Quantitative trait locus analysis of seed germination,seedling vigour and seedling‐regulated hormones in Brassica napus

Good germination and seedling vigour are major breeding targets in winter oilseed rape (Brassica napus), because seedling vigour and prewinter crop establishment are closely associated with postwinter growth and yield. Here, we identified quantitative trait loci (QTL) related to germination, seedling vigour and seedling‐regulated hormones in a doubled haploid (DH) mapping population from a cross between winter oilseed rape parents with high vigour (Express 617) and low vigour (1012‐98). By phenotyping in a climate‐controlled glasshouse, we identified a total of 13 QTL on nine chromosomes for germination and seedling‐related traits at 7 and 14 days after sowing (DAS), explaining up to 11.2% of the phenotypic variation for seedling vigour. Forty‐seven metabolic QTL on 15 chromosomes were identified for auxin, abscisic acid (ABA) and dihydrophaseic acid (DPA) at 5 and 12 DAS, explaining up to 49.4% of phenotypic variation in seedling hormone composition. Multitrait QTL hot spots contribute to our understanding of the genetics and metabolomics of germination and seeding vigour in B. napus, and represent potential targets to breed high‐vigour cultivars.     

Mapping of quantitative trait loci controlling seed longevity of rice (Oryza sativa L.) after various periods of seed storage

Seed longevity varies considerably in cultivated rice (Oryza sativa L.), but the underlying genetic mechanism of longevity has not been well elucidated. Quantitative trait loci (QTL) that control seed longevity after various periods of seed storage were sought using recombinant inbred lines derived from a combination involving ‘Milyang23’(Indica‐type) and ‘Akihikari’ (Japonica‐type). In all, 12 QTLs for germination and normal seedling growth were detected as indices of seed longevity on chromosome 7 (one region) and chromosome 9 (two regions) in treated seeds that had been stored under laboratory conditions for 1, 2 or 3 years.‘Milyang23’ alleles of all QTLs promoted germination and normal seedling growth after all durations of storage. These QTL regions were detected repeatedly in more than one seed condition. Therefore, we infer that these regions control seed longevity.     

Quantitative trait locus mapping for seed artificial aging traits using an F2:3 population and a recombinant inbred line population crossed from two highly related maize inbreds

Quantitative trait locus (QTL) mapping for seed longevity is essential for breeding modern cultivars with resistance to deterioration during postharvest storage. The inbred lines X178 and I178 showed large differences in seed vigour after artificial aging treatment, while they had similar performances in terms of most agronomic traits. An F_2:3 population and a recombinant inbred line (RIL) population were generated to map QTL after 5 days under artificial aging conditions. Positive correlations were observed among all investigated traits including the aging germination rate, relative aging germination rate, aging simple vigour index, aging primary root length, aging shoot length and aging total length. Thirteen QTL were identified to locate on five chromosome regions: Chr.1:297 Mb (chromosome 1 region 297 Mb), Chr.3:205 Mb, Chr.4:240 Mb, Chr.5:205 Mb and Chr.7:155 Mb, with 2 to 4 QTL co‐located on a region. In each region, 3–8 previously identified aging‐related QTL were located, confirming the importance of these regions for controlling seed longevity in different maize populations. Taken together, the results of this work provide a foundation for further QTL fine mapping and the molecular‐assisted breeding of aging tolerant maize.     

Genetic analysis of salt tolerance in a recombinant inbred population of wheat (Triticum aestivum L.)

