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.     

Identification of genomic regions associated with seed dormancy in white-grained wheat

Pre-harvest sprouting (PHS) in developing wheat (Triticum aestivum L.) spikes is stimulated by cool and wet weather and leads to a decline in grain quality. A low level of harvest-time seed dormancy is a major factor for PHS, which generally is a larger problem in white-grained as compared to red-grained wheat. We have in this study analyzed seed dormancy levels at the 92nd Zadok growth stage of spike development in a doubled-haploid (DH) white wheat population and associated variation for the trait with regions on the wheat genome. The phenotypic data was generated by growing the parent lines Argent (non-dormant) and W98616 (dormant) and 151 lines of the DH population in the field during 2002 and 2003, at two locations each year, followed by assessment of harvest-time seed dormancy by germination tests. A genetic map of 2681 cM was constructed for the population upon genotyping 90 DH lines using 361 SSR, 292 AFLP, 252 DArT and 10 EST markers. Single marker analysis of the 90 genotyped lines associated regions on chromosomes 1A, 2B, 3A, 4A, 5B, 6B, and 7A with seed dormancy in at least two out of the four trials. All seven putative quantitative trait loci (QTLs) were contributed by alleles of the dormant parent, W98616. The strongest QTLs positioned on chromosomes 1A, 3A, 4A and 7A were confirmed by interval mapping and markers at these loci have potential use in marker-assisted selection of PHS resistant white-grained wheat.     

QTL analysis for resistance to preharvest sprouting in rice (Oryza sativa)

Preharvest sprouting (PHS) is caused by early breaking of seed dormancy. In Sichuan, a major hybrid rice seed production area of China, PHS in hybrid seeds originated from ‘G46A’ parent may lead to severe yield loss, causing serious damage to agricultural production. To detect quantitative trait loci (QTLs) governing PHS, we developed an F₂ population of 164 plants derived from ‘G46B’ and ‘K81’, a near‐isogenic introgression line of G46B, with high level of resistance to PHS. PHS was evaluated under controlled field and laboratory conditions. Using simple sequence repeat markers, we constructed a linkage map from this population and identified three QTLs for PHS, namely qPSR2, qPSR5 and qPSR8, which were located on chromosomes 2, 5 and 8, respectively. Among these QTLs, qPSR8, residing in the interval between RM447 and RM3754 on chromosome 8, was the major QTL controlling PHS, for it had a relative high logarithm of the odds (LOD) score and explained 43.04% of the phenotypic variation. These results were correspondent to those identified in extreme low germination rate plants (ELGP) using linkage and linkage disequilibrium. At all loci, ‘K81’ was responsible for enhancing the resistance to PHS.     

Detection of QTLs for traits associated with pre-harvest sprouting resistance in bread wheat (Triticum aestivum L.)

Pre-harvest sprouting (PHS) is one of the serious problems for wheat production, especially in rainy regions. Although seed dormancy is the most critical trait for PHS resistance, the control of heading time should also be considered to prevent seed maturation during unfavorable conditions. In addition, awning is known to enhance water absorption by the spike, causing PHS. In this study, we conducted QTL analysis for three PHS resistant related traits, seed dormancy, heading time and awn length, by using recombinant inbred lines from ‘Zenkouji-komugi’ (high PHS resistance) × ‘Chinese Spring’ (weak PHS resistance). QTLs for seed dormancy were detected on chromosomes 1B (QDor-1B) and 4A (QDor-4A), in addition to a QTL on chromosome 3A, which was recently cloned as TaMFT-3A. In addition, the accumulation of the QTLs and their epistatic interactions contributed significantly to a higher level of dormancy. QDor-4A is co-located with the Hooded locus for awn development. Furthermore, an effective QTL, which confers early heading by the Zenkouji-komugi allele, was detected on the short arm of chromosome 7B, where the Vrn-B3 locus is located. Understanding the genetic architecture of traits associated with PHS resistance will facilitate the marker assisted selection to breed new varieties with higher PHS resistance.     

