significant differentiation between lines was found in all the analyzed traits. Seven molecular markers selected as linked to FHB resistance QTLs gave polymorphic products for Zebra and Saar: Xgwm566, Xgwm46, Xgwm389, Xgwm533, Xgwm156, Xwmc238, and Xgwm341. Markers Xgwm389 and Xgwm533 were associated with the rate of Fusarium-damaged kernels (FDK) as well as with kernel weight per spike and thousand kernel weight in control plants. Zebra allele of marker Xwmc238 increased kernel weight per spike and thousand kernel weight both in control and infected plants, whereas Zebra allele of marker Xgwm566 reduced the percentage of FDK and simultaneously reduced the thousand kernel weight in control and infected plants.     

Divergent selection for sprouting resistance in spring wheat

The development of sprouting-resistant spring-wheat ( Triticum aestivum L.) cultivars is a major breeding objective in many wheat-producing regions. Sprouting resistance is thought to be associated with delayed maturity. The primary objective of this study was to measure the reciprocal effects of selection for sprouting resistance and maturity. Two experiments were conducted over a 3–4-year period in Saskatoon, Canada. In the first experiment, two populations of hard red spring wheat were subjected to divergent selection (k = 10%) for maturity. In the second experiment, six populations derived from crosses between two sprouting-resistant, late-maturing, white-grained cultivars ('AUS1293' and 'AUS1408') and three early maturing, red–grained cultivars ('Park', 'PT516' and 'Roblin'), were subjected to divergent selection (k = 10%) for sprouting resistance. Selection for earliness reduced sprouting resistance in one population but had no effect in the second. For both populations, earlier maturity was associated with higher test weight but lower grain yield. In the second experiment, selection for increased sprouting resistance was effective, with realized heritabilities averaging 0.74. Increased sprouting resistance was associated with a slight delay (1–2.5 days) in time to spike emergence in four out of six populations, but had little effect on time to maturity in most populations. There was a trend towards redder grain in the sprouting-resistant selections. The recovery of sprouting-resistant, early maturing segregants was relatively low, averaging less than 10% over the six populations. In conclusion, selection for increased sprouting resistance can result in delayed maturity, but the magnitude of that delay will vary among populations.     

Relationship among planted and harvested kernel weights and grain yield and protein percentage in winter wheat

Summary Wheat (Triticum aestivum L.) kernel weight is an important yield component and seed quality factor that appears to be declining with recent cultivar releases in the major U.S. wheat region. The objectives of experiments were to detmrmine the relationship between planted and harvested 1000-kernel weights and their effect on grain yield and grain protein percentage. Twelve popular hard red winter wheat cultivars were grown with recommended practices at 10 Kansas locations for three years. Rank correlation coefficients between kernel weights and grain yields and protein percentages were calculated.Correlations of planted and harvested kernel weights over all cultivars with grain yield were inconsistent, being positive for some locations and years and negative for other locations and years. Planted kernel weights and grain yields of individual cultivars were not related, but harvested kernel weights and grain yields were correlated positively for eight of the 12 cultivars. Harvested kernel weights and grain protein percentages frequently were correlated positively at individual locations, but always were correlated negatively for individual cumtivars. It was concluded that high stable harvested wheat kernel weight is important as a component of grain yield under a range of environments and that improving this trait would benefit all segments of the wheat industry.Contribution No. 84-395-J, Department of Agronomy, Kansas State University, Manhattan, Kansas 66506 USA.     

红粒小麦Tamyb10单倍型检测及其与穗发芽抗性的关系

穗发芽是影响小麦品质和产量的重要因素之一,通常红粒小麦比白粒小麦具有较高的穗发芽抗性。转录因子Tamyb10是R-1基因的强候选基因,其表达与否和表达水平决定小麦籽粒颜色。为阐明Tamyb10单倍型与红粒小麦穗发芽抗性的关系,利用已开发的Tamyb10基因分子标记,检测119份来自不同麦区的红粒小麦材料,发现Tamyb10基因(Tamyb10-A1、Tamyb10-B1和Tamyb10-D1位点)可分成7类单倍型,分别是baa、aba、bba、aab、bab、abb和bbb。Tamyb10-D1对穗发芽抗性影响最大,Tamyb10-B1次之,Tamyb10-A1作用最小。Tamyb10单倍型没有明显的地域分布特点,但在东北春麦区,Tamyb10单倍型bbb与红粒品种的高穗发芽抗性相关。     