A population of 114 recombinant inbred lines (RILs), derived from the cross Opata85 × W7984, was used to genetically analyze the response of wheat to salt stress. This analysis resulted in the identification of 47 QTL mapping to all wheat chromosomes except 1B, 1D, 4B, 5D and 7D. Of these QTL, 10 were effective during the germination stage, and 37 at the seedling stage. Many of the traits related to salt tolerance mapped to common chromosome intervals, such as Xglk683–Xcdo460 on chromosome 3A, Xfbb168–Xbcd147 on chromosome 3B, Xcdo1081–Xfbb226 on chromosome 4DL and Xpsr106–Xfbb283 on chromosome 6DL. QTL located in the interval Xcdo1081–Xfbb226 (chromosome 4DL) were effective during the germination stage, whereas those in the interval Xfbb231.1–Xmwg916 (chromosome 6DL) were relevant to the seedling stage. The QTL in the intervals Xglk683–Xcdo460 (chromosome 3AS) and Xfbb168–Xbcd147 (chromosome 3BL) were effective at both the germination and seedling stages.     

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.     

Mapping QTLs for salt tolerance with additive,epistatic and QTL × treatment interaction effects at seedling stage in wheat

Quantitative trait loci (QTLs) for salt tolerance with additive, epistatic and QTL × treatment interaction effects at seedling stage in wheat were identified. A set of 131 recombinant inbred lines derived from cross Chuan 35050 × Shannong 483 were evaluated under salt stress and normal conditions. Wide variation was found for all studied traits. A total of 18 additive and 16 epistatic QTLs were detected, among which five and 11 were with significant QTL × treatment effects. Ten QTL clusters were identified, and each may represent a single gene or closely linked genes. The locus controlling shoot K⁺/Na⁺ concentration ratio and shoot Na⁺ concentration on chromosome 5A may be identical to Nax2. The interval Xgwm6‐Xgwm538 on chromosome 4B for total dry weight was also identified in a previous study, both near the marker Xgwm6. The marker Xgwm6 may be useful for marker‐assisted selection. Six pairs of homoeologous QTLs were detected, showing synteny among the A, B and D genomes. These results facilitate understanding the mechanisms and the genetic basis of salt tolerance in wheat.     

Mapping quantitative trait loci in wheat for resistance against greenbug and Russian wheat aphid

Breeding for genetic resistance against greenbug and Russian wheat aphid (RWA) is the most effective way of controlling these widespread pests in wheat. Earlier work had shown that chromosome 7D of a synthetic hexaploid wheat, ‘Synthetic’ (T. dicoccoides × Ae. squarrosa) (AABB × DD) gave resistance when transferred into the genetic background of an aphid‐susceptible cultivar, ‘Chinese Spring’, as the recipient. To map the genes involved, a set of 103 doubled haploid recombinant substitution lines was obtained from crossing the 7D substitution line with the recipient, and used to determine the number and chromosomal location of quantitative trait loci (QTL) controlling antixenosis and antibiosis types of resistance. Antixenosis to RWA was significantly associated with marker loci Xpsr687 on 7DS, and Xgwm437 on 7DL. Antibiosis to greenbug was associated with marker loci Xpsr490, Rc3 (on 7DS), Xgwm44, Xgwm111, Xgwm437, Xgwm121 and D67 (on 7DL). Similarly, antibiosis to RWA was linked to loci Xpsr490, Rc3, Xgwm44, Xgwm437 and Xgwm121. At least two QTL in repulsion phase, one close to the centromere either on the 7DS or 7DL arms, and a second distal on 7DL could explain antibiosis to RWA and, partially, this mechanism against greenbug.     

Detection of quantitative trait loci (QTLs) for seedling traits and drought tolerance in wheat using three related recombinant inbred line (RIL) populations

In order to detect quantitative trait loci (QTLs) for drought tolerance in wheat during seed germination conditional and unconditional QTL analyses of eight seedling traits were conducted under two water regimes using three related F₉ recombinant inbred line populations with a common female parent. A total of 87 QTLs for the eight seedlings traits and 34 specific QTLs related to drought tolerance were detected. Seventy-one of these QTLs were major QTLs with contributions to phenotypic variance of >10 %. Of the 34 QTLs related to drought tolerance only eight were also detected by unconditional analysis of seedling traits under osmotic stress conditions indicating that most of the QTLs related to drought tolerance could not be detected by unconditional QTL analysis. Therefore, conditional QTL analysis of stress-tolerance traits such as drought tolerance was feasible and effective. Of 11 important QTL clusters located on chromosomes 1BL, 1D, 2A, 2B, 2D, 4A, 6B, and 7B, nine were detected in multiple populations and eight were detected by both unconditional and conditional analyses.     