Comparative analysis of Australian and Canadian barleys for seed dormancy and malting quality

The association between high malting quality and pre-harvest sprouting (PHS) susceptibility is a key challenge when developing new malting barley varieties. A new malting barley variety Baudin has successfully combined high malting quality and PHS tolerance. A doubled haploid population was developed for mapping PHS tolerance and seed dormancy from a cross of Baudin?×?AC Metcalfe using 233 molecular markers. Three QTLs were mapped for seed dormancy based on the standard germination test at 24, 48 and 72?h. One major QTL was mapped to the long arm of chromosome 5H controlling seed dormancy and PHS tolerance from Baudin. Two other minor QTLs were identified from Baudin on chromosomes 3 and 7H. QTL/QTL interaction was detected for seed dormancy between chromosomes 3 and 5H. The PHS tolerance allele of the 5H QTL from Baudin contributes to higher malt yield without significant impact on diastatic power, beta-glucan content and wort viscosity. QTL from Baudin provide new sources to integrate PHS tolerance and high malting quality.     

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.     

Grain dormancy QTL identified in a doubled haploid barley population derived from two non-dormant parents

Grain dormancy provides protection against pre-harvest sprouting (PHS) in cereals. Composite interval mapping and association analyses were performed to identify quantitative trait loci (QTL) contributing grain dormancy in a doubled haploid (DH) barley population (ND24260?×?Flagship) consisting of 321 lines genotyped with DArT markers. Harvest-ripe grain collected from three field experiments was germinated over a 7-day period to determine a weighted germination index for each line. DH lines displaying moderate to high levels of grain dormancy were identified; however, both parental lines were non-dormant and displayed rapid germination within the first two?days of testing. Genetic analysis identified two QTL on chromosome 5H that were expressed consistently in each of the three environments. One QTL (donated by Flagship) was located close to the centromeric region of chromosome 5H (qSDFlag), accounting for up to 15% of the phenotypic variation. A second QTL with a larger effect (from ND24260) was detected on chromosome 5HL (qSDND), accounting for up to 35% of the phenotypic variation. qSDFlag and qSDND displayed an epistatic interaction and DH lines that had the highest levels of grain dormancy carried both genes. We demonstrate that qSDND in the ND24260?×?Flagship DH population is positioned proximal and independent to the well-characterised SD2 region that is associated with both high levels of dormancy and inferior malt quality. This indicates that it should be possible to develop cultivars that combine acceptable malting quality and adequate levels of grain dormancy for protection against PHS by utilizing these alternate QTL.     

The rice (Oryza sativa) Rc gene improves resistance to preharvest sprouting and retains seed and milled rice quality

Preharvest sprouting (PHS) remains a severe agricultural problem in rice and other crops. This study investigated the exact effect of Rc gene on PHS resistance along with seed and milled rice quality using two sets of near‐isogenic line‐derived populations, and showed the advantages of the Rc gene over other dormancy‐associated genes in protecting against PHS. Rc gene has a significant effect on PHS resistance, and Rc‐controlled dormancy does not negatively affect next agricultural production cycle. Importantly, Rc has a completely dominant maternal effect on PHS resistance, which confers an advantage to hybrid rice over inbred rice, and the effect fails to be counteracted by GA application in hybrid seed production. Moreover, Rc gene retains the same milled rice qualities of brown‐pericarp rice as that of white‐pericarp rice, and the brown‐pericarp rice possesses a higher antioxidative capacity than the white‐pericarp rice. Thus, wide application of the Rc gene would not only protect against PHS, but would also enhance the production of naturally occurring antioxidants that could make a significant contribution to human health.     