不同Wx蛋白缺失类型小麦穗发芽特性的初步研究

以11份Wx蛋白全部缺失类型（糯性），11份Wx蛋白部分缺失类型（非糯性）和大面积推广品种扬麦158、扬麦12（非糯性）为试验材料，以白皮品种豫麦47为穗发芽对照，研究了它们的穗发芽特性、α-淀粉酶活性和籽粒可溶性糖含量。结果表明，糯小麦的穗发芽率和籽粒可溶性糖含量均极显著高于非糯小麦，而α-淀粉酶活性与非糯小麦差异不显著。α-淀粉酶活性高是非糯小麦穗发芽的原因之一，两者呈指数关系，y=0.4145e^0.4864x(R²=0.3513*)；对于糯小麦，籽粒可溶性糖含量高可能是其穗发芽严重的诱导因素，两者亦呈指数关系，y=0.0001e^2.5599x(R²=0.5338**)。     

Conferring resistance to pre-harvest sprouting in durum wheat by a QTL identified in <Emphasis Type="Italic">Triticum spelta</Emphasis>

Pre-harvest sprouting (PHS) causes significant yield loss and degrade the end-use quality of wheat, especially in regions with prolonged wet weather during the harvesting season. Unfortunately, the gene pool of Triticum durum (tetraploid durum wheat) has narrow genetic base for PHS resistance. Therefore, finding out new genetic resources from other wheat species to develop PHS resistance in durum wheat is of importance. A major PHS resistance QTL, Qphs.sicau-3B.1, was mapped on chromosome 3BL in a recombinant inbred line population derived from ‘CSCR6’ (Triticum spelta), a PHS resistant hexaploid wheat and ‘Lang’, a PHS susceptible Australian hexaploid wheat cultivar. This QTL, Qphs.sicau-3B.1, is positioned between DArT marker wPt-3107 and wPt-6785. Two SCAR markers (Ph3B.1 and Ph3B.2) were developed to track this major QTL and were used to assay a BC₂F₈ tetraploid population derived from a cross between the durum wheat ‘Bellaroi’ (PHS susceptible) and ‘CSCR6’ (PHS resistant). Phenotypic assay and marker-assisted selection revealed five stable tetraploid lines were highly PHS resistant. This study has successfully established that PHS-resistance QTL from hexaploid wheat could be efficiently introgressed into tetraploid durum wheat. This tetraploid wheat germplasm could be useful in developing PHS resistant durum cultivars with higher yield and good end-use quality.     

Field evaluation of resistance to kernel infection and mycotoxin accumulation caused by Fusarium head blight in western Japanese wheat (Triticum aestivum L.) cultivars

Fusarium head blight (FHB) remains a serious problem that causes yield and grain quality losses, and mycotoxin accumulation in wheat production in western Japan. A 3-year field trial with artificial FHB inoculation was conducted to evaluate varietal characteristics of FHB resistance among 31 wheat cultivars/lines cultivated in western Japan, including one standard line. Severity of FHB, frequency of Fusarium-damaged kernels (FDK), deoxynivalenol concentration (DON), nivalenol concentration (NIV), and grain yield showed significant differences among years and among cultivars/lines. Interaction between years and cultivars/lines was also significant in these traits, but F values were larger for cultivars/lines than for the interaction. Correlation analysis showed that cultivars/lines with lower FHB severities tended to have lower FDK, DON and NIV, and a higher yield. Resistance to kernel infection (RKI), residuals calculated by regressing FDK against FHB severity, and resistance to mycotoxin accumulation (RTA), residuals calculated by regressing DON + NIV against FDK, also differed significantly among cultivars/lines. These results indicated that varietal differences in response to FHB symptom development, RKI and RTA exist among wheat cultivars/lines in western Japan. Such information is important to aid producers in controlling the disease and for breeders to improve FHB resistance and reduce mycotoxin accumulation in commercial wheat cultivars.     