Mapping QTLs for seedling root traits in a doubled haploid wheat population under different water regimes

A deep and thick root system has a positive effect on wheat yield, particularly in drought environments. A doubled haploid (DH) population of 150 lines derived from the cross Hanxuan 10?×?Lumai 14 was used to map QTLs for seedling root characteristics. The DH lines were cultivated in an agarose gel-chamber under well-watered (WW) and water-stressed (WS) regimes. Water stress was simulated by adding mannitol to the agarose gel. The seminal root traits, including maximum root length (MRL), seminal root number, total root length, project root area, root surface area, and seminal root angle were measured after 6?days of seedling development. Grain yields (GY) were measured in a field experiment. A total of 29 QTLs were identified for seedlings cultured under WW regimes, and 23 QTLs under WS regimes. Individual QTL accounted for phenotypic variations ranging from 4.98 to 24.31?%. The QTLs were distributed on 17 chromosomes, except 1D, 4D, 6B and 6D. Seven consistently expressed QTLs were detected for all the traits tested except MRL under both water regimes. The QTLs for root traits were unevenly distributed among chromosomes, and clustered in eight loci on seven chromosomes, showing pleiotropic effects on target traits. One region in the interval Xgwm644.2?CP6901.2 on chromosome 3B contained 9 QTLs affecting most root traits. The present data provide an insight into the genetic basis of seedling root development under different water regimes and may benefit breeding programs using marker-assisted selection (MAS) for root traits.     

QTL analysis of quality traits in the spring wheat cross RL4452 × 'AC Domain'

Wheat grain quality is a complex group of traits of tremendous importance to wheat producers, end‐users and breeders. Quantitative trait locus (QTL) analysis studied the genetics of milling, mixograph, farinograph, baking, starch and noodle colour traits in the spring wheat population RL4452/‘AC Domain’. Forty‐seven traits were measured on the population and 99 QTLs were detected over 18 chromosomes for 41 quality traits. Forty‐four of these QTLs mapped to three major QTL clusters on chromosomes 1B, 4D, and 7D. Fourteen QTLs mapped near Glu‐B1, 20 QTLs mapped near a major plant height QTL on chromosome 4D, and 10 QTLs mapped near a major time to maturity QTL on chromosome 7D. Large QTLs were detected for grain and flour protein content, farinograph absorption, mixograph parameters, and dietary fibre on chromosome 2BS. QTLs for yellow alkaline noodle colour parameter L* mapped to chromosomes 5B and 5D, while the largest QTL for the b* parameter mapped to 7AL.     

Quantitative trait loci mapping of seed colour,hairy leaf,seedling anthocyanin,leaf chlorosis and days to flowering in F2 population of Brassica rapa L.