The effect of water and nitrogen availability during grain filling on the timing of dormancy release in malting barley crops

Pre-harvest sprouting (PHS) causes immediate loss of seed viability, making barley (Hordeum vulgare L.) grains worthless for malting purposes. Grain dormancy release rate in barley crops is genetically and environmentally controlled. A 2 year experiment was conducted to evaluate the effect of soil nitrogen and water availability during grain filling on the dormancy release pattern (and then on the PHS susceptibility) for five malting barley commercial cultivars. Drought and well-irrigated control treatments were imposed from anthesis onwards, and contrast nitrogen fertilization treatments were applied at tillering. Nitrogen availability showed no effects on dormancy release. Drought during grain filling accelerated dormancy release with respect to well-irrigated control in 2004, but not in 2005 year. Mean temperatures during the last stages of grain filling were much higher (ca. 6°C) in 2005 than in 2004, indicating that high-dormancy loss promoting temperatures had masked drought effects on dormancy release.     

The effect of introgressions of wheat D-genome chromosomes into 'Presto' triticale

Hexaploid triticale (X Triticosecale Wittmack) (2n= 6x= 42, AABBRR) and wheat (Triticum aestivum L.) (2n= 6x= 42, AABBDD) differ in their R and D-genomes. This produces differences in both agronomic and end-use quality characteristics. Our objective was to determine how introgressions of individual chromosomes from the D-genome of wheat affect these characteristics of a winter triticale 'Presto'. We studied the effects of 18 D-genome chromosome substitution lines, 15 sib-lines as controls, and five check cultivars at Lincoln, NE in 1996, using a randomized complete block design with two replications. The experiment was repeated at Lincoln and Mead, NE in 1997 and 1998 with 15 substitution lines that survived the first winter in Lincoln, along with their 12 control sibs and five check cultivars. Few D-genome chromosomes had positive effects. Chromosomes 2D, 4D, and 6D significantly reduced plant height when substituted for 2R, 4B, and 6R, respectively. No grain yield increases were associated with any of the D-genome chromosomes tested, but three substitutions decreased the grain yield. Depending on the allele of the hardness gene present, chromosome 5D increased or decreased kernel hardness when substituted for 5R or 5A, respectively. Introgressions of chromosomes 1D and 6D improved end-use quality characteristics of Presto. These results suggest that apart from beneficial effects of individual loci located on the D-genome chromosomes, no major benefit can be expected from D-genome chromosome substitutions.     

Mapping and validation of PCR-based markers associated with a major QTL for seed dormancy in wheat

Seed dormancy is one of the important factors controlling pre-harvest sprouting (PHS) resistance in wheat. We identified a major quantitative trait locus (QTL) for seed dormancy on the long arm of wheat chromosome 4A (4AL) via simple sequence repeat (SSR)-based genetic mapping using doubled haploid lines from a cross between Japanese PHS resistant variety ‘Kitamoe’ and the Alpine non-resistant variety “Münstertaler” (K/M). The QTL explained 43.3% of total phenotypic variation for seed dormancy under greenhouse conditions. SSR markers flanking the QTL were assigned to the chromosome long arm fraction length 0.59–0.66 on the basis of chromosome deletion analysis, suggesting that the gene(s) controlling seed dormancy are probably located within this region. Under greenhouse conditions, the QTL explained 28.5 and 39.0% of total phenotypic variation for seed dormancy in Haruyutaka/Leader (HT/L) and OS21-5/Haruyokoi (O/HK) populations, respectively. However, in field conditions, the effect was relatively low or not significant in both the K/M and HT/L populations. These markers were considered to be widely useful in common with various genetic backgrounds for improvement of seed dormancy through the use of marker-assisted selection. Further detailed research using near isogenic lines will be needed to define how this major QTL interacts with environmental conditions in our area.     

QTL analysis of seed storability in rice

A double haploid population, which consists of 127 lines derived from anther culture of a typical indica and japonica hybrid ‘ZYQ8’/‘JX17’, was used in this study. Seed storability was investigated by using the storage property measured by the difference of seed germination rates before and after treatment of the rice seeds under 40°C and 95% relative humidity for 10 days in a phytotron. Three QTLs related to rice seed storability were detected on chromosomes 9, 11 and 12, with the LOD scores 2.76, 4.83 and 2.54, respectively, together explaining 35.4% of the genetic variation. The ‘JX17’ allele at qLS‐9 and the ‘ZYQ8’ alleles at qLS‐11 and qLS‐12 could enhance the rice seed storability. The effects of the ‘ZYQ8’ alleles of qLS‐11 and qLS‐12 were also verified using chromosome segment substitution lines.     