Germplasm improvement for preharvest sprouting resistance in Chinese white-grained wheat: An overview of the current strategy

Most modern white-grained wheat (Triticum aestivum L.) cultivars in China lack sprouting resistance. Some white grain varieties showed lower preharvest sprouting percentage in different years. Tolerance to sprouting in most cultivars comes from the varieties, Wanxian White, Fulingxuxu White, Suiningtuotuo, and Yongchuan White. The α-amylase inhibitor gene under a wheat genetic background inhibited α-amylase activity and delayed sprouting. The falling number of six suspected sources, out of 835Chinese leading varieties since 1950', was independent of late maturityα-amylase. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance,heritability and breeding behaviour of three sources of resistance to Septoria tritici in wheat

Summary We examined the inheritance, heritability, and breeding behaviour of resistance to Septoria tritict in the spring wheat cultivars Seabreeze, Veranopolis, and IAS-20 in crosses to the susceptible Australian spring cultivar Gamenya. Resistance in Seabreeze was higher than that in Veranopolis and IAS-20. Inheritance of resistance in Seabreeze seems to be determined by at least three recessive genes. Resistance in Veranopolis and IAS-20 might be due to a single gene. Similarity of inheritance patterns and breeding behaviour of the resistance from Veranopolis and IAS-20 and the common ancestry of these two cultivars suggest that they may carry the same gene. Standard unit heritabilities of Septoria scores, as measured by correlation of F₂ plant with F₃ family mean data, were high (57–68%). Parent offspring genetic correlations confirmed previously reported associations between resistance and lateness and resistance and tall plant height, but correlations were not sufficiently high to be a major obstacle to selection. Resistance in Veranopolis and IAS-20 were easily recovered in first backcrosses while that in Sea breeze was more difficult to recover.Contribution from the Department of Agriculture Western Australia.     

Determination of the influence of climate on falling number of winter wheat in the dryland production areas of the Free State Province of South Africa

Preharvest sprouting (PHS) has been recognised as the primary cause of low falling numbers (FNs) in wheat. From recent research it is clear that there are a number of additional causes of low FNs, including late maturity ??-amylase. FNs of certain cultivars have varied considerably from year to year and across environments in the wheat producing areas of the Free State Province. In this study we aimed to determine whether climatic conditions could contribute to unexpectedly low FNs. Eleven cultivars were planted over a 5?year period in five different locations in the Free State Province. Total rainfall, minimum and maximum temperatures and growing degree days were determined during six environmental periods. Results from this study indicated that rainfall during the later stages of grain filling and grain maturity had a negative effect on the FNs of 7 of the 11 cultivars, while maximum temperatures during these growing periods were positively correlated with FN in 8 of the 11 cultivars. Minimum temperatures just prior to harvest could also determine the FNs of certain cultivars. Principal component analysis identified three groupings of cultivars varying in the frequency of low FNs experienced over the 5?years of the study. It is clear from this study that rainfall just prior to harvest, and therefore PHS, was not the major factor responsible for the low FNs obtained during this study.     

Grain dormancy in fixed lines of white-grained wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) grown under controlled environmental conditions

Pre-harvest sprouting (PHS) in wheat (Triticum aestivum L.) can be a significant problem, causing deleterious effects on grain quality. However, the adverse impacts of PHS can be reduced by introgressing genes controlling grain dormancy into white-grained bread wheat. Screening for grain dormancy typically involves germination testing of harvest-ripe grain grown in a glasshouse or field. However, the more uniform environmental conditions provided by temperature controlled glasshouses (i.e. controlled environmental conditions—CEC) may provide significant benefits for the assessment of grain dormancy. In this study, the dormancy phenotype of grain grown under CEC incorporating an extended photoperiod, was compared with 2 years of data from field grown material. Four dormant double haploid lines (derived from SW95-50213 and AUS1408) and two locally adapted non-dormant cultivars EGA Gregory and EGA Wills were compared in three replicated experiments grown under CEC (22 ± 3°C and 24 h photoperiod). The germination response of harvest-ripe grain was examined to assess the expression of grain dormancy. Two measures of germination, the predicted time to 50% germination (G ₅₀) and a weighted germination index, both clearly differentiated dormant and non-dormant lines grown under CEC. In addition, levels of grain dormancy were similar to field-grown plants. These results demonstrated that CEC with an extended photoperiod can be used for rapid and reliable characterisation of grain dormancy in fixed lines of bread wheat.