A diversity arrays technology (DArT) map was constructed to identify quantitative trait loci (QTL) affecting seed colour, hairy leaf, seedling anthocyanin, leaf chlorosis and days to flowering in Brassica rapa using a F₂ population from a cross between two parents with contrasting traits. Two genes with dominant epistatic interaction were responsible for seed colour. One major dominant gene controls the hairy leaf trait. Seedling anthocyanin was controlled by a major single dominant gene. The parents did not exhibit leaf chlorosis; however, 32% F₂ plants showed leaf chlorosis in the population. A distorted segregation was observed for days to flowering in the F₂ population. A linkage map was constructed with 376 DArT markers distributed over 12 linkage groups covering 579.7 cM. The DArT markers were assigned on different chromosomes of B. rapa using B. rapa genome sequences and DArT consensus map of B. napus. Two QTL (RSC1‐2 and RSC12‐56) located on chromosome A8 and chromosome A9 were identified for seed colour, which explained 19.4% and 18.2% of the phenotypic variation, respectively. The seed colour marker located in the ortholog to Arabidopsis thaliana Transparent Testa2 (AtTT2). Two QTL RLH6‐0 and RLH9‐16 were identified for hairy leaf, which explained 31.6% and 20.7% phenotypic variation, respectively. A single QTL (RSAn‐12‐157) on chromosome A7, which explained 12.8% of phenotypic variation was detected for seedling anthocyanin. The seedling anthocyanin marker is found within the A. thaliana Transparent Testa12 (AtTT12) ortholog. A QTL (RLC6‐04) for leaf chlorosis was identified, which explained 55.3% of phenotypic variation. QTL for hairy leaf and leaf chlorosis were located 0–4 cM apart on the same chromosome A1. A single QTL (RDF‐10‐0) for days to flowering was identified, which explained 21.4% phenotypic variation.     

不同盐浓度胁迫下小麦苗期苗高和主根长的QTL分析

小麦苗期苗高和主根长是鉴定小麦苗期耐盐性的重要指标。利用小麦品种花培3号×豫麦57获得的DH群体168个株系,在去离子水(对照)以及50,100,200 mmol/L NaCl溶液处理下,进行苗高和主根长的数量性状基因(QTL)定位分析。利用完备区间作图法,共检测到影响苗高和主根长的25个QTL,单个QTL对表型的贡献率为4.19%～23.72%。位于3D染色体区间Xgdm72-Xbarc1119上影响主根长的QTL位点具有最大的遗传效应,贡献率为23.72%;在100 mmol/L和50 mmol/L NaCl处理下,在2D染色体Xwmc170.2-Xgwm539区段,同时检测到影响苗高的2个QTL位点,其贡献率分别为12.59%和8.40%;在100 mmol/L和200 mmol/L NaCl处理下,在4D染色体Xc-fa2173-Xcfe188区段,同时检测到影响主根长的2个QTL位点,其贡献率分别为8.77%和5.70%;在对照和100mmol/L NaCl溶液处理下,在5BL染色体Xgwm213-Xswes861.2区段,同时检测到影响苗高的QTL位点,其贡献率分别为17.49%和6.28%。另外,在50 mmol/L NaCl溶液处理下,4B染色体Xwmc657-Xwmc48区段还定位了1个影响苗高的QTL位点,其贡献率为12.59%;在染色体3A和染色体7D上各检测出与主根长有关的1个不同的QTL;在5A染色体Xbarc358.2-Xgwm186和Xcwem40-Xbarc358.2区间分别检测到1个影响苗高的QTL。这些主效QTL可用于苗高和主根长的分子标记辅助选择。     

落叶松种子活力测定和田间育苗对比试验研究

用直立板发芽器对大兴安岭地区8个种批落叶松(Larix gmelini Rupr)种子活力进行了研究:(1)测定了种子发芽指数、活力指数和幼苗干重;(2)按幼苗高度和幼根、子叶初现状况、再用苗全长的标准差将落叶松幼苗分为5级,最后统计出幼苗健壮率和实验室出苗率(国际发芽率标准),从而改进了幼苗活力分级标准;(3)筛选出提高落叶松种子活力最佳预处理方法——种子预湿冷层积14天。通过8个种批落叶松田间育苗对比试验,得出种子预湿冷层积14天,育苗效果明显优于对照组。各种批的种子活力测定结果、幼苗活力分级结果和田间育苗效能是一致的。     