Effect of the ph1b mutant on chromosome pairing in hybrids between Dasypyrum villosum and Triticum aestivum

Chromosome pairing was analysed in F₁ hybrids of the wheat cultivar ‘Chinese Spring’ (CS) and its ph1b mutant (CSphlb) with Dasypyrum villosum. On average, 1.61 chromosomes per cell paired in the hybrid CS ×D. villosum, but 14.43 in the hybrid CS ph1b×D. villosum. Genomic fluorescence in situ hybridization (GISH) revealed three types of homoeologous association between wheat (W) and D. villosum (D) chromosomes (W‐D, D‐W‐W and D‐W‐D) in pollen mother cells of the CS ph1b×D. villosum hybrid, and only one type (W‐W), in the CS ×D. villosum hybrid. Both F₁ hybrids were self‐sterile. The seed set of the backcross of CS ×D. villosum with CS was 6.67% and that of CS ph1b×D. villosum with CS or CS ph1b was only 0.45%. The chromosome number of BC₁ plants varied from 48 to 72. Translocations of chromosome segments or entire arms between wheat and D. villosum chromosomes were detected by GISH in the BC₁ plants from the backcross of CS ph1b×D. villosum to CS ph1b.     

中国小麦微核心种质及地方品种籽粒休眠特性的分子标记鉴定

为探索我国小麦微核心种质及地方品种籽粒休眠的遗传基础,利用已报道的4个3AS上的SSR标记(Xbarc57、Xbarc294、Xbarc310和Xbarc321)和1个3BL上的Viviparous-1基因标记Vp1-b2对107份我国小麦微核心种质及31份地方品种进行籽粒休眠的分子标记鉴定。结果表明,5个分子标记在试验材料中表现出丰富的等位变异,具有5～6种等位类型,与籽粒萌芽指数(GI)密切相关。根据一般线性模型分析结果,各位点的等位变异显著影响籽粒休眠,其中Vp1-b2和Xbarc294对籽粒休眠作用较其他标记大,可分别解释65.8%和61.2%的表型变异;其次是Xbarc310(56.3%)和Xbarc57(55.8%),最小的是Xbarc321(53.3%)。而5个标记联合可解释95.9%的性状变异,其次是Vp1-b2和Xbarc294的组合(89.1%),解释变异最小的标记组合是Vp1-b2和Xbarc321(79.4%)。5个分子标记即可解释籽粒休眠的绝大部分表型变异,说明我国小麦微核心种质及地方品种籽粒休眠特性受3AS和3BL上的2个主效基因控制。     

杂草稻种子休眠数量性状位点的定位

利用杂草稻与粳稻品种衍生的分离群体对控制种子休眠性的遗传基础进行了研究。检测到4个控制种子休眠性的QTL, 分别位于第1、第2和第6(2个)染色体上, 贡献率分别为7.8%、7.1%、5.5%和4.5%, 其中第2染色体上的QTL可能是一个新的控制种子休眠性的位点, 多项方差分析表明这4个位点的作用具有累加效应。种子发芽率与开花时间存在显著的负相关关系, 检测到的唯一一个控制抽穗期的QTL与位于第6染色体上的一个控制种子休眠性的QTL连锁或具有多效性, 这可能是造成其显著相关的主要原因。     

Chromosomal location of genes affecting tissue-culture response in wheat

Six ‘Chinese Spring/Triticum spelta’ substitution lines for chromosomes 1A, 1D (duplicates), 3D (duplicates), 6D, and one ‘Chinese Spring/ Marquis’ substitution line for chromosome 2B were studied for tissue-culture response (TCR). The results reported here indicate that chromosomes 2B and 6D are critical for TCR, whereas chromosome ID affects callus weight only. Chromosomes 1A and 3D were not found to be critical, however, these chromosomes may carry genes with minor effects.     