普通小麦籽粒黄色素含量的QTL分析

小麦面粉黄色度b*值是反映面粉颜色的重要指标，主要与籽粒黄色素含量有关。利用122对SSR引物、4对贮藏蛋白STS引物和10对AFLP引物组合，分析了中优9507´CA9632的71个DH系，构建了由173个位点组成的遗传连锁图，在小麦21个连锁群上覆盖2 881 cM。将该群体种植2年共计5个地点，测定籽粒黄色素和面粉黄色度b*值含量。采用复合区间作图法（CIM）进行了籽粒黄色素含量和面粉黄色度QTL分析。结果表明，面粉黄色度b*值的QTL位于染色体1DS、2DL、3A、4D、5D、6AL、6D和7AL上，其中7AL的QTL效应最大，贡献率为12.9%~37.6%；籽粒黄色素含量的QTL位于染色体2DL、3DL、4A、5A和7AL，其中7AL的QTL效应最大，贡献率为12.1%~33.9%。面粉黄色度b*值与籽粒黄色素含量共同的QTL位于7AL，与Xgwm264b紧密连锁，遗传距离分别为0~3.9 cM和0~0.9 cM。     

Mapping QTL for stay-green and agronomic traits in wheat under diverse water regimes

Wheat (Triticum aestivum L.) yield is directly proportional to physio-morphological traits. A high-density genetic map consisting of 2575 markers was used for mapping QTL controlling stay-green and agronomic traits in wheat grown under four diverse water regimes. A total of 108 additive QTL were identified in target traits. Among them, 28 QTL for chlorophyll content (CC) were detected on 11 chromosomes, 43 for normalized difference vegetation index (NDVI) on all chromosomes except 5B, 5D, and 7D, five for spikes per plant (NSP) on different chromosomes, nine for plant height (PH) on four chromosomes, and 23 for thousand-kernel weight (TKW) on 11 chromosomes. Considering all traits, the phenotypic variation explained (PVE) ranged from 3.61 to 41.62%. A major QTL, QNDVI.cgb-5A.7, for NDVI with a maximum PVE of 20.21%, was located on chromosome 5A. A stable and major PH QTL was observed on chromosome 4D with a PVE close to 40%. Most distances between QTL and corresponding flanking markers were less than 1 cM, and approximately one-third of the QTL coincided with markers. Each of 16 QTL clusters on 10 chromosomes controlled more than one trait and therefore could be regarded as pleiotropic regions in response to different water regimes. Forty-one epistatic QTL were identified for all traits having PVE of 6.00 to 25.07%. Validated QTL closely linked to flanking markers will be beneficial for marker-assisted selection in improving drought-tolerance in wheat.     

QTLs for Fusarium head blight response in a wheat DH population of Wangshuibai/Alondra‘s’

Summary A doubled haploid (DH) wheat population derived from the cross Wangshuibai/Alondra‘s’ was developed through chromosome doubling of haploids generated by anther culture of hybrids. Fusarium head blight (FHB) was evaluated for three years from 2001 to 2003 in Jianyang, Fujian Province, China, where epidemics of FHB have been consistently severe. After 307 pairs of simple sequence repeat (SSR) primers were screened, 110 pairs were polymorphic between Wangshuibai and Alondra`s’, and used to construct a genetic linkage map for detection of quantitative trait loci (QTLs). A stable QTL for low FHB severity was detected on chromosomes 3B over all three years, and QTLs on chromosomes 5B, 2D, and 7A were detected over two years. Additional QTLs on chromosomes 3A, 3D, 4B, 5A, 5D, 6B and 7B showed marginal significance in only one year. Six QTLs were detected when phenotypic data from three years were combined. In addition, significant additive-by-additive epistasis was detected for a QTL on 6A although its additive effect was not significant. Additive effects (A) and additive-by-additive epistasis (AA) explained a major portion of the phenotypic variation (76.5%) for FHB response. Xgwm533-3B and Xgwm335-5B were the closest markers to QTLs, and have potential to be used as selectable markers for marker-assisted selection (MAS) in wheat breeding programs.     

Upland rice breeding in Brazil: a simultaneous genotypic evaluation of stability, adaptability and grain yield

A durum wheat recombinant inbred line population developed from PDW 233 × Bhalegaon 4 cross was analyzed in five environments to understand the genetic network responsible for test weight (TW), thousand kernel weight (TKW), grain yield (YLD), spike length (SL), spikelets per spike (SPS), kernels per spike (KER) and kernel weight per spike (KWS). Genotype, environment and their interactions were main sources of variance for all the traits. TW and TKW were influenced by 11 main effect QTL and 6 digenic epistatic interactions detected on chromosomes 2A, 2B, 4B and 7A. Grain yield was influenced by three epistatic interactions and five main effect QTL, of which two on chromosome 2A were most consistent. A major QTL for spike length was observed on chromosome 3B. QTL for spike characters were distributed over 9 chromosomes. All the traits showed significant influence of digenic epistasis (QQ) and, to a certain extent, QTL × environment interactions (QQE). Therefore, while breeding for complex traits like kernel characters and grain yield components, these interactions should also be considered important. The consistent QTL on chromosome 2A between the marker interval Xgwm71.2–Xubc835.4 with pleiotropic effect on TW and TKW, may be utilized in early generation selection to improve TW and TKW and thereby the milling potential of the durum wheat.     

Seed Size Variation and its Effects on Germination and Seedling Vigour in Rice

The extent of seed size variations within a rice variety and its effect on germination and seedling vigour were investigated. Seed size was determined by weighing individual grains of randomly selected one hundred panicles. Grains were sorted into four size classes: 11–15, 16–20, 21–25 and 25–30 mg. There was nearly a three-fold variation between the smallest and the largest seeds, which ranged from 11 to 30 mg. Germination rate and seedling vigour index values increased with the increase of seed size suggesting the selection of larger seeds for good stand establishment.     

Genome-wide association study reveals loci associated with seed longevity in common wheat (Triticum aestivum L.)

Seed longevity could significantly determine seed regeneration cycle and greatly affect wheat production. With the 90 K chip assays, a genome-wide association study was performed to identify seed longevity-related markers and loci in common wheat. Seed germination ratios (GR) under artificially ageing of 166 wheat accessions across three environments were evaluated to assess seed longevity. Totally, 23 longevity-related loci were identified in the study, explaining 6.7%–11.4% of the phenotypic variations. Of these, QlgGR.cas-1A and QlgGR.cas-2B.2 were deemed as stable loci associated with wheat seed longevity. Fifteen loci were found overlapped with known quantitative trait loci or genes. Besides, QlgGR.cas-1A, QlgGR.cas-2B.2, QlgGR.cas-3D.1, QlgGR.cas-3D.2, QlgGR.cas-4A.2, QlgGR.cas-5A.1, QlgGR.cas-5A.2 and QlgGR.cas-6A.1 were colocated with seed dormancy-related loci. Significant additive effects were obtained for seed longevity by pyramiding favourable alleles. Several candidate genes were found involved in signal transduction and stress resistance pathways by sequencing analysis of significantly longevity-related molecular markers. These results might provide new sights into the genetic architecture of seed longevity.     

Use of barley seed vigour to discriminate drought and cold tolerance in crop years with high seed vigour and low trait variation

Seed vigour is a precondition for early and homogenous field emergence of barley, in addition to effective malting. This study aimed to assess the selection of barley varieties by using seed vigour as the indicator. Seed vigour of barley (quantified as the germination percentage) was evaluated under drought (?0.2 MPa) and temperature stress (10°C). At two locations over a 3‐year period, 1 population of 133 Derkado × B83‐12/21/5 doubled haploid (DH) lines (and parents) was evaluated for seed vigour, of which 108 DH lines were assessed for three malting parameters. The relatively high values of vigour during the 3‐year period (overall average 94–95%) probably impeded high variations in genetic potential. A total of 27 DH lines of the 133 evaluated showed transgression for vigour (up to 98%) in comparison with the parents (Derkado: 96%; B83: 92%). In conclusion, caution should be applied when selecting for seed vigour, even in good crop years with high levels of seed vigour and low trait variations. Such selection might improve vigour, particularly in crop years with unsuitable weather conditions.