Genetic and environmental control of dormancy in white-grained wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Grain dormancy in wheat is an important component of resistance to preharvest sprouting and hence an important trait for wheat breeders. The significant influence of environment on the dormancy phenotype makes this trait an obvious target for marker-assisted-selection. Closely related breeding lines, SUN325B and QT7475, containing a major dormancy QTL derived from AUS1408 located on chromosome 4A, but substantially different in dormancy phenotype, were compared with a non-dormant cultivar, Hartog, in a range of controlled environments. As temperature increased, dormancy at harvest-ripeness decreased particularly for QT7475. The dormancy phenotypes of reciprocal F₁ grains involving all possible combinations of Hartog, QT7475 and SUN325B were also compared in two environments with different temperatures. The results were consistent with the presence of QTL in addition to 4A in SUN325B, compared with QT7475, at least one of which was associated with the seed coat. Genetic analysis of a doubled haploid population derived from SUN325B × QT7475 identified a highly significant QTL located on chromosome 3BL, close to the expected position of the mutant allele of the red seed coat colour gene in white-grained wheat, R-B1a. When the lines in the population were grouped according to the parental alleles at marker loci flanking the 3B QTL, the dormancy phenotype frequency distribution for the SUN325B group was shifted towards greater dormancy compared with the QT7475 group. However, significant variation for dormancy phenotype remained within each group. Lines representing the extremes of the range of phenotypes within each group maintained their relative ranking across seven environments consistent with the presence of another unidentified QTL contributing to dormancy in SUN325B.     

Genetic Control of Vernalization, Day-length Response, and Earliness per se by Homoeologous Group-3 Chromosomes in Wheat

To identify homoeologous group-3 chromosomes that carry genes for vernalization, day-length responses, and earliness per se, a series of aneuploid lines (mono-somics and tetrasomics) and chromosome-substitution lines in ‘Chinese Spring’ (CS) were surveyed under different vernalization and day-length regimes in controlled environments. The results indicated that genes on all three chromosomes of group 3 can have striking effects on ear-emergence time. The replacement of CS 3B by its homologues in ‘Lutescens 62’ and ‘Cheyenne’ produced an increased insensitivity to vernalization, while 3B homologues from ‘Ceska Presivka’ gave CS a remarkable sensitivity to vernalization. This provided evidence for multiple allelism at a new Vrn locus on chromosome 3B. A negative association between gene dosage and day-length response was found in CS 3D which was thought to carry a gene for promoting insensitivity to day-length. The behaviour of CS monosomic 3A and CS (Timstein 3A), in reducing numbers of days to heading independently of environmental stimuli, suggested the presence of earliness per se genes on this chromosome.     

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.     

Reciprocal substitutions analysis of freezing resistance in wheat (Triticum aestivum L.)

To investigate the effects of individual chromosomes on freezing resistance, as well as their interactions with the genetic background, reciprocal sets of chromosome substitution lines between two hard red winter wheat cultivars, ‘Cheyenne’ and ‘Wichita’, were used in this study. Duplicate lines for each chromosome were included to check background homogeneity. Two experiments were carried out in complete block designs with two replications for each duplicate. Crown and leaf water content and leaf wet weight were measured in the field experiments. Crown survival, electrolyte leakage and 50% lethality temperature (LT50) were measured in the laboratory. The results showed that ‘Cheyenne’ was more resistant than ‘Wichita’. Crown survival was significantly correlated with crown water content, crown wet weight and electrolyte leakage. Chromosomes 6A, 3B and 5D substituted from ‘Wichita’ into ‘Cheyenne’ (‘CNN‐WI’), decreased the crown survival, and increased membrane stability, crown water content and crown wet weight of ‘Cheyenne’. Thus, these chromosomes from ‘Wichita’ decreased freezing resistance in ‘Cheyenne’. Reciprocally, chromosomes 5A, 5D, 3B and 4D from ‘Cheyenne’ into ‘Wichita’ increased crown survival and decreased crown water content and crown wet weight of ‘Wichita’. It was concluded that these chromosomes from ‘Cheyenne’ cause freezing resistance in ‘Wichita’ and carry freezing‐resistance genes.